Clinical trial NCT03507686
Conditions: Choroideremia
Keywords: NightstaRx NSR-REP1 CHM Gene Therapy AAV REP1 Timrepigene Emparvovec
Sponsor: Biogen
Start date: 2017-11-29; Completion date: 2022-06-29
Enrollment: 66 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
The objective of the study is to evaluate the safety of bilateral, sequential sub-retinal administration of a single dose of BIIB111 in adult male participants with Choroideremia (CHM).

Detailed description:
This study was previously posted by NightstaRx Ltd. In October 2020, sponsorship of the trial was transferred to Biogen.

Arms: arm 1: Participants will receive a single dose of sub-retinal injection of BIIB111 in each eye at Day 0 separated by an interval of \<6 months, 6-12 months, or \>12 months.
Interventions: intervention 1: BIIB111
Intervention descriptions: intervention 1: Administered as specified in the treatment arm.

Protocol documents:
Prot_000.pdf: 
CLINICAL STUDY PROTOCOL 
NSR-REP-02 (273CH203)
AAV2-REP1
An Open-Label Safety Study of Retinal Gene Therapy for 
Choroideremia with Bilateral, Sequential Administration of Adeno-
Associated Viral Vector (AAV2) Encoding Rab Escort Protein 1 
(REP1) 
GEMINI Study
INDICATION:
Choroideremia (CHM)
STUDY PHASE:
2
EUDRACT NUMBER:
2017-002395-75
CLINICAL TRIAL NUMBER: 
NCT03507686
INN:
Timrepigene emparvovec
SPONSOR:
NightstaRx Ltd. (A Biogen Company)
Biogen Idec Research Limited
Innovation House
70 Norden Road
Maidenhead, Berkshire
SL6 4AY
United Kingdom
Final Protocol Date:
4 Jul 2017
Amendment 1, Version 2
17 May 2018
Amendment 2, Version 3
4 February 2019 (Internal Only Version 3.0)
Amendment 2.1, Version 3.1
19 February 2019 
Amendment 3.0, Version 4.0
30 October 2019
Amendment 4.0, Version 5.0
30 Apr 2020
Amendment 5.0, Version 6.0
12 Nov 2020
Version 6 of protocol NSR-REP-02 supersedes Version 5, dated 30 Apr 2020. 
CONFIDENTIALITY STATEMENT
This protocol is the property of Biogen/NightstaRx Ltd.  It is not to be transmitted, copied or published without 
written permission from Biogen/NightstaRx and must be kept in a confidential manner.  Persons to whom the 
information is disclosed must be informed that the information is CONFIDENTIAL and may not be further 
disclosed by them.


AAV2-REPI 
Protoool: NSR-REI'-02 
SPONSOR APPROVAL PAGE 
Version 6.0 
12 Nov2020 
Clinical Study Protocol Number: NSR-REP-02 (273CH203) 
l'rotocol Title: 
Protocol Date: 
Approved By: 
An Open-Label Safety Study of Retinal Gene Therapy 
lor Choroideremia with Bilateral, Sequential 
Administration of Adcno-Associatcd Viral Vector 
(AA 
Encoding Rab Escort Protein 1 (REP I) 
12 Nov2020 
, MD 
 
Protocol No. NSR-REP-02 was approved by: 
 
Biogen 
CONFIDENTIAL 


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INVESTIGATOR’S SIGNATURE PAGE
Clinical Study Protocol Number:
NSR-REP-02 (273CH203)
Protocol Title:
An Open-Label Safety Study of Retinal 
Gene Therapy for Choroideremia with 
Bilateral, Sequential Administration of 
Adeno-Associated Viral Vector (AAV2) 
Encoding Rab Escort Protein 1 (REP1)
Protocol Date:
12 Nov 2020
I have read the Investigator’s Brochure for AAV2-REP1 and I have read Protocol NSR-REP-
02 and agree to conduct the study as outlined and in compliance with the Declaration of 
Helsinki (where required), the International Council on Harmonisation (ICH) guideline for 
Good Clinical Practice (GCP), and all applicable local and federal regulatory requirements 
and state/local laws.  I agree to maintain confidentiality of my subjects and all information 
received or developed in relation to this protocol.
Signed:
Date:
Name
Title
Institution
City, State (as applicable), Country


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CLINICAL RESEARCH ORGANIZATION CONTACT INFORMATION
Name
Address
Telephone 
Number
Covance (Formerly known 
as Chiltern International, 
Ltd)
Chiltern International Limited
Covance Clinical and Periapproval Services, 
Ltd
Osprey House,
Maidenhead Office Park,
Westacott Way, Littlewick Green,
Maidenhead SL6 3QH, UK
+44 (0) 1628 
548000
Additional contact information is available in the Site Operations Manual.


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2
PROTOCOL SYNOPSIS
Name of Sponsor/Company: NightstaRx, Ltd
Name of Test Product: AAV2-REP1
Protocol Title: An Open-Label Safety Study of Retinal Gene Therapy for Choroideremia with 
Bilateral, Sequential Administration of Adeno-Associated Viral Vector (AAV2) Encoding Rab 
Escort Protein 1 (REP1)
Protocol Number: NSR-REP-02
Study centres: The study will be conducted at approximately 5 study centres in Europe and 
United States.
Study period (years): 
Estimated overall study duration: 12 months of follow-up 
for each eye
Phase of development: Phase 2
Study Objective: The objective of the study is to evaluate the safety of bilateral, sequential sub-
retinal administration of a single dose of AAV2-REP1 in adult male subjects with choroideremia 
(CHM).
Primary Endpoint: The safety of bilateral administration of AAV2-REP1 will be evaluated 
with the following safety measures:

Best corrected visual acuity (BCVA) as measured by the Early Treatment of Diabetic 
Retinopathy Study (ETDRS) chart

Ophthalmic examination assessments (including intraocular pressure [IOP], slit lamp 
examination, lens opacity grading and dilated ophthalmoscopy)

Spectral domain optical coherence tomography (SD-OCT)

Fundus autofluorescence (AF)

Fundus photography

Microperimetry

Adverse event (AE) reporting 

Vector shedding post-treatment

Immunogenicity sampling post-treatment 

Vital signs
Secondary Efficacy Endpoints will include:

Change from Baseline in BCVA as measured by the ETDRS chart 

Change from Baseline in AF

Change from Baseline in SD-OCT

Change from Baseline in microperimetry
Study Design: This is a multi-centre, open-label, prospective, 2-period, bilateral interventional 
safety study of AAV2-REP1 in adult male subjects with genetically confirmed CHM.  The study 
will consist of a Screening Period followed by 2 treatment periods (Period 1 and Period 2) with up 
to 9 visits per period.  During the Screening Period, subjects will be assessed for eligibility of both 
eyes.  The Investigator will assign the order in which the eyes are treated (i.e., study eye 1 [SE1]-
Period 1 and study eye 2 [SE2]- Period 2, respectively) with a number assigned to each eye.  This 
will be done in collaboration with the subject; however, the worse eye will generally be selected 
for first treatment. Study eye selection and eligibility for surgery will then be confirmed 
collectively by the Patient Eligibility Review Committee, which includes the Investigator, a 
consulting surgeon, and the Sponsor (see the Patient Eligibility Review Plan). The estimated 
target interval between the surgical procedures of SE1 and SE2 will be determined at the 
Screening Visit.
Each study eye will be followed for at least 12 months post-treatment, for up to 9 visits per 
treatment period: Visit 1 (Day 0, Injection Day Visit); Visit 2 (Day 1, post-operative follow-up 
visit); Visit 3 (Day 3 + 2 days, post-operative follow-up visit); Visit 4 (Day 7 -1/+2 days); Visit 5 


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(Day 14 ± 3 days); Visit 6 (Month 1 ± 7 days); Visit 7 (Month 3 ± 14 days); Visit 8 (Month 6 ± 14 
days) and Visit 9 (Month 12 ± 14 days; end of study visit).
At Period 1, Visit 1 (Day 0, Injection Day Visit for SE1), subjects will undergo a vitrectomy with 
retinal detachment and receive a sub-retinal injection of AAV2-REP1 in SE1.  Visits 2-9 will then 
be conducted according to the Schedule of Study Procedures, unless Period 2 commences during 
this time, in which case, the study visits will follow the schedule for SE2.  Ophthalmic 
assessments during Period 1, Visits 2-9 will be performed on both eyes. 
The interval between SE1 and SE2 treatment is expected to vary among subjects from weeks to 
many months.  While the interval will be decided on a case-by-case basis, an effort will be made 
to schedule varying treatment intervals in order to better characterise the immunological and 
safety profile of sequential treatment administration.  Thus, approximately 20 subjects each will 
be treated with a short (<6 month), medium (6-12 months), or long (>12 month) surgery window 
between treatment of the first and second eye.  Because the timing of the second surgery visit 
(Period 2, Visit 1) will vary among subjects, the duration of study Period 1 will also be variable.  
The post-operative outcome and visual function of SE1 will inform the scheduling of Period 2.  
Signs of post-operative inflammation, or other post-operative sequelae, as judged by the 
Investigator, should be resolved in SE1 for the subject to continue to Period 2.  After 
comprehensive evaluation of all safety assessments and visual functional tests of SE1 and SE2, 
the Investigator will determine the eligibility of the subject and decide the appropriate time to 
schedule SE2, Visit 1.  
All baseline ophthalmic assessments must be conducted within 10 weeks prior to SE2 surgery 
(Period 2, Visit 1).  Eligibility for SE2 surgery will then be confirmed collectively by the Patient 
Eligibility Review Committee, which includes the Investigator, a consulting surgeon, and the 
Sponsor (see the Patient Eligibility Review Plan). If, at that time, the BCVA in SE2 is <34 
ETDRS letters, surgery may be performed when, in the opinion of the Patient Eligibility Review 
Committee: 1) the subject is found eligible by all other inclusion and exclusion criteria, and 2) the 
subject is considered an appropriate candidate who has potentially modifiable disease. 
At Period 2, Visit 1 (Day 0, Injection Day Visit for SE2), subjects will undergo a vitrectomy with 
retinal detachment and sub-retinal injection of AAV2-REP1 in the contralateral, untreated eye 
(SE2).  Post-treatment, subjects will no longer follow the Period 1 visit schedule, but will instead 
attend Period 2, Visits 2-9 according to the Schedule of Study Procedures. Ophthalmic 
assessments scheduled for Period 2, Visits 2-9 will be performed on both eyes. 
Subjects will be assessed for safety and efficacy throughout the study; assessments are outlined in 
the Schedule of Study Procedures.  Subjects who develop cataracts may undergo cataract surgery 
if deemed clinically necessary.  If cataract surgery is performed, it should be carried out at least 
4 weeks before Month 12 (Visit 9) for the respective eye.  The cataract surgery timing must be 
discussed with the medical monitor prior to the procedure.
Number of Subjects (Planned): Approximately 60 subjects are planned for the study.
Inclusion Criteria: During the Screening Period, subjects will be found eligible for study 
participation if they meet all of the following inclusion criteria. 
1.
Are willing and able to give informed consent for participation in the study to have both 
eyes treated
2.
Are male and ≥18 years of age
3.
Have documentation of a genetically-confirmed diagnosis of CHM 
4.
Have active disease clinically visible within the macular region of both eyes
5.
Have a BCVA of ≥34 ETDRS letters (20/200 or better Snellen acuity) in both eyes, or in 
the untreated eye, if the other eye was previously treated with AAV2-REP1*
*If previously treated with AAV2-REP1 in an antecedent study, subjects may be eligible 
for participation following Sponsor approval. 
6.
For subjects who received treatment with AAV2-REP1 in an antecedent study, have 
biological samples available to complete an adequate immunology profile
Exclusion Criteria: At the Screening Visit, subjects will be found not eligible for study 
participation if they meet any of the following exclusion criteria. 


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1.
Have a history of amblyopia or inflammatory disorder in either eye
2.
Are unwilling to use barrier contraception methods, or abstain from sexual intercourse 
for a period of 3 months following treatment with AAV2-REP1 in either eye
3.
Have had previous intraocular surgery performed within 3 months of the Screening Visit 
in either eye
4.
Have any other significant ocular or non-ocular disease/disorder which, in the opinion of 
the Patient Eligibility Review Committee or Investigator, may either put the subjects at 
risk because of participation in the study, or may influence the results of the study, or the 
subject’s ability to participate in the study.  This includes but is not limited to a potential 
subject:

with a contraindication to oral corticosteroid (e.g. prednisolone/prednisone)

with clinically significant cataract in either eye

who, in the clinical opinion of the Patient Eligibility Review Committee or 
Investigator, is not an appropriate candidate for sub-retinal surgery (see the 
Patient Eligibility Review Plan)
5.
Have participated in another research study involving an investigational product in the 
past 12 weeks or received a gene/cell-based therapy at any time previously, except if 
treated within in an antecedent study with AAV2-REP1.
Test Product, Dosage, and Mode of Administration: For each eye, subjects will undergo 
vitrectomy and retinal detachment and receive a sub-retinal injection of up to 0.1 mL of study 
drug containing 1×10^11 AAV2-REP1 genome particles (gp).  
Reference Therapy (Comparator), Dosage, and Mode of Administration: Not Applicable.
Criteria for Evaluation:
Safety: The safety evaluation will be based on BCVA (as measured by the ETDRS chart); full 
ophthalmic examination (including IOP, slit lamp examination, lens opacity grading and dilated 
ophthalmoscopy); SD-OCT; fundus AF; fundus photography; microperimetry; AE reporting; 
vector shedding and immunogenicity; and vital signs.
Efficacy: The efficacy evaluation will be based on BCVA (as measured by the ETDRS chart); 
fundus AF; SD-OCT and microperimetry.
Statistical Methodology: No formal sample size calculation will be performed.
Continuous variables will be summarised over time using descriptive statistics (i.e., mean, 
standard deviation, 95% confidence intervals [CI], median, first and third quartiles, fifth and 
ninety-fifth percentiles, minimum, and maximum).  Categorical variables will be described over 
time using counts, percentages, and 95% CIs.  Summaries will be tabulated by visit and eye.  No 
formal statistical testing will be performed. 
AEs will be summarised by system organ class, preferred term, and eye.  The number of eyes with 
an AE, as well as the number of events, will be summarised, by period and eye.    Similar 
summaries will be produced for study drug/procedure-related AEs, AEs leading to 
discontinuation, and serious AEs.  AEs will also be summarised by maximum severity, 
relationship to study drug/procedure, and time to onset and resolution.  Vector shedding and 
immune response profiles will be described.
The remaining safety evaluations will be analysed using descriptive statistics.


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3
TABLE OF CONTENTS, LIST OF TABLES, AND LIST OF FIGURES
2
PROTOCOL SYNOPSIS .......................................................................................5
3
TABLE OF CONTENTS, LIST OF TABLES, AND LIST OF FIGURES...........8
4
ABBREVIATIONS AND DEFINITIONS...........................................................11
5
INTRODUCTION................................................................................................12
5.1
Choroideremia.......................................................................................................12
5.2
Gene Therapy with Adeno-Associated Viral (AAV) Vectors ..............................12
5.3
Study Rationale.....................................................................................................13
5.4
Risk / Benefit Assessment ....................................................................................14
6
STUDY OBJECTIVES AND ENDPOINTS........................................................16
6.1
Objectives .............................................................................................................16
6.2
Endpoints ..............................................................................................................16
7
INVESTIGATIONAL PLAN...............................................................................17
7.1
Overall Study Design............................................................................................17
7.2
Discussion of Design ............................................................................................19
8
SELECTION AND WITHDRAWAL OF SUBJECTS........................................21
8.1
Inclusion Criteria ..................................................................................................21
8.2
Exclusion Criteria .................................................................................................21
8.3
Subject Withdrawal Criteria .................................................................................22
9
STUDY TREATMENT........................................................................................23
9.1
Treatments Administered......................................................................................23
9.2
Description of Study Drug....................................................................................23
9.3
Packaging, Labeling, and Storage.........................................................................23
9.4
Vitrectomy Procedure and Injection of AAV2-REP1 ..........................................23
9.5
Randomisation ......................................................................................................25
9.6
Study Masking ......................................................................................................25
9.7
Study Drug Accountability ...................................................................................25
9.8
Concomitant Therapy............................................................................................25
9.9
Treatment Compliance..........................................................................................26
10
STUDY VISITS AND PROCEDURES...............................................................27
10.1
Screening Period ...................................................................................................27
10.1.1
Baseline Visit, Period 2 (<10 Weeks from Visit 1, Period 2 [SE2]) ....................28
10.2
Period 1 and 2, Visit 1 (Day 0, Injection Day Visit).............................................29
10.3
Period 1 and 2, Visit 2 (Post-operative Day 1) .....................................................30
10.4
Period 1 and 2, Visit 3 (Post-operative Day 3 + 2 Days)......................................30
10.5
Period 1 and 2, Visit 4 (Post-operative Day 7 -1/+2 Days) ..................................30


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10.6
Period 1 and 2, Visit 5 (Post-operative Day 14 ± 3).............................................31
10.7
Period 1 and 2, Visit 6 (Month 1 ± 7 Days) and Visit 7 (Month 3 ± 14 Days).....31
10.8
Period 1 and 2, Visit 8 (Month 6 ± 14 Days) and Visit 9 (Month 12 ± 14 Days)*
...............................................................................................................................32
10.9
Early Termination (ET Visit)................................................................................32
10.10
Unscheduled Visits ...............................................................................................33
11
ASSESSMENT OF SAFETY...............................................................................34
11.1
Evaluation, Recording, and Reporting Adverse Events........................................34
11.1.1
Definitions ............................................................................................................34
11.1.2
Recording of Adverse Events...............................................................................35
11.1.3
Follow-up of Adverse Events...............................................................................36
11.1.4
Reporting of Serious Adverse Events...................................................................37
11.1.5
Data Monitoring Committee.................................................................................37
11.2
Pregnancy..............................................................................................................37
11.3
Stopping Criteria...................................................................................................38
11.4
Laboratory Assessments .......................................................................................38
11.4.1
Vector Shedding ...................................................................................................38
11.4.2
Immunogenicity....................................................................................................38
11.5
Vital Signs.............................................................................................................38
11.6
Best-Corrected Visual Acuity...............................................................................38
11.7
Microperimetry .....................................................................................................39
11.8
Fundus Autofluorescence......................................................................................39
11.9
Spectral Domain Optical Coherence Tomography...............................................39
11.10
Full Ophthalmic Examination...............................................................................39
11.11
7-Field Colour Fundus Photography.....................................................................39
12
ASSESSMENT OF EFFICACY ..........................................................................41
12.1
Best-Corrected Visual Acuity...............................................................................41
12.2
Fundus Autofluorescence......................................................................................41
12.3
Spectral Domain Optical Coherence Tomography...............................................41
12.4
Microperimetry .....................................................................................................41
13
STATISTICAL CONSIDERATIONS .................................................................42
13.1
Sample Size...........................................................................................................42
13.2
Procedure for Accounting for Missing Data.........................................................42
13.3
Analysis Sets.........................................................................................................42
13.4
Descriptive Statistics and Conventions.................................................................42
13.5
Demographics and Baseline Characteristics.........................................................42
13.6
Safety Analyses.....................................................................................................42


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13.6.1
Visual Acuity........................................................................................................42
13.6.2
Ophthalmic Examination......................................................................................43
13.6.3
Spectral Domain Optical Coherence Tomography...............................................43
13.6.4
Fundus Autofluorescence .....................................................................................43
13.6.5
7-Field Colour Fundus Photography ....................................................................43
13.6.6
Microperimetry.....................................................................................................43
13.6.7
Adverse Events.....................................................................................................43
13.6.8
Immunogenicity and Vector shedding..................................................................43
13.6.9
Vital Signs ............................................................................................................43
13.7
Efficacy Analyses .................................................................................................43
13.7.1
Visual Acuity........................................................................................................43
13.7.2
Spectral Domain Optical Coherence Tomography...............................................44
13.7.3
Fundus Autofluorescence .....................................................................................44
13.7.4
Microperimetry.....................................................................................................44
14
INFORMED CONSENT, ETHICAL REVIEW AND REGULATORY 
CONSIDERATIONS............................................................................................45
14.1
Informed Consent..................................................................................................45
14.2
Ethical/Regulatory Review ...................................................................................45
14.3
Regulatory Considerations....................................................................................45
15
ADMINISTRATIVE PROCEDURES.................................................................46
15.1
Data Quality Control and Assurance ....................................................................46
15.2
Data Handling and Records Management ............................................................46
15.3
Access to Source Documentation and Subject Privacy ........................................47
15.4
Time and Schedule of the Study ...........................................................................47
15.5
Policy for Publication and Presentation of Data...................................................47
16
APPENDIX...........................................................................................................48
17
REFERENCE LIST..............................................................................................52
LIST OF TABLES
Table 1:
Schedule of Study Procedures ..............................................................................49
LIST OF FIGURES
Figure 1
Overall Study Schematic ......................................................................................17
Figure 2
Vitrectomy and Sub-Retinal Injection of AAV2 Vector......................................24


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4
ABBREVIATIONS AND DEFINITIONS
Abbreviation or Term
Definition
AAV
adeno-associated virus
AAV2
AAV serotype 2
AAV2-REP1
adeno-associated viral vector (AAV2) encoding rab escort protein 1
AE
adverse event
AF
autofluorescence
BCVA
best corrected visual acuity
BGH-polyA
bovine growth hormone polyadenylation
BSS
balanced salt solution
CBA
chicken β actin
cDNA
complementary deoxyribonucleic acid
CHM
choroideremia
CI
confidence interval
CRC
central reading centre
CRO
contract research organisation
eCRF
electronic case report form
DMC
data monitoring committee
EDC
electronic data capture
ELISA
enzyme-linked immunosorbent assay
ELISPOT
enzyme-linked immunospot
ET
early termination
ETDRS
Early Treatment of Diabetic Retinopathy Study
GCP
good clinical practice
gp
genome particle
GTMP
gene therapy medicinal product
ICH
International Council on Harmonisation
IEC
Independent Ethics Committee
IOP
intraocular pressure
IRB
Institutional Review Board
LOCS
lens opacities classification system
REP1
Rab escort protein 1
SAE
serious adverse event
SAP
statistical analysis plan
SD
study day
SD-OCT
spectral domain optical coherence tomography
SE1/SE2
study eye 1/study eye 2
SOP
standard operating procedure
SUSAR
suspected unexpected serious adverse reaction
UK
United Kingdom
VA
visual acuity
WPRE
woodchuck hepatitis post-transcriptional regulatory element


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5
INTRODUCTION
5.1
Choroideremia
Choroideremia (CHM) is a rare, bilateral, untreatable retinal degeneration that begins in 
childhood with loss of night vision, and gradually progresses to blindness by middle age.  
CHM is caused by loss of function of the gene encoding Rab escort protein 1 (REP1), which 
is located on the X-chromosome (Cremers, van de Pol et al. 1990, Seabra, Brown et al. 1993).  
The disease has an X-linked recessive mode of inheritance, affects approximately 1 in 50,000 
people, and is mostly caused by loss of function (null) mutations (Sankila, Tolvanen et al. 
1992, MacDonald, Russell et al. 2009).
5.2
Gene Therapy with Adeno-Associated Viral (AAV) Vectors
Gene therapy is rapidly emerging as a promising approach for the treatment of incurable 
retinal diseases (Bryant, Duker et al. 2012, Boye, Boye et al. 2013).  Gene therapy medicinal 
products (GTMPs) are a type of advanced therapy medicinal product defined as medicines 
derived from genes and cells (EMA 2015).  GTMPs contain or consist of recombinant nucleic 
acid delivered via a viral vector to targeted cells.  The central goal of gene therapy is to 
replace non-functional or defective genes with new genes that are fully functional so that the 
level of genetic expression can return to normal.
The eye is particularly well suited for gene therapy because of its accessibility, relative 
immune privilege status, small size, compartmentalisation and the presence of a fellow 
control eye (Boye, Boye et al. 2013).  The eye is one of the few relatively immunologically-
privileged sites in the body, so vectors used in gene therapy are unlikely to cause a significant 
systemic immune response.  It has long been established that to maintain the transparent 
structures required for vision, the eye has conserved a number of adaptations that selectively 
diminish the immune response (Streilein 2003).  For example, complement-fixing antibodies, 
neutrophils and macrophages are generally excluded from mitigation of an inflammatory 
response to limit the formation of opacities.  Anatomically, the retina is protected by the 
blood-retinal barrier to minimise the circulation of immune cells (Streilein 2003).  Studies 
have demonstrated that antigens introduced to the vitreous and sub-retinal space exhibit a 
reduced immune response due to the varied cytokine environment and the presence of 
molecules that suppress the immune system (Stein-Strelein and Watte 2007). 
Given the defined volume of the eye, the amount of viral vector needed to achieve a 
therapeutic effect will likely be small, reducing the risk of toxicity and increasing the 
likelihood of being able to manufacture quantities of vector sufficient to treat the retina.  The 
eye also allows for localised treatment without intravenous delivery, thus decreasing the 
chance of systemic absorption and toxicity.  Finally, the effects of localised ocular treatments 
can be easily observed and monitored for efficacy and safety, something that cannot be 
readily done with systemic conditions (Bryant, Duker et al. 2012).
The most commonly used delivery system for retinal gene therapy is the adeno-associated 
virus (AAV), and numerous early phase clinical trials using AAV vectors are currently 
underway for potential treatment of various retinal diseases (Boye, Boye et al. 2013, 
MacLaren, Groppe et al. 2014, Edwards, Jolly et al. 2016).  AAVs remain in the nucleus as 
episomes with no integration into the human genome, thereby decreasing the risk of 
insertional oncogenesis.  AAVs allow for stable and long term transgene expression in 
different retinal cells, including photoreceptors, retinal pigment epithelium cells, ganglion 
and Müller cells (Vanderberghe and Auricchio 2012, Day, Byrne et al. 2014).  Lastly, AAVs 


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elicit a minimal immune response as supported by emerging data from non-clinical and 
clinical studies.  
One non-clinical study evaluated immunological and functional consequences of adeno-
associated viral vector type 2 (AAV2) vector re-administration to the contralateral eye in 
dogs and non-human primates (Amado, Mingozzi et al. 2010).  In this study, rAAV2-
hRPE65v2 administered to the untreated, contralateral eye 15 days after the first injection 
was found to be both efficacious and safe.  The study demonstrated that cell-mediated 
immune responses were benign, with only 1 of 10 animals in the study developing a 
persistent T cell immune response to AAV2, a response that was mediated by CD4+ T cells.  
Sequential bilateral injection caused minimal inflammation and improved visual function in 
affected animals.  The authors concluded that sub-retinal re-administration of AAV2 is safe 
even in the setting of pre-existing immunity to the vector, a parameter that has been used to 
exclude patients from gene therapy trials.  In another non-clinical study in rats, lack of 
apparent immunogenicity was observed following same-day bilateral administration of 
6×10^9 genome particles (gp)/eye of adeno-associated viral vector (AAV2) encoding Rab 
escort protein 1 (AAV2-REP1) (CharlesRiverLabs 2015).  This total antigen exposure 
following bilateral dosing is comparable to a unilateral dose of 1.2×10^10 gp, which equates 
to a 1.2×10^12 gp dose in humans. This provides a 12-fold immunogenic safety margin 
compared to the planned clinical dose of 1×10^11 gp.
In the clinical setting, repeat administration of AAV2-RPE65 to the contralateral eye of 
children and adults was also shown to be safe, irrespective of baseline immune status 
(Bennett, Wellman et al. 2016). In this study, 11 children and adults with Leber’s congenital 
amaurosis received a single sub-retinal administration of AAV2-RPE65 at a dose of 
1.5×10^11 gp (1.7-4.6 years after initial sub-retinal injection) to the contralateral, previously 
untreated eye.  No adverse events (AEs) related to AAV2 vector were reported, and humoral 
and cell-mediated responses to AAV2 capsid and RPE65 transgene were benign in all 
patients.  Two individuals who had high pre-existing neutralizing AAV2-antibodies did not 
experience an inflammatory response in either eye following contralateral eye administration.  
AAV2-RPE65, voretigene neparvovec-rzyl (LUXTURNA™, Spark Therapeutics, Inc), has 
been approved in the US and the EU as the first adeno-associated virus vector type 2 
(AAV2)-based gene therapy indicated for the treatment of patients with confirmed bi-allelic 
RPE65 mutation-associated retinal dystrophy.
5.3
Study Rationale
CHM is incurable and treatment is supportive at best.  NightstaRx is developing AAV2-REP1 
as a potential GTMP for the treatment of CHM.  AAV2-REP1 is a recombinant  AAV2 
particle encapsulating 1.962kB complementary deoxyribonucleic acid (cDNA) of the wild-
type human REP1 gene.  Non-clinical studies with AAV2-REP1 have been very promising 
(Tolmachova, Tolmachov et al. 2013).  AAV2-REP1 expresses high levels of human REP1 
protein, restores REP1 to human CHM fibroblasts, provides functional rescue of human 
CHM cells, expresses protein in the retina of CHM mice in vivo, and is non-toxic when over-
expressed by one log unit.  Further, over-expression of the human REP1 protein does not 
significantly affect retinal function (Tolmachova, Tolmachov et al. 2013).
In 2011, an investigator-sponsored, first-in-human study of AAV2-REP1 was initiated in 
adult male patients with CHM.  The first 6 patients were treated with a single sub-retinal 
injection of 1×10^10 gp in 1 eye (MacLaren, Groppe et al. 2014).  Six months after receiving 
AAV2-REP1, 2 patients with impaired vision at baseline gained 21 letters and 11 letters in 
best corrected visual acuity (BCVA) (i.e., >4 and >2 lines of vision, respectively).  All other 


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patients who received the full dose of vector, and all of whom had near normal BCVA at 
baseline, did not suffer any clinically significant deterioration in vision post-treatment.  From 
a safety perspective, the GTMP and surgical application method appeared to be generally 
well tolerated.  A second cohort of 8 patients was treated with a single sub-retinal injection of 
1×10^11 gp in 1 eye.  Following favorable results, 3 other investigator-sponsored studies 
were then initiated in which a total of 26 adult male CHM patients received a single sub-
retinal injection of 1×10^11 gp AAV2-REP1 in 1 eye .  The initial results of these trials are 
consistent with improved rod and cone function, despite any negative effects of retinal 
detachment, and the gene therapy was generally well tolerated.  AAV2-REP1 gene therapy 
has been shown to positively affect visual acuity (VA), considering both maintenance and 
stabilization of vision in the majority of subjects, as well as improvement of vision in subsets 
of CHM subjects for up to 2 years and beyond.  This clinical evidence supports the durability 
of the therapeutic effect of AAV2-REP1 for treating CHM.  
These findings lend support to further assessment of AAV2 gene therapy in the treatment of 
CHM.  NightstaRx is currently sponsoring a multi-centre, global Phase 3 study of a single 
subretinal administration of AAV2-REP1 for the treatment of CHM  (NSR-REP-01, the 
STAR study, NCT03496012).
Considering that CHM affects both eyes, exploration of bilateral AAV2-REP1 administration 
is desirable.  As noted, emerging data from non-clinical and clinical studies with AAV2 
vectors demonstrate that these vectors illicit a minimal immune response, including after 
bilateral administration.  The aim of this study is to provide important insight into the safety 
and tolerability of sequential, bilateral treatment with AAV2-REP1.
5.4
Risk / Benefit Assessment
No treatment currently exists for CHM.
The non-clinical studies conducted with AAV2-REP1 showed that it provides efficient and 
functional transgene expression in CHM mouse and human cells, as well as in mouse and 
human RPE and photoreceptors, without overt toxicity.  Results from the 26-week single-
dose combined toxicity/biodistribution study conducted in rats indicate that administration of 
AAV2-REP1 by single sub-retinal injection to both eyes is well tolerated at dose levels of 
1×10^9 and 6×10^9 gp/eye (equal to 1×10^11 and 6×10^11 gp/eye in humans) when 
evaluated 4 and 26 weeks after injection.  Minor reductions in some ERG parameters, ocular 
inflammation, and microscopic signs of retinal/corneal degeneration were observed and were 
considered procedure-related or not biologically significant.
In humans, application of AAV2-REP1 to the surface of the retina requires retinal 
detachment via vitrectomy.  The 2-step procedure employed in delivering AAV2-REP1 sub-
retinally (see Section 9.4), allows the management of unexpected surgical complications of 
retinal detachment before the AAV2-REP1 is applied. Furthermore, since the volume of fluid 
required to detach the fovea is variable, by excluding the vector from the first step, and 
administering the vector after successful retinal detachment has been achieved, a precise, 
consistent dose in terms of genome particles can be applied into the sub-retinal space.  
Nevertheless, sub-retinal injection of AAV2-REP1 carries the risks associated with 
vitrectomy and retinal detachment, which include intra-operative and post-operative 
complications; infection (most notably infectious endophthalmitis); low and elevated IOP; 
choroidal detachment; persistent retinal detachment, retinal tears, holes and breaks; macular 
hole and macular oedema; vitreous haemorrhage; visual impairment; metamorphopsia; and 
photopsia (Park, Marcus et al. 1995, Thompson, Sjaarda et al. 1996, Banker, Freeman et al. 


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1997, Cheng, Azen et al. 2001, Anderson, Fineman et al. 2006, Stein, Zacks et al. 2009, 
Recchia, Scott et al. 2010).  Post-operative intraocular inflammation caused by vitrectomy is 
often associated with transient or sometimes permanent visual impairment.  Another 
complication of vitrectomy is cataract formation, which may require an additional surgical 
procedure (cataract extraction) (Park, Marcus et al. 1995, Cheng, Azen et al. 2001, Recchia, 
Scott et al. 2010).  In addition, the surgical procedure is performed under general anesthesia, 
which includes the risks of dizziness, confusion, nausea and vomiting.
Loss of VA has been observed post-treatment with AAV2-REP1.  This VA loss was 
determined by the Investigators to be plausibly related to the administration of AAV2-REP1, 
however without definitively attributing the cause to either the study procedure or the GTMP.  
Complications from the surgical procedure are expected and are not considered reportable as 
suspected unexpected serious adverse reactions (SUSARs) unless also assessed to be at least 
possibly related to AAV2-REP1.  The Reference Safety Information described in the current 
version of the Investigator’s Brochure for AAV2-REP1 defines visual acuity reduced as an 
expected serious adverse drug reaction, with a frequency of 4/79 (5.1%), defined as common 
(≥1/100 to <1/10).  Loss of VA is considered an identified risk of AAV2-REP1. 
Inflammation is a potential risk of AAV2-REP1. 
See the Investigator’s Brochure for further details.
Thus, although there are risks associated with the administration of the study treatment via 
vitrectomy and retinal detachment, the potential for benefit in the form of improved VA that 
may be provided to subjects with CHM following treatment with AAV2-REP1 provides an 
acceptable risk-benefit profile for participation in this study.


