Conditions: Osteopetrosis
Allocation: NA | Intervention model: SINGLE_GROUP
Enrollment: 5
Healthy volunteers: no

Brief summary:
This study evaluates the effects of ACTIMMUNE (IFN-γ1b) in children and adults with intermediate osteoporosis. All participants will receive treatment with ACTIMMUNE for 12 months. The investigators hypothesize that ACTIMMUNE will be tolerated by participants for the full 12 months and result in decreased disease severity.

Detailed description:
Osteopetrosis is a rare inherited metabolic bone disease characterized by impaired osteoclast function resulting in defective bone resorption and generalized high bone mass and mineral density (BMD). In patients with severe disease, this high bone mass compromises bone marrow space leading to marrow failure and frequent infections, along with hepatosplenomegaly from extramedullary hematopoiesis. Currently, the only treatment for individuals with severe forms of osteopetrosis is hematopoietic cell transplantation (HCT), however survival in patients with osteopetrosis treated with HCT is only around 55%. Therefore, this treatment is only indicated in select individuals with life-threatening complications of their disease. Thus additional treatments for osteopetrosis are needed both for individuals who are not candidates for HCT and to prolong the time until HCT is needed.

Interferon gamma (IFN-γ) is a naturally occurring cytokine that has been shown to have anti-microbial and anti-viral immunomodulatory effects, and is a potent stimulator of superoxide anion production which in turn promotes the formation and activation of osteoclasts. Two previous studies of IFN-γ1b in a small group of individuals with osteopetrosis found a decrease in trabecular bone area, an increase in marrow space, a decrease in the number of severe infections requiring antibiotic therapy, and an increase in superoxide generation by granulocyte-macrophage colonies.

Therefore, the investigators will conduct an early phase 2, multi-center, open-label, 12-month clinical trial of ACTIMMUNE (IFN-γ1b) treatment of patients with intermediate osteopetrosis to determine the following:

1. The feasibility and tolerability of interferon gamma-1b treatment for 1 year in patients with intermediate osteopetrosis. Specifically, i) the ability to enroll patients, and ii) continued treatment throughout the 1-year observational period.
2. Change in immunologic and hematologic function, bone mineral density and osteoclast function, physical function and quality of life.

Arms:
arm 1: Gamma interferon-1b 100 mcg subcutaneous (SC) 3 times weekly for 12 months

Interventions: intervention 1: Interferon gamma-1b
Intervention descriptions: intervention 1: gamma interferon-1b dose escalation over first 4 weeks of study to 100 mcg SC 3 times weekly

Protocol document:
Prot_SAP_000.pdf: 
Open-label Phase 1/2 Study with a Single Arm of Interferon Gamma-1b 
Treatment of Osteopetrosis 
 
Short title: 
ACTIMMUNE in OP 
 
Regulatory Sponsor: 
Lynda Polgreen, MD, MS 
 
Assistant Professor 
 
Pediatric Endocrinology 
 
LA BioMed at Harbor-UCLA  
 
1124 West Carson Street, RB-1, Box 446 
 
Torrance, CA 90502 
 
Telephone: 310-222-1961 
 
Facsimile: 310-222-3887 
 
lpolgreen@labiomed.org 
 
Funding Sponsor: 
Horizon Pharma 
 
Study Product: 
Interferon gamma-1b (ACTIMMUNE®) 
 
 
IND Number: 
127052 
 
Trial Registration: 
NCT02666768 
 
IRB Number: 
21549-01 
 
Version / Date: 
version 1.8 / 13-Dec-2017 
 
 
 
 
 
 
 
 
 
 
CONFIDENTIAL 
This document is confidential and the property of the LABioMed. No part of it may be transmitted, reproduced, published, 
or used by other persons without prior written authorization from the study sponsor. 
 