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6
STUDY OBJECTIVES AND ENDPOINTS
6.1
Objectives
The objective of the study is to evaluate the safety of bilateral, sequential sub-retinal 
administration of a single dose of AAV2-REP1 in adult male subjects with CHM.
6.2
Endpoints
The safety of bilateral administration of AAV2-REP1 will be evaluated with the following 
safety measures:

BCVA as measured by the Early Treatment of Diabetic Retinopathy Study (ETDRS) 
chart

Ophthalmic examination assessments (including intraocular pressure [IOP], slit lamp 
examination, lens opacity grading and dilated ophthalmoscopy)

Spectral domain optical coherence tomography (SD-OCT)

Fundus autofluorescence (AF)

Fundus photography

Microperimetry

AE reporting 

Vector shedding post-treatment

Immunogenicity sampling post-treatment 

Vital signs
The following will also be considered secondary efficacy endpoints:

Change from Baseline in BCVA as measured by the ETDRS chart

Change from Baseline in AF

Change from Baseline in SD-OCT

Change from Baseline in microperimetry


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7
INVESTIGATIONAL PLAN
7.1
Overall Study Design
This is a multi-centre, open-label, prospective, 2-period, bilateral interventional safety study 
of AAV2-REP1 in adult male subjects with genetically confirmed CHM.  The study will 
consist of a Screening Period followed by 2 treatment periods (Period 1 and Period 2) with up 
to 9 visits per treatment period.  A second baseline visit prior to surgery of SE2 (Period 2, 
Visit 1) may be required.  A study schematic is presented in Figure 1.
Figure 1
Overall Study Schematic
During the Screening Period, subjects will be assessed for eligibility of both eyes.  The 
Investigator will assign the order in which the eyes are treated (i.e., study eye 1 [SE1]-Period 
1 and study eye 2 [SE2]-Period 2, respectively) with a number assigned to each eye.  This 
will be done in collaboration with the subject; however, the worse eye will generally be 
selected for first treatment.  Study eye selection and eligibility for surgery will then be 
confirmed collectively by the Patient Eligibility Review Committee, which includes the 
Investigator, a consulting surgeon, and the Sponsor (see the Patient Eligibility Review Plan). 
The estimated target interval between the surgical procedures of SE1 and SE2 will be 
determined at the Screening Visit.
Each study eye will be followed for at least 12 months post-treatment, for up to 9 visits per 
treatment period: Visit 1 (Day 0, Injection Day Visit); Visit 2 (Day 1, post-operative follow-
up visit); Visit 3 (Day 3 + 2 Days); Visit 4 (Day 7 -1/+2 Days); Visit 5 (Day 14 ± 3 Days); 
Visit 6 (Month 1 ± 7 Days); Visit 7 (Month 3 ± 14 Days); Visit 8 (Month 6 ±14 Days) and 
Visit 9 (Month 12 ± 14 Days; end of study visit).
At Period 1, Visit 1 (Day 0, Injection Day Visit for SE1), subjects will undergo a vitrectomy 
with retinal detachment and receive a sub-retinal injection of AAV2-REP1 in SE1.  Visits 2-9 
will then be conducted according to the Schedule of Study Procedures, unless Period 2 
commences during the time of follow-up for SE; in which case, the study visits will continue 
following the schedule for SE2.  Ophthalmic assessments during Period 1, Visits 2-9 will be 
performed on both eyes.
The interval between SE1 and SE2 treatment is expected to vary among subjects from  weeks 
to many months.  While the interval will be decided on a case-by-case basis, an effort will be 
made to schedule varying treatment intervals in order to better characterise the 
immunological and safety profile of sequential treatment administration.  Thus, 
approximately 20 subjects each will be treated with a short- (<6 month), medium- (6-12 
months), or long- (>12 month) surgery window between treatment of the first and second 


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eye.  Because the timing of the second surgery visit (Period 2, Visit 1) will vary among 
subjects, the duration of study Period 1 will also be variable.  
The post-operative outcome and visual function of SE1 will inform the scheduling of 
Period 2. Signs of post-operative inflammation or other post-operative sequelae, as judged by 
the Investigator, should be resolved in SE1 for the subject to continue to Period 2.  After 
comprehensive evaluation of all safety assessments and visual functional tests of SE1 and 
SE2, the Investigator will determine the eligibility of the subject and decide the appropriate 
time to schedule SE2, Visit 1.
All baseline ophthalmic assessments must be conducted within 10 weeks prior to SE2 surgery 
(Period 2, Visit 1).  If these assessments are not available within the required timeframe, a 
Baseline Visit must be added to collect current baseline data on SE2.  Study eye selection and 
eligibility for surgery will then be confirmed collectively by the Patient Eligibility Review 
Committee, which includes the Investigator, a consulting surgeon, and the Sponsor (see the 
Patient Eligibility Review Plan).  If, at that time, BCVA in SE2 is <34 ETDRS letters, 
surgery may be performed when, in the opinion of the Patient Eligibility Review Committee: 
1) the subject is found eligible by all other inclusion and exclusion criteria, and 2) the subject 
is considered an appropriate candidate who has potentially modfiable disease.
At Period 2, Visit 1 (Day 0, Injection Day Visit for SE2) subjects will undergo a vitrectomy 
with retinal detachment and sub-retinal injection of AAV2-REP1 in the contralateral, 
untreated eye (SE2).  Post-treatment, subjects will no longer follow the Period 1 visit 
schedule, but will instead attend Period 2, Visits 2-9 according to the Schedule of Study 
Procedures.  Ophthalmic assessments scheduled for Period 2, Visits 2-9 will be performed on 
both eyes. 
Study data will thus be collected for both eyes.  Subjects will be assessed for safety and 
efficacy throughout the study, as outlined in the Schedule of Study Procedures (Table 1).  
The safety evaluation will be based on a full ophthalmic examination (indirect 
ophthalmoscopy, slit lamp examination, IOP, anterior chamber and vitreous inflammation 
grading and lens opacities classification system [LOCS] III cataract grading); fundus 
photography; AE reporting; vector shedding and immunogenicity assessment and vital signs.  
The efficacy evaluation will be based on BCVA (as measured by the ETDRS chart), AF, SD-
OCT and microperimetry.  Any safety information collected as a result of the efficacy 
assessments (e.g., BCVA) will also be used in the overall safety evaluation, as applicable.
Subjects who develop cataracts may undergo cataract surgery if deemed clinically necessary.  
If cataract surgery is performed, it should be carried out at least 4 weeks before Month 12 
(Visit 9) for the respective eye.  The cataract surgery timing must be discussed with the 
medical monitor prior to the procedure.
A subject is considered to have completed the study if he completes Period 2, Visit 9 
(Month 12).  The end of the trial is the date the last subject completes Period 2, Visit 9 
(Month 12) assessments (or early termination [ET] assessments in the event of premature 
discontinuation) or the date of last data collection if the last subject is lost to follow-up.  After 
study completion, subjects will be invited to participate in a long-term safety follow-up study 
that will permit continued safety and efficacy monitoring over an extended period of time 
post-treatment.


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Subjects Previously Treated with AAV2-REP1
Subjects who have previously participated in a clinical trial of AAV2-REP1 for the treatment 
of CHM and have received AAV2-REP1 in one eye may be eligible for participation in this 
study.  In these cases, only the subject’s untreated eye will be assessed for eligibility per 
Section 8.  Following the Screening Visit and determination of eligibility, the subject will 
move directly into Period 2 for treatment of SE2. 
For subjects who received previous treatment with AAV2-REP1, biological samples must be 
available to obtain an adequate immunology profile related to treatment of the first eye in the 
previous study.  These subjects will require Sponsor approval before being enrolled into the 
study.
7.2
Discussion of Design
Since CHM is a bilateral disease, this open-label study is being conducted to assess the safety 
of AAV2-REP1 administered to both eyes of subjects with CHM.  All enrolled subjects will 
receive treatment with AAV2-REP1 at a dose of 1×10^11 AAV2-REP1 gp/eye. Bilateral sub-
retinal administration of AAV2-REP1 was explored in a pre-clinical toxicology study in rats 
and was found to be well tolerated and not associated with drug-related AEs.  Furthermore, 
sequential sub-retinal administration of an AAV2 vector to the contralateral eye in children 
and adults with Leber's congenital amaurosis was demonstrated to be safe and efficacious, 
irrespective of baseline immune status (Bennett, Wellman et al. 2016).
The planned sample size (approximately 60 subjects, with a total of 120 treated eyes) is 
considered sufficient to characterise the immunological, vector shedding, and safety profiles 
of bilateral retinal gene therapy with AAV2-REP1.  A prospective trial period of at least 
12 months following the administration of AAV2-REP1 in each eye is considered a sufficient 
period of time to observe any treatment related safety concerns.  
The dose of vector being employed in this study is based on previous clinical trials using the 
AAV2 vector with a chicken β actin (CBA) promoter (Maguire, High et al. 2009, MacLaren, 
Groppe et al. 2014), and the Phase 1 / 2 investigator-sponsored clinical studies in which 
AAV2-REP1 was administered to subjects with CHM (MacLaren, Groppe et al. 2014) 
(Dimopolous, Hoang et al. 2018, Fischer, Ochakovski et al. 2018, Lam, Davis et al. 2019).
Application of AAV2-REP1 to the retina requires retinal detachment following vitrectomy.  
The risks of vitrectomy and retinal detachment are summarized in Section 5.4.
In line with guidance from regulatory agencies, the treatment intervals were chosen to 
comprehensively characterise the immunological profile following bilateral, sequential 
AAV2-REP1 treatment, and to reflect real-world application of gene therapy.  As noted, both 
preclinical data with AAV2-REP1, and clinical data with sub-retinal administration of an 
AAV2 vector in subjects with Leber's congenital amaurosis, support bilateral administration 
of AAV2-REP1 in humans.
To minimise inflammation resulting from surgery, and potential or unexpected immune 
responses to vector/transgene, all subjects will be given a course of oral 
prednisone/prednisolone, initiated 2 days prior to surgery on both SE1 and SE2.  The regimen 
has been modified from the 17-day protocol established in the Philadelphia AAV gene 
therapy clinical trial (Maguire, Simonelli et al. 2008), with the allowance of an extra 4 days 
for tapering the dose at the end of the course.  The corticosteroid regimen is 1 mg/kg/day (for 
a maximum of 80 mg/daily) prednisone/prednisolone for a total of 10 days (beginning 2 days 
before the vector injection, on the day of injection, and then for 7 days); followed by 


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0.5 mg/kg/day for 7 days; 0.25 mg/kg/day for 2 days; and 0.125 mg/kg/day for 2 days 
(21 days in total).  See Section 9.8 for details.


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8
SELECTION AND WITHDRAWAL OF SUBJECTS
The study will enroll approximately 60 subjects with CHM, for a total of 120 treated eyes 
(see Section 13.1 for discussion of the sample size determination).
8.1
Inclusion Criteria
During the Screening Period, subjects will be found eligible for study participation if they 
meet all of the following inclusion criteria.
1. Are willing and able to give informed consent for participation in the study to have 
both eyes treated
2. Are male and ≥18 years of age
3. Have documentation of a genetically-confirmed diagnosis of CHM
4. Have active disease clinically visible within the macular region of both eyes  
5. Have a BCVA of ≥34 ETDRS letters (20/200 or better Snellen acuity) in both eyes, or 
in the untreated eye, if the other eye was previously treated with AAV2-REP1*
*If previously treated with AAV2-REP1 in an antecedent study, subjects may be 
eligible for participation following Sponsor approval. 
6. For subjects who received treatment with AAV2-REP1 in an antecedent study, have 
biological samples available to complete an adequate immunology profile
8.2
Exclusion Criteria
During the Screening Period, subjects will be found not eligible for study participation if they 
meet any of the following exclusion criteria.
1. Have a history of amblyopia or inflammatory disorder in either eye
2. Are unwilling to use barrier contraception methods or abstain from sexual intercourse 
for a period of 3 months following treatment with AAV2-REP1 in either eye
3. Have had previous intraocular surgery performed within 3 months of the Screening 
Visit in either eye
4. Have any other significant ocular or non-ocular disease/disorder which, in the opinion 
of the Patient Eligibility Review Committee or Investigator, may either put the 
subjects at risk because of participation in the study, or may influence the results of 
the study or the subject’s ability to participate in the study.  This includes but is not 
limited to a potential subject:

with a contraindication to oral corticosteroid (e.g., prednisolone/prednisone)

with clinically significant cataract in either eye

who, in the clinical opinion of the Patient Eligibility Review Committee or 
Investigator, is not an appropriate candidate for sub-retinal surgery (see the Patient 
Eligibility Review Plan)
5. Have participated in another research study involving an investigational product in the 
past 12 weeks or received a gene/cell-based therapy at any time previously, except if 
treated within an antecedent study with AAV2-REP1


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8.3
Subject Withdrawal Criteria
Each subject has the right to withdraw from the study at any time without prejudice.  In 
addition, the Investigator may discontinue a subject from the study at any time if the 
Investigator considers it necessary for any reason, including:

Significant protocol deviation

Significant non-compliance with study requirements

AE which results in an inability to continue to comply with study assessments

Lost to follow up

Other (to be specified on the electronic case report form [eCRF]).
o
If severe ocular inflammation (e.g. endophthalmitis) or any other post-
operative sequela that is unresponsive to treatment occurs in SE1, the second 
eye should not be treated.  However, Period 1 should be continued for 
monitoring of safety for the duration of the study.
o
At the discretion of the Investigator, if a persistent post-operative complication 
or poor visual function outcome presents post-operatively in SE1, the second 
eye may not be treated. However, Period 1 should be continued for monitoring 
of safety for the duration of the study.
In the event that a subject discontinues the study, the reason for withdrawal is to be recorded 
in the eCRF, and the site should use every reasonable effort to ensure that an end of trial (ET) 
Visit is conducted as outlined in the Schedule of Study Procedures (Table 1).  If the subject is 
withdrawn due to an AE, the Investigator will arrange for follow-up until the event has 
resolved, subsided, stabilised, or the subject withdraws consent or is lost to follow-up.  For 
subjects who withdraw consent, data will be collected through their last available study visit.  
Subjects withdrawn from the study may possibly be replaced.  Subjects who withdraw from 
the study must undergo the ET Visit assessments. See Table 1.
Withdrawal from the study will not result in the exclusion of a subject’s data acquired up to 
the point of withdrawal.


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9
STUDY TREATMENT
9.1
Treatments Administered
Eligible subjects will undergo vitrectomy and retinal detachment in each eye.  At Period 1, 
Visit 1 (Day 0, the Injection Day Visit for SE1), subjects will receive in SE1 a sub-retinal 
injection of up to 0.1 mL of study drug containing 1×10^11 AAV2-REP1 gp.  At Period 2, 
Visit 1 (Day 0, the Injection Day Visit for SE2), subjects will receive in SE2 up to 0.1 mL of 
AAV2-REP1 via sub-retinal injection.
9.2
Description of Study Drug
The AAV2 vector contains recombinant human cDNA encoding REP1 (AAV2-REP1).  The 
vector genome (AAV2-CBA-hREP1-WPRE-BGH) is comprised of a strong constitutive 
expression cassette, a hybrid CBA promoter, the human cDNA encoding REP1, a modified 
woodchuck hepatitis post-transcriptional regulatory element (WPRE) sequence, and a bovine 
growth hormone polyadenylation (BGH-polyA) sequence flanked by AAV2 inverted 
terminal repeats.  The cDNA fragment was originally isolated from a human retinal cDNA 
library from unaffected individuals.
The AAV2-REP1 drug product is formulated in a sterile, 20 mM Tris-buffered solution, 
pH 8.0, and contains 1 mM MgCl2, 200 mM NaCl and 0.001% PF68.  The drug product is a 
clear to slightly opalescent, colourless, sterile-filtered suspension with a target concentration 
of 1×10^12 gp/mL.
9.3
Packaging, Labeling, and Storage
AAV2-REP1 is currently supplied in sterile single-use vials, stoppered and capped.  A total 
of 0.3 mL vector suspension will be supplied for each eye to be treated.  Prior to shipment, 
each vial will be placed in a labeled secondary container.  The drug product is to be stored at 
<−60oC (<−76oF) in a controlled access, temperature monitored freezer.
The GTMP will be labeled in compliance with regulatory standards. 
9.4
Vitrectomy Procedure and Injection of AAV2-REP1
Injection of AAV2-REP1 is to be performed by an appropriately qualified and experienced 
retinal surgeon.  All surgeons must have completed surgical training and obtained 
certification by NightstaRx to perform the study procedure, before treating a study 
participant.  Training conducted as part of a previous AAV2-REP1 clinical trial or alternative 
AAV NightstaRx-sponsored gene therapy trial, can be used as the basis for approval.
Due to the complexity and unpredictability of detaching the retina in CHM, in which the 
retina and choroid can be extremely thin and fused in places, a modification to the technique 
of sub-retinal gene therapy has been developed.  This involves performing the vector delivery 
in 2 steps after vitrectomy.  An advantage of a 2-step procedure is that any unexpected 
complications of retinal detachment can be managed conservatively, minimising concerns 
about the vector escaping into the vitreous.  Further, the injection could be deferred until a 
later date if, for instance, a macular hole was created which required treatment with gas.  
Also, since the volume of fluid required to detach the fovea is variable, by removing the 
vector from the first step, a precise consistent dose in terms of genome particles can still be 
applied into the sub-retinal space.


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Initially, subjects will undergo a standard vitrectomy and detachment of the posterior hyaloid 
in the respective study eye (Figure 2).  All surgeries will be conducted using the standard 
BIOM vitrectomy system.  A 23-gauge sutured approach is usually favored to avoid any 
potential risks of wound leakage.  The retina will be detached with 0.1-0.5 mL of balanced 
salt solution (BSS) injected through a 41-gauge sub-retinal cannula connected to a vitreous 
injection set.
In the second step of the procedure, the BSS cannula is removed from the eye and AAV2-
REP1 is prepared for injection.  A dose of up to 1.0×10^11 gp of AAV2-REP1 is injected 
into the sub-retinal space through the same entry site.  To improve visualization of the vector 
and facilitate dosing, surgeons are given the option of adding a minute quantity of trypan blue 
ophthalmic solution (~6 µL) to the vector solution.  See the GEMINI Surgical Manual for 
further details.
Prior to administration, the vector needs to be primed in the 1-mL syringe to avoid formation 
of air bubbles, and a connector is used so that the 1-mL syringe can be connected to the 
constant pressure line of the vitrectomy machine.  The sub-retinal injection will target any 
area of the macula, but also include the fovea, if possible.  In each case, the vector is injected 
so that the sub-retinal fluid overlies all edge boundaries of the central region that has yet to 
undergo chorioretinal degeneration, as identified by fundus AF.  After wound closure, care 
will be taken to dispose of all irrigating fluids that may have passed through the eye to limit 
potential vector spread.  See the GEMINI Surgical Manual for further details.
Subjects will be carefully monitored for the occurrence of AEs peri- and post-operatively.  
All AEs, irrespective of relationship to the study drug and/or the surgical procedure, will be 
captured in the subject’s medical record and reported in the eCRF.
Air fluid exchange during pars plana vitrectomy is not required for this surgical procedure.  
However, if an air fluid exchange is considered necessary, patients must be instructed to 
avoid air travel, travelling to high elevations or scuba diving until the air bubble has 
completely dissipated from the eye.  It may take one week or more following injection for the 
air bubble to dissipate.  A change in altitude while the air bubble is still present can result in 
irreversible vision loss.  The dissipation of the air bubble should be verified through 
meticulous ophthalmic examination.
Figure 2
Vitrectomy and Sub-Retinal Injection of AAV2 Vector
A
B
(A) A standard vitrectomy through the BIOM operating system to remove the vitreous gel is followed by (B) a 
2- step procedure: 1) retinal detachment by injection of BSS; 2) injection of a volume of up to 0.1 mL vector 
suspension through a 41-gauge cannula into the sub-retinal space.


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9.5
Randomisation
Not applicable for this study.
9.6
Study Masking
Not applicable for this study.
9.7
Study Drug Accountability
Records of the receipt and dispensing of study drug will be kept by each study centre until the 
end of the study to provide complete accounting of all used and unused study drug.  
Dispensation logs will be checked by the Sponsor (or its designee).  Study centres will 
destroy all used vials in accordance with local regulations, and will return all unused study 
drug to the Sponsor (or its designee) at the end of the study.  Final drug accountability will be 
verified by the Sponsor (or its designee).
9.8
Concomitant Therapy
Subjects cannot have participated in another research study involving an investigational 
product in the past 12 weeks or received a gene/cell-based therapy at any time previously, 
except if treated within another study with AAV2-REP1.
Throughout the study, subjects may be prescribed any concomitant medications, procedures, 
or treatments deemed necessary to provide adequate supportive care.  Details of concomitant 
medications, procedures and/or treatments will be collected during the Screening Period and 
updated at every study visit (including the ET Visit, if applicable).  Concomitant medications 
(including oral corticosteroid), procedures and/or treatments taken during the study are to be 
recorded in the subject’s medical records and eCRF; an exception to this is any medication 
used in the course of conducting study assessments (e.g., ophthalmic dyes, topical 
anaesthesia, dilating eye drops).
In addition, all subjects will be prescribed a course of oral corticosteroids. 
The oral prednisone / prednisolone regimen has been modified from the 17-day protocol 
established in the Philadelphia voretigene neparvovec-rzyl gene therapy clinical trial for 
treatment of patients with confirmed bi-allelic RPE65 mutation-associated retinal dystrophy 
(Maguire, Simonelli et al. 2008), with the allowance of an extra 4 days for tapering the dose 
at the end of the course.  The corticosteroid regimen is 1 mg/kg/day prednisone/prednisolone 
(not to exceed 80 mg/daily) for a total of 10 days (beginning 2 days before the vector 
injection, on the day of injection, and then for 7 days); followed by 0.5 mg/kg/day for 7 days; 
0.25 mg/kg/day for 2 days; and 0.125 mg/kg/day for 2 days (21 days in total; Study Day (SD) 
-2 through SD18).  This regimen is outlined in detail below. 
Full Corticosteroid Regimen
Prednisone or prednisolone administered as follows:
SD -2 through SD 7 (10 days): 1 mg/kg by mouth daily (not to exceed 80 mg/daily)
SD 8 through SD 14 (7 days): 0.5 mg/kg/day by mouth daily
SD 15 through SD 16 (2 days): 0.25 mg/kg/day by mouth daily
SD 17 through SD 18 (2 days): 0.125 mg/kg/day by mouth daily
For SE1, the Full Corticosteroid Regimen should be initiated as described.  


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For SE2 surgeries that occur on day 21 or later, the Full Corticosteroid Regimen should be 
initiated as described, beginning 2 days before the scheduled SE2 surgery.
If SE2 is to be performed prior to Study Day 21, then the Full Corticosteroid Regimen should 
be initiated 2 days prior to the scheduled surgery for SE2, and this will supersede the steroid 
taper in progress for SE1.  The full 21 days of treatment will then be completed for SE2.  
Details of corticosteroid usage will be captured by each subject in a diary card. 
If inflammation is observed in the study eye, and in the opinion of the Investigator additional 
treatment with corticosteroid medication is indicated, corticosteroid therapy may be increased 
during the taper period (to a maximum of 1 mg/kg/day), may be reinitiated following 
completion of the taper, and/or may be supplemented by intraocular corticosteroids.  
9.9
Treatment Compliance
This study involves a single sub-retinal injection of a volume of up to 0.1 mL AAV2-REP1 in 
each eye.  Measure of treatment compliance with AAV2-REP1 is therefore not necessary.
Compliance with the use of prednisone/prednisolone will be captured in the eCRF; subject 
diary cards will be provided to participants prior to each surgery.


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10 STUDY VISITS AND PROCEDURES
The schedule of study procedures is presented in Table 1.
At each study visit, an attempt should be made to perform all procedures in both eyes.
10.1 Screening Period
The Investigator will explain the study purpose, procedures and subject responsibilities to 
each potential study subject.  The subject's willingness and ability to meet the protocol 
requirements will be determined.
Prior to any study-specific procedure, written informed consent will be obtained.  The subject 
will sign and date 1 copy of the consent form in the presence of the Investigator or his/her 
designee.  The original signed form will be retained at the study site and an additional copy 
will remain in the subject’s medical records; a copy will also be given to the subject.
After informed consent has been obtained, the subject will be allocated a subject identifier 
and evaluated to determine study eligibility.  
Screening procedures will consist of the following (all ophthalmic assessments will be 
conducted on both eyes):

Demography, medical and ocular history

Vital signs

Weight

Vector shedding sampling – blood, tears (both eyes), urine, saliva

Immunogenicity sampling

BCVA 

Microperimetry

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT 

AF

7-field colour fundus photography (including stereo photographs for fields 1, 2, and 3)

AE and serious adverse event (SAE) monitoring

Concomitant medication, procedures, and treatment review
Subjects who meet all of the inclusion criteria and none of the exclusion criteria will be 
assigned a subject number and will be enrolled into the study.  The Investigator will assign 
the order in which the eyes are treated (i.e., SE1 and SE2, respectively).  This will be done in 
collaboration with the subject; however, the worse eye will generally be selected for 
treatment first.  Study eye selection and eligibility for surgery will then be confirmed 
collectively by the Patient Eligibility Review Committee, which includes the Investigator, a 
consulting surgeon, and the Sponsor (see the Patient Eligibility Review Plan). The target 
interval between the surgical procedures of SE1 and SE2 will be determined at this visit.
For each subject, the interval between SE1 and SE2 treatment is expected to range from  
weeks to months.  While this interval will be decided on a case-by-case basis, an effort will 


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be made to schedule varying treatment intervals in order to better characterise the 
immunological and safety profile of sequential treatment administration and to reflect the 
anticipated real-world application of the study drug.  Thus, approximately 20 subjects will be 
treated with a short- (<6 month), medium- (6-12 months), or long- (>12 month) duration 
surgery window between treatment of the first and second eye.  Because the timing of the 
second surgery visit (Period 2, Visit 1) will vary among subjects, the duration of study Period 
1 will also be variable.  To avoid unnecessary study visits, every effort should be made by the 
Investigator to schedule Period 2, Visit 1 to coincide with a planned Period 1 post-treatment 
visit. 
The next study visit (i.e., surgery and dosing, Period 1, Visit 1) is to be scheduled within 
10 weeks of the Screening Period.  Subjects will attend Period 1, Visits 2-9 unless the second 
surgery visit (Period 2, Visit 1) begins, which will mark the start of study Period 2.  Because 
the timing of the second surgery visit (Period 2, Visit 1) will vary among subjects, the 
duration of study Period 1 will also be variable.
Subjects will be treated with oral corticosteroids (e.g., prednisolone / prednisone) during the 
peri-operative period for each eye.  They will be instructed to start taking the drug 2 days 
before their surgery at Period 1, Visit 1 for SE1.  For SE1, the Full Corticosteroid Regimen 
should be initiated, as described in Section 9.8.
For SE2 surgeries that occur on Day 21 or later, the Full Corticosteroid Regimen should be 
initiated as described, begining 2 days before the scheduled surgery. 
If SE2 is to be performed prior to Study Day 21, then the Full Corticosteroid Regimen should 
be initiated 2 days prior to the scheduled surgery for SE2, and this will supersede the steroid 
taper in progress for SE1.  The full 21 days of treatment will then be completed for SE2. 
Subjects will be issued a diary card to capture corticosteroid compliance.  Subjects will also 
be instructed to use barrier contraception or abstain from sexual intercourse for a period of 
3 months from the time they are treated.
The schedule of procedures is identical for Periods 1 and 2; during each period, procedures 
are to be conducted in both SE1 and SE2.
If a subject has previously participated in a clinical trial of AAV2-REP1 for the treatment of 
CHM and has received AAV2-REP1 in one eye, the untreated eye will be assigned as SE2 at 
the Screening Visit and the subject will immediately enter Period 2.
10.1.1 Baseline Visit, Period 2 (<10 Weeks from Visit 1, Period 2 [SE2])
The following baseline assessments for Period 2 must be performed within 10 weeks prior to 
surgery for SE2 (Visit 1, Period 2).  If the assessments have not been performed within a 
regularly scheduled visit within this window, an additional visit is required. 

Vital signs

Weight

Vector shedding sampling – blood, tears (both eyes), urine, saliva

Immunogenicity sampling

BCVA 

Microperimetry


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
Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT 

AF

7-field colour fundus photography (including stereo photographs for fields 1, 2, and 3)

AE and SAE monitoring

Concomitant medication, procedures and treatment review
Eligibility for SE2 surgery will then be confirmed collectively by the Patient Eligibility 
Review Committee, which includes the Investigator, a consulting surgeon, and the Sponsor 
(see the Patient Eligibility Review Plan). 
If BCVA in SE2 is <34 ETDRS letters, surgery may be performed when, in the opinion of the 
Patient Eligibility Review Committee: 
1. the subject is found eligible by all other inclusion and exclusion criteria, and
2. the subject is considered an appropriate candidate who has potentially modifiable 
disease.
10.2 Period 1 and 2, Visit 1 (Day 0, Injection Day Visit)
At Visit 1 (Day 0, Injection Day Visit), the following assessments will be performed:

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination 

AE/SAE monitoring

Concomitant medication, procedures, and treatment review (including review of the 
corticosteroid diary card)
It may be necessary for subjects to undergo a pre-surgical workup according to local hospital 
procedures (e.g., blood draw for anesthetist) which are outside of the protocol-defined 
assessments.  Any assessments performed outside of the protocol will not be collected as part 
of the study analysis.
Subjects will then undergo vitrectomy and receive a volume of up to 0.1 mL sub-retinal 
injection of AAV2-REP1, containing 1×10^11 gp (see Section 9.4 for details).  Subjects will 
be carefully monitored for the occurrence of AEs during the procedure.  Subjects will return 
to the site 1, 3, 7 and 14 days after surgery for post-operative follow-up (Visit 2 [Day 1], 
Visit 3 [Day 3 + 2 Days], Visit 4 [Day 7 -1/+2 Days] and Visit 5 [Day 14 ± 3 Days], 
respectively).
The post-operative outcome and visual function of SE1 will inform the scheduling of 
Period 2.  Significant signs of post-operative inflammation or other post-operative sequelae, 
as judged by the Investigator, should be resolved in SE1 for the subject to continue to 
Period 2.  After a comprehensive evaluation of all safety assessments and visual functional 
tests of SE1 and SE2, the Investigator will decide the appropriate time to schedule SE2, 
Visit 1. If severe ocular inflammation or any other post-operative sequela that is unresponsive 
to treatment occurs in SE1, the second eye should not be treated. However, Period 1 should 
be continued for monitoring of safety for the duration of the study.


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To allow accurate characterisation of AAV2-REP1 safety and immunogenicity profile, 
treatment of SE2 should not occur if a previous intraocular surgery was performed on the 
same eye within 3 months of the planned treatment date.  If intraocular surgery has occurred 
within 3 months, treatment of SE2 should be delayed until such a time that the 3-month 
interval has elapsed and there is complete post-operative recovery of the eye.
10.3 Period 1 and 2, Visit 2 (Post-operative Day 1)
For Visit 2, ocular assessments and procedures will be performed on each eye.  At Visit 2 
(Post-operative Day 1), subjects will return to the site for their first post-operative visit for the 
same eye that underwent surgery at Visit 1 (Period 1 [SE1] or Period 2 [SE2]).  The 
following assessments will be performed:

Vital signs

Vector shedding sampling – blood, tears (both eyes), urine, saliva

Immunogenicity sampling 

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AE/SAE monitoring

Concomitant medication, procedures, and treatment review (including review of the 
corticosteroid diary card)
10.4 Period 1 and 2, Visit 3 (Post-operative Day 3 + 2 Days)
For Visit 3 (Post-operative Day 3 + 2 Days), ocular assessments and procedures will be 
performed on each eye.  At Visit 3 (Day 3+ 2 Days), subjects will return to the site for their 
second post-operative visit for the same eye that underwent surgery at Visit 1 (Period 1 [SE1] 
or Period 2 [SE2]).  The following assessments will be performed:

Vital signs

Vector shedding sampling – blood, tears (both eyes), urine, saliva

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

AE/SAE monitoring

Concomitant medication, procedures and treatment review (including review of the 
corticosteroid diary card)
10.5 Period 1 and 2, Visit 4 (Post-operative Day 7 -1/+2 Days)
For Visit 4 (Post-operative Day 7 -1/+2 days), ocular assessments and procedures will be 
performed on each eye.  At Visit 4 (Day 7 -1/+2 days), subjects will return to the site for their 
third post-operative visit for the same eye that underwent surgery at Visit 1 (Period 1 [SE1] 
or Period 2 [SE2]).  The following assessments will be performed:

Vector shedding sampling – blood, tears (both eyes), urine, saliva


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
Immunogenicity sampling

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AF

AE/SAE monitoring

Concomitant medication, procedures and treatment review (including review of the 
corticosteroid diary card)
10.6 Period 1 and 2, Visit 5 (Post-operative Day 14 ± 3)
For Visit 5 (Post-operative Day 14 ± 3 days), ocular assessments and procedures will be 
performed on each eye.  At Visit 5 (Day 14 ± 3 days), subjects will return to the site for their 
fourth post-operative visit for the same eye that underwent surgery at Visit 1 (Period 1 [SE1] 
or Period 2 [SE2]).  The following assessments will be performed:

Vector shedding sampling – blood, tears (both eyes), urine, saliva 

Immunogenicity sampling 

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AF 

AE/SAE monitoring

Concomitant medication, procedures and treatment review (including review of the 
corticosteroid diary card)
10.7 Period 1 and 2, Visit 6 (Month 1 ± 7 Days) and Visit 7 (Month 3 ± 14 Days)
For Visit 6 (Month 1 ± 7 days) and Visit 7 (Month 3 ± 14 days), ocular assessments and 
procedures will be performed on each eye.  The following assessments will be performed:

Vector shedding sampling – blood, tears (both eyes), urine, saliva

Immunogenicity sampling 

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AF

Microperimetry

AE/SAE monitoring


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
Concomitant medication, procedures and treatment review (including review/return of 
the corticosteroid diary card at Visit 6)
10.8 Period 1 and 2, Visit 8 (Month 6 ± 14 Days) and Visit 9 (Month 12 ± 14 Days)*
For Visit 8 (Month 6 ± 14 days) and Visit 9 (Month 12 ± 14 days), ocular assessments and 
procedures will be performed on each eye.  The following assessments will be performed:

Vital signs (Visit 9, Month 12 only)

Immunogenicity sampling 

BCVA

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AF

Microperimetry

7-field colour fundus photography (including stereo photographs for fields 1, 2, and 3) 
(Visit 9, Month 12 only)

AE/SAE monitoring

Concomitant medication, procedures and treatment review
10.9 Early Termination (ET Visit)
In the event that a subject discontinues the study at any time, the site should use every 
reasonable effort to ensure that an ET Visit is conducted.  The following assessments should 
be performed (all ophthalmic assessments will be conducted on both eyes):

Vital signs

Vector shedding sampling – blood, tears (both eyes), urine, saliva (only if the ET 
Visit occurs within 3 months post-treatment)

Immunogenicity sampling 

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AF

Microperimetry

7-field colour fundus photography (including stereo photographs for fields 1, 2, and 3) 

AE/SAE monitoring

Concomitant medication, procedures and treatment review (including review/ return 
of the corticosteroid diary card, if applicable)


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10.10 Unscheduled Visits
If clinically indicated, subjects may need to return to the site for an unscheduled visit.  At a 
minimum, the following assessments will be performed (ophthalmic assessments will be 
conducted on any eye deemed appropriate by the Investigator):

BCVA 

Full ophthalmic examination, including IOP, a slit-lamp examination, lens opacity, 
and dilated fundus examination

SD-OCT

AE/SAE monitoring

Concomitant medication, procedures and treatment review (including review / return 
of the corticosteroid diary card, if applicable)


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11 ASSESSMENT OF SAFETY
11.1 Evaluation, Recording, and Reporting Adverse Events
11.1.1 Definitions
11.1.1.1 Adverse Event
An AE is any untoward medical occurrence in a clinical investigation subject, which does not 
necessarily have a causal relationship with the study drug/surgical procedure.  An AE can 
therefore be any unfavourable and unintended sign (including an abnormal laboratory 
finding), symptom, or disease temporally associated with the use of the study drug/surgical 
procedure, whether or not related to the investigational product or with the surgical procedure 
described in this protocol.
AEs are to also include any pre-existing condition (other than CHM) or illness that worsens 
during the study (i.e., increases in frequency or intensity).
Worsening of the signs, symptoms and visual function related to CHM is not to be considered 
an AE unless, in the opinion of the Investigator, the changes are not consistent with the 
natural progression of the disease. 
11.1.1.2 Serious Adverse Event
An SAE is defined as any untoward medical occurrence that:

Results in death

Is life-threatening

Requires inpatient hospitalisation or prolongation of existing hospitalisation

Results in persistent or significant disability/incapacity

Is a congenital anomaly/birth defect

Results in vision loss or is vision threatening

Is another important medical event(s)
The term ‘life-threatening’ in the definition of ‘serious’ refers to an event in which the 
subject is at risk of death at the time of the event.  It does not refer to an event that 
hypothetically might cause death if it were more severe.
Hospitalisation that was pre-scheduled prior to obtaining the consent to participate in the 
study or for an elective procedure or routinely scheduled treatment for a pre-existing 
condition which has not worsened, does not constitute an SAE.
A prescheduled or elective procedure or a routinely scheduled treatment will not be 
considered an SAE, even if the subject is hospitalized.  The study site must document all of 
the following:

The prescheduled or elective procedure or routinely scheduled treatment was 
scheduled (or was on a waiting list to be scheduled) prior to obtaining the subject’s 
consent to participate in the study.

The condition requiring the prescheduled or elective procedure or routinely scheduled 
treatment was present before and did not worsen or progress in the opinion of the 
Investigator between the subject’s consent to participate in the study and the time of 
the procedure or treatment.


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
The prescheduled or elective procedure or routinely scheduled treatment is the sole 
reason for the intervention or hospital admission.
If a subject is hospitalized due to local requirements for administration of the study treatment, 
the hospitalization should not be considered an SAE unless one of the requirements in 
Section 11.1.1.2 is met.
Other events that may not result in death, are not life-threatening or do not require 
hospitalisation, may be considered an SAE when, based upon appropriate medical judgment, 
the event may jeopardise the subject and may require medical or surgical intervention to 
prevent 1 of the outcomes listed above.
Vision Loss to be Reported as Serious Adverse Events
Vision Loss not to be Reported as an SAE:

Surgery-related BCVA decrease of ≥15 letters on ETDRS chart occurring within 1 
day of surgery, but recovering / resolving at post-operative Days 7 and 14. 
Vision Loss or Vision-Threatening Event to be Reported as an SAE:

Surgery-related BCVA decrease of ≥15 letters on ETDRS chart that occurs within 1 
day of surgery and that has not recovered* by the 1-Month Visit.

A decrease in BCVA of ≥15 letters on ETDRS chart that occurs within 1 day of 
surgery, however, in the Investigator’s opinion:
o
Has an evolution not consistent with the expected post-operative course; 
o
May be attributable to a complication that occurred during surgery, or another 
untoward event, or the study drug; 
o
Actually or potentially requires any surgical or medical intervention to prevent 
permanent loss of vision.

Non-surgery-related, sustained (>48 hours duration) decrease from baseline in BCVA 
of ≥15 letters on ETDRS chart.
*Recovery / Resolution of BCVA is defined as a return to baseline BCVA within 5 letters on 
the ETDRS chart.
11.1.2 Recording of Adverse Events
AEs/SAEs will be collected from the time the subject provides written informed consent 
(Screening Visit) through Period 2, Visit 9 (Month 12 ± 14 Days or ET Visit, if applicable). 
Subjects will be questioned on the occurrence of an AE at every visit including any 
unscheduled visit, by using non-leading questioning such as ‘How have you been since the 
last visit?’  
All AEs occurring during the study observed by the Investigator or reported by the subject, 
whether or not attributed to study drug or the surgical procedure, will be recorded in the 
subject’s medical records and in the eCRF.  Any clinically significant changes in laboratory 
results or vital sign measurements (as determined by the Investigator) are to be recorded as an 
AE.
Ocular AEs are to be recorded on a per-eye basis.


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The following information will be recorded in the eCRF for each AE: description, date of 
onset and end date, outcome, severity, assessment of relatedness to study drug/study 
procedure, the action taken and confirmation of whether the event is considered serious (see 
Section 11.1.1.2 for the definition of seriousness).  Follow-up information should be provided 
as necessary (see Section 11.1.3 for specifics on follow-up procedures).
AE severity and relationship to the study drug or the surgical procedure will be assessed at 
the site by the Investigator or a medically qualified designee.
AE Severity
The severity of events will be assessed on the following scale:
1 = mild (awareness of sign or symptom, but easily tolerated) 
2 = moderate (discomfort sufficient to cause interference with normal activities)
3 = severe (incapacitating, with inability to perform normal activities).
AE Relationship
When assigning relatedness of the AE, consideration will be given to whether there is a 
plausible relationship to either the study drug or the surgical procedure.  The following are 
definitions of relatedness that will be used in this study: 
Unrelated: 
The event is not reasonably related in time to the administration 
of the study drug/surgical procedure, or exposure of the study 
drug/surgical procedure has not yet occurred, or biologic 
plausibility does not exist
Related: 
A reasonable possibility exists that the study drug / study 
procedure caused the AE. A suspected AE can be further 
defined by:
Possibly related: 
a relationship is clinically or biologically 
reasonable relative to the administration of the study 
drug/surgical procedure, but the event could have been due to 
another equally likely cause
Probably related: 
a relationship is clinically/biologically 
reasonable relative to the administration of the study 
drug/surgical procedure, and the event is more likely explained 
by exposure to/administration of the study drug/surgical 
procedure than by other factors and causes
Definitely related: 
there is a causal relationship of the onset 
of the event, relative to administration of the study 
drug/surgical procedure and there is no other cause to explain 
the event.
When a relationship is determined to exist, the Investigator or medical designee will further 
define if that relationship is to the study drug, the study procedure, both, or unknown.
11.1.3 Follow-up of Adverse Events
AEs will be followed until the subject has recovered or the subject’s participation in the study 
is complete.


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Subjects who are withdrawn from the study as a result of a drug-related AE will be followed 
up until the event has resolved, subsided, stabilised, or the subject withdraws consent or is 
lost to follow-up.
All SAEs, regardless of attribution to study drug or the surgical procedure, should be 
followed-up until the event has resolved, subsided, stabilised, or the subject withdraws 
consent or is lost to follow-up.  The Sponsor (or designee) will follow up on SAE reports to 
completion.  Investigators are expected to provide the requested additional information for a 
complete assessment and documentation of the SAE reports in a timely manner.
11.1.4 Reporting of Serious Adverse Events
The Investigator shall immediately (within 24 hours of learning of the event) report any SAE 
to the Sponsor (or its designee) by completing and emailing the SAE form.  For reporting 
purposes, the date of SAE form submission by the Investigator to the Sponsor will be 
designated as Day 0.. The initial report shall be promptly followed up with a more detailed 
report providing specifics about the subject and the event.  Copies of hospital reports, autopsy 
reports, and other documents should be provided (if applicable).
The Sponsor will report SAEs and SUSARs to investigative sites, the Institutional Review 
Board (IRB)/Independent Ethics Committee (IEC), and regulatory authorities in compliance 
with current regulations.  All cases that are fatal or life-threatening will be reported 
immediately after the Sponsor received the initial report from the Investigator.  All non-fatal 
or non-life-threatening cases will be reported within a maximum of 15 days after the initial 
Investigator’s report.  The Sponsor will also provide periodic safety reports to IRBs/IECs and 
regulatory authorities, as applicable.  Follow-up SAE reports will be submitted within 15 
days of receiving the information.
A sample SAE form is provided in the Study Operations Manual, along with the SAE 
reporting contact information.
11.1.5 Data Monitoring Committee
An independent Data Monitoring Committee (DMC) will be used in this study to safeguard 
the safety and interests of study subjects and assess the safety and risk/benefit of the gene 
therapy intervention during the trial.
At regular intervals during the study, the DMC will review the progress and accrued study 
data and provide advice to the Sponsor on the efficacy and safety aspects of the study.  The 
DMC will also inform the Sponsor if there is a consensus that the ongoing data show that the 
gene therapy, its method of administration, and/or the study design are no longer in the best 
interests of study subjects.  Details of DMC organisation and responsibilities are included 
within a DMC charter.
11.2 Pregnancy
Any pregnancy that occurs during the clinical study in a female partner of a study subject 
should be recorded on a Pregnancy Notification Form.  Consent from the pregnant partner is 
required prior to the collection of personal data.  However, the Investigator shall immediately 
(within 24 hours of learning of the event) report the pregnancy, with at least preliminary data, 
to the Sponsor (or its designee) by completing and emailing the Pregnancy Notification Form.  
Follow-up information including delivery, termination, and any congenital abnormality 
should be reported within 24 hours of the study site staff becoming aware. 


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11.3 Stopping Criteria

Severe ocular inflammation (e.g. endophthalmitis) that is unresponsive to treatment 
should result in the affected subject not being treated in the second eye. However, the 
subject will continue to be followed for safety for the duration of the study. 

At the request of the DMC, the trial may be stopped based on specific safety concerns 
(i.e., SUSARs), not defined in the Investigator’s Brochure (Section 6.3).

The study may be discontinued if the Sponsor deems it necessary for medical, safety, 
regulatory, business, or other reasons consistent with applicable laws or regulations.
11.4 Laboratory Assessments
11.4.1 Vector Shedding
Blood, tears (both eyes), urine and saliva samples will be collected at the times indicated in 
Table 1 and tested using an appropriate assay for evidence of vector shedding and dispersion.  
Samples will be sent to a central laboratory for analysis.  Refer to the Study Operations 
Manual for details on the shipping and handling of samples.
11.4.2 Immunogenicity
For the evaluation of immunogenicity, blood will be collected at the times indicated in Table 
1.  Selected samples will be analysed.  
Immunoassays are planned to assess cell-based and antibody-based immune responses 
against AAV2-REP1.  Enzyme-linked immunospot (ELISPOT) assays will be used for the 
assessment of T-cell-mediated immune responses to transgene product (REP1) and the 
capsid.  Antibody responses against the capsid and transgene product will be measured using 
a cell-based neutralization assay and an enzyme-linked immunosorbent assay (ELISA), 
respectively.
Refer to the Study Operations Manual for details on the shipping and handling of samples.
Remaining samples may be stored for up to 15 years or per local regulations. 
11.5 Vital Signs
Vital signs (pulse and systolic and diastolic blood pressure) will be taken at the times 
indicated in Table 1.  Vital signs should be taken after the subject is seated for at least 
5 minutes.
11.6 Best-Corrected Visual Acuity
To evaluate changes in BCVA over the study period, BCVA will be assessed using the 
ETDRS VA chart and performed for both eyes at the times indicated in Table 1.
The BCVA test should be performed prior to pupil dilation, and distance refraction should be 
carried out before BCVA is measured.  Initially, letters are read at a distance of 4 metres from 
the chart.  If <20 letters are read at 4 metres, testing at 1 metre should be performed.  BCVA 
is to be reported as number of letters read correctly by the subject.
For BCVA, assessors will be appropriately qualified for conducting the assessment.
If a subject is not able to perform an assessment due to poor VA, it will be documented 
accordingly in the eCRF and will not be recorded as a protocol deviation.


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11.7 Microperimetry
Microperimetry will be conducted on both eyes at the times indicated in Table 1.
Microperimetry will be conducted by certified technicians to assess changes in retinal 
sensitivity within the macula.  All microperimetry images will be sent by the sites to a 
Central Reading Centre (CRC) for review.  For complete technical specifications for 
microperimetry, refer to the Study Operations Manual (which will include procedures from 
the CRC regarding how measurements are to be taken).
11.8 Fundus Autofluorescence
To assess changes in the area of viable retinal tissue, fundus AF will be performed on both 
eyes at the times indicated in Table 1.
All fundus AF images will be performed by certified technicians at the site after dilation of 
the subject’s pupil and sent to a CRC for review; the CRC will enter the data into the 
electronic data capture (EDC) system.  For complete technical specifications for AF, refer to 
the Study Operations Manual (which will include procedures from the CRC regarding how 
measurements are to be taken).
11.9 Spectral Domain Optical Coherence Tomography 
SD-OCT will be performed on both eyes at the times indicated in Table 1.
SD-OCT measurements will be taken by certified technicians at the site after dilation of the 
subject’s pupil.  All OCT scans will be submitted by the sites to a CRC where the scans will 
be evaluated; the CRC will enter the data into the EDC system.  SD-OCT will be used to 
quantify integrity of the ellipsoid zone and reduction in the signal from the outer nuclear 
layer and choroid. In addition, foveal changes will be assessed.  For complete technical 
specifications for SD-OCT, refer to the Study Operations Manual (which will include 
procedures from the CRC regarding how measurements are to be taken).
11.10 Full Ophthalmic Examination
A full ophthalmic examination will be performed for both eyes at the times indicated in Table 
1.
Each ophthalmic examination will include IOP, slit lamp examination, lens opacity grading, 
and dilated ophthalmoscopy.  The same slit lamp machine and lighting conditions should be 
used across study visits for any given subject.
In addition to the parameters listed above, subjects will be carefully examined for the 
presence of intraocular inflammation after vector administration.  Cataract can also develop 
as a result of the vitrectomy procedure and can potentially affect VA.  Pre-operative grading 
of lens opacity and colour should therefore be documented by the established clinical LOCS 
III.  A recent study has shown that cataract surgery is effective in subjects with CHM and 
without any specific risks (Edwards, Jolly et al. 2016).  Subjects who develop cataracts may 
undergo cataract surgery if deemed clinically necessary.  If cataract surgery is performed, it 
should be carried out at least 4 weeks before Month 12 (Visit 9) for the respective eye.
11.11 7-Field Colour Fundus Photography
Seven-field colour fundus photography will be performed for both eyes at the times indicated 
in Table 1.


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Fundus photography will be performed by certified technicians following pupil dilation.  
Stereo photos should be performed for fields 1, 2 and 3.  All fundus photographs will be sent 
by the sites to the CRC for review; the CRC will enter the data into the EDC system.  For 
complete technical specifications for fundus photography, refer to the Study Operations 
Manual.


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12 ASSESSMENT OF EFFICACY
The following assessments are primary safety assessments that will also be considered 
secondary efficacy endpoints.  
12.1 Best-Corrected Visual Acuity
See Section 11.6. 
12.2 Fundus Autofluorescence
See Section 11.8. 
12.3 Spectral Domain Optical Coherence Tomography 
See Section 11.9. 
12.4 Microperimetry
See Section 11.7. 


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13 STATISTICAL CONSIDERATIONS
13.1 Sample Size
Approximately 60 subjects are planned to be enrolled in the study.  A total of 120 treated 
eyes is considered sufficient to characterise the immunological profile of bilateral AAV2-
REP1 administration, which is the key concern of gene therapy re-administration.
13.2 Procedure for Accounting for Missing Data
All reasonable efforts will be made to obtain complete data for both eyes on all subjects.  
However, missing observations may occur.  Management of dropout and missing 
observations will depend on their nature and frequency and will be discussed as part of the 
Data Review meeting before database lock.
Safety data will be analysed on observed data (missing data will not be imputed).  The 
analysis of efficacy endpoints will be performed using the last observation carried forward 
approach for imputation of missing data.
13.3 Analysis Sets
The ‘All Treated Subjects’ Analysis Set will consist of all subjects who attend the ‘Day of 
Surgery’ visit of Period 1 (or, of Period 2, for subjects who received AAV2-REP1 treatment 
in SE1 in an antecedent study).  The All Treated Subjects Analysis Set will be the primary 
population for demographics, baseline characteristics, and safety and efficacy analyses.
13.4 Descriptive Statistics and Conventions
Detailed specifications of the planned analyses will be included in a separate Statistical 
Analysis Plan (SAP).
Continuous variables (including changes from Baseline) will be summarised over time using 
descriptive statistics (i.e., mean, standard deviation, 95% confidence interval [CI], median, 
first and third quartiles, fifth and ninety-fifth percentiles, minimum, and maximum).  
Categorical variables (including shifts from Baseline) will be described over time using 
counts, percentages, and 95% CIs.
Baseline is defined, for each eye and each period, as the last non-missing value prior to 
treatment of the respective period, in the respective eye.
13.5 Demographics and Baseline Characteristics
Demographics will be summarised for all subjects combined. Age at the date of the Period 1 
informed consent will be calculated. Baseline ocular characteristics will be summarised by 
eye and period.
13.6 Safety Analyses
No statistical tests will be performed.
13.6.1 Visual Acuity
See Section 13.7.1. 


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13.6.2 Ophthalmic Examination
IOP and change from Baseline in IOP will be summarised by eye, period and visit.  
Abnormal slit lamp examination findings and dilated ophthalmoscopy findings will be 
summarised by eye, period and visit. Lens opacity categories and shifts from baseline will be 
summarised by eye, period and visit.
13.6.3 Spectral Domain Optical Coherence Tomography
See Section 13.7.2.  
13.6.4 Fundus Autofluorescence
See Section 13.7.3. 
13.6.5 7-Field Colour Fundus Photography
Categories of colour fundus photography findings will be summarised by eye, period and 
visit.
13.6.6 Microperimetry 
See Section 13.7.4. 
13.6.7 Adverse Events
AEs will be coded using the Medical Dictionary for Regulatory Affairs, version 21.0 or 
higher.  Events will be summarised by system organ class and preferred term and eye.  The 
number of eyes with an AE, as well as the number of events, will be summarised, by period 
and eye.  Similar summaries will be produced for study drug/procedure-related AEs, AEs 
leading to discontinuation, and SAEs. AEs will also be summarised by maximum severity, 
relationship to study drug/procedure, and time to onset and resolution.
13.6.8 Immunogenicity and Vector shedding
Vector shedding and immune response profiles will be described.
13.6.9 Vital Signs
Vital signs will be summarised by period and visit.
13.7 Efficacy Analyses
13.7.1 Visual Acuity
The proportion of subjects with a ≥10- and ≥15-letter improvement from baseline in BCVA 
will be summarised by eye, period and visit.  Mean BCVA and mean change from Baseline in 
BCVA will be tabulated by eye, period and visit. The proportion of subjects with a ≥10- and 
≥15-letter improvement from baseline in BCVA will be summarised by eye, period and visit.  
Mean BCVA and mean change from Baseline in BCVA will be tabulated by eye, period and 
visit.


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13.7.2 Spectral Domain Optical Coherence Tomography
Vitreo-macular Interface Disease, Intraretinal Hyper-Reflective Spots, Cystoid Macular 
Edema, Subretinal Fluid, Subretinal Hyper-reflective Material and Pigment Epithelial 
Detachment will be summarised by eye, period and visit, as well as shifts from baseline.
13.7.3 Fundus Autofluorescence
Total Area of Preserved AF and Distance from Foveal Center to nearest Border of Preserved 
AF, and their change from baseline, will be summarized by eye, period and visit.
13.7.4 Microperimetry
Fixation Location and Stability, and their shift from baseline, will be summarised by eye, 
period and visit. Bivariate Contour Ellipse Area and Mean Sensitivity, and their change from 
baseline, will be summarised by eye, period and visit. An analysis specific to the Sensitivity 
Points will be performed, the details of which will be described in the GEMINI SAP.Any 
additional categorical/continuous efficacy endpoints will be summarised using descriptive 
statistics, with 95% 2-sided CIs calculated where appropriate.  Shift from baseline will be 
summarised by eye and by visit, where applicable.  


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14 INFORMED CONSENT, ETHICAL REVIEW AND REGULATORY 
CONSIDERATIONS
14.1 Informed Consent
Male subjects with CHM who meet all of the entry criteria will be invited to take part in the 
study.  Subjects must personally sign and date the latest IEC / IRB approved version of the 
informed consent form before any study-specific procedures are performed.
Written and verbal versions of the subject information and informed consent will be 
presented to the subjects detailing no less than: the exact nature of the study; the implications 
and constraints of the protocol; and the known side effects and any risks involved in taking 
part.  It will be clearly stated that the subject is free to withdraw from the study at any time 
for any reason without prejudice to future care, and with no obligation to give the reason for 
withdrawal.
The subject will be allowed as much time as needed to consider the information and the 
opportunity to question the Investigator, their primary care physician/general practitioner or 
other independent parties to decide whether they will participate in the study.  Written 
informed consent will then be obtained by means of subject dated signature and dated 
signature of the person who presented and obtained the informed consent.  The person who 
obtained the consent must be suitably qualified and experienced and have been authorised to 
do so by the Investigator.  A copy of the signed informed consent will be given to each 
subject.  The original signed form will be retained at the study site and an additional copy 
will remain in the subject’s medical records.
14.2 Ethical/Regulatory Review
The protocol, informed consent form, subject information sheet and any proposed advertising 
material will be submitted to an appropriate Research Ethics Committee (IEC or IRB), 
regulatory authorities and host institution(s) for written approval.  If there are any changes to 
the approved protocol (with the exception of emergency modifications required for the 
subject’s safety), a protocol amendment will be issued by the Sponsor.  When required by 
local law, the IEC/IRB and Competent Authority must give written approval of any 
amendments likely to affect the safety of subjects or study conduct.  Each site must maintain 
accurate and updated records of all correspondence with the IEC/IRB.
14.3 Regulatory Considerations
The study will be conducted in full conformity with all applicable laws and regulations, 
including the International Council on Harmonisation (ICH) Guidelines for Good Clinical 
Practice (GCP) (CPMP/ICH/135/95).
The study, where permissible, will be conducted in accordance with the relevant articles of 
the Declaration of Helsinki as adopted by the 18th World Medical Assembly in 1964 and as 
revised in Tokyo (1975), Venice (1983), Hong Kong (1989), South Africa (1996), Scotland 
(2000), Washington (2002), Tokyo (2004), Seoul (2008), and Brazil (2013) 


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15 ADMINISTRATIVE PROCEDURES
15.1 Data Quality Control and Assurance
The study will be conducted in accordance with the current approved protocol, ICH GCP, 
relevant regulations and standard operating procedures.
The Sponsor and its selected vendors have systems in place for implementing and 
maintaining quality assurance and quality control systems, with written standard operating 
procedures (SOPs) to ensure that all aspects of the trial will be conducted in compliance with 
this protocol and data will be generated, documented and reported in compliance with this 
protocol.
Data will be entered into a validated clinical study database and subject to programmed 
validation checks and manually verified for accuracy and completeness by a Sponsor’s 
representative, both remotely and during on-site monitoring visits.  Any discrepancies will be 
resolved with the Investigator or designee, as appropriate.
Regular monitoring will be performed by the Sponsor or its designee according to ICH GCP.  
Data will be evaluated for compliance with the protocol and accuracy in relation to source 
documents.  Following written SOPs, the monitors will verify that the study is conducted and 
data are generated, documented and reported in compliance with the protocol, ICH GCP and 
applicable regulatory requirements.
In addition, this study will be subject to quality assurance audits in order to independently 
verify compliance with the protocol.
15.2 Data Handling and Records Management
The Investigator must maintain adequate and accurate source documents, which will be the 
basis of information for the eCRFs.  The source documents are to be separate and distinct 
from the eCRFs.  All study data will be entered on an encrypted EDC system with pass-codes 
known to all Investigators and appropriately delegated study team members.  This electronic 
data entry system has been validated. Incomplete or inconsistent data will result in data 
queries that require resolution by the Investigator or designee.
The Investigator must ensure that clinical study records are retained according to national 
regulations.  The Investigator must immediately inform the Sponsor if any documents are to 
be destroyed, transferred to another facility or transferred to a different owner.
In addition, files containing photos or digital outputs will be electronically transmitted to the 
reading centre for centralised, standardised review.  One reading centre is anticipated. Data 
from the reading centre will either be entered into the EDC system or provided as an external 
data set that will be loaded directly into the study database.
Samples will also be provided to a central laboratory for analysis of vector shedding and 
immunogenicity.  Results from these analyses will also be provided as an external data set 
that will be loaded directly into the study database.
The Investigator must retain sufficient documentation that these images, outputs and samples 
were handled and transmitted appropriately.


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15.3 Access to Source Documentation and Subject Privacy
Direct access will be granted to authorised representatives from the Sponsor (or designee), 
host institution, the IEC/IRB, and regulatory authorities to permit trial-related monitoring, 
audits and inspections.
The trial staff will ensure that the subject’s anonymity is maintained.  All documents will be 
stored securely and only accessible by trial staff and authorised personnel.  Subjects will be 
identified by a subject ID number on the eCRF and any electronic database.  The subject’s 
name and any other identifying detail will NOT be included in any study data electronic file.  
The study will comply with the data protection laws which require data to be anonymised. 
Subject medical information obtained in this study is confidential, and disclosure to third 
parties other than those noted below is prohibited.  As required by Personal Information 
Protection and Electronics Documents Act and Personal Health Information Protection Act, 
upon the subject’s permission, medical information may be given to his or her personal 
physician or other appropriate medical personnel responsible for his or her welfare.  Data 
generated by this study must be available for inspection by regulatory agencies, national and 
local health authorities, the Sponsor or their representative, and the IEC/IRB.
15.4 Time and Schedule of the Study
The estimated overall study duration is 12 months of follow-up per eye.
15.5 Policy for Publication and Presentation of Data
The detailed procedures for publications and data presentations are set out in the clinical trial 
agreement entered into with the Sponsor (or designee) in connection with this study.


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16 APPENDIX
Table 1 presents a schedule of study procedures.


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Table 1:
Schedule of Study Procedures
Study Period 1 / Period 2
Visit
V1
V2
V3
V4
V5
V6
V7
V8
V9
Study Day/Month
Visit Window
Period 1 
Screeninga 
Period 2
Baselineb
(<10 Weeks 
from Visit 1, 
Period 2)
Day 0 
Injection 
Dayc
(≤10 
weeks 
Screening) 
Day 1 
Post 
op
Day 3 
+2 d
Day 7 -
1/+2 d
Day 14 
± 3d
M 1
 ± 7d
M 3 
± 14d 
M6 ± 
14d
M 12 
± 14d
ET Visitd
Unscheduled 
Visite
Assessment/Procedures
(All subjects/eyes)
Informed Consent
X
Demography, medical and 
ocular history 
X
Vital signs (pulse, blood 
pressure)
X
X
X
X
X
X
Weightf
X
X
Vector shedding 
samplingg
X
X
X
X
X
X
X
X
Xh
Immunogenicity 
samplingi 
X
X
X
X
X
X
X
X
X
X
BCVA
X
X
X
X
X
X
X
X
X
X
X
X
Microperimetry
X
X
X
X
X
X
X
Full ophthalmic examj
X
X
X
X
X
X
X
X
X
X
X
X
X
SD-OCT
X
X
X
X
X
X
X
X
X
X
X
Autofluorescence
X
X
X
X
X
X
X
X
X
7-field colour fundus 
photosk
X
X
X
X
Study drug / sub-retinal 
injection / vitrectomy / 
retinal detachment
X
AE/SAE monitoringl
X
X
X
X
X
X
X
X
X
X
X
X
X


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Study Period 1 / Period 2
Visit
V1
V2
V3
V4
V5
V6
V7
V8
V9
Study Day/Month
Visit Window
Period 1 
Screeninga 
Period 2
Baselineb
(<10 Weeks 
from Visit 1, 
Period 2)
Day 0 
Injection 
Dayc
(≤10 
weeks 
Screening) 
Day 1 
Post 
op
Day 3 
+2 d
Day 7 -
1/+2 d
Day 14 
± 3d
M 1
 ± 7d
M 3 
± 14d 
M6 ± 
14d
M 12 
± 14d
ET Visitd
Unscheduled 
Visite
Assessment/Procedures
(All subjects/eyes)
Concomitant medication, 
procedures, and treatment 
reviewm
X
X
X
X
X
X
X
X
X
X
X
X
X
Abbreviations: AE=adverse event; BCVA=best corrected visual acuity; ET=early termination; IOP=intraocular pressure; LOCS=lens opacities classification system; SAE=serious adverse 
event; SD-OCT=spectral domain optical coherence tomography; SE1=Study Eye 1; SE2=Study Eye 2
a
Screening Visit/Period must be performed <10 weeks of Period 1, Visit 1. Selection of SE1 and SE2 will be determined by the Investigator in collaboration with the subject. 
b
Baseline assessments must occur before the SE2 is dosed. These can occur at a regularly scheduled visit if that visit is within 10 weeks prior to surgery in SE2. If it has been more 
than 10 weeks since a visit, the subject must return for assessments at this Baseline Period 2 visit.  
c
Subjects will undergo vitrectomy and AAV2-REP1 administration to SE1 in Period 1; SE2 in Period 2. 
d
ET visit is to be performed if a subject discontinues at any time. 
e
If clinically indicated, subjects may need to return to the site for an unscheduled visit. At a minimum, the following assessments will be performed: BCVA, full ophthalmic 
examination, SD-OCT, AF, concomitant medication, procedures, and treatment review and AE/SAE monitoring.
f
Weight is collected for dose calculation of the corticosteroid regimen.
g
Blood, tears (both eyes), urine and saliva samples. 
h
If ET visit occurs within 3 months post-treatment.
i
Immunoassays are planned to assess cell-based and antibody-based immune responses against AAV2-REP1. Enzyme-linked immunospot (ELISPOT) assays will be used for the 
assessment of T-cell-mediated immune responses to transgene product (REP1) and the capsid.  Antibody responses against the capsid and transgene product will be measured 
using a cell-based neutralization assay and an enzyme-linked immunosorbent assay (ELISA), respectively. All immunogenicity samples will be sent to a central laboratory for 
analysis.
j
The ophthalmic examination will include IOP, slit lamp examination, lens opacity grading, and dilated ophthalmoscopy. The same slit lamp machine and lighting conditions 
should be used across the study for each subject.
k
Stereo photos for fields 1, 2, 3.
l
AEs/SAEs will be collected from the time the subject provides written informed consent (Screening Visit) through Period 2, Visit 9 (or ET Visit, if applicable). 
m
Subjects will be given a course of oral corticosteroid before each surgical visit, and instructed to start taking the drug 2 days prior to SE1/SE2 treatment, unless the SE2 surgical 
treatment coincides with the SE1 steroid treatment period, when an extended course of steroid can cover both surgical treatments.  This regimen is outlined in detail in Section 9.8.


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For SE1, the Full Corticosteroid Regimen should be initiated as described.  For SE2, and surgeries that occur on day 21 or later, the Full Corticosteroid Regimen should be initiated 
as described, beginning 2 days before the scheduled SE2 surgery.  If SE2 is to be performed prior to Study Day 21, then the Full Corticosteroid Regimen should be initiated 2 days 
prior to the scheduled surgery for SE2, and this will supersede the steroid taper in progress for SE1.  The full 21 days of treatment will then be completed for SE2.


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17 REFERENCE LIST
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development for gene therapy in the retina." Adv Exp Med Biol 801: 687-693.
Dimopolous, I. S., S. C. Hoang, A. Radziwon, N. M. Binczyk, M. C. Seabra, R. E. MacLaren, R. 
Somani, M. T. S. Tennant and I. M. MacDonald (2018). "Two-Year Results After AAV2-
Mediated Gene Therapy for Choroideremia: The Alberta Experience." Am J Ophthalmol 193: 
130-142.


AAV2-REP1 
Protocol: NSR-REP-02
CONFIDENTIAL
53
Edwards, T. L., J. K. Jolly, M. Groppe, A. R. Barnard, C. L. Cottriall, T. Tolmachova, G. C. 
Black, A. R. Webster, A. J. Lotery, G. E. Holder, K. Xue, S. M. Downes, M. P. Simunovic, M. 
C. Seabra and R. E. MacLaren (2016). "Visual Acuity after Retinal Gene Therapy for 
Choroideremia." N Engl J Med 374(20): 1996-1998.
EMA (2015). Guideline on the quality, non-clinical and clinical aspects of gene therapy 
medicinal products.
Fischer, M. D., G. A. Ochakovski, B. Beier, I. P. Seitz, Y. Vaheb, C. Korteum, F. F. L. Reichel, 
L. Kuehlewein, N. A. Kahle, T. Peters, A. Girach, E. Zrenner, M. Ueffing, R. E. MacLaren, K. 
Bartz-Schmidt and W. B. (2018). "Changes in retinal sensitivity after gene therapy in 
choroideremia." Retina 40(1): 160-168.
Lam, B. L., J. L. Davis, N. Z. Gregori, R. E. MacLaren, A. Girach, J. D. Verriotto, B. Rodrigues, 
P. R. Rosa, X. Zhang and W. J. Feuer (2019). "Choroideremia gene therapy phase 2 clinical trial: 
24-month results." Am J Ophthalmol 197: 65-73.
MacDonald, I. M., L. Russell and C. C. Chan (2009). "Choroideremia: new findings from ocular 
pathology and review of recent literature." Surv Ophthalmol 54(3): 401-407.
MacLaren, R. E., M. Groppe, A. R. Barnard, C. L. Cottriall, T. Tolmachova, L. Seymour, K. R. 
Clark, M. J. During, F. P. Cremers, G. C. Black, A. J. Lotery, S. M. Downes, A. R. Webster and 
M. C. Seabra (2014). "Retinal gene therapy in patients with choroideremia: initial findings from 
a phase 1/2 clinical trial." Lancet 383(9923): 1129-1137.
Maguire, A. M., K. A. High, A. Auricchio, J. F. Wright, E. A. Pierce, F. Testa, F. Mingozzi, J. L. 
Bennicelli, G. S. Ying, S. Rossi, A. Fulton, K. A. Marshall, S. Banfi, D. C. Chung, J. I. Morgan, 
B. Hauck, O. Zelenaia, X. Zhu, L. Raffini, F. Coppieters, E. De Baere, K. S. Shindler, N. J. 
Volpe, E. M. Surace, C. Acerra, A. Lyubarsky, T. M. Redmond, E. Stone, J. Sun, J. W. 
McDonnell, B. P. Leroy, F. Simonelli and J. Bennett (2009). "Age-dependent effects of RPE65 
gene therapy for Leber's congenital amaurosis: a phase 1 dose-escalation trial." Lancet 
374(9701): 1597-1605.
Maguire, A. M., F. Simonelli, E. A. Pierce, E. N. Pugh, Jr., F. Mingozzi, J. Bennicelli, S. Banfi, 
K. A. Marshall, F. Testa, E. M. Surace, S. Rossi, A. Lyubarsky, V. R. Arruda, B. Konkle, E. 
Stone, J. Sun, J. Jacobs, L. Dell'Osso, R. Hertle, J. X. Ma, T. M. Redmond, X. Zhu, B. Hauck, O. 
Zelenaia, K. S. Shindler, M. G. Maguire, J. F. Wright, N. J. Volpe, J. W. McDonnell, A. 
Auricchio, K. A. High and J. Bennett (2008). "Safety and efficacy of gene transfer for Leber's 
congenital amaurosis." N Engl J Med 358(21): 2240-2248.
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D. J. D'Amico (1995). "Posterior segment complications after vitrectomy for macular hole." 
Ophthalmology 102(5): 775-781.
Recchia, F. M., I. U. Scott, G. C. Brown, M. M. Brown, A. C. Ho and M. S. Ip (2010). "Small-
gauge pars plana vitrectomy: a report by the American Academy of Ophthalmology." 
Ophthalmology 117(9): 1851-1857.


AAV2-REP1 
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CONFIDENTIAL
54
Sankila, E. M., R. Tolvanen, J. A. van den Hurk, F. P. Cremers and A. de la Chapelle (1992). 
"Aberrant splicing of the CHM gene is a significant cause of choroideremia." Nat Genet 1(2): 
109-113.
Seabra, M. C., M. S. Brown and J. L. Goldstein (1993). "Retinal degeneration in choroideremia: 
deficiency of rab geranylgeranyl transferase." Science 259(5093): 377-381.
Stein-Strelein, J. and C. Watte (2007). Cross-talk among cells promoting anterior chamber-
associated immune deviation. Immune Response and the Eye J. Y. Niederkorn and H. J. Kaplan. 
Basel, Chem Immunol Allergy Karger. 92: 115-130.
Stein, J. D., D. N. Zacks, D. Grossman, H. Grabe, M. W. Johnson and F. A. Sloan (2009). 
"Adverse events after pars plana vitrectomy among medicare beneficiaries." Arch Ophthalmol 
127(12): 1656-1663.
Streilein, J. W. (2003). "Ocular immune privilege: the eye takes a dim but practical view of 
immunity and inflammation." J Leukoc Biol 74(2): 179-185.
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pressure after macular hole surgery." Am J Ophthalmol 121(6): 615-622.
Tolmachova, T., O. E. Tolmachov, A. R. Barnard, S. R. deSilva, D. M. Lipinski, N. J. Walker, R. 
E. MacLaren and M. C. Seabra (2013). "Functional expression of Rab escort protein 1 following 
AAV2-mediated gene delivery in the retina of choroideremia mice and human cells ex vivo." J 
Mol Med (Berl)(7): 825-837.
Vanderberghe, L. H. and A. Auricchio (2012). "Novel adeno-associated viral vectors for retinal 
gene therapy." Gene Ther 19(2): 162-168.


CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
1
Biogen MA Inc.
225 Binney Street
Cambridge, MA  02142
United States
Biogen Idec Research Limited
Innovation House
70 Norden Road
Maidenhead, Berkshire
SL6 4AY
United Kingdom
______________________________________________________________________
AMENDMENT SUMMARY
Biogen Protocol 273CH203 (Formerly NSR-REP-02)
An Open-Label Safety Study of Retinal Gene Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-Associated Viral Vector (AAV2) Encoding Rab Escort 
Protein 1 (REP1)
GEMINI Study
For Global Protocol Amendment: Version 6.0
Date:  12 November 2020
EUDRA CT Number:  2017-002395-75
Version 6.0 of the protocol has been prepared for this amendment, which supersedes Version 5.0 
dated 30 April 2020.


Amendment Summary for Protocol 273CH203 (NSR-REP-02), Version 6.0
CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
2
PRIMARY REASON FOR AMENDMENT
The primary reason for this amendment to Protocol 273CH203 (NSR-REP-02) is for safety of 
participants: a maximum dose of 80 mg daily of corticosteroid has been added to the 
corticosteroid regimen.
New text is shown in bold type; deleted text is shown with a strikethrough.
The synopsis was revised to reflect changes made throughout the protocol.
Section 7.2 Discussion of Design
Now reads:
The corticosteroid regimen is 1 mg/kg/day (for a maximum of 80 mg/daily) 
prednisone/prednisolone for a total of 10 days (beginning 2 days before the vector injection, on 
the day of injection, and then for 7 days); followed by 0.5 mg/kg/day for 7 days; 0.25 mg/kg/day 
for 2 days; and 0.125 mg/kg/day for 2 days (21 days in total).
Rationale: A maximum dose of 80 mg daily of corticosteroid has been added to the 
corticosteroid regimen.
This change also affects:
Section 9.8, Concomitant Therapy
The corticosteroid regimen is 1 mg/kg/day prednisone/prednisolone (not to exceed 80 mg/daily) 
for a total of 10 days (beginning 2 days before the vector injection, on the day of injection, and 
then for 7 days); followed by 0.5 mg/kg/day for 7 days; 0.25 mg/kg/day for 2 days; and 0.125 
mg/kg/day for 2 days (21 days in total; Study Day (SD) -2 through SD18). 
Full Corticosteroid Regimen
Prednisone or prednisolone administered as follows:
SD -2 through SD 7 (10 days): 1 mg/kg by mouth daily (not to exceed 80 mg/daily)
SD 8 through SD 14 (7 days): 0.5 mg/kg/day by mouth daily
SD 15 through SD 16 (2 days): 0.25 mg/kg/day by mouth daily
SD 17 through SD 18 (2 days): 0.125 mg/kg/day by mouth daily


Amendment Summary for Protocol 273CH203 (NSR-REP-02), Version 6.0
CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
3
Rationale: For safety reasons a maximum of 80 mg daily of corticosteroid has been added to the 
corticosteroid regimen.


Amendment Summary for Protocol 273CH203 (NSR-REP-02), Version 6.0
CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
4
SUMMARY OF MAJOR CHANGES TO THE PROTOCOL
Not Applicable


Amendment Summary for Protocol 273CH203 (NSR-REP-02), Version 6.0
CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
5
SUMMARY OF MINOR CHANGES TO THE PROTOCOL
The following minor changes were made to the protocol, as appropriate:

The version number and date were updated throughout the protocol.

Biogen protocol number was added.

Sponsor information was updated.

Sponsor approval page personnel was updated.

Patient Eligibility Review Committee was added to the Synopsis Exclusion Criteria


Amendment Summary for Protocol 273CH203 (NSR-REP-02), Version 6.0
CONFIDENTIAL
The information contained herein may not be used, disclosed, or published without the written consent of 
Biogen MA Inc.
6
LIST OF ABBREVIATIONS
Not Applicable


AAV2-REP1 
 
Version 5.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 Apr 2020 
 
CONFIDENTIAL 
Page 1 of 5 
 
SUMMARY OF CHANGES FOR CLINICAL STUDY PROTOCOL  
NSR-REP-02  
 
AAV2-REP1 
 
An Open-Label Safety Study of Retinal Gene Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-Associated Viral Vector (AAV2) Encoding Rab Escort 
Protein 1 (REP1) 
GEMINI Study 
Indication: 
Choroideremia 
Study Phase: 
2 
Sponsor: 
NightstaRx Ltd 
2nd Floor 
10 Midford Place 
London W1T 5BJ, UK 
Telephone:  +44 (0) 020 7062 2777 
Summary of Changes: 
Protocol Amendment 4.0, Version 5.0 
30 Apr 2020 
CONFIDENTIALITY STATEMENT 
This protocol is the property of NightstaRx Ltd. It is not to be transmitted, copied or published 
without written permission from NightstaRx and must be kept in a confidential manner. 
Persons to whom the information is disclosed must be informed that the information is 
CONFIDENTIAL and may not be further disclosed by them. 
 
 


AAV2-REP1 
 
Version 5.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 Apr 2020 
 
CONFIDENTIAL 
Page 2 of 5 
 
 
SUMMARY OF CHANGES SPONSOR APPROVAL PAGE 
 
Clinical Study Protocol Number: 
NSR-REP-02 
Protocol Title: 
An Open-Label Safety Study of Retinal Gene 
Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-
Associated Viral Vector (AAV2) Encoding 
Rab Escort Protein 1 (REP1) 
Summary of Changes for Protocol:  
Version 5.0 30 Apr 2020 
Approved By:  
 
 
 
 
 
The person listed below is authorised to sign the summary of changes on behalf of NightstaRx 
Ltd/Biogen. 
 
Date: 
 
, MD, PhD  
 
 
Digitally signed by 
 
DN: cn=
, o=Biogen, 
ou=Ophthalmology DU, 
email=
 c=US 
Date: 2020.04.30 21:10:37 -04'00'


AAV2-REP1 
 
Version 5.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 Apr 2020 
 
CONFIDENTIAL 
Page 3 of 5 
 
Overview / Rationale:  
The change to this protocol triggering Version 5.0 was to reference the Patient Eligibility Review 
Process.  The protocol now clearly describes that the Investigator as well as the Sponsor and a 
surgical consultant through a collective decision-making process outlined in the Patient 
Eligibility Review Plan (version 2.0, dated 03 April 2020) ultimately determine the subject’s 
eligibility for surgery.  This process takes places subsequent to the subject’s eligibility 
determination based on meeting inclusion and exclusion criteria and undergoing baseline 
assessments.  The aim of the Patient Eligibility Review is to determine if the subject is an 
appropriate candidate for surgery through examination of baseline imaging (optical coherence 
tomography, fundus autofluorescence, fundus photography), best-corrected visual acuity, and 
microperimetry data, as well as genetic test results, taking into consideration disease progression 
and the status of the choroideremic retina.  Both the Data Monitoring Committee for the 
GEMINI study, and the principal investigators have been consulted regarding this amendment, 
and both parties did not recommend further amending the protocol to include specific surgical 
eligibility criteria, preferring the current collaborative Patient Eligibility Review process.   
The Patient Eligibility Review Plan and each subject’s signed GEMINI surgery authorization 
form lie within the Trial Master File for the GEMINI study.  
 
 


AAV2-REP1 
 
Version 5.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 Apr 2020 
 
CONFIDENTIAL 
Page 4 of 5 
 
Summary of Changes 
Protocol NSR-REP-02 
Version 5.0, 30 Apr 2020 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
Header 
Version 4.0 
30 Oct 2019 
Version 5.0 
30 Apr 2020 
Updated text to 
latest protocol 
Title Page 
 
 
Amendment 4.0, Version 5.0 
30 Apr 2020 
Added latest 
protocol 
Sponsor 
Approval, 
Page 2 
19 February 
30 Apr 2020 
Updated approval 
page date and 
personnel to 
version 5.0 
  
 
, MD, PhD 
 
The person listed above is 
authorized to sign the protocol on 
behalf of NightstaRx Ltd. The wet 
ink signature is on file and 
available upon request.  
The person listed above is 
authorized to approve the protocol 
on behalf of NightstaRx 
Ltd/Biogen. A signature is on file 
and available upon request.  
The current 
approval process 
is electronic. 
 
 
 
Investigator 
Signature Page, 
Page 3 
19 February 
30 Apr 2020 
Updated signature 
page date to 
version 5.0 
Synopsis 
Study Design, 
Pages 5 & 6 
 
Study eye selection and eligibility 
for surgery will then be confirmed 
collectively by the Patient 
Eligibility Review Committee, 
which includes the Investigator, a 
consulting surgeon, and the 
Sponsor (see the Patient Eligibility 
Review Plan).    
Added text to 
include study eye 
selection and 
eligibility for 
study is defined by 
the Patient 
Eligibility Review 
Committee  
Committee investigator and after 
consultation with the sponsor 
Eligibility for SE2 surgery will 
then be confirmed collectively by 
the Patient Eligibility Review 
Committee, which includes the 
Investigator, a consulting surgeon, 
and the Sponsor (see the Patient 
Eligibility Review Plan). 
If, at that time, the BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the Investigator and after 
consultation with the sponsor: 
If, at that time, the BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the Patient Eligibility Review 
Committee: 
Synopsis 
Exclusion 
Criteria 
Inclusion #4 
Page 7 
 
• who, in the clinical opinion of the 
Patient Eligibility Review 
Committee or Investigator, is not 
an appropriate candidate for sub-
retinal surgery 
Added text to 
reference the 
Patient Eligibility 
Review 
Committee 
Section 7.1 
Overall Study 
Design 
Page 17 
 
Study eye selection and eligibility 
for surgery will then be confirmed 
collectively by the Patient 
Eligibility Review Committee, 
which includes the Investigator, a 
consulting surgeon, and the 
Added text to 
include study eye 
selection and 
eligibility for 
study is defined by 
the Patient 


AAV2-REP1 
 
Version 5.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 Apr 2020 
 
CONFIDENTIAL 
Page 5 of 5 
 
Summary of Changes 
Protocol NSR-REP-02 
Version 5.0, 30 Apr 2020 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
Sponsor (see the Patient Eligibility 
Review Plan).  
Eligibility Review 
Committee 
Section 7.1 
Overall Study 
Design 
Page 18 
 
Study eye selection and eligibility 
for surgery will then be confirmed 
collectively by the Patient 
Eligibility Review Committee, 
which includes the Investigator, a 
consulting surgeon, and the 
Sponsor (see the Patient Eligibility 
Review Plan). 
Added text to 
include study eye 
selection and 
eligibility for 
study is defined by 
the Patient 
Eligibility Review 
Committee 
If, at that time, the BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the Investigator and after 
consultation with the sponsor 
If, at that time, the BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the Patient Eligibility Review 
Committee 
Section 8.2 
Exclusion 
Criteria #4 
Page 20 
 
Patient Eligibility Review 
Committee or Investigator 
Added text to 
include Patient 
Eligibility Review 
Committee 
• who, in the clinical opinion of the 
Patient Eligibility Review 
Committee or Investigator, is not 
an appropriate candidate for sub-
retinal surgery (see the Patient 
Eligibility Review Plan) 
Section 10.1 
Screening Period 
Page 26 & 28 
 
Study eye selection and eligibility 
for surgery will then be confirmed 
collectively by the Patient 
Eligibility Review Committee, 
which includes the Investigator, a 
consulting surgeon, and the 
Sponsor (see the Patient Eligibility 
Review Plan). 
Added text to 
include study eye 
selection and 
eligibility for 
study is defined by 
the Patient 
Eligibility Review 
Committee 
If BCVA in SE2 is <34 ETDRS 
letters, surgery may be performed 
when, in the opinion of the 
investigator and after consultation 
with the sponsor: 
Eligibility for SE2 surgery will 
then be confirmed collectively by 
the Patient Eligibility Review 
Committee, which includes the 
Investigator, a consulting surgeon, 
and the Sponsor (see the Patient 
Eligibility Review Plan).  
If BCVA in SE2 is <34 ETDRS 
letters, surgery may be performed 
when, in the opinion of the Patient 
Eligibility Review Committee  
 


AAV2-REP1 
 
Version 4.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 OCT 2019 
 
CONFIDENTIAL 
Page 1 of 6 
 
SUMMARY OF CHANGES FOR CLINICAL STUDY PROTOCOL  
NSR-REP-02  
 
AAV2-REP1 
 
An Open-Label Safety Study of Retinal Gene Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-Associated Viral Vector (AAV2) Encoding Rab Escort 
Protein 1 (REP1) 
GEMINI Study 
Indication: 
Choroideremia 
Study Phase: 
2 
Sponsor: 
NightstaRx Ltd 
2nd Floor 
10 Midford Place 
London W1T 5BJ, UK 
Telephone:  +44 (0) 020 7062 2777 
Summary of Changes: 
Protocol Amendment 3.0, Version 4.0 
30 OCT 2019 
CONFIDENTIALITY STATEMENT 
This protocol is the property of NightstaRx Ltd. It is not to be transmitted, copied or published 
without written permission from NightstaRx and must be kept in a confidential manner. 
Persons to whom the information is disclosed must be informed that the information is 
CONFIDENTIAL and may not be further disclosed by them. 
 
 




AAV2-REP1 
 
Version 4.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 OCT 2019 
 
CONFIDENTIAL 
Page 3 of 6 
 
Overview / Rationale:  
The main changes to this protocol triggering Version 4.0 were: 
Visit windows changed for Visit 3 (Day 3 + 2 days, post-operative follow-up visit) and Visit 4 
(Day 7 -1/+2 day); added to decrease patient burden and minimize weekend post-operative 
follow-ups. 
New data derived from the STAR study caused a change in the Reference Safety Information 
and an update to the Investigator’s Brochure.  This information has been added to the 
Risk/Benefit section of the protocol. 
Additional information was added to better define serious adverse events. 
 
 


AAV2-REP1 
 
Version 4.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 OCT 2019 
 
CONFIDENTIAL 
Page 4 of 6 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 4.0, 30 Oct 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
Header 
Version 3.1 
19 Feb 
Version 4.0 
30 Oct 2019 
Updated text to 
latest protocol 
Title Page 
 
 
Amendment 3.0, Version 4.0 
30 October 2019 
Added latest 
protocol 
Sponsor 
Approval, 
Page 2 
19 February 
30 October 
Updated approval 
page date to 
version 4.0 
Investigator 
Signature Page, 
Page 3 
19 February 
30 October 
Updated signature 
page date to 
version 4.0 
Contact 
Information, 
Page 4 
 
CLINICAL RESEARCH 
ORGANIZATION 
Added to header 
Ashfield Pharmacovigilance 
5003 S. Miami Blvd, Suite 500 
Durham NC 27703 
Email: 
APV.AENightstar@ashfieldpv.com 
+1-919-401-8003 
Covance (Formerly known as 
Chiltern International, Ltd) 
Chiltern International Limited 
Covance Clinical and Periapproval 
Services, Ltd 
Osprey House, 
Maidenhead Office Park, 
Westacott Way, Littlewick Green, 
Maidenhead SL6 3QH, UK 
+44 (0) 1628 548000 
Updated the 
pharmacovigilance 
information 
Serious Adverse Event (SAE) and 
Pregnancy Notification Forms 
should be emailed to 
safetyGEMINI@nightstartx.com.  
For further information refer to 
Section 11, Assessment of Safety. 
 
Deleted 
information no 
longer necessary 
Synopsis 
Study Design, 
Pages 5 & 6 
 
Each study eye will be followed for 
at least 12 months post-treatment, 
for up to 9 visits per treatment 
period: Visit 1 (Day 0, Injection 
Day Visit); Visit 2 (Day 1, post-
operative follow-up visit); Visit 3 
(Day 3 + 2 days, post-operative 
follow-up visit); Visit 4 (Day 
7 -1/+2 days);  Visit 5 (Day 14 ± 3 
days); Visit 6 (Month 1 ± 7 days); 
Visit 7 (Month 3 ± 14 days); Visit 
8 (Month 6 ± 14 days) and Visit 9 
(Month 12 ± 14 days; end of study 
visit). 
Added to visit 
window to 
decrease patient 
burden and 
minimize weekend 
post-operative 
follow-ups.  
Section 5.4 
Risk/Benefit 
Assessment 
Page 15 
Loss of VA is, therefore, 
considered a Potential Risk of 
AAV2-REP1 treatment, in addition 
to the above described potential 
and anticipated surgical risks. 
See the Investigator’s Brochure for 
AAV2-REP1 for further details. 
Complications from the surgical 
procedure are expected and are not 
considered reportable as suspected 
unexpected serious adverse 
reactions (SUSARs) unless also 
assessed to be at least possibly 
related to timrepigene emparvovec.  
New data derived 
from the STAR 
study caused a 
change in the 
Reference Safety 
Information This 
new information 


AAV2-REP1 
 
Version 4.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 OCT 2019 
 
CONFIDENTIAL 
Page 5 of 6 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 4.0, 30 Oct 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
The Reference Safety Information 
described in the current version of 
the Investigator’s Brochure for 
AAV2-REP1 defines visual acuity 
reduced as an expected serious 
adverse drug reaction, with a 
frequency of 4/79 (5.1%), defined 
as common (≥1/100 to <1/10). Loss 
of VA is considered an identified 
risk of AAV2-REP1.  
Inflammation is a potential risk of 
AAV2-REP1. 
has been added to 
the Benefit-Risk 
Section. 
Section 7.1 
Overall Study 
Design, 
Page 19 
 
Each study eye will be followed for 
at least 12 months post-treatment, 
for up to 9 visits per treatment 
period: Visit 1 (Day 0, Injection 
Day Visit); Visit 2 (Day 1, post-
operative follow-up visit); Visit 3 
(Day 3 + 2 Days); Visit 4 (Day 7 -
1/+2 Days); Visit 5 (Day 14 ± 3 
Days); Visit 6 (Month 1 ± 7 Days); 
Visit 7 (Month 3 ± 14 Days); Visit 
8 (Month 6 ±14 Days) and Visit 9 
(Month 12 ± 14 Days; end of study 
visit). 
Added to decrease 
patient burden and 
minimize weekend 
post-operative 
follow-ups. 
Section 9.4 
Vitrectomy 
Procedure and 
Injection of 
AAV2-REP1, 
Page 24 
 
In the second step of the procedure, 
the BSS cannula is removed from 
the eye and AAV2-REP1 is 
prepared for injection.  A dose of 
up to 1.0×10^11 gp of AAV2-
REP1 is injected into the sub-
retinal space through the same 
entry site.   
Clarified text 
Section 10.4 
Period 1 and 2, 
Visit 3, (Post-
operative Day 3 + 
2), 
Page 29 
 
Subjects will return to the site 1, 3, 
7 and 14 days after surgery for 
post-operative follow-up (Visit 2 
[Day 1], Visit 3 [Day 3 + 2 Days], 
Visit 4 [Day 7 -1/+2 Days] and 
Visit 5 [Day 14 ± 3 Days], 
respectively). 
Added to decrease 
patient burden and 
minimize weekend 
post-operative 
follow-ups. 
Sections 10.5  
Period 1 and 2, 
Visit 4 (Post-
operative Day 7 -
1/+2 days),  
Page 29 
10.5 
Period 1 and 2, Visit 4 
(Post-operative Day 7 ± 1-Days) 
For Visit 4 (Post-operative Day 7 ± 
1 -day), ocular assessments and 
procedures will be performed on 
each eye.  At Visit 4 (Day 7 ± 1 -
1/+2 days), 
10.5 
Period 1 and 2, Visit 4 
(Post-operative Day 7 -1/+2 Days) 
For Visit 4 (Post-operative Day 7 -
1/+2 days), ocular assessments and 
procedures will be performed on 
each eye.  At Visit 4 (Day 7 -1/+2 
days), 
Added to decrease 
patient burden and 
minimize weekend 
post-operative 
follow-ups. 
Section 11.1.1.2 
Hospitalisation, including elective 
procedures… 
Hospitalisation that was pre-
scheduled prior to obtaining the 
Added text to 
better define 


AAV2-REP1 
 
Version 4.0 
Summary of Changes: Protocol: NSR-REP-02 
 
30 OCT 2019 
 
CONFIDENTIAL 
Page 6 of 6 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 4.0, 30 Oct 2019 
Section, Page 
Previous Text 
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Rationale 
Serious Adverse 
Event, Pages 33-
34 
consent to participate in the study 
or for an elective procedure or 
routinely scheduled treatment for a 
pre-existing condition, which has 
not worsened, does not constitute 
an SAE. 
A prescheduled or elective 
procedure or a routinely scheduled 
treatment will not be considered an 
SAE, even if the subject is 
hospitalized.  The study site must 
document all of the following: 
• 
The prescheduled or elective 
procedure or routinely 
scheduled treatment was 
scheduled (or was on a waiting 
list to be scheduled) prior to 
obtaining the subject’s consent 
to participate in the study. 
• 
The condition requiring the 
prescheduled or elective 
procedure or routinely 
scheduled treatment was 
present before and did not 
worsen or progress in the 
opinion of the Investigator 
between the subject’s consent 
to participate in the study and 
the time of the procedure or 
treatment. 
• 
The prescheduled or elective 
procedure or routinely 
scheduled treatment is the sole 
reason for the intervention or 
hospital admission. 
 
If a subject is hospitalized due to 
local requirements for 
administration of the study 
treatment, the hospitalization 
should not be considered an SAE 
unless one of the requirements in 
Section 11.1.1.2 is met. 
serious adverse 
events 
Table 1 
Schedule of 
Study Procedures 
V3 Day 3 Post op 
V4 Day 7 ±1 d 
V3 Day 3 +2 d 
V4 Day 7 -1/+2 d 
Updated Table 1 
headers 
 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 1 of 42 
 
SUMMARY OF CHANGES FOR CLINICAL STUDY PROTOCOL  
NSR-REP-02  
 
AAV2-REP1 
 
An Open-Label Safety Study of Retinal Gene Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-Associated Viral Vector (AAV2) Encoding Rab Escort 
Protein 1 (REP1) 
GEMINI Study 
Indication: 
Choroideremia 
Study Phase: 
2 
Sponsor: 
NightstaRx Ltd 
2nd Floor 
10 Midford Place 
London W1T 5BJ, UK 
Telephone:  +44 (0) 020 7062 2777 
Summary of Changes: 
Protocol Version 3.0 
4 Feb 2019 
CONFIDENTIALITY STATEMENT 
This protocol is the property of NightstaRx Ltd. It is not to be transmitted, copied or published 
without written permission from NightstaRx and must be kept in a confidential manner. 
Persons to whom the information is disclosed must be informed that the information is 
CONFIDENTIAL and may not be further disclosed by them. 
 
 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 2 of 42 
 
SUMMARY OF CHANGES SPONSOR APPROVAL PAGE 
Clinical Study Protocol Number: 
NSR-REP-02 
Protocol Title: 
An Open-Label Safety Study of Retinal Gene 
Therapy for Choroideremia with Bilateral, 
Sequential Administration of Adeno-
Associated Viral Vector (AAV2) Encoding 
Rab Escort Protein 1 (REP1) 
Summary of Changes for Protocol:  
Version 3.0   4 Feb 2019 
Approved By: 
, MD 
 
The person listed above is authorised to sign the summary of changes on behalf of NightstaRx 
Ltd.  A wet ink signature is on file and available upon request.  
 
 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 3 of 42 
 
Overview / Rationale of Major Changes:  
This protocol amendment was triggered primarily by recommendations and interactions from 
regulatory bodies that have occurred in 2018. Five key revisions were made:  1) the sample size 
was increased from 15 to 60; 2) specific language was added regarding the allocation of these 60 
subjects into 3 surgery windows of varying length, as recommended by FDA, to provide the 
broadest safety profile available on subjects treated with short, medium and long time periods 
between surgeries; 3) the corticosteroid regimen was revised to clarify administration required 
with bilateral treatment performed with very short windows between surgeries; 4) the visual 
acuity inclusion criterion was revised to allow for the eligibility of subjects with more significant 
visual impairment; and lastly, 5) new requirements have been added to mandate baseline 
assessments prior to treating the second eye in the case of long surgery windows, to assure that 
the second eye remains eligible and that the investigator still recommends treatment based on 
safety assessments and the visual function of both eyes at that time, as well as to establish 
accurate baseline values. In addition, a new inclusion criterion has been added mandating that 
subjects who enter this study from an antecedent study must have biological samples available to 
complete an adequate immunogenicity profile, if needed. 
A new benefit-risk assessment section has also been added to the Introduction, based on 
information collected from ongoing safety reporting of the Phase 3 STAR study.  This new 
section outlines the expected risks of AAV2-REP1 treatment to include visual acuity loss post-
treatment. The definition of vision loss as a serious adverse event was clarified, without changing 
the original meaning. Adverse event reporting was also modified to a binary system that still 
allowed the investigator to choose 3 levels of relatedness under “related to study drug or 
procedure”.  
Minor changes were made solely for clarification. 
 
 
 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 4 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
Title Page 
 
Version 2.0 
17 May 2018 
Version 3.0 
1 Feb 2019 
Updated document 
header 
  
CT Number: NCT03507686  
INN: Timrepigene emparvovec 
Amendment 2 
1 February 2019 
Added to title page 
Wellcome Gibbs Building, 
215 Euston Road 
London NW1 2BE, UK 
207 611 2077 
 
2nd Floor 
10 Midford Place 
London W1T 5BJ UK 
020 7062 2777 
Change of address 
and telephone 
Sponsor Approval, 
Page 2 
17 May 2018 
1 February 2019 
Updated to date of 
protocol version 
3.0 
, MD  
 
, MD 
 
 
Personnel change 
The person listed below is 
authorised to sign the 
protocol on behalf of 
NightstaRx Ltd. 
Signed: Date: 
, MD  
 
The person listed above is 
authorised to sign the protocol on 
behalf of NightstaRx Ltd.  The wet 
ink signature is on file and available 
upon request 
Removed signature 
line from sponsor 
approval page and 
added referral for 
wet signature 
Investigator 
Signature Page, 
Page 3 
17 May 2018 
1 February 2019 
Updated protocol 
date to version 3.0 
 
 (where required) 
Clarified text 
Contact 
Information, 
Page 4 
 
NightstaRx Ltdr 
Wellcome Gibbs Building, 
Euston Road 
London NW1 2BE, UK 
 
 
The contact page 
within the protocol 
now includes only 
pharmacovigilance 
contact and the 
responsible CRO. 
The complete 
study contact list 
will be included in 
the study manual. 
 
 
NightstaRx Ltd 
Wellcome Gibbs Building, 
Euston Road 
London NW1 2BE, UK 
 
 
 
 
Medical Monitor  
, MD 
NightstaRx Ltd 
Wellcome Gibbs Building, 
215 Euston Road 
London NW1 2BE, UK 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 5 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
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Revised Text 
(Added Text Shown as Red) 
Rationale 
Pharmacovigilance 
, MD 
NightstaRx Ltd 
Wellcome Gibbs Building, 
215 Euston Road 
London NW1 2BE, UK 
Pharmacovigilance 
Ashfield Pharmacovigilance 
5003 S. Miami Blvd, Suite 500 
Durham NC 27703 
Email: 
APV.AENightstar@ashfieldpv.com 
+1-919-401-8003 
Contact 
information 
changed 
Contract Research 
Organization 
Chiltern International, Ltd 
171 Bath Road, Slough, 
Berkshire, SL1 4AA, UK 
+44 (0) 175 351 2000 
Contract Research Organization 
(CRO) 
Covance Clinical and Periapproval 
Services, Ltd 
Osprey House 
Maidenhead Office Park, 
Westacott Way, Littlewick Green 
Maidenhead SL6 3QH, UK 
 
Additional contact information is 
available in the Site Operations 
Manual. 
For further information refer to 
Section 11, Assessment of Safety. 
Added text for 
contact 
information  
Synopsis 
Study Period 
(years), 
Page 5 
 
Approximately 24 months 
(12 months of follow-up for each 
eye) 
Clarified text 
Synopsis 
Study Objective, 
Page 5 
 
in adult male subjects…. 
Added text to 
clarify subject 
criteria in objective 
Synopsis   
Primary Endpoints, 
Page 5 
The primary endpoint is the 
evaluation of safety 
following of bilateral 
administration of AAV2-
REP1. 
 
The safety of bilateral 
administration of AAV2-REP1 will 
be evaluated with the following 
safety measures: 
•Best corrected visual acuity 
(BCVA) as measured by the Early 
Treatment of Diabetic Retinopathy 
Study (ETDRS) chart 
•Ophthalmic examination 
assessments (including intraocular 
pressure [IOP], slit lamp 
examination, lens opacity grading 
and dilated ophthalmoscopy) 
•Spectral domain optical coherence 
tomography (SD-OCT) 
•Fundus autofluorescence (AF) 
•Fundus photography 
•Microperimetry 
•Adverse event (AE) reporting  
•Vector shedding post-treatment 
•Immunogenicity sampling post-
treatment  
The original 
protocol did not 
specify the exact 
measures to use in 
evaluating the 
safety of bilateral 
AAV2-REP1. Here 
we list al measures 
that will comprise 
the safety 
evaluation 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 6 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
•Vital signs. 
Synopsis 
Secondary 
Endpoints, 
Page 5 
•Change from Baseline in 
best corrected visual acuity 
(BCVA) as measured by the 
Early Treatment of Diabetic 
Retinopathy Study 
(ETDRS) chart  
•Change from Baseline in 
autofluorescence (AF) 
•Change from Baseline in 
spectral domain optical 
coherence tomography (SD-
OCT) 
Secondary Efficacy Endpoints 
include: 
 
Removed 
definitions of 
abbreviations 
previously defined 
in text 
Synopsis 
Study Design, 
Pages 5 & 6 
The study will consist of a 
Screening Visit followed by 
2 treatment periods (Period 
1 and Period 2) with up to 9 
visits per period.  At the 
Screening Visit, subjects 
will be assessed for 
eligibility of both eyes. 
This is a multi-centre, open-label, 
prospective, 2 period, …, 
The study will consist of a 
Screening Period followed by 2 
treatment periods (Period 1 and 
Period 2) with up to 9 visits per 
period. During the Screening 
Period, subjects will be assessed for 
eligibility of both eyes. 
Added text, 
Clarified text 
 
The investigator will assign the 
order in which the eyes are treated 
(i.e., study eye 1 [SE1]-Period 1] 
and study eye 2 [SE2]- Period 2], 
respectively with a number assigned 
to each eye.  This will be done in 
collaboration with the subject; 
however, the worse eye will 
generally be selected for first 
treatment. The estimated target 
interval between the surgical 
procedures of SE1 and SE2 will be 
determined at the Screening Visit. 
Added text to 
specify period 1 
and period 2, and 
target interval 
 
 
…Visit 4 (Day 7 ±  1 day); Visit 5 
(Day 14 ±  3 days); Visit 6 (Month 
1 ±  7  days); Visit 7 (Month 3 ±  14 
days); Visit 8 (Month 6 ±  14 days) 
and Visit 9 (Month 12 ±  14 days; 
end of study visit). 
Added text to 
clarify visit 
windows 
 
…, in which case, the study visits 
will follow the schedule for SE2. 
Clarified procedure 
The interval between SE1 
and SE2 treatment is 
expected to vary subject 
from a few weeks to many 
months.   
The interval between SE1 and SE2 
treatment is expected to vary among 
subjects from weeks to many 
months.   
Clarified text 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 7 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Revised Text 
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Rationale 
Each site will work with the 
Sponsor and the assigned 
Contract Research 
Organisation (CRO) to try 
and ensure an even spread 
of subjects across the 
intervals. 
Thus, approximately 20 subjects 
each will be treated with a short (<6 
month), medium (6-12 months), or 
long (>12 month) surgery window 
between treatment of the first and 
second eye.   
Specified 
allocation of 
subjects into 3 
surgery windows 
as recommended 
by FDA to mimic 
real-world 
application of drug 
and to provide a 
broad assessment 
of safety with 
bilateral treatment. 
Because the timing of the 
second surgery visit (Period 
2, Visit 1) will vary 
between subjects, the 
duration of study Period 1 
will also be variable.  To 
avoid unnecessary study 
visits, every effort should 
be made by the investigator 
to schedule Period 2, Visit 1 
to coincide with a planned 
Period 1 post-treatment 
visit.   
Because the timing of the second 
surgery visit (Period 2, Visit 1) will 
vary among subjects the duration of 
study Period 1 will also be variable. 
Clarified text, 
removed excess 
text in synopsis 
  
or other post-operative sequelae, 
Expanded text 
 
…the Investigator will determine 
the eligibility of the subject and 
decide the appropriate time to 
schedule SE2, Visit 1.   
 
 All baseline ophthalmic 
assessments must be conducted 
within 3 months of SE2 surgery 
(Period 2, Visit 1).  If, at that time, 
the BCVA in SE2 is <34 ETDRS 
letters, surgery may be performed 
when, in the opinion of the 
investigator and after consultation 
with the sponsor: 1) the subject is 
found eligible by all other inclusion 
and exclusion criteria, and 2) the 
subject is considered an appropriate 
candidate who has a potentially 
modifiable disease . 
Mandated that 
baseline 
assessments be 
conducted if the 
second eye is done 
with a long time 
period between 
surgeries to assure 
that the eligibility 
and safety of 
treating the second 
eye and the overall 
safety and visual 
function of the 
subject. 
 
Post treatment subjects will no 
longer follow the Period 1 visit 
schedule…. 
Clarified text, 
added text to 
clarify procedure 
for cataract surgery 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 8 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Revised Text 
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Rationale 
The cataract surgery timing must be 
discussed with the medical monitor 
prior to the procedure. 
Synopsis 
Number of 
Subjects, 
Page 6 
At least 15 subjects are 
planned for the study. 
Approximately 60 subjects are 
planned for the study. 
 
Updated number of 
subjects planned 
Synopsis  
Inclusion Criteria, 
Pages 6 & 7 
At the Screening Visit, 
subjects are found eligible 
for study participation if 
they meet all of the 
following inclusion criteria 
During the Screening Period, 
subjects will be found eligible for 
study participation if they meet all 
of the following inclusion criteria 
Clarified text 
≥34 ETDRS letters 
(equivalent to better than or 
equal 20/40 or better 
Snellen acuity) and ≤≥74 73 
ETDRS letters (equivalent 
to better than or equal to 6/9 
or 20/32 Snellen, decimal 
0.63, LogMar 0.2) in both 
eyes1 or both untreated 
eyes*, if no prior treatment 
with AAV2-REP1 
o≥74 ETDRS letters 
(equivalent to better than or 
equal to 6/9 or 20/32 
Snellen, decimal 0.63, 
LogMar 0.2) in the 
untreated eye, if prior 
treatment with AAV2-
REP1 was received in an 
alternative study* 
3.Have documentation of a 
genetically-confirmed diagnosis of 
CHM 
5. Have a BCVA of ≥34 ETDRS 
letters (20/200 or better Snellen 
acuity) in both eyes, or in the 
untreated eye, if the other eye was 
previously treated with AAV2-
REP1* 
* If previously treated with AAV2-
REP1 in an antecedent study, 
subjects may be eligible for 
participation following Sponsor 
approval.  
6. For subjects who received 
treatment with AAV2-REP1 in an 
antecedent study, have biological 
samples available to complete an 
adequate immunology profile. 
Updated inclusion 
criteria to specify 
criteria for CHM 
diagnosis and eye 
for BCVA, added 
criteria for subjects 
in a previous 
study. 
See Rationale. 
Synopsis 
Exclusion Criteria, 
Page 7 
At the Screening Visit, 
subjects are found not 
eligible… 
At the Screening Visit, subjects will 
be found not eligible…, 
2. …or abstain from sexual 
intercourse… 
5. …in an antecedent study with 
AV2-REP1. 
Added text to 
further explain 
exclusion criteria 
Synopsis 
Test Product, 
Dosage, and Mode 
of Administration, 
Page 7 
…receive 0.1 mL sub-
retinal injection of study 
drug containing 1 × 10^11 
AAV2-REP1 genome 
particles. 
…receive a sub-retinol injection of 
up to 0.1 mL of study drug 
containing 1 × 10^11 AAV2-REP1 
genome particles (gp) 
Clarified text 
Synopsis 
Criteria for 
Evaluation, 
Page 7 
 
The safety evaluation will 
be based full ophthalmic 
examination (including 
intraocular pressure [IOP], 
slit lamp examination, lens 
opacity grading and dilated 
The safety evaluation will be based 
on BCVA (as measured by the 
ETDRS chart); full ophthalmic 
examination (including IOP, slit 
lamp examination, lens opacity 
grading and dilated 
Updated criteria to 
include BCVA, 
SD-OCT, AF, and 
microperimetry 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 9 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Revised Text 
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Rationale 
ophthalmoscopy), fundus 
photography; adverse event 
(AE) reporting; vector 
shedding and 
immunogenicity sampling; 
and vital signs. 
ophthalmoscopy), SD-OCT; fundus 
AF; fundus photography; 
microperimetry; adverse event (AE) 
reporting; vector shedding and 
immunogenicity; and vital signs. 
Synopsis 
Statistical 
Methodology, 
Page 7 
median, Q1, Q3, P05, P95, 
min, and max). 
median, first and third quartiles, 
fifth and ninety-fifth percentiles, 
min, and max).   
Descriptive 
statistics clarified 
Both the number of subjects 
and the number of eyes 
experiencing an AE, as well 
as the number of events, 
will be summarised. 
The number of eyes with an AE, as 
well as the number of events, will 
be summarised, by period and eye.   
Clarified text for 
number of eyes 
and events 
summarised 
Abbreviations, 
Page 11 
(c)DNA  (complementary) 
deoxyribonucleic acid 
cDNA Complementary 
deoxyribonucleic acid 
Clarified text 
AF autofluorescence 
AAV2-REP1 recombinant… 
SAP Statistical Analysis Plan 
SD study day 
SUSAR Suspected unexpected 
serious adverse reaction 
Added 
abbreviations 
Section 5.2 
Gene Therapy with 
Adeno-Associated 
AAV, 
Page 13 
Considering same-day 
bilateral administration, 
there is total antigen 
exposure following the 
6×10^9 genome particles 
(gp)/eye dose administered 
is comparable to a 
1.2×10^10 gp unilateral 
dose, which equates to a 
1.2×10^12 gp dose in 
humans, . providing a 12-
fold immunogenicity safety 
margin compared to the 
maximum clinical dose of 
1×10^11 gp. 
In another non-clinical study in rats, 
lack of apparent immunogenicity 
was observed following same-day 
bilateral administration of 6×10^9 
genome particles (gp)/eye AAV2-
REP1 (Charles River Study No. 
5700437).  This total antigen 
exposure following bilateral dosing 
is comparable to a unilateral dose of 
1.2×10^10 gp, which equates to a 
1.2×10^12 gp dose in humans. This 
provides a 12-fold immunogenic 
safety margin compared to the 
planned clinical dose of 1×10^11 
gp. 
Clarified dosing 
text  
In this study, 11 children 
and adults with Leber’s 
congenital amaurosis 
received one single sub-
retinal administration of 
AAV2-RPE65 at 1.5×10^11 
In the clinical setting, repeat 
administration…, 
In this study, 11 children and adults 
with Leber’s congenital amaurosis 
received a single sub-retinal 
administration of AAV2-RPE65 at a 
dose of 1.5×10^11…, 
AAV2-RPE65, voretigene 
neparvovec-rzyl (LUXTURNA™, 
Spark Therapeutics, Inc) has been 
approved in the US- and the EU as 
the first adeno-associated virus 
Clarified text, 
Added text for 
further information 
on AAV2 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 10 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
vector type 2 (AAV2)-based gene 
therapy indicated for the treatment 
of patients with confirmed bi-allelic 
RPE65 mutation-associated retinal 
dystrophy. 
Section 5.3 
Study Rational, 
Page 13 & 14 
In defective lymphocytes 
and fibroblasts isolated 
from patients with CHM, 
delivery of full-length 
human REP1 via AAV2-
REP1 restored RabGGTase 
activity.  In addition, 
AAV2-REP1 expresses 
high levels of human REP1 
protein… 
AAV2-REP1 is a recombinant 
AAV2 particle…, 
 
Clarified text 
In 2012, an investigator-
sponsored, first-in-human 
study of AAV2-REP1 was 
initiated in patients with 
CHM (MacLaren 1 study).   
In 2011, an investigator-sponsored, 
first-in-human study of AAV2-
REP1 was initiated in adult male 
patients with CHM.   
Specified males 
Based upon these favorable 
results tThree other 
investigator-sponsored 
studies were then initiated 
in which a total of 32 CHM 
patients have received a 
single sub-retinal injection 
of 1×10^11 gp in 1 eye with 
AAV2-REP1 (the 
MacDonald, Fischer and 
Lam studies).  The initial 
(unpublished) results of 
these trials are consistent 
with improved rod and cone 
function that overcome any 
negative effects of retinal 
detachment, and the gene 
therapy appears to be 
generally well tolerated 
following a single treatment 
with AAV-REP1.   
 
Emerging data from non-
clinical and clinical studies 
with AAV2 vectors 
demonstrate that AAV2 
vectors illicit a minimal 
immune response, including 
after bilateral 
administration.   
Following favorable results 3 other 
investigator-sponsored studies were 
then initiated in which a total of 26 
adult male CHM patients received a 
single sub-retinal injection of 
1×10^11 gp AAV2-REP1 in 1 eye.  
The initial results of these trials are 
consistent with improved rod and 
cone function despite any negative 
effects of retinal detachment, and 
the gene therapy was generally well 
tolerated.  AAV2 REP1 gene 
therapy has been shown to 
positively affect visual acuity (VA), 
considering both maintenance and 
stabilization of vision in the 
majority of subjects, as well as 
improvement of vision in subsets of 
CHM subjects for up to 2 years and 
beyond.  This clinical evidence 
supports the durability of the 
therapeutic effect of AAV2-REP1 
for treating CHM.   
These findings lend support to 
further assessment of AAV2 gene 
therapy in the treatment of CHM.  
NightstaRx is currently sponsoring 
a multi-centre, global Phase 3 study 
of a single subretinal administration 
of AAV2-REP1 for the treatment of 
Clarified text, 
updated # of adult 
male CHM 
patients, added text 
to show clinical 
evidence of 
efficacy and safety 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 11 of 42 
 
Summary of Changes 
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Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
CHM (NSR-REP-01, the STAR 
study, NCT03496012). 
As noted, emerging data from non-
clinical and clinical studies with 
AAV2 vectors demonstrate that 
these vectors illicit a minimal 
immune response, including after 
bilateral administration. 
Section 5.4 
Risk/Benefit 
Assessment 
Page 14 
 
5.4 Risk / Benefit Assessment 
No treatment currently exists for 
CHM. 
The non-clinical studies conducted 
with AAV2-REP1 showed that it 
provides efficient and functional 
transgene expression in CHM 
mouse and human cells, as well as 
in mouse and human RPE and 
photoreceptors, without overt 
toxicity.  Results from the 26-week 
single-dose combined 
toxicity/biodistribution study 
conducted in rats indicate that 
administration of AAV2-REP1 by 
single sub-retinal injection to both 
eyes is well tolerated at dose levels 
of 1×10^9 and 6×10^9 gp/eye 
(equal to 1×10^11 and 6×10^11 
gp/eye in humans) when evaluated 4 
and 26 weeks after injection.  Minor 
reductions in some ERG 
parameters, ocular inflammation, 
and microscopic signs of 
retinal/corneal degeneration were 
observed and were considered 
procedure-related or not 
biologically significant. 
In humans, application of AAV2-
REP1 to the surface of the retina 
requires retinal detachment via 
vitrectomy.  The 2-step procedure 
employed in delivering AAV2-
REP1 sub-retinally (see Section 
9.4), allows the management of 
unexpected surgical complications 
of retinal detachment before the 
AAV2-REP1 is applied. 
Furthermore, since the volume of 
fluid required to detach the fovea is 
variable, by excluding the vector 
from the first step, and 
Added section for 
Risk Benefit 
Assessment with 
new definition of 
vision loss as 
possible risk, as 
recommended by 
regulatory body 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 12 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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administering the vector after 
successful retinal detachment has 
been achieved, a precise, consistent 
dose in terms of genome particles 
can be applied into the sub-retinal 
space.  Nevertheless, sub-retinal 
injection of AAV2-REP1 carries the 
risks associated with vitrectomy and 
retinal detachment, which include 
intra-operative and post-operative 
complications; infection (most 
notably infectious endophthalmitis); 
low and elevated IOP; choroidal 
detachment; persistent retinal 
detachment, retinal tears, holes and 
breaks; macular hole and macular 
oedema; vitreous haemorrhage; 
visual impairment; 
metamorphopsia; and photopsia 
(Park et al., 1995; Thompson et al., 
1996; Banker et al., 1997; Cheng et 
al., 2001; Anderson et al., 2006; 
Stein et al., 2009; Recchia et al., 
2010).  Post-operative intraocular 
inflammation caused by vitrectomy 
is often associated with transient or 
sometimes permanent visual 
impairment.  Another complication 
of vitrectomy is cataract formation, 
which may require an additional 
surgical procedure (cataract 
extraction) (Park et al., 1995; Cheng 
et al., 2001; Recchia et al., 2010).  
In addition, the surgical procedure 
is performed under general 
anesthesia, which includes the risks 
of dizziness, confusion, nausea and 
vomiting. Loss of visual acuity has 
been observed post-treatment with 
AAV2-REP1.; this visual acuity 
loss was determined by the 
investigators to be plausibly related 
to the administration of AAV2-
REP1, however without definitively 
attributing the cause to either the 
study procedure or the GTMP.  Loss 
of visual acuity is, therefore, 
considered a Potential Risk of 
AAV2-REP1 treatment, in addition 
to the above described potential and 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 13 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
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anticipated surgical risks.  See the 
Investigator’s Brochure for AAV2-
REP1 for further details. 
Thus, although there are risks 
associated with the administration 
of the study treatment via 
vitrectomy and retinal detachment, 
the potential for benefit in the form 
of improved visual acuity that may 
be provided to subjects with CHM 
following treatment with AAV2-
REP1 provides an acceptable risk-
benefit profile for participation in 
this study. 
Section 6.0 
Study Objectives 
and Endpoints, 
Page 15 
 
 
The objective of the study is to 
evaluate the safety of bilateral, 
sequential sub-retinal administration 
of a single dose of AAV2-REP1 in 
adult male subjects with CHM. 
Added clarification 
to subject criteria 
The primary endpoint is the 
evaluation of safety 
following bilateral 
administration of AAV2-
REP1.   
The safety of bilateral 
administration of AAV2-REP1will 
be evaluated with the following 
safety measures: 
•BCVA as measured by the ETDRS 
chart 
•Ophthalmic examination 
assessments (including intraocular 
pressure [IOP], slit lamp 
examination, lens opacity grading 
and dilated ophthalmoscopy) 
•Spectral domain optical coherence 
tomography (SD-OCT) 
•Autofluorescence (AF) 
•Fundus photography 
•Microperimetry 
•Adverse event (AE) reporting  
•Vector shedding post-treatment 
•Immunogenicity sampling post-
treatment  
•Vital signs 
Added safety 
assessments to 
endpoints. These 
were already in 
protocol but were 
not specified as 
part of the 
endpoint analyses 
Secondary endpoints 
include: 
The following will also be 
considered secondary efficacy 
endpoints: 
•Change from Baseline in BCVA as 
measured by the ETDRS chart 
•Change from Baseline in AF 
•Change from Baseline in SD-OCT 
•Change from Baseline in 
microperimetry. 
Clarified text, used 
abbreviations 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 14 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
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Section 7.1  
Overall Study 
Design, 
Pages 17 & 18 
 
 
This is a multi-centre, open-
label, prospective, two-
period… 
The study will consist of a 
Screening Visit followed by 
2 treatment periods (Period 
1 and Period 2) with up to 9 
visits per treatment period.  
This is a multi-centre, open-label, 
prospective, 2-period, bilateral 
interventional safety study of 
AAV2-REP1…, 
The study will consist of a 
Screening Period followed by 2 
treatment periods (Period 1 and 
Period 2) with up to 9 visits per 
treatment period.  A second baseline 
visit prior to surgery of SE2 (Period 
2, Visit 1) may be required.   
Clarified text, 
added bilateral to 
design, added 
second baseline  
At the Screening Visit, …,  
 
During the Screening Period…, 
…(i.e., study eye 1 [SE1]-Period 1] 
and study eye 2 [SE2]-Period 2], 
respectively)… 
The estimated target interval 
between the surgical procedures of 
SE1 and SE2 will be determined at 
the Screening Visit. 
Clarified text 
 
Visit 4 (Day 7 ± 1 Day); Visit 5 
(Day 14 ± 3 Days); Visit 6 (Month 
1 ± 7 Days); Visit 7 (Month 3 ± 14 
Days); Visit 8 (Month 6 ±14 Days) 
and Visit 9 (Month 12 ± 14 Days; 
end of study visit). 
Added text to 
clarify visit 
window 
 
The estimated target interval 
between the surgical procedures in 
of SE1 and SE2 will be determined 
at the Screening Visit unless Period 
2 commences during the time of 
follow-up for SE1; in which case, 
the study visits will continue 
following the schedule for SE2.   
Updated text 
The interval between SE1 
and SE2 treatment is 
expected to vary for each 
subject from a few weeks to 
many months.   
Each site will work with the 
Sponsor and the assigned 
Contract Research 
Organisation (CRO) to try 
and ensure an even spread 
across the intervals.  Given 
that the timing of the 
second surgery visit (Period 
2, Visit 1) will vary 
between subjects, the 
duration of study Period 1 
will also be variable.  To 
The interval between SE1 and SE2 
treatment is expected to vary among 
subjects from weeks to many 
months.   
Thus, approximately 20 subjects 
each will be treated with a short- 
(<6 month), medium- (6-12 
months), or long- (>12 month) 
surgery window between treatment 
of the first and second eye.  Because 
the timing of the second surgery 
visit (Period 2, Visit 1) will vary 
among subjects, the duration of 
study Period 1 will also be variable.   
Clarified text, 
added text for 
surgery window as 
recommended by 
FDA. See 
Rationale. 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 15 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
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avoid unnecessary study 
visits, every effort should 
be made by the investigator 
to schedule Period 2, Visit 1 
to coincide with a planned 
Period 1 post-treatment 
visit. 
 
Signs of post-operative 
inflammation or other post-
operative sequelae, as judged by the 
investigator, should be resolved in 
SE1 for the subject to continue to 
Period 2.  After comprehensive 
evaluation of all safety assessments 
and visual functional tests of SE1 
and SE2, the Investigator will 
determine the eligibility of the 
subject and decide the appropriate 
time to schedule SE2, Visit 1. 
Clarified text to be 
more inclusive 
 
All baseline ophthalmic assessments 
must be conducted within 3 months 
prior to SE2 surgery (Period 2, Visit 
1).  If, at that time, BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the investigator and after 
consultation with the sponsor: 1) the 
subject is found eligible by all other 
inclusion and exclusion criteria, and 
2) the subject is considered an 
appropriate candidate who has 
potentially modifiable disease. 
Updated to include 
baseline 
assessment 
requirements if 
second eye is 
treated with a long 
time period 
between surgeries 
Subjects Previously 
Treated with 
AAV2-REP1, 
Pages 18 & 19 
In these subjects, the period 
between SE1 (from 
previous study) and SE2 
(current study) may exceed 
6 months; prolonged 
intervals will not preclude 
subjects from being 
considered for study 
participation. 
Of note, subjects who 
received previous treatment 
with AAV2-REP1 in an 
investigator-sponsored trial, 
an adequate immunology 
profile must be available 
from the previous study for 
the treated eye.   
For subjects who received previous 
treatment with AAV2-REP1 
biological samples must be 
available to obtain an adequate 
immunology profile related to 
treatment of the first eye in the 
previous study.   
For subjects whose first eye was 
dosed in an antecedent study, and 
who do not have the required 
baseline assessments within 12 
weeks prior to surgery, a Baseline 
Visit (see Section 10.1.1 will occur 
be necessary within 12 weeks prior 
to second eye dosing at Visit 1, 
Period 2. 
Updated criteria 
for previously 
treated subjects in 
an antecedent 
study. This new 
language assures a 
proper baseline 
assessment has 
been conducted 
prior to treated 
second eye. 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 16 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Section 7.2 
Discussion of 
Design, 
Page 19 
Bilateral sub-retinal 
administration of AAV2-
REP1 was explored in a 
pre-clinical toxicology 
study in rats and was found 
to be well tolerated and 
safe.   
All enrolled subjects will receive 
treatment with AAV2-REP1 at a 
dose of 1×10^11 AAV2-REP1 
gp/eye. Bilateral sub-retinal 
administration of AAV2-REP1 was 
explored in a pre-clinical toxicology 
study in rats and was found to be 
well tolerated and not associated 
with drug-related AEs.   
Added text for 
dose 
The planned sample size (at 
least 15 subjects/30 eyes)  
The planned sample size 
(approximately 60 subjects, with a 
total of 120 treated eyes) … 
Updated sample 
size 
This open-label safety study 
has no formal control arm. 
The inclusion criterion of 
BCVA ≥74 ETDRS letters 
ensures that the study eyes 
are optimal surgical 
candidates (i.e. earlier phase 
of the disease with larger 
area of viable retina).  This 
will improve evaluation of 
immunogenicity of AAV2-
REP1, including correlation 
between immunogenicity 
and AEs, by limiting 
surgery-related AEs and 
confounding variables. 
 
Deleted text 
The dose of vector being 
employed in this study is 
based on previous clinical 
trials using the AAV2 
vector with a chicken β 
actin (CBA) promoter 
(Maguire 2009; MacLaren 
2014), and the ongoing 
investigator-driven clinical 
studies in which AAV2-
REP1 is being administered 
to subjects with CHM 
(MacLaren 2014; 
MacDonald, Lam and 
Fischer, unpublished data). 
The dose of vector being employed 
in this study is based on previous 
clinical trials using the AAV2 
vector with a chicken β actin (CBA) 
promoter (Maguire 2009; MacLaren 
2014), and the Phase 1 / 2 
investigator sponsored clinical 
studies in which AAV2-REP1 is 
was administered to subjects with 
CHM (MacLaren 2014; 
MacDonald, Lam and Fischer, 
unpublished data). 
Clarified Text 
Application of AAV2-
REP1 to the retina requires 
retinal detachment 
following vitrectomy.  As 
such, sub-retinal injection 
of AAV2-REP1 carries the 
risks associated with 
The risks of vitrectomy and retinal 
detachment are summarised in 
Section 5.4. 
Moved the rsisks 
to the new risk 
benefit assessment 
section 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 17 of 42 
 
Summary of Changes 
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vitrectomy and retinal 
detachment, which include 
intra-operative and post-
operative complications; 
infection (most notably 
infectious endophthalmitis); 
low and elevated IOP; 
choroidal detachment; 
macular oedema; vitreous 
haemorrhage; visual 
impairment; 
metamorphopsia; and 
photopsia (Park 1995; 
Thompson 1996; Banker 
1997; Cheng 2001; 
Anderson 2006; Stein 2009; 
Recchia 2010).  Post-
operative intraocular 
inflammation caused by 
vitrectomy is often 
associated with transient 
visual impairment.  A long-
term complication of 
vitrectomy is cataract 
formation, which may 
require an additional 
surgical procedure (cataract 
extraction) (Park 1995; 
Cheng 2001; Recchia 
2010). 
In line with guidance from 
regulatory agencies, the 
treatment intervals ranging 
from a few weeks to ≥ 6 
months apart were chosen 
to comprehensively 
characterise the 
immunological profile 
following bilateral, 
sequential AAV2-REP1 
treatment.  As noted in the 
first paragraph above, both 
preclinical data with 
AAV2-REP1, … 
In line with guidance from 
regulatory agencies, the treatment 
intervals were chosen to 
comprehensively characterise the 
immunological profile following 
bilateral, sequential AAV2-REP1 
treatment, and to reflect real-world 
application of gene therapy.  As 
noted both preclinical data with 
AAV2-REP1, and clinical data with 
sub-retinal administration of an 
AAV2 vector in subjects with 
Leber's congenital amaurosis, 
support bilateral administration of 
AAV2-REP1 in humans. 
Removed text not 
used in study 
design discussion 
To minimise inflammation 
resulting from potential 
immune responses to 
vector, including treatment 
of the second eye (SE2), 
To minimise inflammation resulting 
from surgery and potential or 
unexpected immune responses to 
vector/transgene, all subjects will be 
given a course of oral 
Added text to 
update clarify 
procedure/ regimen 
for all subjects to 
minimize 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
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Summary of Changes 
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subjects will be given 
ainitiate a  21-day course of 
oral corticosteroid before 
each surgical visit, and will 
be instructed to start taking 
the drug medication 2 days 
prior to SE1/SE2 treatment.  
If the SE2 steroid treatment 
coincides overlaps with the 
SE1 steroid treatment 
period, an extended course 
of steroid treatment will be 
implemented to will cover 
both surgical treatments . 
prednisone/prednisolone, initiated 2 
days prior to surgery on both SE1 
and SE2.  The regimen has been 
modified from the 17-day protocol 
established in the Philadelphia AAV 
gene therapy clinical trial (Maguire 
2008), with the allowance of an 
extra 4 days for tapering the dose at 
the end of the course.  The 
corticosteroid regimen is 1 
mg/kg/day prednisone/prednisolone 
for a total of 10 days (beginning 2 
days before the vector injection, on 
the day of injection, and then for 7 
days); followed by 0.5 mg/kg/day 
for 7 days; 0.25 mg/kg/day for 2 
days; and 0.125 mg/kg/day for 2 
days (21 days in total).  See Section 
9.8 for details. 
inflammation from 
surgery. This was 
necessary due to 
the inclusion of 
short time frames 
between surgeries 
in which case 
steroid treatments 
will overlap. 
Section 8 
Selection and 
Withdrawal of 
Subjects, 
Page 21 
The study will enroll at 
least 15 subjects/30 eyes 
with CHM… 
 
The study will enroll approximately 
60 subjects with CHM, for a total of 
120 treated eyes… 
 
Clarified number 
of subjects and 
eyes enrolled in 
study 
At the Screening Visit, 
subjects are eligible… 
During the Screening Period, 
subjects will be found eligible… 
Clarified text 
Section 8.1 
Inclusion Criteria 
Page 21 
≥34 ETDRS letters 
(equivalent to better than or 
equal 20/40 or better 
Snellen acuity) and ≤≥74 73 
ETDRS letters (equivalent 
to better than or equal to 6/9 
or 20/32 Snellen, decimal 
0.63, LogMar 0.2) in both 
eyes1 or both untreated 
eyes*, if no prior treatment 
with AAV2-REP1 
o≥74 ETDRS letters 
(equivalent to better than or 
equal to 6/9 or 20/32 
Snellen, decimal 0.63, 
LogMar 0.2) in the 
untreated eye, if prior 
treatment with AAV2-
REP1 was received in an 
alternative study* 
3.Have documentation of a 
genetically-confirmed diagnosis of 
CHM 
5. Have a BCVA of ≥34 ETDRS 
letters (20/200 or better Snellen 
acuity) in both eyes, or in the 
untreated eye, if the other eye was 
previously treated with AAV2-
REP1* 
* If previously treated with AAV2-
REP1 in an antecedent study, 
subjects may be eligible for 
participation following Sponsor 
approval.  
6. For subjects who received 
treatment with AAV2-REP1 in an 
antecedent study, have biological 
samples available to complete an 
adequate immunology profile. 
Updated inclusion 
criteria to study 
parameters, added 
criteria for subjects 
previously treated. 
See Section 7 in 
protocol for 
discussion of 
design and 
rationale for 
inclusion criteria. 
Section 8.2  
Exclusion Criteria 
Page 21 
At the Screening Visit, 
subjects are found not 
eligible… 
During the Screening Period, 
subjects will be found not eligible… 
Clarified text 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
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Summary of Changes 
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2. Are unwilling to use barrier 
contraception methods or abstain 
from sexual intercourse for a period 
of 3 months following treatment 
with AAV2-REP1 in either eye. 
Added text for 
clarity 
Section 8.3 
Subject 
Withdrawal 
Criteria 
Pages 22 
Death 
 
Removed text 
Severe ocular inflammation 
(e.g. endophthalmitis) that 
is unresponsive to 
treatment, should result in 
the termination of the trial 
for the affected subject. 
At the discretion of the 
investigator, if a persistent 
post-operative complication 
or poor visual function 
outcome presents post-
operatively in SE1, the 
second eye may not be 
treated and the subject may 
be discontinued from the 
study. 
If severe ocular inflammation (e.g. 
endophthalmitis) or any other post-
operative sequela that is 
unresponsive to treatment occurs in 
SE1 the second eye should not be 
treated.  However, Period 1 should 
be continued for monitoring of 
safety for the duration of the study. 
At the discretion of the investigator, 
if a persistent post-operative 
complication or poor visual function 
outcome presents post-operatively 
in SE1, the second eye may not be 
treated. However, Period 1 should 
be continued for monitoring of 
safety for the duration of the study. 
Updated/clarified 
other withdrawal 
criteria 
 
Subjects who withdraw from the 
study must undergo the Early 
Termination Visit assessments. See 
Table 1. 
Added text for ET 
visit requirement. 
Section 9.1 
Treatments 
Administered, 
Page 23 
At Period 1, Visit 1 (Day 0, 
the Injection Day Visit for 
SE1), subjects will receive a 
volume of up to 0.1 mL of 
study drug containing 
1×10^11 AAV2-REP1 gp.  
At Period 2, Visit 1 (Day 0, 
the Injection Day Visit for 
SE2), subjects will receive 
the same  dose via a sub-
retinal injection. 
At Period 1, Visit 1 (Day 0, the 
Injection Day Visit for SE1), 
subjects will receive in SE1 a sub-
retinal injection up to 0.1 of study 
drug containing 1×10^11 AAV2-
REP1 gp.  At Period 2, Visit 1 (Day 
0, the Injection Day Visit for SE2), 
subjects will receive in SE2 up to 
0.1 mL of AAV2-REP1 via a sub-
retinal injection. 
Clarified text for 
treatments 
Section 9.3 
Packaging, 
Labeling, and 
Storage, Page 23 
(on either the primary or 
secondary container) and 
include the protocol study 
number, Sponsor’s name, 
product name, titre, vial and 
lot number, expiration date, 
storage conditions, and 
caution statement.   
AAV2-REP1 is currently supplied 
in sterile single- use vials… 
Clarified text 
Section 9.4 
Vitrectomy 
Procedure and 
 
To improve visualization of the 
vector and facilitate dosing, 
surgeons are given the option of 
adding a minute quantity of trypan 
Added text to 
include blue dye 
option for 
surgeons, clarified 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
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Summary of Changes 
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Version 3.0, 4 Feb 2019 
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Injection of AAV2-
REP1, 
Page 24 
blue ophthalmic solution (~6 µL) to 
the vector solution.  See the AAV2-
REP1 Surgical Manual for further 
details. 
Prior to administration the vector 
needs to be primed… 
See the GEMINI Surgical Manual 
for further details. 
text, added 
references. 
Section 9.8 
Concomitant 
Therapy, 
Page 25 
Throughout the study, 
investigators may prescribe 
any concomitant 
medications or treatments 
deemed necessary to 
provide adequate supportive 
care.  Details of 
concomitant medications 
will be collected at the 
Screening Visit and updated 
at every study visit 
(including the ET Visit, if 
applicable).  Concomitant 
medications (including oral 
corticosteroid) taken during 
the study are to be recorded 
in the subject’s medical 
records and eCRF; an 
exception to this is any 
medication used in the 
course of conducting a 
study procedure (e.g., 
anaesthesia, dilating eye 
drops). 
To minimise inflammation 
resulting from surgery and 
potential or unexpected 
immune responses to 
vector/transgene, all 
subjects will be given a 21-
day course of oral 
prednisone/prednisolone 
prior to each surgery2 .   
 
Administration of oral 
prednisone/prednisolone is 
to closely follow the 17-day 
protocol established in the 
Philadelphia AAV gene 
therapy clinical trial 
(Maguire 2008), except for 
the allowance of an extra 4 
Throughout the study, subjects may 
be prescribed any concomitant 
medications, procedures, or 
treatments deemed necessary to 
provide adequate supportive care.  
Details of concomitant medications, 
procedures and/or treatments will be 
collected during the Screening 
Period and updated at every study 
visit (including the ET Visit, if 
applicable).  Concomitant 
medications (including oral 
corticosteroid) taken during the 
study are to be recorded in the 
subject’s medical records and 
eCRF; an exception to this is any 
medication used in the course of 
conducting study assessments (e.g., 
ophthalmic dyes, topical anesthesia, 
dilating eye drops). 
In addition, all subjects will be 
prescribed a course of oral 
corticosteroids.  
 
The oral prednisone / prednisolone 
regimen has been modified from the 
17-day protocol established in the 
voretigene neparvovec-rzyl 
Philadelphia AAV gene therapy 
clinical trial for treatment of 
patients with confirmed biallelic 
RPE65 mutation-associated retinal 
dystrophy (Maguire 2008), with the 
allowance of an extra 4 days for 
tapering the dose at the end of the 
course.  The corticosteroid regimen 
is 1 mg/kg/day 
prednisone/prednisolone for a total 
of 10 days (beginning 2 days before 
the vector injection, on the day of 
injection, and then for 7 days); 
followed by 0.5 mg/kg/day for 7 
This revision was 
made to include 
more than just 
concomitant 
medications, but 
also procedures or 
treatments, in the 
collection of 
medical status and 
updates of 
subjects. 
 
 
Also the details of 
the corticosteroid 
regimen are 
provided, and how 
these change 
depending on 
when the second 
eye is treated. 


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da ys i n t otal; St u d y Da y ( S D) -2 
t hr o u g h S D 1 8).  T his re gi me n is 
o utli ne d i n detail bel o w.  
F ull C ortic oster oi d Re gi me n  
Pre d nis o ne or pre d nis ol o ne 
a d mi nistere d as f oll o ws:  
S D -2 t hr o u g h S D 7 ( 1 0 da ys): 1 
m g/ k g b y m o ut h dail y  
S D 8 t hr o u g h S D 1 4 ( 7 da ys): 0. 5 
m g/ k g/ da y b y m o ut h dail y  
S D 1 5 t hr o u g h S D 1 6 ( 2 da ys): 
0. 2 5 m g/ k g/ da y b y m o ut h dail y  
S D 1 7 t hr o u g h S D 1 8 ( 2 da ys): 
0. 1 2 5 m g/ k g/ da y b y m o ut h dail y  
F or S E 1 t he F ull C ortic oster oi d 
Re gi me n s h o ul d be i nitiate d as 
descri be d.   
F or S E 2 s ur gerie s t hat occ ur o n da y 
2 1 or later, t he F ull C ortic oster oi d 
Re gi me n s h o ul d be i nitiate d as 
descri be d, be gi n ni n g 2 da ys b ef ore 
t he sc he d ule d S E 2 s ur ger y. 
If S E 2 is t o be perf or me d pri or t o 
St u d y Da y 2 1, t he n t he F ull 
C ortic oster oi d Re gi me n s h o ul d be 
i nitiate d 2 da ys pri or t o t he 
sc he d ule d s ur ger y f or S E 2, a n d t his 
will s u perce de t he ster oi d ta per i n 
pr o gress f or S E 1.  T he f ull 2 1 da ys 
of treat me nt will t he n be c o m plete d 
f or S E 2.   
If i nfla m mati o n is o bser ve d i n t he 
st u d y e ye, a n d i n t he o pi ni o n of t he 
i n vesti gat or a d diti o nal treat me nt 
wit h c ortic oster oi d me dicati o n is 
i n dicate d, c ortic oster oi d t hera p y 
ma y be i ncrease d d uri n g t he ta per 
peri o d (t o a ma xi m u m of 1 
m g/ k g/ da y), ma y be rei nitiate d 
f oll o wi n g c o m pleti o n of t he ta per, 
a n d/ or ma y be s u p ple me nte d b y 
i ntra ocular c ortic oster oi ds.   
Secti o n 1 0. 1  
Scree ni n g Peri o d  
Scree ni n g  Visit  
 
(e nz y me-li n ke d 
i m m u n os or be nt assa y 
[ E LI S A] a n d e nz y me-
 Scree ni n g  P eri o d  
 
 
U p date d Hea der,  
Re m o ve d te xt  


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 22 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
linked immunospot 
[ELISPOT]) 
 
• Concomitant medication, 
procedures, and treatment review 
Added text  
Subjects will be given two 
21-day courses of oral 
corticosteroid (e.g., 
prednisolone / prednisone) 
and instructed to start 
taking the drug days before 
their next surgery, at Period 
1, Visit 1 for SE1 and 
Period 2, Visit 1 for SE2.  
 
 Subjects will be issued a 
diary card to capture 
corticosteroid compliance 
throughout each 21-day 
period.  Subjects will also 
be instructed to use barrier 
contraception for a period 
of 3 months from the time 
they are treated. 
For each subject, the interval 
between SE1 and SE2 treatment is 
expected to range from weeks to 
months.  While this interval will be 
decided on a case-by-case basis, an 
effort will be made to schedule 
varying treatment intervals in order 
to better characterise the 
immunological and safety profile of 
sequential treatment administration 
and to reflect the anticipated real-
world application of the study drug.  
Thus, approximately 20 subjects 
will be treated with a short- (<6 
month), medium- (6-12 months), or 
long- (>12 month) duration surgery 
window between treatment of the 
first and second eye.  Because the 
timing of the second surgery visit 
(Period 2, Visit 1) will vary among 
subjects, the duration of study 
Period 1 will also be variable. 
The next study visit (i.e., surgery 
and dosing, Period 1, Visit 1) is to 
be scheduled within 10 weeks of the 
Screening Period.   
Subjects will be treated with oral 
corticosteroids (e.g., prednisolone / 
prednisone) during the peri-
operative period for each eye.  They 
will be instructed to start taking the 
drug 2 days before their surgery at 
Period 1, Visit 1 for SE1.  For SE1, 
the Full Corticosteroid Regimen 
should be initiated, as described in 
Section 9.8.   
For SE2 surgeries that occur on Day 
21 or later, the Full Corticosteroid 
Regimen should be initiated as 
described, beginning 2 days before 
the scheduled surgery.  
If SE2 is to be performed prior to 
Study Day 21, then the Full 
Corticosteroid Regimen should be 
initiated 2 days prior to the 
scheduled surgery for SE2, and this 
Updated screening 
visit to include 
discussion on 
subject duration of 
surgery window, 
corticosteroid 
treatment and 
regimen and 
baseline 
assessments. 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 23 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
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(Added Text Shown as Red) 
Rationale 
will supercede the steroid taper in 
progress for SE1.  The full 21 days 
of treatment will then be completed 
for SE2. 
Subjects will be issued a diary card 
to capture corticosteroid 
compliance.  Subjects will also be 
instructed to use barrier 
contraception or abstain from sexual 
intercourse for a period of 3 months 
from the time they are treated. 
All baseline ophthalmic assessments 
must be conducted within 3 months 
prior to SE2 surgery (Period 2, Visit 
1).  If, at that time, BCVA in SE2 is 
<34 ETDRS letters, surgery may be 
performed when, in the opinion of 
the investigator and after 
consultation with the sponsor:  
1. the subject is found eligible by all 
other inclusion and exclusion 
criteria, and 
2. the subject is considered an 
appropriate candidate who will 
potentially benefit from treatment. 
Section 10.1.1 
 
Baseline Visit, 
Period 2 (<12 
Weeks from Visit 1, 
Period 2 [SE2]), 
Page 29 
 
10.1.1 
Baseline Visit, Period 2 
(<12 Weeks from Visit 1, Period 2 
[SE2]) 
The following baseline assessments 
for Period 2 must be performed 
within 12 weeks prior to surgery for 
SE2 (Visit 1, Period 2).  If the 
assessments have not been 
performed within a regularly 
scheduled visit within this window, 
an additional visit is required.  
• Demography, medical and ocular 
history update 
• Vital signs 
• Weight 
• Vector shedding sampling – blood, 
tears (both eyes), urine, saliva 
• Immunogenicity sampling 
• BCVA  
• Microperimetry 
• Full ophthalmic examination, 
including IOP, a slit-lamp 
examination, lens opacity, and 
dilated fundus examination 
• SD-OCT  
Added section 
10.1.1 for Baseline 
Assessments. This 
is a new visit that 
might be necessary 
for subjects whose 
second eye is 
treated an extended 
period of time 
from the first eye. 
This assures 
continued 
eligibility and 
safety of the 
subject for re-
treatment. 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 24 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
(Deleted Text Shown by 
Strikethrough) 
Revised Text 
(Added Text Shown as Red) 
Rationale 
• AF 
• 7-field colour fundus photography 
(including stereo photographs for 
fields 1, 2, and 3) 
• AE) and SAE monitoring 
• Concomitant medication, 
procedures and treatment review 
If BCVA in SE2 is <34 ETDRS 
letters, surgery may be performed 
when, in the opinion of the 
investigator and after consultation 
with the sponsor:  
1. the subject is found eligible by all 
other inclusion and exclusion 
criteria, and 
2. the subject is considered an 
appropriate candidate who has 
potentially modifiable disease. 
Section 10.2  
Period 1 and 2, 
Visit 1 (Day 0, 
Injection Day 
Visit), 
Page 30 
 
• Concomitant medication, 
procedures, and treatment review 
(including review of the 
corticosteroid diary card) 
Updated text 
Subjects will then undergo 
vitrectomy and receive a 
volume to 0.1 mL sub-
retinal injection of AAV2-
REP1, containing 1 × 10^11 
gp in their first (SE1) eye 
Subjects will then undergo 
vitrectomy and receive a volume of 
up to 0.1 mL sub-retinal injection of 
AAV2-REP1, containing 1 × 10^11 
gp. 
Clarified text 
 
or other post-operative sequelae 
Added text 
 
If severe ocular inflammation or any 
other post-operative sequela that is 
unresponsive to treatment occurs in 
SE1, the second eye should not be 
treated. However, Period 1 should 
be continued for monitoring of 
safety for the duration of the study. 
Added text for 
severe ocular 
inflammation or 
other post-
operative sequela 
Sections 10.3 – 10.9 
Pages 31-33 
(ELISA and ELISPOT) 
At Visit X (Post -operative Day X), 
• Concomitant medication, 
procedures, and treatment review 
Clarified text for  
post-operative 
Visits 3-8, and ET 
Visit in sections 
10.3 – 10.9 
Section 10.9 
Early Termination 
(ET Visit), 
Page 31 
 
10.9 Early Termination (ET Visit) 
Added to header 
Section 10.10 
Unscheduled Visits, 
Page 34 
 
(including review and return of the 
corticosteroid diary card) 
Added text 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 25 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
Section 11.1.1.1 
Adverse Event, 
Page 35 
 
Worsening of the signs, symptoms 
and visual function related to CHM 
is not to be considered an AE 
unless, in the opinion of the 
investigator, the changes are not 
consistent with the natural 
progression of the disease. 
Text added to 
clarify that 
worsening of CHM 
is not considered 
an AE unless the 
PI considers it is 
inconsistent with 
the normal course 
of the disease. 
Surgery-related events of 
VA decrease are defined as 
VA decreases occurring in 
close temporal association 
(within <24 hours) with the 
surgical administration of 
the study medication, and 
which are resolving at Day 
7 (Period 1/2, Visit 4) post-
surgery.  These events are 
not to be reported as an AE 
or SAE. However, they 
should be reported as an AE 
if in the investigator's 
opinion, their evolution in 
terms of duration or 
severity cannot be 
explained by the procedure.  
This would include, but not 
be limited to instances 
where the abnormal course 
of post-surgery VA 
decrease is associated with 
another complication 
attributable to the surgery 
or the study medication, or 
where the abnormal course 
of post-surgery VA 
decrease can be attributed 
to another identifiable 
cause. 
• AEs that in the opinion of 
the investigator, actually or 
potentially require any 
surgical or medical 
intervention to prevent 
permanent loss of sight. 
Vision Loss to be Reported as 
Serious Adverse Events 
Vision Loss  not to be Reported as 
an SAE: 
• Surgery-related BCVA decrease of 
≥15 letters on ETDRS chart 
occurring within 1 day of surgery, 
but recovering / resolving at post-
operative Days 7 and 14.  
Vision Loss or Vision-Threatening 
Event to be Reported as an SAE: 
• Surgery-related BCVA decrease of 
≥15 letters on ETDRS chart that 
occurs within 1 day of surgery and 
that has not recovered* by the 1-
Month Visit. 
• A decrease in BCVA of ≥15 letters 
on ETDRS chart that occurs within 
1 day of surgery, however, in the 
investigator’s opinion: 
o Has an evolution not consistent 
with the expected post-operative 
course;  
o May be attributable to a 
complication that occurred during 
surgery, or another untoward 
event, or the study drug;  
o Actually or potentially requires 
any surgical or medical 
intervention to prevent permanent 
loss of vision. 
• Non-surgery-related, sustained 
(>48 hours duration) decrease from 
baseline in BCVA of ≥15 letters on 
ETDRS chart. 
*Recovery / Resolution of BCVA is 
defined as a return to baseline 
BCVA within 5 letters on the 
ETDRS chart. 
Clarified vision 
loss procedure for 
AE/SAE reporting. 
Without changing 
the meaning of the 
original language, 
we have revised 
this section to 
clarify what does 
and what does not 
need to be reported 
as an SAE 
regarding vision 
loss. 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 26 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Rationale 
Section 11.1.2 
 
Recording of 
Adverse Events, 
Page 36 
Non-serious AEs will be 
collected from Period 1, 
Visit 1 (Injection Day Visit 
for SE1) through Period 2, 
Visit 9 (or ET Visit, if 
applicable). 
AEs/SAEs  will be collected from 
the time the subject provides written 
informed consent (Screening Visit) 
through Period 2, Visit 9 (Month 12 
± 14 Days or ET Visit, if 
applicable).  
Clarified text 
Unlikely to be Related: 
there are factors (evidence) 
explaining the occurrence 
of the event (e.g. 
progression of the 
underlying disease or 
concomitant medication 
more likely to be associated 
with the event) or a 
convincing alternative 
explanation for the event 
AE severity and relationship to the 
study medication or the surgical 
procedure will be assessed at the 
site by the investigator or a 
medically qualified designee. 
Added text to 
clarify AE severity 
and relationship 
assessments 
 
AE Severity 
AE Relationship 
Added headers 
 
Related:  A reasonable possibility 
exists that the study medication / 
study procedure caused the AE. A 
suspected AE can be further defined 
by: … 
Added text to 
clarify suspected 
related AE 
definitions 
AE severity and 
relationship to the study 
medication or the surgical 
procedure will be assessed 
at the site by the 
investigator or a medically 
qualified designee. 
When a relationship is determined 
to exist, the investigator or medical 
designee will further define if that 
relationship is to the study 
medication, the study procedure, 
both, or unknown. 
Clarified text 
Section 11.1.3 
 
Follow-up of 
Adverse Events, 
Page 38 
Investigators are expected 
to timely provide the 
requested additional 
information for a complete 
assessment and 
documentation of the SAE 
reports. 
Investigators are expected to  
provide the requested additional 
information for a complete 
assessment and documentation of 
the SAE reports within a timely 
manner 
Clarified text 
Section 11.1.4 
Reporting of 
Serious Adverse 
Events, 
Page 39 
All cases that are fatal or 
life-threatening will be 
reported immediately, and 
not later than 7 days… 
 
…via phone to the Chiltern 
Medical Monitor.  The NSR 
Medical Monitor is the first 
escalation point when the 
Chiltern/NSR Medical 
Monitor is not available.  
For reporting purposes, this time 
will be designated as Day 1. 
 
The Sponsor will report SAEs and 
Suspected, Unexpected, Serious 
Adverse Reactions (SUSARs) to 
investigative sites, the Institutional 
Review Board (IRB)s/Independent 
Ethics Committee (IEC)s, and 
regulatory authorities in compliance 
with current regulations.  All cases 
Updated the 
adverse event 
reporting, what 
defines Day 1, and 
when serious life-
threatening events 
should be reported 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 27 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Rationale 
The second escalation point 
is the NSR Chief Medical 
Officer. 
that are fatal or life-threatening will 
be reported immediately after the 
Sponsor received the initial report 
from the Investigator.   
 
Emergency contact information is 
provided below.  Any urgent 
queries relating to subject safety 
should be communicated 
immediately via phone using the 
contact information provided below.  
Additional contact information will 
be available in the Site Operations 
Manual. 
, MD 
 
Medical Monitor 
NightstaRx, Ltd 
Contract Research 
Organisation 
Chiltern Medical Monitor 
(out-of-hours) EU 
, MD 
Chiltern Ltd. 
Direct: 
 
 
Mobile: 
 
 
E mail: 
 
Pharmacovigilance 
5003 S. Miami Blvd, Suite 500 
Durham, NC  27703 
Email:  
APV.AENightstar@ashfieldpv.com 
+1 919 401 8003 
 
Contract Research Organisation 
Covance Clinical and Periapproval 
Services, Ltd 
Osprey House 
Maidenhead Office Park, 
Westacott Way, Littlewick Green 
Maidenhead SL6 3QH, UK  
+44 (0) 1628 548000 
Updated Contact 
information  
Section 11.2 
Pregnancy, 
Page 40 
 
In addition, if possible, outcome of 
the pregnancy (birth or spontaneous 
abortion)  fathered by the subject 
should be recorded  and any 
incidents of  congenital abnormality 
or birth defect should be reported. 
Clarified text to 
align with 
informed consent 
Section 11.3 
Stopping Criteria, 
Page 40 
• Severe ocular 
inflammation (e.g. 
endophthalmitis) that is 
unresponsive to treatment 
should result in termination 
of the trial for the affected 
subject. 
• Severe ocular inflammation (e.g. 
endophthalmitis) that is 
unresponsive to treatment should 
result in the affected subject not 
being treated in the second eye. 
However, the subject will continue 
to be followed for safety for the 
duration of the study. 
Clarified text so 
that it is clear that 
first eye should be 
followed until 
event has resolved 
even if the second 
eye will no longer 
be treated  
 
At the request of the Data 
Monitoring Committee, the trial 
may be stopped based on specific 
safety concerns (ie, SUSARs), not 
Clarified text 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 28 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
Previous Text 
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Rationale 
defined in the Investigator’s 
Brochure (Section 6.3). 
The study may be discontinued if 
the Sponsor deems it necessary for 
medical, safety, regulatory, 
business, or other reasons consistent 
with applicable laws or regulations. 
Section 11.4.2 
Immunogenicity 
 
 
 
All immunogenicity 
samples will be sent to a 
central laboratory for 
analysis. 
Selected samples will be analysed.  
 
 
 
 
 
 
 
 
Remaining samples may be stored 
for up to 15 years or per local 
regulations.  
Removed test 
stating that all 
samples will be 
analysed. Added 
text that selected 
samples will be 
analysed. Added 
text regarding the 
storing of samples.  
Section 11.6 
Best-Corrected 
Visual Acuity, 
Page 41 
 
11.6 Best-Corrected Visual Acuity 
To evaluate changes in BCVA over 
the study period, BCVA will be 
assessed using the ETDRS VA chart 
and performed for both eyes at the 
times indicated in Table 1. 
The BCVA test should be 
performed prior to pupil dilation, 
and distance refraction should be 
carried out before BCVA is 
measured.  Initially, letters are read 
at a distance of 4 metres from the 
chart.  If <20 letters are read at 4 
metres, testing at 1 metre should be 
performed.  BCVA is to be reported 
as number of letters read correctly 
by the subject. 
For BCVA, assessors will be 
appropriately qualified for 
conducting the assessment. 
BCVA is also used for evaluations 
of efficacy.  
If a subject is not able to perform an 
assessment due to poor VA, it will 
be documented accordingly in the 
eCRF and will not be recorded as a 
protocol deviation. 
Re-ordered 
assessment to 
appear first in 
safety and then 
linked the 
respective 
assessments 
repeated in 
efficacy 
 
 
Section 11.7 
Microperimetry, 
 
11.7 Microperimetry 
Re-ordered 
assessment to 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 29 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
Pages 41 
Microperimetry will be conducted 
on both eyes at the times indicated 
in Table 1. 
Microperimetry will be conducted 
by certified technicians to assess 
changes in retinal sensitivity within 
the macula.  All microperimetry 
images will be sent by the sites to a 
Central Reading Centre (CRC) for 
review. For complete technical 
specifications for microperimetry, 
refer to the Study Operations 
Manual (which will include 
procedures from the CRC regarding 
how measurements are to be taken). 
appear first in 
safety and then 
linked the 
respective 
assessments 
repeated in 
efficacy 
Section 11.8 
Fundus 
Autofluorescence, 
Page 48 
 
11.8 Fundus Autofluorescence 
To assess changes in the area of 
viable retinal tissue, fundus 
autofluorescence will be performed 
on both eyes at the times indicated 
in Table 1. 
All AF images will be performed by 
certified technicians at the site after 
dilation of the subject’s pupil and 
sent to a CRC for review; the CRC 
will enter the data into the EDC 
system.  For complete technical 
specifications for AF, refer to the 
Study Operations Manual (which 
will include procedures from the 
CRC regarding how measurements 
are to be taken). 
 
Re-ordered 
assessment to 
appear first in 
safety and then 
linked the 
respective 
assessments 
repeated in 
efficacy 
Section 11.9 
Spectral Domain 
Optical Coherence 
Tomography, 
Page 42 
 
11.9 Spectral Domain Optical 
Coherence Tomography  
SD-OCT will be performed on both 
eyes at the times indicated in Table 
1. 
SD-OCT measurements will be 
taken by certified technicians at the 
site after dilation of the subject’s 
pupil.  All OCT scans will be 
submitted by the sites to a CRC 
where the scans will be evaluated; 
the CRC will enter the data into the 
EDC system.  SD OCT will be used 
to quantify integrity of the ellipsoid 
zone and reduction in the signal 
from the outer nuclear layer and 
choroid. In addition, foveal changes 
Re-ordered 
assessment to 
appear first in 
safety and then 
linked the 
respective 
assessments 
repeated in 
efficacy 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
Page 30 of 42 
 
Summary of Changes 
Protocol NSR-RPGR-02 
Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
will be assessed.  For complete 
technical specifications for SD-
OCT, refer to the Study Operations 
Manual (which will include 
procedures from the CRC regarding 
how measurements are to be taken). 
Sections 11.10 & 
11.11,  
Page 42 
11.6 Full Ophthalmic 
Examination 
11.7 7-Field Colour Fundus 
Photography 
11.10 Full Ophthalmic Examination 
11.11 7-Field Colour Fundus 
Photography 
Header numbers 
renumbered to 
account for 
additional sections 
above. 
Section 11.11 
Field Colour 
Fundus 
Photography, 
Page 42 
(which will include 
procedures from the CRC 
regarding how 
measurements are to be 
taken). 
 
Text removed 
Section 12 
Assessment of 
Efficacy, 
Page 43 
All efforts should be made 
to conduct the efficacy 
assessments; if a subject is 
not able to perform an 
assessment due to poor 
visual acuity (VA), it will 
be documented accordingly 
in the eCRF and will not be 
recorded as a protocol 
deviation. 
 
To evaluate changes in VA 
over the study period, 
BCVA will be assessed 
using the ETDRS VA chart 
and performed for both eyes 
at the times indicated in 
Table 1. 
The BCVA test should be 
performed prior to pupil 
dilation, and distance 
refraction should be carried 
out before BCVA is 
measured.  Initially, letters 
are read at a distance of 4 
metres from the chart.  If 
<20 letters are read at 4 
metres, testing at 1 metre 
should be performed.  
BCVA is to be reported as 
number of letters read 
correctly by the subject. 
For BCVA, assessors will 
be appropriately qualified 
The following assessments are 
primary safety assessments that will 
also be considered secondary 
efficacy endpoints.   
 
Best-Corrected Visual Acuity 
 
See Section 11.6. 
 
See Section 11.7. 
 
See Section 11.8. 
 
See Section 11.9. 
Removed text from 
sections 12.1 – 
12.4 and added 
references to safety 
sections for 
procedure 
information, added 
best-corrected to 
VA section header 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
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Version 3.0, 4 Feb 2019 
Section, Page 
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Rationale 
for conducting the 
assessment. 
 
To assess changes in the 
area of viable retinal tissue, 
fundus autofluorescence 
will be performed on both 
eyes at the times indicated 
in Table 1. 
All fundus autofluorescence 
images will be performed 
by certified technicians at 
the site after dilation of the 
subject’s pupil and sent to a 
CRC for review; the CRC 
will enter the data into the 
EDC system.  For complete 
technical specifications for 
autofluorescence, refer to 
the Study Operations 
Manual (which will include 
procedures from the CRC 
regarding how 
measurements are to be 
taken). 
 
SD-OCT will be performed 
on both eyes at the times 
indicated in Table 1. 
12.4 
SD-OCT 
measurements will be taken 
by certified technicians at 
the site after dilation of the 
subject’s pupil.  All OCT 
scans will be submitted by 
the sites to a CRC where 
the scans will be evaluated; 
the CRC will enter the data 
into the EDC system.  SD 
OCT will be used to 
quantify integrity of the 
ellipsoid zone and reduction 
in the signal from the outer 
nuclear layer and choroid. 
In addition, foveal changes 
will be assessed.  For 
complete technical 
specifications for SD-OCT, 
refer to the Study 
Operations Manual (which 


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4 FEB 2019 
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will include procedures 
from the CRC regarding 
how measurements are to 
be taken). 
 
Microperimetry will be 
conducted on both eyes at 
the times indicated in Table 
1. 
Microperimetry will be 
conducted by certified 
technicians to assess 
changes in retinal 
sensitivity within the 
macula.  All 
microperimetry images will 
be sent by the sites to a 
CRC for review; the CRC 
will enter the data into the 
EDC system.  For complete 
technical specifications for 
microperimetry, refer to the 
Study Operations Manual 
(which will include 
procedures from the CRC 
regarding how 
measurements are to be 
taken). 
Section 13 
Statistacal 
Considerations, 
Page 45 
Details of the statistical 
analyses will be described 
separately in the Statistical 
Analysis Plan. 
 
Removed reference 
to SAP 
Section 13.1 
Sample Size, 
Page 45 
At least 15 subjects 30 eyes 
are planned to be enrolled 
in the study.  This sample 
sizeis considered sufficient 
Approximately 60 subjects are 
planned to be enrolled in the study.  
A total of 120 treated eyes is 
considered sufficient … 
Updated number of 
subjects and eyes 
Section 13.2 
Missing Data, 
Page 45 
 
Management of dropout and 
missing observations will depend on 
their nature and frequency and will 
be discussed as part of the Data 
Review meeting before database 
lock. 
Added text for 
procedure 
Section 13.3 
Analysis Sets, 
Page 45 
 
The ‘Safety Analysis Set 
will consist of all subjects 
who receive study treatment 
(vitrectomy/AAV2-REP1) 
in the second (study) eye.  
The Safety Subjects 
Analysis Set will be the 
primary population for 
The “All Treated Subjects’ Analysis 
Set will consist of all subjects who 
attend the ‘Day of Surgery’ visit of 
Period 1 (or, of Period 2, for 
subjects who received AAV2-REP1 
treatment in an antecedent study). 
The All Treated Subjects Analysis 
Set will be the primary population 
Updated text for 
All Treated 
Subjects analysis  


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4 FEB 2019 
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demographics, baseline 
characteristics, and safety 
and efficacy analyses. 
for demographics, baseline 
characteristics, and safety and 
efficacy analyses. 
Section 13.4 
Descriptive 
Statistics and 
Conventions, 
Page 45 
13.4 Descriptive Statistics 
and Conventions 
Continuous variables 
(including changes from 
Baseline) will be 
summarised over time using 
descriptive statistics (i.e., 
mean, standard deviation, 
95% confidence interval 
[CI], median, Q1, Q3, P05, 
P95, minimum, and 
maximum).   
Data of subjects with no 
surgery in the second eye 
will be listed. 
13.4 Descriptive Statistics and 
Conventions 
Detailed specifications of the 
planned analyses will be included in 
a separate Statistical Analysis Plan 
(SAP). 
Continuous variables (including 
changes from Baseline) will be 
summarised over time using 
descriptive statistics (i.e., mean, 
standard deviation, 95% confidence 
interval [CI], median, first and third 
quartiles,Q1, fifth and ninety-fifth 
percentiles, minimum, and 
maximum).   
Baseline is defined, for each eye 
and each period, as the last non-
missing value prior to treatment of 
the respective period, in the 
respective eye. 
Added to header, 
Added text for 
SAP, 
Added text to 
clarify baseline 
descriptive 
statistics 
Section 13.5 
Demographics and 
Baseline 
Characteristics, 
Page 45 
 
Demographics will be summarised 
for all subjects combined. Age at 
the date of the Period 1 informed 
consent will be calculated. Baseline 
ocular characteristics will be 
summarised by eye and period. 
Updated to include 
age at informed 
consent calculation 
Section 13.6  
Safety Analysis, 
Pages 45 & 46 
Statistical tests will not be 
performed. Alternatively, 
95% 2-sided CIs will be 
calculated where 
appropriate. 
 
No statistical tests will be 
performed. Safety assessments that 
are also considered efficacy 
assessments are presented in this 
section and not repeated in the 
efficacy section. 
Updated text for no 
statistical test 
being performed 
Section 13.6.1 
Visual Acuity, 
Page 46 
The proportion of subjects 
with a ≥5, ≥10- and ≥15-
letter decrease from 
baseline in BCVA will be 
summarised by eye, period 
and visit. 
The proportion of subjects 
with a ≥10- and ≥15-letter 
improvement from baseline 
in BCVA will be 
summarised by eye, period 
and visit.  Mean BCVA and 
mean change from Baseline 
in BCVA will be tabulated 
by eye, period and visit. 
13.6.1 
Visual Acuity 
See Section 13.7.1 
Added text to 
reference VA 
safety analysis 
section in 
Statistical 
Considerations 


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4 FEB 2019 
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Version 3.0, 4 Feb 2019 
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Section 13.6.2 
Ophthalmic 
Examination, 
Page 46 
Full Ophthalmic 
Examination 
IOP and change from 
Baseline in IOP will be 
summarised by visit and 
eye.  Abnormal slit lamp 
examination findings and 
dilated ophthalmoscopy 
findings will be summarised 
by visit and eye. 
Lens opacity categories and 
shifts from baseline will be 
summarised by visit and 
eye. 
13.6.2 
Ophthalmic Examination 
IOP and change from Baseline in 
IOP will be summarised by eye, 
period and visit.  Abnormal slit 
lamp examination findings and 
dilated ophthalmoscopy findings 
will be summarised by eye, period 
and visit. Lens opacity categories 
and shifts from baseline will be 
summarised by eye, period and 
visit. 
Updated text for 
ophthalmic 
examination 
Section 13.6.3 
Spectral Domain 
Optical Coherence 
Tomography, 
Page 46 
Spectral Domain Optical 
Coherence Tomography 
See Section 13.7.2 
Added reference  
to efficacy 
description for SD-
OCT analysis 
Section 13.6.4 
Fundus 
Autofluorescence, 
Page 46 
 
See Section 13.7.3 
Added reference to 
efficacy 
description for AF 
analysis 
Section 13.6.5 
 
7-Field Colour 
Fundus 
Photography, 
Page 46 
7-Field Colour Fundus 
Photography 
Categories of colour fundus 
photography findings will 
be summarised by visit and 
eye. 
Categories of colour fundus 
photography findings will be 
summarised by eye, period and 
visit. 
Updated text to 
clarify summaries 
Section 13.6.6 
Microperimetry, 
Page 46 
Microperimetry 
See Section 13.7.4 
Added reference to 
efficacy 
description for 
microperimetry 
analysis 
Section 13.6.7 
Adverse Events, 
Page 46 
Spectral Domain Optical 
Coherence Tomography 
Microperimetry 
Full Ophthalmic 
Examination 
IOP and change from 
Baseline in IOP will be 
summarised by visit and 
eye.  Abnormal slit lamp 
examination findings and 
dilated ophthalmoscopy 
findings will be summarised 
by visit and eye. 
Lens opacity categories and 
shifts from baseline will be 
AEs will be coded using the 
Medical Dictionary for Regulatory 
Affairs, version 21.0 or higher .  
Events will be summarised by 
system organ class and preferred 
term and eye.  The number of eyes 
with an AE, as well as the number 
of events, will be summarised, by 
period and eye.  Similar summaries 
will be produced for study 
drug/procedure-related AEs, AEs 
leading to discontinuation, and 
SAEs. AEs will also be summarised 
by maximum severity, relationship 
Addeded text for 
AE coding and 
summaries 


NightstaRx, Ltd 
 
4 FEB 2019 
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Version 3.0 
 
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Version 3.0, 4 Feb 2019 
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summarised by visit and 
eye. 
7-Field Colour Fundus 
Photography 
Categories of colour fundus 
photography findings will 
be summarised by visit and 
eye. 
to study drug/procedure, and time to 
onset and resolution. 
Section 13.6.8 
Immunogenicity 
and Vector 
Shedding, 
Page 47 
 
 
Heading number 
changed to 13.6.8 
Section 13.6.9 
Vital Signs, 
Page 47 
14.6.3Adverse Events 
AEs will be coded using the 
Medical Dictionary for 
Regulatory Affairs, version 
XX.  The version of the 
dictionary current at the 
time of database lock will 
be used.  Events will be 
summarised by system 
organ class and preferred 
term.  The number of 
subjects and the number of 
eyes with an AE, as well as 
the number of events, will 
be summarised.  Similar 
summaries will be produced 
for study drug/procedure-
related AEs, AEs leading to 
discontinuation, and SAEs. 
AEs will also be 
summarised by maximum 
severity, relationship to 
study drug/procedure, and 
time to onset and resolution. 
14.6.42.1.1Full Ophthalmic 
Examination 
IOP and change from 
Baseline in IOP will be 
summarised by visit and 
eye.  Abnormal slit lamp 
examination findings and 
dilated ophthalmoscopy 
findings will be summarised 
by visit and eye. 
Lens opacity categories and 
shifts from baseline will be 
13.6.9 
Vital Signs 
Vital signs will be summarised by 
period and visit. 
Moved all deleted 
text to safety 
section, Added 
vital sign to safety 
analysis 


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Version 3.0, 4 Feb 2019 
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summarised by visit and 
eye. 
14.6.52.1.17-Field Colour 
Fundus Photography 
Categories of colour fundus 
photography findings will 
be summarised by visit and 
eye. 
14.6.62.1.1Vital Signs 
Vital signs will be 
summarised in a descriptive 
manner. 
Section 13.7.1 
Visual Acuity 
Page 48 
 
13.7.1 
Visual Acuity 
The proportion of subjects with a 
≥10- and ≥15-letter improvement 
from baseline in BCVA will be 
summarised by eye, period and 
visit.  Mean BCVA and mean 
change from Baseline in BCVA will 
be tabulated by eye, period and 
visit. 
These descriptions 
were moved from 
safety to efficacy 
in the statistical 
section 
Section 13.7.2 
Spectral Domain 
Optical Coherence 
Tomography, 
Page 48 
See Section 13.6.3. 
13.7.2 
Spectral Domain Optical 
Coherence Tomography 
Vitreo-macular Interface Disease, 
Intraretinal Hyper-Reflective Spots, 
Cystoid Macular Edema, Subretinal 
Fluid, Subretinal Hyper-reflective 
Material and Pigment Epithelial 
Detachment will be summarised by 
eye, period and visit, as well as 
shifts from baseline. 
These descriptions 
were moved from 
safety to efficacy 
in the statistical 
section 
Section 13.7.3 
 
Fundus 
Autofluorescence, 
Page 48 
See Section 13.6.4.   
13.7.3 
Fundus Autofluorescence 
Total Area of Preserved AF and 
Distance from Foveal Center to 
nearest Border of Preserved AF, and 
their change from baseline, will be 
summarized by eye, period and 
visit. 
These descriptions 
were moved from 
safety to efficacy 
in the statistical 
section 
Section 13.7.4 
Microperimetry, 
Page 48 
Formal statistical tests will 
not be performed.The 
proportion of subjects with 
a ≥10- and ≥15-letter 
improvement from baseline 
in BCVA will be 
summarised by eye and by 
visit. 
Mean BCVA and mean 
change from Baseline in 
BCVA will be tabulated by 
eye and by visit. 
13.7.4 
Microperimetry 
Fixation Location and Stability, and 
their shift from baseline, will be 
summarised by eye, period and 
visit. Bivariate Contour Ellipse Area 
and Mean Sensitivity, and their 
change from baseline, will be 
summarised by eye, period and 
visit. An analysis specific to the 
Sensitivity Points will be 
performed, the details of which will 
be described in the GEMINI SAP 
These descriptions 
were moved from 
safety to efficacy 
in the statistical 
section 


NightstaRx, Ltd 
 
4 FEB 2019 
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Version 3.0, 4 Feb 2019 
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Section 14 
Informed Consent, 
Page 47 
Independent Ethics 
Committee (IEC) / 
Institutional Review Board 
(IRB) 
 
Clarified text 
 
Male subjects…… 
Added Male to 
clarify subject 
criteria 
Section 14.3 
Regulatory 
Considerations, 
Page 47 
 
The study, where permissible, … 
Clarified data 
Section 15.2 
Data Handling and 
Records 
Management, 
Page 48 
database (by KCT Data). 
database (by KCT Data). 
 
Deleted text 
Section 15.3 
Access to source 
Documentation and 
Subject Privacy, 
Page 49 
The study will comply with 
the data protection laws 
which require data to be 
anonymised. as soon as it is 
practical to do so. 
 
Deleted text 
Section 15.4 
 
Time and Schedule 
of the Study, 
Page 49 
 
The estimated overall study 
duration is 12 months of follow-up 
per eye. 
Clarified text 
Reference List, 
Page 50 
 
Charles River Final Report Study 
Number 5700437; Sponsor 
Reference Number NSRP0914. A 4 
and 24 week (single dose) ocular 
tolerance study following subretinal 
injection of a rAAV2.REP-1 vector 
in the Brown Norway rat. IND 
SN0000, Module 4.2.3.1. 
 
Streilein JW. Ocular immune 
privilege: the eye takes a dim but 
practical view of immunity and 
inflammation. J Leukoc Biol 
2003;74(2):179-85. 
 
Stein-Streilein J, Watte C. Cross-
talk among cells promoting anterior 
chamber-associated immune 
deviation. Niederkorn JY, Kaplan 
HJ (eds): Immune Response and the 
Eye. Chem Immunol Allergy. Basel, 
Karger, 2007, vol 92: 115-130. 
Added references 
Section 17  
Appendix,  
 
 
Changes reflect 
changes in 


NightstaRx, Ltd 
 
4 FEB 2019 
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Version 3.0 
 
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Version 3.0, 4 Feb 2019 
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Table 1 
Schedule of Study 
Procedures 
Page 53 & 54 
assessments 
detailed above 
under individual 
visits, 
Abbreviations 
updated 
 
 
 
Immunogenicity sampling 
(ELISA and ELISPOT) 
 
ELISA=enzyme-linked 
immunosorbent assay; 
ELISPOT=enzyme-linked 
immunospot 
a. Screening Visit/Period must be 
performed <10 weeks of Period 1, 
Visit 1. Selection of SE1 and SE2 
will be determined by the 
investigator in collaboration with 
the subject.  
b. Baseline assessments must occur 
before the SE2 is dosed. These can 
occur at a regularly scheduled visit 
if that visit is within 12 weeks prior 
to surgery in SE2. If it has been 
more than 12 weeks since a visit, 
the subject must return for 
assessments at this Baseline Period 
2 visit.  Similarly, for subjects who 
have previously been treated 
unilaterally with AAV2-REP1 in an 
antecedent study, these baseline 
assessments must be performed ≤12 
weeks prior to surgery on Visit 1, 
Period 2. A separate visit is only 
required if the assessments have not 
been performed within 12 weeks 
prior of surgery in SE2.  
c. Subjects will undergo vitrectomy 
and AAV2-REP1 administration to 
SE1 in Period 1; SE2 in Period 2.  
d. ET visit is to be performed if a 
subject discontinues at any time.  
e. If clinically indicated, subjects 
may need to return to the site for an 
unscheduled visit. At a minimum, 
the following assessments will be 
performed: BCVA, full ophthalmic 
examination, SD-OCT, 
autofluorescence, concomitant 
medication, procedures and 
treatment review and AE/SAE 
monitoring. 
f .Weight is collected for dose 
calculation of the corticosteroid 
regimen. 
Updated text in all 
footnotes to reflect 
changes in 
schedule of 
assessments 


NightstaRx, Ltd 
 
4 FEB 2019 
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g. Blood, tears (both eyes), urine 
and saliva samples.  
h. If ET visit occurs within 3 
months post-treatment. 
i. Immunoassays are planned to 
assess cell-based and antibody-
based immune responses against 
AAV2-REP1. Enzyme-linked 
immunospot (ELISPOT) assays will 
be used for the assessment of T-
cell-mediated immune responses to 
transgene product (REP1) and the 
capsid.  Antibody responses against 
the capsid and transgene product 
will be measured using a cell-based 
neutralization assay and an enzyme-
linked immunosorbent assay 
(ELISA), respectively. All 
immunogenicity samples will be 
sent to a central laboratory for 
analysis. 
j. The ophthalmic examination will 
include IOP, slit lamp examination, 
lens opacity grading, and dilated 
ophthalmoscopy. The same slit 
lamp machine and lighting 
conditions should be used across the 
study for each subject. 
k. Stereo photos for fields 1, 2, 3. 
l. AEs/SAEs will be collected from 
the time the subject provides written 
informed consent (Screening Visit) 
through Period 2, Visit 9 (or ET 
Visit, if applicable).  
m. Subjects will be given a course 
of oral corticosteroid before each 
surgical visit, and instructed to start 
taking the drug 2 days prior to 
SE1/SE2 treatment, unless the SE2 
surgical treatment coincides with 
the SE1 steroid treatment period, 
when an extended course of steroid 
can cover both surgical treatments.  
This regimen is outlined in detail in 
Section 9.8. 
For SE1, the Full Corticosteroid 
Regimen should be initiated as 
described.  For SE2, and surgeries 
that occur on day 21 or later, the 
Full Corticosteroid Regimen should 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
CONFIDENTIAL 
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Version 3.0, 4 Feb 2019 
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be initiated as described, beginning 
2 days before the scheduled SE2 
surgery.  If SE2 is to be performed 
prior to Study Day 21, then the Full 
Corticosteroid Regimen should be 
initiated 2 days prior to the 
scheduled surgery for SE2, and this 
will supercede the steroid taper in 
progress for SE1.  The full 21 days 
of treatment will then be completed 
for SE2 
Section 17 
Table 1 
Headers 
Screening 
Period 1 Screeninga 
Period 2 Screeninga Baselineb 
Added column to 
separate Screening 
Visit to 2 
columns;, Period 1 
Screening & 
Period 2 Baseline, 
Clarified text in 
superscript to 
reference footnotes 
Day 0 Injection Dayb 
Day 0 Injection Dayc 
ET Visitc 
Unscheduled Visitd 
ET Visitd 
Unscheduled Visite 
Table 1, 
Screening Column 
 
Period 1 Screeninga 
Added column to 
present Period 1 
Screening 
assessment 
procedures 
Table 1  
Assessment/ 
Procedures Column 
 
Weightf 
Vector shedding samplingg 
Immunogenicity samplingi  
Full ophthalmic examj 
7-field colour fundus photosk 
AE/SAE monitoringl 
Concomitant medication, 
procedures, and treatment reviewm 
Updated 
superscript to 
footnote changes 
Weighte 
Vector shedding samplingf 
(ELISA and ELISPOT) 
Full ophthalmic examh 
7-field colour fundus 
photosi 
AE/SAE monitoringj 
Concomitant medicationk 
a- Screening Visit/Period must be 
performed … 
b- Baseline assessments must occur 
before the SE2 is dosed. These can 
occur at a regularly scheduled visit 
if that visit is within 12 weeks prior 
to surgery in SE2. If it has been 
more than 12 weeks since a visit, 
the subject must return for 
assessments at this Baseline Period 
2 visit.  Similarly, for subjects who 
have previously been treated 
unilaterally with AAV2-REP1  in an 
antecedent study, these baseline 
assessments must be performed ≤12 
weeks prior to surgery on Visit 1, 
Period 2. A separate visit is only 
Updated footnote 
references, added 
footnotes b and i, 
clarified text, 
removed non-
serious AE 
collection from 
footnote l, 
Added SE1 &SE2 
corticosteroid 
regimen paragraph 
to end of footnotes 
 


NightstaRx, Ltd 
 
4 FEB 2019 
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Version 3.0 
 
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Version 3.0, 4 Feb 2019 
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required if the assessments have not 
been performed within 12 weeks 
prior of surgery in SE2.  
f- Weight is collected for dose 
calculation of the corticosteroid 
regimen. 
i- Immunoassays are planned to 
assess cell-based and antibody-
based immune responses against 
AAV2-REP1. Enzyme-linked 
immunospot (ELISPOT) assays will 
be used for the assessment of T-
cell-mediated immune responses to 
transgene product (REP1) and the 
capsid.  Antibody responses against 
the capsid and transgene product 
will be measured using a cell-based 
neutralization assay and an enzyme-
linked immunosorbent assay 
(ELISA), respectively. All 
immunogenicity samples will be 
sent to a central laboratory for 
analysis. 
j- The ophthalmic examination will 
include IOP, slit lamp examination, 
lens opacity grading, and dilated 
ophthalmoscopy. The same slit 
lamp machine and lighting 
conditions should be used across the 
study for each subject. 
l- AEs/SAEs will be collected from 
the time the subject provides written 
informed consent (Screening Visit) 
through Period 2, Visit 9 (or ET 
Visit, if applicable).  
m- Subjects will be given a course 
of oral corticosteroid before each 
surgical visit, and instructed to start 
taking the drug 2 days prior to 
SE1/SE2 treatment, unless the SE2 
surgical treatment coincides with 
the SE1 steroid treatment period, 
when an extended course of steroid 
can cover both surgical treatments.  
This regimen is outlined in detail in 
Section 9.8. 
For SE1, the Full Corticosteroid 
Regimen should be initiated as 
described.  For SE2, and surgeries 
that occur on day 21 or later, the 


NightstaRx, Ltd 
 
4 FEB 2019 
Summary of Changes: Protocol: NSR-REP-02 
 
Version 3.0 
 
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Version 3.0, 4 Feb 2019 
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Full Corticosteroid Regimen should 
be initiated as described, beginning 
2 days before the scheduled SE2 
surgery.  If SE2 is to be performed 
prior to Study Day 21, then the Full 
Corticosteroid Regimen should be 
initiated 2 days prior to the 
scheduled surgery for SE2, and this 
will supercede the steroid taper in 
progress for SE1.  The full 21 days 
of treatment will then be completed 
for SE2. 
 
Table 1 
Footnotes 
l- AEs/SAEs will be 
collected from the time the 
subject provides written 
informed consent 
(Screening Visit) through 
Period 2, Visit 9 (or ET 
Visit, if applicable). Non-
serious AEs will be 
collected from Period 1, 
Visit 1 (Injection Day Visit 
for SE1) through Period 2, 
Visit 9 (or ET Visit, if 
applicable). 
 
Updated the 
adverse event 
collection to all 
AEs and not just 
SAEs 
 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 1 of 14 
SUMMARY OF CHANGES 
CLINICAL STUDY PROTOCOL  
NSR-REP-02  
AAV2-REP1 
An Open-Label Safety Study of Retinal Gene Therapy for 
Choroideremia with Bilateral, Sequential Administration of Adeno-
Associated Viral Vector (AAV2) Encoding Rab Escort Protein 1 (REP1)  
GEMINI Study 
INDICATION: 
 
 
Choroideremia 
STUDY PHASE: 
 
 
2 
EUDRACT NUMBER: 
 
2017-002395-75 
SPONSOR: 
NightstaRx Ltd 
Wellcome Gibbs Building, 215 Euston Road 
London NW1 2BE, UK 
Telephone:  +44 (0) 207 611 2077 
 
Final Protocol Date: 
 4 Jul 2017 
Amendment 1 
17 May 2018 
CONFIDENTIALITY STATEMENT 
This protocol is the property of NightstaRx Ltd. It is not to be transmitted, copied or published 
without written permission from NightstaRx and must be kept in a confidential manner. Persons 
to whom the information is disclosed must be informed that the information is CONFIDENTIAL 
and may not be further disclosed by them. 
 
 




AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 3 of 14 
 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
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Rationale 
Page 4, 
Contact 
Information 
 
 
Personnel change 
Synopsis 
Study 
Design 
The target interval 
between the surgical 
procedure in SE1 and 
SE2 will be determined 
at the Screening Visit. 
 
The time interval 
between 
surgeries is more 
clearly explained 
in subsequent 
paragraph. 
 
The interval between SE1 and 
SE2 treatment is expected to 
vary for each subject from a 
few weeks to many months. 
While the interval will be 
decided on a case-by-case 
basis, an effort will be made to 
schedule varying treatment 
intervals in order to better 
characterise the 
immunological and safety 
profile of sequential treatment 
administration. Each site will 
work with the Sponsor and the 
assigned Contract Research 
Organisation (CRO) to ensure 
an even spread across the 
intervals. Given that the 
timing of the second surgery 
New text to 
clarify time 
period between 
surgeries and 
requirement for 
an assessment of 
overall ocular 
health and 
function before 
scheduling of 
second eye. 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 4 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
visit (Period 2, Visit 1) will 
vary between subjects, the 
duration of study Period 1 will 
also be variable. To avoid 
unnecessary study visits, every 
effort should be made by the 
investigator to schedule Period 
2, Visit 1 to coincide with a 
planned Period 1 post-
treatment visit. 
The post-operative outcome 
and visual function of SE1 
will inform the scheduling of 
Period 2. Significant signs of 
post-operative inflammation, 
as judged by the investigator, 
should be resolved in SE1 for 
the subject to continue to 
Period 2. After comprehensive 
evaluation of all safety 
assessments and visual 
functional tests of SE1 and 
SE2, the Investigator will 
decide the appropriate time to 
schedule SE2, Visit 1. 
Page 14, 
Section 5.3, 
Study 
Rationale 
 
A multi-centre, global Phase 3 
study of a single subretinal 
administration of AAV2-REP1 
for the treatment of CHM is 
ongoing. 
Added test 
referring to 
STAR Phase 3 
study ongoing. 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 5 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
Section 7.1 
Overall Study 
Design, Page 
16 
 
The interval between SE1 and 
SE2 treatment is expected to 
vary for each subject from a 
few weeks to many months. 
While the interval will be 
decided on a case-by-case 
basis, an effort will be made to 
schedule varying treatment 
intervals in order to better 
characterise the 
immunological and safety 
profile of sequential treatment 
administration. Each site will 
work with the Sponsor and the 
assigned Contract Research 
Organisation (CRO) to ensure 
an even spread across the 
intervals. Given that the 
timing of the second surgery 
visit (Period 2, Visit 1) will 
vary between subjects, the 
duration of study Period 1 will 
also be variable. To avoid 
unnecessary study visits, every 
effort should be made by the 
investigator to schedule Period 
2, Visit 1 to coincide with a 
planned Period 1 post-
treatment visit.  
The post-operative outcome 
and visual function of SE1 
New text to 
clarify time 
period between 
surgeries and 
requirement for 
an assessment of 
overall ocular 
health and 
function before 
scheduling of 
second eye. 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 6 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
will inform the scheduling of 
Period 2. Significant signs of 
post-operative inflammation, 
as judged by the investigator, 
should be resolved in SE1 for 
the subject to continue to 
Period 2. After comprehensive 
evaluation of all safety 
assessments and visual 
functional tests of SE1 and 
SE2, the Investigator will 
decide the appropriate time to 
schedule SE2, Visit 1. 
Section 7.1 
Overall Study 
Design, Page 
17 
For each subject, the 
interval between SE1 
and SE2 treatment is 
expected to range from a 
few weeks to many 
months. While the 
interval will be decided 
on a case-by-case basis, 
an effort should be made 
to schedule varying 
treatment intervals (e.g. 
1, 3 or 6+ months apart) 
in order to better 
characterise the 
immunological and 
safety profile of 
sequential treatment 
administration. Each site 
 
This text has 
been moved up 
and altered 
slightly as above 
to clarify time 
period between 
surgeries and 
requirement for 
an assessment of 
overall ocular 
health and 
function before 
scheduling of 
second eye. 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 7 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
will work with the 
assigned Contract 
Research Organisation 
to ensure an even spread 
across the intervals. 
Given that the timing of 
the second surgery visit 
(Period 2, Visit 1) will 
vary between subjects, 
the duration of study 
Period 1 will also be 
variable. To avoid 
unnecessary study visits, 
every effort should be 
made by the investigator 
to schedule Period 2, 
Visit 1 to coincide with a 
planned Period 1 post-
treatment visit. 
Section 7.2 
Discussion of 
Design, Page 
18-19 
To minimise 
inflammation resulting 
from potential immune 
responses to vector, 
including a secondary 
immune response 
following administration 
of AAV2-REP1 in the 
second eye (SE2), 
subjects will be given  a 
course of oral 
corticosteroid (e.g., 
To minimise inflammation 
resulting from potential 
immune responses to vector, 
including treatment of the 
second eye (SE2), subjects 
will be given a 21-day course 
of oral corticosteroid before 
each surgical visit, and 
instructed to start taking the 
drug 2 days prior to SE1/SE2 
treatment. If the SE2 steroid 
treatment coincides with the 
Clarified the 
corticosteroid 
regimen, also 
when the two 
surgeries are 
planned close in 
time 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 8 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
prednisolone/prednisone
) which will commence 
prior to each surgery 
SE1 steroid treatment period, 
an extended course of steroid 
will cover both surgical 
treatments. 
Section 8.3 
Subject 
Withdrawal 
Criteria 
 
•  Other (to be specified on the 
electronic case report form 
[eCRF]). 
• Severe ocular inflammation 
(e.g., endophthalmitis) that is 
unresponsive to treatment 
should result in the 
termination of the trial for 
the affected subject. 
• At the discretion of the 
investigator, if a persistent 
post-operative complication 
or 
poor 
visual 
function 
outcome 
presents 
post-
operatively 
in 
SE1, 
the 
second eye may not be 
treated and the subject may 
be discontinued from the 
study. 
Text now 
clarifies that if 
the post-
operatively 
severe ocular 
inflammation is 
present, the 
subject will be 
terminated from 
the trial, and that 
any post-
operative 
complication or 
poor visual 
outcome will 
prevent the 
second eye from 
continuing to 
surgery. 
Section 8.3 
Subject 
Withdrawal 
Criteria, Page 
20 
The study may be 
discontinued if the 
Sponsor deems it 
necessary for medical, 
safety, regulatory or 
other reasons consistent 
 
This sentence has 
been moved to 
Section 11.3 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 9 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
with applicable laws or 
regulations. 
Setion 9.4 
Vitrectomy 
Procedure 
A single dose of AAV2-
REP1 will be injected 
into the sub-retinal fluid 
through the same entry 
site. 
 
This sentence is 
repeated in 
paragraph below 
 
 
Air fluid exchange during pars 
plana vitrectomy is not 
required for this surgical 
procedure. However, if an air 
fluid exchange is considered 
necessary, patients must be 
instructed to avoid air travel, 
travelling to high elevations or 
scuba diving until the air 
bubble has completely 
dissipated from the eye. It may 
take one week or more 
following injection for the air 
bubble to dissipate. A change 
in altitude while the air bubble 
is still present can result in 
irreversible vision loss. The 
dissipation of the air bubble 
should be verified through 
meticulous ophthalmic 
examination. 
Details of the 
vitrectomy 
procedure have 
been added for 
clarification.  
Section 9.8, 
Concomitant 
 
If the SE2 steroid treatment 
coincides with the SE1 steroid 
treatment period, an extended 
Added this 
sentence to 
clarify 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 10 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
Therapy, Page 
25 
course of steroid will cover 
both surgical treatments. 
corticosteroid 
regimen when 
surgeries are 
planned close in 
time. 
Section 10.2 
Period 1 and 
2, Visit 1 
(Day 0, 
Injection Day 
Visit) Page 26 
 
The post-operative outcome 
and visual function of SE1 
will inform the scheduling of 
Period 2. Significant signs of 
post-operative inflammation, 
as judged by the Investigator, 
should be resolved in SE1 for 
the subject to continue to 
Period 2. After a 
comprehensive evaluation of 
all safety assessments and 
visual functional tests of SE1 
and SE2, the Investigator will 
decide the appropriate time to 
schedule SE2, Visit 1. 
New text to 
clarify time 
period between 
surgeries and 
requirement for 
an assessment of 
overall ocular 
health and 
function before 
scheduling of 
second eye. 
Section 11.1.4 
Reporting of 
Serious 
Adverse 
Events Page 
32 
 
Emergency contact 
information is provided below. 
Any urgent queries relating to 
subject safety should be 
communicated immediately 
via phone to the Chiltern 
Medical Monitor.  The NSR 
Medical Monitor is the first 
escalation point when the 
Chiltern/NSR Medical 
Monitor is not available.  The 
Emergency 
contact 
information has 
been added. 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 11 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
second escalation point is the 
NSR Chief Medical Officer.   
 
Contact Name and Title 
Telephone, Email 
, MD 
 
Medical Monitor 
NightstaRx, Ltd 
Direct: 
 
Mobile: 
 
E mail: 
 
 
Chiltern Medical Monitor 
(out-of-hours) EU 
, MD 
Chiltern Ltd. Direct: 
 
 
Mobile: 
 
E mail: 
 
Chiltern Medical Monitor 
(out-of-hours) US 
, MD 
Chiltern Ltd. Direct: 
 
 
Mobile: 
 


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 12 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
E mail: 
11.3 Stopping 
Criteria  
 
11.3 Stopping Criteria 
• Severe ocular inflammation 
(e.g., endophthalmitis) that is 
unresponsive to treatment 
should result in termination of 
the trial for the affected 
subject. 
• At the request of the Data 
Monitoring Committee, the 
trial may be stopped based on 
specific safety concerns (ie, 
SUSARs), not defined in the 
Investigator’s Brochure 
(Section 6.3).  
• The study may be 
discontinued if the Sponsor 
deems it necessary for 
medical, safety, regulatory or 
other reasons consistent with 
applicable laws or regulations. 
New Section 
added for 
additional safety 
gating and 
stopping criteria 
11.4.2 
Immunogenic
ity 
 
For the evaluation of 
immunogenicity, blood will be 
collected at the times indicated 
in Table 1. Immunoassays are 
planned to assess cell-based 
and antibody-based immune 
responses against AAV2-
REP1. ELISPOT assays will 
Immunogenicity 
testing has been 
specified.  


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 13 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
be used for the assessment of 
T-cell- mediated immune 
responses to transgene product 
(REP1) and the capsid.  
Antibody responses against 
the capsid and transgene 
product will be measured 
using a cell-based 
neutralization assay and an 
ELISA, respectively. 
All immunogenicity samples 
will be sent to a central 
laboratory for analysis. Refer 
to the Study Operations 
Manual for details on the 
shipping and handling of 
samples. 
Section 13.7 
Efficacy 
Analysis Page 
38 
Formal statistical tests 
will not be performed  
and 95% CIs will be 2-
sided. 
Any additional 
categorical/continuous 
efficacy endpoints will be 
summarised using descriptive 
statistics, with 95% 2-sided 
CIs will be calculated where 
appropriate. 
The text has been 
aligned with that 
of safety 
analyses.  
Section 13.7 
Efficacy 
Analysis Page 
38 
Any additional 
continuous efficacy 
endpoints will be 
analysed using 
descriptive statistics. 
 
The text has been 
aligned with that 
of safety 
analyses.  
Table of 
Procedures 
 
k. Subjects will be given two 
21-day courses of oral 
Steroid therapy 
in the peri-


AAV2-REP1  
Protocol: NSR-REP-02 
Version 2.0 
17 MAY 2018 
 
CONFIDENTIAL 
Page 14 of 14 
SUMMARY OF CHANGES  
PROTOCOL NSR-REP-02 
Version 2.0, 17 MAY 2018 
Overview / Rationale 
This protocol amendment has clarified the requirement for an evaluation by the investigator of 
the status of the first operated eye, regarding post-operative inflammation and visual function, 
and of the visual function and health overall of both eyes before scheduling the second eye 
surgery.  It also specifies stopping criteria if there are signs of inflammation in the first eye. 
Immunogenicity testing is clarified. Other minor changes and contact information have been 
updated. 
Page, 
Section 
Previous Text  
(Deleted Text Shown 
by Strikethrough) 
Revised Text  
(Added Text Shown as Red) 
Rationale 
and Visits 
Page 46 
Footnote k 
corticosteroid before each 
surgical visit, and instructed to 
start taking the drug 2 days 
prior to SE1/SE2 treatment, 
unless the SE2 surgical 
treatment coincides with the 
SE1 steroid treatment period 
when an extended course of 
steroid can cover both surgical 
treatments.  
operative period 
is clarified for 
when surgeries 
are temporally 
close. 
 

SAP_001.pdf: 
Protocol: 273CH203 / NSR-REP-02 
Confidential 
Biogen / NightstaRx, Ltd 
Final 2.0                                                             Page 1 of 36 
03MAR2022 
VERSION HISTORY 
 
SAP Version 
Date 
Primary Reasons for Amendment 
Version 1 
31JAN2020 
Original version 
Version 2 
03MAR2022 
Removed planned analyses on Fixation 
Location due to data availability 
Removed planned analyses on time to 
resolution of AEs to keep consistency with 
other studies in the BIIB111 program.  
Changed planned analyses on microperimetry to 
exploratory to be performed as post-hoc 
analyses if needed 
Changed efficacy analyses to use observed case 
method as the main analyses. Added additional 
supportive analyses of BCVA-related endpoints, 
including LOCF imputation 
Adjusted subgroups for subgroup analyses 
Added summary on impact of COVID-19, and 
on protocol deviations 
Added details for immunogenicity and vector 
shedding 
Removed lists of table figure listing 
Removed inflammation related MedDRA 
Preferred Terms 
Provided additional clarification and 
information on definitions 
 
 
 


Protocol: 273CH203 / NSR-REP-02 
Confidential 
Biogen / NightstaRx, Ltd 
Final 2.0                                                             Page 2 of 36 
03MAR2022 
 
Statistical Analysis Plan 
GEMINI Study 
PROTOCOL:  
273CH203 / NSR-REP-02 (Timrepigene emparvovec) 
An Open-Label Safety Study of Retinal Gene Therapy for 
Choroideremia with Bilateral, Sequential Administration of 
Adeno-Associated Viral Vector (AAV2) Encoding Rab Escort 
Protein 1 (REP1) (GEMINI) 
INDICATION: 
Choroideremia 
STUDY PHASE: 
2 
EUDRACT NUMBER: 
2017-002395-75 
CLINICAL TRIALS NUMBER 
NTC03507686 
SPONSOR: 
Biogen 
                                                                      225 Binney Street 
                                                                     Cambridge, MA 02142 
 
SAP for GEMINI Protocol Version
Amendment 3, Version 4
Document Status:
Final Version 2.0
Version 1 
Version 2 
31JAN2020 
03MAR2022 
 
 
 




Protocol: 273CH203 / NSR-REP-02 
Confidential 
Biogen / NightstaRx, Ltd 
Final 2.0                                                             Page 4 of 36 
03MAR2022 
Table of Contents 
VERSION HISTORY .............................................................................................................. 1 
ABBREVIATIONS .................................................................................................................. 6 
1 
INTRODUCTION............................................................................................................ 8 
2 
STUDY OBJECTIVES AND ENDPOINTS .................................................................. 9 
2.1 
OBJECTIVE .................................................................................................................. 9 
2.2 
ENDPOINTS ................................................................................................................. 9 
2.2.1 
Safety Endpoints................................................................................................. 9 
2.2.2 
Secondary Efficacy Endpoints ........................................................................... 9 
3 
STUDY DESIGN ............................................................................................................ 10 
3.1 
DISCUSSION OF STUDY DESIGN ................................................................................. 10 
3.2 
STUDY TREATMENT .................................................................................................. 11 
3.3 
STUDY SCHEDULE ..................................................................................................... 11 
3.4 
RANDOMISATION ...................................................................................................... 12 
3.5 
BLINDING .................................................................................................................. 12 
3.6 
SAMPLE SIZE ............................................................................................................. 12 
4 
STATISTICAL METHODOLOGY............................................................................. 13 
4.1 
ANALYSIS SETS ......................................................................................................... 13 
4.2 
CONVENTIONS .......................................................................................................... 13 
4.3 
DEFINITIONS ............................................................................................................. 13 
4.4 
DATA OF SUBJECTS WITH FIRST EYE TREATED IN AN ANTECEDENT STUDY ............. 14 
4.5 
SCHEDULE OF ANALYSES .......................................................................................... 14 
4.6 
ADJUSTMENT FOR MULTIPLICITY.............................................................................. 14 
4.7 
SUBGROUP ANALYSIS ............................................................................................... 14 
4.8 
HANDLING OF MISSING VALUES ............................................................................... 15 
4.9 
STATISTICAL ANALYSES HANDLING OF DATA IMPACTED BY COVID-19 ................. 15 
4.10 
DISPOSITION OF SUBJECTS ........................................................................................ 16 
4.11 
PROTOCOL DEVIATIONS ............................................................................................ 16 
4.12 
DEMOGRAPHIC AND BASELINE CHARACTERISTICS ................................................... 16 
4.13 
MEDICAL AND OCULAR HISTORY ............................................................................. 16 
4.14 
PRIOR AND CONCOMITANT MEDICATION .................................................................. 16 
4.15 
EXPOSURE ................................................................................................................. 17 
4.16 
EFFICACY ANALYSIS ................................................................................................. 17 
4.16.1 
Visual Acuity .................................................................................................... 17 
4.16.2 
Fundus Autofluorescence ................................................................................. 19 
4.16.3 
Spectral-Domain Optical Coherence Tomography ......................................... 19 
4.16.4 
Macular Integrity Assessment Microperimetry ............................................... 20 
4.17 
SAFETY ANALYSIS .................................................................................................... 21 
4.17.1 
Visual Acuity .................................................................................................... 21 
4.17.2 
Intraocular Pressure ........................................................................................ 21 
4.17.3 
Slit Lamp Examination ..................................................................................... 21 
4.17.4 
Lens Opacity Grades........................................................................................ 22 
4.17.5 
Dilated Ophthalmoscopy ................................................................................. 22 
4.17.6 
SD-OCT............................................................................................................ 23 
4.17.7 
Fundus Autofluorescence ................................................................................. 23 
4.17.8 
Fundus Photography ........................................................................................ 23 


Protocol: 273CH203 / NSR-REP-02 
Confidential 
Biogen / NightstaRx, Ltd 
Final 2.0                                                             Page 5 of 36 
03MAR2022 
4.17.9 
Microperimetry ................................................................................................ 23 
4.17.10 Adverse Events ................................................................................................. 23 
4.17.11 Immunogenicity ................................................................................................ 25 
4.17.11.1 Baseline Definition........................................................................................... 26 
4.17.11.2 Assay Positivity by Visit ................................................................................... 26 
4.17.11.3 Assay Treatment-emergent Analysis by Visit ................................................... 26 
4.17.11.4 Relationship Between Immunogenicity Assay and Safety ................................ 27 
4.17.11.5 Data Listing ..................................................................................................... 27 
4.17.12 Vector Shedding ............................................................................................... 28 
4.17.13 Vital Signs ........................................................................................................ 28 
4.18 
PROTOCOL VIOLATIONS OR DEVIATIONS .................................................................. 28 
4.19 
DEVIATIONS FROM THE STATISTICAL ANALYSIS PLAN ............................................. 28 
4.20 
CHANGES IN PLANNED ANALYSES FROM THE PROTOCOL ......................................... 28 
4.21 
ALGORITHMS/SAS CODES ........................................................................................ 28 
5 
TABLES AND LISTINGS ............................................................................................ 30 
5.1 
FORMAT .................................................................................................................... 30 
5.2 
CONVENTIONS FOR OUTPUT DISPLAY ........................................................................ 30 
6 
REFERENCES ............................................................................................................... 31 
7 
APPENDICES ................................................................................................................ 32 
7.1 
APPENDIX 1  IMPUTATION OF PARTIAL DATES ........................................................ 32 
7.2 
SCHEDULE OF STUDY PROCEDURES .......................................................................... 32 
List of Tables 
Table 1 Subgroups Defined at Baseline ................................................................................ 15 
Table 2 Visit Assessment Windows...................................................................................... 18 
Table 3 Schedule of Study Procedures ................................................................................. 33 
List of Figures 
Figure 1 Overall Study Schematic.......................................................................................... 10 
 
 


Protocol: 273CH203 / NSR-REP-02 
Confidential 
Biogen / NightstaRx, Ltd 
Final 2.0                                                             Page 6 of 36 
03MAR2022 
Abbreviations 
AAV 
adeno-associated viral vector 
ADA
anti-drug antibodies
AE
adverse event
ATC
anatomical therapeutic chemical
AF
autofluorescence
BCVA 
best corrected visual acuity 
BIIB111
timrepigene emparvovec
CHM
choroideremia
CI
confidence interval
CM
concomitant medication
CSR
clinical study report
eCRF
electronic case report form
ELISA 
enzyme-linked immunosorbent assay 
ELISpot
enzyme-linked immunospot
ERM
epiretinal membrane
ET
early termination
ETDRS
Early Treatment Diabetic Retinopathy Study
FC 
foveal center 
IOP
intraocular pressure
LOCF
last observation carried forward
LOCS
lens opacities classification system
MAIA
Macular Integrity Assessment
MedDRA
Medical Dictionary for Regulatory Activities
MH 
macular hole 
Nabs
neutralizing timrepigene emparvovec antibodies
OD
oculus dextrus (right eye)
OU
oculus uterque (both eyes)
OS
oculus sinister (left eye)
P05 
fifth percentile 
P95 
ninety fifth percentile 
PT 
preferred term 
Q1
first quartile
Q3
third quartile
REP1
rab escort protein 1
RPE
retinal pigment epithelium
SAE 
serious adverse event 
SAP
statistical analysis plan
SAS
statistical analysis system
SD
standard deviation
SD-OCT
spectral domain optical coherence tomography
SE1 
study eye 1 
SE2 
study eye 2 
SFU
spot forming units
SOC
system organ class
SRF
subretinal fluid


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TEAE
treatment-emergent adverse event
USAN
United States adopted name
vg
vector genomes
VMT
vitreomacular traction
WHODD
World Health Organisation drug dictionary


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1 Introduction 
This document presents the statistical analysis plan (SAP) for the NightstaRx-sponsored 
Protocol No. 273CH203 / NSR-REP-02: An Open-Label Safety Study of Retinal Gene 
Therapy for Choroideremia with Bilateral, Sequential Administration of Adeno-Associated 
Viral Vector (AAV2) Encoding Rab Escort Protein 1 (REP1) (GEMINI Study).  
AAV2-REP1 is denoted hereafter by the USAN timrepigene emparvovec (BIIB111).   
This analysis plan is based on the final protocol dated 30 Oct 2019, incorporating the 
following versions/ amendments: 
 Original protocol, dated 4 Jul 2017 
 Amendment No. 1, version 2, dated 17 May 2018 
 Amendment No. 2, version 3, dated 04 Feb 2019 (internal amendment only) 
 Amendment No. 2.1, version 3.1, dated 19 Feb 2019 
 Amendment No. 3, version 4, dated 30 Oct 2019 
The SAP provides the description of the final analyses for the generation of the clinical study 
report. 


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2 Study Objectives and Endpoints 
2.1 Objective 
The objective of the study is to evaluate the safety of bilateral, sequential sub-retinal 
administration of a single dose of timrepigene emparvovec in adult male subjects with 
choroideremia (CHM). 
2.2 Endpoints 
2.2.1 Safety Endpoints 
The safety of bilateral administration of timrepigene emparvovec will be evaluated with the 
following safety measures: 
 Best Corrected Visual Acuity (BCVA) as measured by the Early Treatment of 
Diabetic Retinopathy Study (ETDRS) chart 
 Ophthalmic examination assessments (including intraocular pressure [IOP], slit lamp 
examination, lens opacity grading and dilated ophthalmoscopy) 
 Spectral domain optical coherence tomography (SD-OCT) 
 Fundus autofluorescence 
 Fundus photography 
 Microperimetry 
 Adverse Event (AE) reporting 
 Vector shedding post-treatment 
 Immunogenicity sampling post-treatment 
 Vital signs 
2.2.2 Secondary Efficacy Endpoints 
 Change from Baseline in BCVA as measured by the ETDRS chart 
 Change from Baseline in autofluorescence 
 Change from Baseline in SD-OCT 
 Change from Baseline in microperimetry  
 
 


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3 Study Design 
3.1 Discussion of Study Design 
This is a Phase 2, multi-centre, open-label, prospective, 2-period, bilateral interventional safety 
study of timrepigene emparvovec in adult male subjects with genetically confirmed CHM. 
The study will consist of a Screening Period followed by 2 treatment periods (Period 1 and 
Period 2) with up to 9 visits per treatment period. A second baseline visit prior to surgery of 
Study Eye 2 (SE2) (Period 2, Visit 1) may be required. A study schematic is presented in 
Figure 1. 
Figure 1 
Overall Study Schematic 
 
 
During the Screening Period, subjects will be assessed for eligibility of both eyes. The 
investigator will assign the order in which the eyes are treated (i.e., study eye 1 [SE1] period 
1 and study eye 2 [SE2] period 2, respectively) with a number assigned to each eye. This will 
be done in collaboration with the subject; however, the worse eye will generally be selected 
for first treatment. The estimated target interval between the surgical procedures of SE1 and 
SE2 will be determined at the Screening Visit. 
Each study eye will be followed for at least 12 months post-treatment, for up to 9 visits per 
treatment period: Visit 1 (Day 0, Injection Day Visit); Visit 2 (Day 1, post-operative follow 
up visit); Visit 3 (Day 3 + 2 Days); Visit 4 (Day 7 -1/+2 Days); Visit 5 (Day 14 ± 3 Days); 
Visit 6 (Month 1 ± 7 Days); Visit 7 (Month 3 ± 14 Days); Visit 8 (Month 6 ± 14 Days) and 
Visit 9 (Month 12 ± 14 Days). 
The interval between SE1 and SE2 treatment is expected to vary among subjects from weeks 
to many months. While the interval will be decided on a case-by-case basis, an effort will be 
made to schedule varying treatment intervals in order to better characterise the 
immunological and safety profile of sequential treatment administration. Thus, approximately 
20 subjects each will be treated with a short- (<6 month), medium- (6-12 months), or long- 
(>12 month) surgery window between treatment of the first and second eye. Because the 
timing of the second surgery visit (Period 2, Visit 1) will vary among subjects, the duration of 
study Period 1 will also be variable. 
All baseline ophthalmic assessments must be conducted within 3 months prior to SE2 surgery 
(Period 2, Visit 1). If these assessments are not available within the required timeframe, a 
Baseline Visit must be added to collect current baseline data on SE2. If, at that time, BCVA 
in SE2 is <34 ETDRS letters, surgery may be performed when, in the opinion of the 


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investigator and after consultation with the sponsor: 1) the subject is found eligible by all 
other inclusion and exclusion criteria, and 2) the subject is considered an appropriate 
candidate who has potentially modifiable disease. 
The post-operative outcome and visual function of SE1 will inform the scheduling of Period 
2. Signs of post-operative inflammation or other post-operative sequelae, as judged by the 
investigator, should be resolved in SE1 for the subject to continue to Period 2. After 
comprehensive evaluation of all safety assessments and visual functional tests of SE1 and 
SE2, the Investigator will determine the eligibility of the subject and decide the appropriate 
time to schedule SE2, Visit 1.  
Subjects who develop cataracts may undergo cataract surgery if deemed clinically necessary. 
If cataract surgery is performed, it should be carried out at least 4 weeks before Month 12 
(Visit 9) for the respective eye. The cataract surgery timing must be discussed with the 
medical monitor prior to the procedure.  
Subjects Previously Treated with Timrepigene Emparvovec 
Subjects who have previously participated in a clinical trial of AAV2-REP1 for the treatment 
of CHM and have received AAV2-REP1 in one eye may be eligible for participation in this 
Following the Screening Visit and determination of eligibility, the subject will move directly 
into Period 2 for treatment of SE2. 
3.2 Study Treatment 
Eligible subjects will undergo vitrectomy and retinal detachment in each eye. At Period 1, 
Visit 1 (Day 0, the Injection Day Visit for SE1), subjects will receive in SE1 a sub-retinal 
injection of up to 0.1 mL of study drug containing 1×10^11 timrepigene emparvovec vg. At 
Period 2, Visit 1 (Day 0, the Injection Day Visit for SE2), subjects will receive in SE2 up to 
0.1 mL of timrepigene emparvovec via sub-retinal injection. 
3.3 Study Schedule 
At Period 1, Visit 1 (Day 0, Injection Day Visit for SE1), subjects will undergo a vitrectomy 
with retinal detachment and receive a sub-retinal injection of timrepigene emparvovec in 
SE1. Visits 2-9 will then be conducted according to the Schedule of Study Procedures, unless 
Period 2 commences during the time of follow-up for SE1; in which case, the study visits will 
continue following the schedule for SE2. Ophthalmic assessments during Period 1, Visits 2-9 
will be performed on both eyes.  
At Period 2, Visit 1 (Day 0, Injection Day Visit for SE2), subjects will undergo a vitrectomy 
with retinal detachment and sub-retinal injection of timrepigene emparvovec in the 
contralateral, untreated eye (SE2). Post-treatment, subjects will no longer follow the Period 1 
visit schedule, but will instead attend Period 2, Visits 2-9 according to the Schedule of Study 
Procedures. Ophthalmic assessments scheduled for Period 2, Visits 2-9 will be performed on 
both eyes.  


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Study data will thus be collected for both eyes in each subject. Subjects will be assessed for 
safety and efficacy throughout the study, as outlined in the Schedule of Study Procedures 
(Section 7.2).  
A subject is considered to have completed the study if he completes Period 2, Visit 9 (Month 
12). The end of the trial is the date the last subject completes Period 2, Visit 9 (Month 12) 
assessments (or early termination [ET] assessments in the event of premature 
discontinuation) or the date of last data collection if the last subject is lost to follow-up.  
3.4 Randomisation 
Not applicable for this study.  
3.5 Blinding 
Not applicable for this study.  
3.6 Sample Size 
Approximately 60 subjects are planned to be enrolled in the study. A total of 120 treated eyes 
is considered sufficient to characterise the immunological profile of bilateral timrepigene 
emparvovec administration, which is the key concern of gene therapy re-administration. 
 
 


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4 Statistical Methodology 
4.1 Analysis Sets 
All Treated Subjects analysis set will consist of all subjects who complete 
timrepigene emparvovec 
treatment in SE1 in an antecedent study). All Treated Subjects analysis set will be the primary 
population for demographics, baseline characteristics, safety and efficacy analyses. 
Immunogenicity analysis set 
analysis set with baseline sample and at least one post-surgery sample evaluable for 
immunogenicity. The Immunogenicity analysis set will be used for the immunogenicity 
analyses. 
4.2 Conventions 
Summary statistics will be presented for continuous variables, by way of number of subjects, 
mean, standard deviation (SD), median, first and third quartiles (Q1 and Q3, respectively), 5th 
and 95th percentiles (P05 and P95, respectively), minimum and maximum. For categorical 
variables including binary variables, counts and percentages will be presented. Percentages will 
be calculated using the total number of eyes in the respective column or the total number of 
eyes with an assessment, the total number of treated subjects, depending on the nature of the 
analysis. 
In addition, 95% 2-sided confidence intervals (CIs) will be presented, where specified. The 
95% CI of the mean value of an assessment will be calculated using the t-test formula. The 
95% CI of the mean of change from baseline of an assessment will be calculated using the t-
test formula. In case of percentages, the CI will be computed using the Clopper-Pearson 
method. (See Section 4.21 for the corresponding SAS code). 
No formal statistical comparison will be performed. 
Summary statistics will be presented by period and overall for all endpoints. For selected 
endpoints (see details below), summary statistics will also be presented by Surgery Window 
(Short [<6 months], Medium [6-12 months] and Long [>12 months]) and overall. Where 
appropriate, summary statistics will be displayed by eye (SE1 and SE2), and by period. 
  
4.3 Definitions 
Study Eye 1 (SE1) is defined as the first treated eye (within GEMINI or in an antecedent 
clinical trial). Study Eye 2 (SE2) is defined as the second treated eye (irrespective of whether 
the first eye was treated in GEMINI or in an antecedent clinical trial). Subjects whose first eye 
is treated in an antecedent study will contribute to Period 2 only (except for the immunogenicity 
analyses), where Study Eye 1 is the eye treated in the antecedent study and Study Eye 2 is the 
eye treated in GEMINI.  


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Period 1 starts on the day of the surgery of the first treated eye and ends on the day prior to the 
day of the surgery of the second treated eye. Period 2 starts on the day of the surgery of the 
second treated eye. 
Baseline Value of Period 1/2 is defined as the last non-missing value prior to treatment of the 
respective period. Baseline of each period for each eye will be used to calculate the change 
from baseline for the data in the same period and same eye. For example, period 1 SE1 baseline 
will be used to calculate the change from baseline for all the data of SE1 in period 1, whereas 
period 2 SE1 baseline will be used to calculate the change from baseline for all the data of SE1 
in period 2.  
For immunogenicity, baseline will be defined separately in Section 4.17.11. 
Analysis day for all the post-baseline visits will be calculated as the date of the post-baseline 
visits minus the corresponding injection date, depending on which baseline that was used.  
For the 6 subjects whose first eye was treated in the antecedent study (Subjects 
), only period 2 data are collected in GEMINI. 
Therefore, only period 2 data will be presented for these 6 patients.  
Change from Baseline is defined as the difference of the value of an assessment at the 
considered visit from the baseline value of the corresponding period. 
Duration is measured in days and is calculated as end date  onset date + 1. 
Injection dates of SE1 and SE2 will be coded Day 0 for the Period 1 and Period 2 respectively. 
4.4 Data of Subjects with First Eye Treated in an Antecedent Study 
For the 6 subjects who received timrepigene emparvovec treatment in their first eye in an 
antecedent study, the data from the antecedent study will not be included in the analyses 
presented in the Clinical Study Report (CSR). 
4.5 Schedule of Analyses 
At regular intervals during the study, the Data Monitoring Committee will review the progress 
and accrued study data and provide advice to the Sponsor on the efficacy and safety aspects of 
the study. 
A final analysis will be conducted after all subjects complete the study. 
4.6 Adjustment for Multiplicity 
Not applicable for this study. 
4.7 Subgroup Analysis 
Demographic and baseline characteristics will be summarised by Surgery Site. The efficacy 
endpoints of BCVA will be summarised by the subgroups defined in Table 1. 




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4.10 Disposition of Subjects 
The number of subjects who sign Informed Consent, who receive study treatment, who 
complete the study and the reasons for any premature discontinuation from the study will be 
presented. The number of subjects included in the All Treated Subjects analysis set and the 
Immunogenicity analysis set will also be presented. For each study visit, the number of subjects 
who attend the visit will be summarised.  
4.11 Protocol Deviations  
A summary table of protocol deviation will be provided which contains the number (%) of 
subjects with at least one protocol deviation. The table will also present the summary statistics 
for important protocol deviations (if defined). Similar table will be generated for COVID-19 
related protocol deviation only. 
4.12 Demographic and Baseline Characteristics 
Demographic (
50 and > 50 years, 
60 and > 
60 years,  median age and > median age]) and baseline characteristics (including weight) will 
be summarised in the All Treated Subjects analysis set. Baseline ocular characteristics will also 
be presented by eye (SE1 and SE2) and Period. 
Demographic and baseline characteristics will also be summarised by Surgery Site in the All 
Treated Subjects analysis set. 
Age is calculated as (Informed Consent date  date of birth + 1)/365.25 and presented to 1 
decimal place. No rounding will be carried out prior to summarising age. Where the day and 
month of birth are not known, the date of birth will be assigned the value 30JUNYYYY. 
4.13 Medical and Ocular History 
Ocular History will be summarised by eye, by System Organ Class (SOC) and Preferred Term 
(PT), in the All Treated Subjects analysis set. 
Where data 
 (both eyes), the event will be attributed to both eyes (SE1 
and SE2) for the purpose of the summary tables. 
Medical History will be listed, with no summary table created. 
Medical and ocular history will be coded according to the latest version (at the time of the data 
cut-off) of the Medical Dictionary for Regulatory Activities (MedDRA). 
4.14 Prior and Concomitant Medication 
Prior and concomitant (Ocular and Non-Ocular) medications will be presented in the All 
Treated Subjects analysis set by Anatomical Therapeutic Chemical (ATC) Level 4 and 
Preferred Term, and by Period for concomitant medications. Ocular medications will also be 


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presented by eye (SE1 and SE2). 
counted once for each eye (SE1 and SE2). 
Prior medications are those that start and stop before exposure to the treatment of SE1; 
concomitant medications are all medications taken from the day of first surgery, including 
those started before but ongoing at the time of the surgery. 
The medication start and stop dates will be used to assign the medication to Period 1 or Period 
2. 
Imputation rules for partial dates are presented in Appendix 7.1. 
Where a medication start date is partially or fully missing, and after imputation it is still 
unclear as to whether the medication is prior or concomitant, it will be assumed that it is 
concomitant and will be attributed to SE1. 
An eye with more than one occurrence of the same medication in a particular ATC class will 
be counted only once in the total of those with a report of medication in that particular ATC 
class. 
Corticosteroid Treatment/Tapering will be listed. No summary table will be generated. 
Medications will be coded according to WHO-Drug Dictionary (WHODD). The WHODD 
version will be the latest available at the time of the data cut-off. 
4.15 Exposure 
As the treatment consists of a single injection, exposure to study medication will not be 
evaluated. 
4.16 Efficacy Analysis 
Efficacy analysis will be performed in the All Treated Subjects analysis set. 
4.16.1 Visual Acuity 
Best-corrected visual acuity (BCVA) and change from baseline will be summarised by visit, 
by eye, by period as well as overall. The corresponding 95% CIs will be calculated (See Section 
4.2).  
 increase, 
, and an 
increase or a loss <5 letters (i.e. change from baseline >-5 letters) will be summarised by visit, 
by eye, by period as well as overall. The corresponding 95% CI will be presented (See Section 
4.2).  
The above main analyses will use the observed case method only. Imputation using LOCF 
method regardless of the reasons of missing data will be performed for supportive analyses 
(See Section 4.8).  
In addition to the by-period, by-eye presentation mentioned above, as supportive analyses, for 
both continuous and categorical endpoints, the period 2 Study Eye 1 data will be presented by 


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remapping them to visits after their last visit in period 1 and calculating change from baseline 
using baseline at period 1 for the same eye (Study Eye 1). This way a full picture of Study Eye 
1 treatment journey during the entire study will be presented.   
For all assessments of periods 1 and 2 of Study Eye 1, the data records will be assigned to 
calculated visit window (using study day) as described in Table 2 below. The window of the 
visits following baseline (including unscheduled visits) will be constructed in such a way that 
the upper limit of the interval falls half way between the two visits. 
If a patient has more than one assessment included within a window, the assessment closest to 
the target day will be used. In case of ties between observations located on different sides of 
the target date, the value with the later date will be used. 
Table 2 
    Visit Assignment Windows  
Day / Month 
Target Day 
Day Range included in the visit 
Baseline 
0 
The last assessment on or before day 0 
Day 1 
1 
Day 1 
Day 3 
3 
Day 2 to 5 
Day 7 
7 
Day 6 to 10 
Day 14 
14 
Day 11 to 22 
Month 1 
30 
Day 23 to 61 
Month 3 
91 
Day 62 to 136 
Month 6 
182 
Day 137 to 272 
Month 9 
273 
Day 228 to 318 
Month 12 
365 
Day 319 to 410 
Month 15 
456 
Day 411 to 501 
Month 18 
548 
Day 502 to 593  
Month 21 
639 
Day 594 to 684 
Month 24 
731 
Day 685 onwards  
 
Both continuous and categorical endpoints for BCVA will be summarised by subgroups 
mentioned in Section 4.7. 


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4.16.2  
Fundus Autofluorescence 
Observed value, change from baseline, and percent change from baseline for Total Area of 
Preserved AF, square root of the Total Area of Preserved AF and Distance from Foveal Center 
(FC) to nearest Border of Preserved AF will be summarised by visit, by eye, and by period. 
The number and percentage of subjects with Subfoveal or non-Subfoveal Lesion in Relation to 
FC will be summarised by visit, by eye, and by period. 
Shift from baseline in Lesion will be summarised by calculating the number of subjects falling 
in each of the following categories: 
 Not subfoveal at Baseline to Not subfoveal 
 Not subfoveal at Baseline to Subfoveal 
 Subfoveal at Baseline to Not subfoveal 
 Subfoveal at Baseline to Subfoveal 
Those summaries will be presented by visit, by eye and by period. 
For Subfoveal or non-Subfoveal Lesion in Relation to FC
 
Image quality will be listed only. This includes image completeness, focus/clarity, field 
Is the total area of preserved AF fully captured within 
. 
w
. 
4.16.3 Spectral-Domain Optical Coherence Tomography 
Observed value, change from baseline, and percent change from baseline for Foveal Subfield 
Thickness, Total Macular Volume, Central Horizontal Ellipsoid Width, Central Ellipsoid Area, 
the square root of the Central Ellipsoid Area and Choroidal Thickness at Foveal Center will be 
summarised by visit, by eye, and by period.  
Presence of Vitreomacular Traction (VMT), Epiretinal Membrane (ERM), Macular Hole 
(MH), Intraretinal Hyper-Reflective Spots, Cystoid Macular Edema, Subretinal Fluid (SRF), 
Subretinal Hyper-Reflective Material and Pigment Epithelial Detachment will be summarised 
by visit, by eye, and by period. Shift from baseline will be evaluated for each of these 
assessments.  
The categories for the shift from baseline for VMT, ERM, MH and Intraretinal Hyper-
Reflective Spots assessments are defined as follow: 
 Absent at Baseline to Absent 
 Absent at Baseline to Present 
 Present at Baseline to Absent 


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 Present at Baseline to Present 
The categories for the shift from baseline for Cystoid Macular Edema, SRF, Subretinal Hyper-
Reflective Material and Pigment Epithelial Detachment assessments are defined as follows: 
 None at Baseline to None 
 None at Baseline to Yes, foveal/Yes, non-foveal 
 Yes, foveal/Yes, non-foveal at Baseline to None 
 Yes, foveal/Yes, non-foveal at Baseline to Yes, foveal/Yes, non-foveal 
For VMT, ERM, MH and Intraretinal Hyper-
SRF, Subretinal Hyper-Reflective 
Material and Pigment Epithelial Detachme
Yes, foveal/Yes, non-foveal  will be assigned 
 
Image quality will be listed only. This includes the Image Completeness, Centration, Tilt, 
Motion Artifacts and Signal Strength. 
4.16.4 Macular Integrity Assessment Microperimetry 
Mean Sensitivity, Bivariate Contour Ellipse Area 63%, Bivariate Contour Ellipse Area 95% 
and Fixation Losses will also be summarised by visit, by eye, by period as well as overall. The 
corresponding 95% CIs will be calculated (See Section 4.2). 
The number and percentage of subjects within each category of Fixation Stability will be 
summarised by visit, by eye, by period and by surgery window as well as all surgery windows 
combined. Shift from baseline will also be evaluated. 
The categories for the shift from baseline 
follows: 
 Stable at Baseline to Stable 
 Stable at Baseline to Relatively Unstable/Unstable 
 Relatively Unstable/Unstable at Baseline to Stable 
 Relatively Unstable/Unstable at Baseline to Relatively Unstable/Unstable 
Where microperimetry data are collected using the automated text files, the following 
algorithm will be used to define the value of Fixation Stability. 
 
 
 
 
 If P1<75% and P2<75%, then fixation stability is considered as unstable 
Image quality will be listed only. This includes image completeness, focus/clarity and field 
definition. 
 


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4.17 Safety Analysis 
Safety analysis will be performed on the All Treated Subjects analysis set using observed 
cases only. 
4.17.1 Visual Acuity 
T
decrease from baseline 
-5, -10 and -15 letters, respectively) will be summarised by visit, 
by eye, by period as well as overall in the same categorical summary tables mentioned in 
Section 4.16.1. The corresponding 95% CI will be presented (See Section 4.2).  
4.17.2 Intraocular Pressure 
Intraocular pressure and change from baseline will be summarised by visit, by eye, by period 
as well as overall.  
4.17.3 Slit Lamp Examination 
The number and percentage of subjects within each category of Slit Lamp Examinations 
outcomes will be summarised by visit, by eye, by period as well as overall. Shift from baseline 
will also be evaluated. 
The categories for the shift 
s: 
 Normal/Clinically insignificant abnormality at Baseline to Normal/Clinically 
insignificant abnormality 
 Normal/Clinically insignificant abnormality at Baseline to Clinically significant 
abnormality 
 Clinically significant abnormality at Baseline to Normal/Clinically insignificant 
abnormality 
 
Clinically significant abnormality at Baseline to Clinically significant abnormality  
The categories for the shift 
mation 
s: 
 Absent at Baseline to absent 
 Absent at Baseline to present 
 Present at Baseline to absent 
 Present at Baseline to present 
is defined as any other category (except missing). 


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and present is defined as any other category (except missing). 
defined as any other category (except missing). 
4.17.4 Lens Opacity Grades 
The number and percentage of subjects within each category of Nuclear Opalescence Grade, 
Nuclear Colour Grade, Cortical Cataract Grade and Posterior Cataract Grade will be 
summarised by visit, by eye, by period as well as overall. Shift from baseline will also be 
evaluated. 
The categories for Lens Opacity Grades are defined as Category 1, 2, 3 and 4. Category 1 
includes values 1, 1.0 and 0.x. Category 2 includes values 2, 2.0 and 1.x. Category 3 includes 
values 3, 3.0 and 2.x. Category 4 includes values 4, 4.0, 3.x an any values above 4. 
The categories for the shift from baseline are defined as follows: 
 No change in category 
 Decrease in category 
 Category 1 at Baseline to Category 2, 3 or 4 
 Category 2 at Baseline to Category 3 or 4 
 Category 3 at Baseline to Category 4 
4.17.5 Dilated Ophthalmoscopy 
The number and percentage of subjects within each category of each Dilated Ophthalmoscopy 
outcomes will be summarised by visit, by eye, by period as well as overall. Shift from baseline 
will also be evaluated. 
The categories for the shift from baseline for 
s: 
 Normal/Clinically insignificant abnormality at Baseline to Normal/Clinically 
insignificant abnormality 
 Normal/Clinically insignificant abnormality at Baseline to Clinically significant 
abnormality 
 Clinically significant abnormality at Baseline to Normal/Clinically insignificant 
abnormality 
 Clinically significant abnormality at Baseline to Clinically significant abnormality  
The categories for the shift from baseline 
assessments are defined as follows: 
 Absent at Baseline to absent 


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 Absent at Baseline to present 
 Present at Baseline to absent 
 Present at Baseline to present 
4.17.6 SD-OCT 
See Section 4.16.3. 
4.17.7 Fundus Autofluorescence 
See Section 4.16.2. 
4.17.8 Fundus Photography 
Presence and severity of Retinal Pigment Epithelium (RPE) Hyperplasia, Retinal Arteriolar 
Narrowing, Retinal Vessel Sheathing, Optic Atrophy/Pallor and Optic Disc Swelling will be 
summarised by visit, by eye, and by period. Shift from baseline will also be evaluated. 
The categories for the shift from baseline are defined as follows: 
 Absent at Baseline to absent 
 Absent at Baseline to present 
 Present at Baseline to absent 
 Present at Baseline to present 
where Absent corresponds to None and Present to Mild, Moderate or Severe. 
Image quality will be listed only. This includes image completeness, focus/clarity, field 
definition, stereopsis, exposure and contrast. 
4.17.9 Microperimetry 
See Section 4.16.4. 
4.17.10 Adverse Events 
Treatment Emergent Adverse Events (TEAEs) are defined as AEs starting on or after the 
day of the first surgery. AEs reported in subjects who received timrepigene emparvovec 
treatment in their first eye in an antecedent study will be considered as TEAEs. Where an AE 
start date is partially or fully missing, and it is unclear as to whether the AE is treatment 
emergent, it will be assumed that it is. Summary tables will include TEAEs only. TEAEs will 
be summarised by eye (for ocular events), by period, and overall. Each by-period summary 
table will be repeated by surgery window instead. 
The number and percentage of subjects reporting any event, any non-ocular event, any ocular 
event, any ocular event in SE1 and any ocular event in SE2 will be summarised. Percentage 
will be calculated based on the number of subjects in the corresponding period or surgery 
window or overall. The 95% CI will be calculated using the Clopper-Pearson method. 


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Overall summary table of TEAE by period (or by surgery window) will be provided for the 
following categories:  
 Number and percentage of subjects with a TEAE 
 Number and percentage of subjects with a serious TEAE 
 Number and percentage of subjects with a Treatment-related (study drug or study 
procedure) TEAE 
 Number and percentage of subjects with a Treatment-related (study drug or study 
procedure) serious TEAE 
 Number and percentage of subjects with a TEAE leading to death 
 Number and percentage of subjects who discontinued due to a TEAE 
 Number and percentage of subjects with a TEAE by severity 
 Number and percentage of subjects with a TEAE by outcome 
 Number and percentage of subjects with a TEAE by action taken 
Adverse events will be summarized by period (or by surgery window), and overall.  The 
following summaries of TEAEs will be provided: 
 TEAEs by SOC and PT 
 TEAEs by PT with percentage 
5% 
 Treatment-related (study drug or study procedure) TEAEs by SOC and PT 
 Study procedure-related TEAEs by SOC and PT 
 Study drug-related TEAEs by SOC and PT 
 TEAEs by SOC, PT, and maximum severity (missing severity, if any, will be counted 
as severe). 
 Serious TEAEs by SOC and PT 
 Treatment-related (study drug or study procedure) serious TEAEs by SOC and PT 
 Study procedure-related serious TEAEs by SOC and PT 
 Study drug-related serious TEAEs by SOC and PT 
 TEAEs leading to death by SOC and PT 
 TEAEs reported by subjects who discontinued due to SAE by SOC and PT 
Both counts of subjects and events will be reported. A subject with more than one occurrence 
of the same AE in a particular SOC will be counted only once in the total of those experiencing 
AEs in that particular SOC.  
If a subject experiences the same AE at more than one severity, the most severe rating will be 
given precedence. 
If a subject experiences the same AE at more than one relationship to study drug/procedure, 
causal relationship will be given precedence over no causal relationship. However, where a 
causal relationship is identified by the Investigator, the subject will be counted once within 
each sub-category (Study Drug, Study Procedure, Study Drug and Study Procedure, unknown). 


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In the summary tables, related events are defined as events that are assessed by the investigator 
to be related to study drug/procedure/both or with an unknown relationship. 
AEs will be assigned to Period 1 or Period 2 based on the AE Start Date. An AE with a start 
date on or after the day of surgery of Study Eye 2 will be attributed to Period 2; otherwise, the 
AE will be attributed to Period 1. 
An ocular event is an event where the site of the event is reported as OU, OD or OS. Where an 
r each eye (SE1 and SE2). 
Separate additional summaries will be provided for adverse events classified as Ocular 
Inflammation and Visual Acuity Reduced. The lists of PTs assigned to Ocular Inflammation 
and Visual Acuity Reduced are provided in a separate document (BIIB111 Signal Detection 
Plan). The following summaries will be provided by eye, by period (or by surgery window), 
and overall: 
 Ocular Inflammation-related TEAEs by SOC and PT 
 Ocular Inflammation-related TEAEs by SOC and PT, and by time 
>30 days) 
 Visual Acuity Reduced-related TEAEs by SOC and PT 
 Visual Acuity Reduced-related TEAEs 
>30 days) 
Imputation rules for partial dates are presented in Appendix 7.1. 
The following listings will be provided: 
 All AEs 
 AEs leading to death 
AEs will be coded according the MedDRA version 23.0 or higher. 
4.17.11 Immunogenicity 
Immunogenicity analyses will be performed on the Immunogenicity Analysis Set. Three 
immunogenicity data sets will be analyzed for the study: Anti-Drug Antibodies (ADA) to the 
transgenic product (REP1), neutralizing timrepigene emparvovec antibodies (Nabs) and 
ELISpot.  
In the ELISpot assay, a subject will be tested using 4 different analytes peptide pools (REP1 
Pool 1, REP1 Pool 2, REP2 Pool 3, AAV2) at each visit. All 4 pools together are used to 
evaluate the immune response to the whole therapeutic product, BIIB111, in which, the 3 REP1 
pools are used to evaluate the immune response to the therapeutic gene product, and the AAV2 
pool is used to evaluate the immune response to the AAV vector. A subject will be determined 
to have a positive result to the overall treatment with BIIB111 if any of the 4 analytes is 
positive. A subject will be determined to have a positive result with respect to the REP1 if any 
of the 3 REP1 result is positive. A subject will be determined to have a positive result with 
respect to the AAV2 if the AVV2 analyte is positive. 


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Samples with a numeric titer are considered as positive for ADA or Nabs respectively. For 
ELISpot, only samples which have spot counts (presented as SFU=spots per 1 mio cells) 3-fold 
of control (which is SFU of untreated cells) will present as positive. 
All the immunogenicity analyses specified in this section will be performed by assays (ADA, 
Nabs, ELISpot). 
4.17.11.1 
Baseline Definition 
As the second eye treatment in Period 2 in GEMINI may potentially increase the risk of a 
subject developing an immune response against the therapeutic product, BIIB111, we define 
two different types of baselines to evaluate the treatment emergent immune response, anchored 
to the time when the first eye was treated, and when the second eye was treated, respectively. 
The definition is as below. 
Period 1 baseline is defined as the last available value prior to surgery of the first eye. For 
subjects without any assessments prior to surgery of the first eye, Day 1 sample will be used 
as baseline.  If neither assessment prior to surgery of the first eye nor Day 1 sample are 
available, the period 1 baseline is set missing. 
Period 2 baseline is defined as the last available value within 10 weeks prior to the surgery of 
the second eye. For subjects without any assessments within 10 weeks prior to surgery of the 
second eye, Day 1 sample post-second eye surgery will be used as baseline for period 2. If 
neither assessment prior to surgery of the second eye nor Day 1 sample are available, the period 
2 baseline is set missing. 
Note that the immunogenicity data is collected as one record per subject per visit. 
4.17.11.2 
Assay Positivity by Visit 
The number and percentage of subjects with positive results at each visit will be presented by 
baseline status (such as screen positive, negative etc) and by treatment group. The positivity 
analysis for ELISpot will be summarized for the overall treatment of BIIB111 as well as with 
respect to either REP1 or AAV2. 
4.17.11.3 
Assay Treatment-emergent Analysis by Visit 
The number and percentage of subjects with a positive result will be further tabulated by 
treatment-emergent status (see definition below, treatment-emergent and non-treat-emergent 
positive) by, visit, period, surgery windows, and surgery windows combined.  
A treatment-emergent positive result is defined as follows for ADA and Nabs:  
 A post-baseline result that is positive when the baseline result is negative. 
 A post-baseline result that has a titer greater than or equal to 4 times the baseline titer 
when the baseline result is positive, while titer values are available at both baseline 
and post-baseline visits.  
For ELISpot, the treatment-emergent positive for each analyte is defined as a post-baseline 
result that is positive when the baseline result is negative. If any of the 3 REP1 analytes is 
treatment-emergent positive, it is a treatment-emergent positive for REP1. The treatment-
emergent positive for AAV2 is defined as a post-baseline result that is positive when the 


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baseline result is negative for the AAV2 analyte. If either REP1 or AAV2 is treatment-
emergent positive, it is a treatment-emergent positive for the overall treatment of BIIB111. The 
treatment-emergent analysis for ELISpot will be summarized for the overall treatment of 
BIIB111 as well as with respect to either REP1 or AAV2. 
If either the valid baseline samples (see definition above, prior to surgery or Day 1) or valid 
post-baseline samples at a particular visit are not available, the subject will be assigned as 
missing for treatment-emergent status at that particular visit and excluded from the treatment-
emergent assessment. The remaining available valid baseline samples and post-baseline 
assessments will be tabulated and contribute to the positive/negative overtime summary 
mentioned in 14.15.11.2. Invalid sample results include but are not limited to Quantity Not 
tc and will be deemed missing. 
For analyses on treatment-emergent immunogenicity, period 1 baseline will be used to derive 
the change from baseline for data of period 1 visits (before 2nd eye injection) and period 2 
baseline will be used to derive the change from baseline for data of period 2 visits (post 2nd eye 
injection).  
4.17.11.4 
Relationship Between Immunogenicity Assay and Safety 
The relationship between the immunogenicity assays (ADA, Nab and ELISpot) and selected 
adverse events will be assessed. These adverse events include ocular inflammation-related 
adverse events, visual acuity reduced-related adverse events and hypersensitivity-related 
adverse events. The number and percentage of post-treatment (treatment-emergent) ocular 
inflammation-related adverse events, visual acuity reduced-related adverse events and 
hypersensitivity-related adverse events (based on the adverse event data), as well as the number 
(based on the visual acuity assessment data) will be summarized by (1) subjects with negative 
assay at period 1 baseline; and (2) subjects with positive assay at period 1 baseline; and (3) 
subjects with at least one treatment-emergent positive assay at any visit during the study; and 
(4) subjects with no treatment-emergent positive assay during the study. In (3) and (4), to 
determine the treatment-emergent positivity, the period 1 baseline will be used to calculate the 
change from baseline for data in period 1 and period 2 baseline will be used to calculate the 
change from baseline for data in period 2.  
The analyses will be conducted by eye, by period and surgery window. It will be repeated by 
AE severity (mild, moderate 
-
 
4.17.11.5 
Data Listing 
Individual subject data will be listed with their positive/negative status, titer information (spot 
forming units (SFU) for ELISpot assay) and AEs of interests (as mentioned above) where 
available. The impact of assay status (positive/negative, treatment-emergent positive yes/no) 
on clinical efficacy may be further evaluated if needed.    


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4.17.12 Vector Shedding 
Samples of vector shedding will be summarized by categorical results and will be tabulated 
by visit, by period and eyes. Categorical results include but not limit to Positive, Negative 
and Below the Level of Quantification. The data of vector shedding will be listed. 
4.17.13 Vital Signs 
Blood pressure and pulse rate and change from baseline will be summarised by visit and by 
period. 
4.18 Protocol Violations or Deviations 
Deviations from the protocol will be documented on an ongoing basis by the study 
monitors/project manager throughout the study period.  
4.19 Deviations from the Statistical Analysis Plan 
Any deviations from the original statistical plan will be described and justified in the final 
clinical study report. 
4.20 Changes in Planned Analyses from the Protocol 
There are some changes to the planned analyses described in the protocol. 
1. The protocol refers to the assessment of Fixation Location. Because of a change in the 
way microperimetry data are collected, fixation location is not available in the database. 
Therefore, Fixation Location was removed from the SAP. 
2. The protocol mentions that specific analyses of the sensitivity points (microperimetry) 
will be performed. These analyses are not included in the SAP as they are considered 
as exploratory across the BIIB111 program and will be performed as post-hoc analyses, 
if needed to be consistent with other studies in the same program. 
3. For consistency of other studies in the program, time to resolution of AEs will not be 
summarized.  
3. The protocol specifies that LOCF will be used as imputation method for efficacy 
analyses. The main analyses of categorical and continuous efficacy endpoints will use 
the observed case method, as the main purpose of the study is the evaluation of BCVA 
from a safety perspective. Additional supportive analyses of BCVA-related endpoints, 
including LOCF imputation, will be applied and are described in detail in Section 
4.16.1. 
 
4.21 Algorithms/SAS Codes 
 Descriptive statistics for continuous variables: 




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5 Tables and Listings 
5.1 Format 
All outputs will be produced using SAS version 9.4 or a later version. 
A header will include the following text: NightstaRx, Ltd  NSR-REP-02  <FINAL ANALYSIS>  
In the top left portion of each table/listing, a table/listing number followed by the title of the 
table/listing will be presented. After the title line, optional sub-title or population information 
can be presented. Horizontal lines will appear before and after the column heading of the 
table/listing. Footnotes will be put under the main body of text at the bottom of the page.  
The sponsor name, protocol number, programmers User ID, status of the table/listing (i.e. 
draft or final) and SAS program name will appear bottom left in a string and the page number 
will appear on the bottom right corner of each table/listing. The date and time of creation of 
table/listing will appear bottom left under the sponsor name. The source listing number will 
appear bottom left.  
A landscape layout is proposed for both table and listing presentations. 
appropriate identification (e.g., the subject ID number) must be presented at the beginning of 
that page. 
Outputs will be produced as rtf files. The output file name will include the table number and 
name. 
5.2 Conventions for output display 
Unless otherwise specified, in summary tables of continuous variables, the minimum and 
maximum values will be displayed to the same number of decimal places as the raw data, the 
mean and median will be presented to one extra decimal place compared to the raw data, and 
the SD will be displayed to two extra decimal places compared to the raw data. Wherever 
possible data will be decimal aligned. 
Unless otherwise specified frequency tabulations will be presented by number and 
percentage, where the percentage is presented in brackets to 1 decimal place. 
Any date information in the listing will use the date9. format, for example, 07MAY2002. In 
the listing, a unit associated with a variable will be presented only once within parentheses 
either below or next to that variable in the heading portion. If a parameter has multiple units, 
each unit will be displayed only once, as applicable. 
All tables will have their source listing referenced in a footnote. Listings should be sorted 
by subject number, eye (starting with Study Eye or Study Eye 1) and visit. Listings 
should have the source data received from data management referenced in a footnote. 


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6 References 
MAIA, Macular Integrity Assessment, Microperimetry Handbook, First Edition, Centervue. 
 
 


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7 Appendices 
7.1 Appendix 1  Imputation of Partial Dates 
These imputation rules are limited to AE and CM partial dates only.  
Partial start dates (of intervention or event) 
 Case 1, day is missing (only month and year are present): 
o If year and month are same as treatment period start date then assign the day 
of treatment period start date to the partial date. 
o However, if end date of event or intervention is clearly before treatment period 
 
o 
e. 
 Case 2, only year is present: 
o If year is same as treatment period start date then assign the month and day of 
the treatment period start date to the partial date. 
o However, if end date of event or intervention is clearly before treatment period 
start dat
 
o 
 
 Case 3, completely missing date, no imputation is performed. 
Partial end dates (of intervention or event) 
 Case 1, day is missing (only month and year are present): 
o If year and month are same as study end date then assign the day of the study 
end date to the partial date. 
o Otherwise, assign day of last day of the month (28, 29, 30 or 31) to the partial 
end date. 
 Case 2, only year is present: 
o If year is same as study end date then assign the month and day of the study 
end date to the partial date. 
o 
 
 Case 3, completely missing date, no imputation is performed.      
Note: If any partial dates have missing month, with day present, then day is ignored and also 
considered missing. Similarly, if year is missing, with day or month present, we handle as if 
the entire date is missing. 
Note: If the study is ongoing and study end date is not available then the cut-off date will be 
used in the place of study end date.  If both a cut-off date and study end date are present for a 
patient then the minimum of the two dates will be used as the study (or reference) end date. 
7.2 Schedule of Study Procedures 
Table 3 
 presents a schedule of study procedures.