 


Interferon gamma-1b in Osteopetrosis 
 
 
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Version History 
 
Version # 
Approval 
Date 
Significant Changes from Previous Version 
1.1 
 
1. Added anti-drug antibodies (see Sections 6.2.4 and 6.3.2, and 
Tables 1 and 2). 
2. Added specific genotypes that will be tested if an osteopetrosis 
related mutation has not been previously identified (see Section 
6.3.2 and Tables 1 and 2).  
3. Added justification for maximum dose and criteria to de-
escalate dose (see Sections 3 and 6.3.6) 
4. Added the following withdrawal criteria: subject contracts an 
intercurrent illness that requires treatment not consistent with 
the protocol requirements. (see Section 5.4.8) 
5. Added specification that the method used to calculate body 
surface area (BSA) is the Mosteller method (see Section 6.3.6). 
6. Added exclusion criteria of history of hematopoietic cell 
transplantation (HCT) 
7. Additional details on ex vivo assays were added to Section 
6.3.5.5. 
 
1.2 
22Mar2016 
Study subject compensation was clarified in section 6.9. 
1.3 
12Apr2016 
1. Added that screening will be within 3 months of baseline visit 
2. Added 25-hydroxy vitamin D level at 6 week lab visit 
3. Added additional instructions for processing blood and urine 
specimens (see Table 2 and sections 6.2.2, 6.2.3, and 6.2.4) 
4. Safety monitor was changed due to conflict of interest for Dr. 
Peter Liu (see Appendix A) 
1.4 
7Sept2016 
Added collection of a PAXgene tube for RNA extraction and buffy 
coat to be stored for future DNA studies (see Table 2, and Sections 
6.3.2 and 9.6). 
1.5 
22Nov2016 
Added “thalassemia or other hemoglobinopathy” to the exclusion 
criteria. 
1.6 
8Mar2017 
1. Updated PI contact info 
2. Changed UMN site PI due to Dr. Petryk leaving UMN. 
3. Section 5.4.4: Language was clarified to indicate that HCT 
cannot be expected to occur within 12 months of staring 
this study. 
4. Section 5.4.6:  Added exclusion criteria of a left ventricular 
ejection fraction of <30% to ensure consistency in using the 
screening ECHO results in determining enrollment in 
study. 
5. Section 6.3.2: Corrected labs to be collected at weeks 4 and 
6 and month 9.  This paragraph is now consistent with 
informed consent document and other areas in this 
protocol. 
6. Section 6.3.2:  Added adults ≥18 years old to the vitamin D 
and calcium treatment plans.   


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1.7 
10Aug2017 
1. Section 6.3.4.2: Language was clarified to add 3% and 38% 
sites of the non-dominant radius. 
2. Clarification of timing of RNA and DNA sample collection 
was added to Table 1. 
3. Section 6.9: Clarification of subject compensation between 
LA Biomed and University of Minnesota sites. 
1.8 
13Dec2017 
1. Section 6.2.2: Clarification of DNA, RNA and Urine 
collection, processing, shipping and Storage. 
2. Section 6.9: Correction of subject compensation which will 
be the same at each site. 
3. Section 6.3.6:  Extended duration for study drug titration to 
maximum 
dose 
and 
added 
recommendations 
for 
medications that may be used to treat adverse effects of the 
study drug. 
4. Changed site PI and clinical coordinator at UMN, and 
clinical coordinator at LA BioMed. 
5. Improved language in sections 5.4.4, 6.2.2, 6.2.3, and 8.2 
to improve clarity. 
6. Section 6.2.3: Changed the shipping contact and address for 
samples sent to UMN. 
 


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Table of Contents 
Version History ....................................................................................................................................................... 2 
Table of Contents .................................................................................................................................................... 4 
Study Summary ...................................................................................................................................................... 7 
List of Abbreviations .............................................................................................................................................. 8 
1 
Study Contact Information ............................................................................................................................. 10 
1.1 
Principal Investigator (PI) / Sponsor ....................................................................................................... 10 
1.2 
UMN Site PI ............................................................................................................................................ 10 
1.3 
Co-Investigators ...................................................................................................................................... 10 
1.4 
Study Personnel ....................................................................................................................................... 10 
2 
Introduction / Background and Rationale....................................................................................................... 11 
2.1 
Description of the disease. ........................................................................................................................... 11 
2.2 
Interferon gamma-1b in osteopetrosis.. ....................................................................................................... 12 
2.3 
Significance of research .............................................................................................................................. 12 
3 
Interferon gamma-1b (ACTIMMUNE) Description/Indications ............................................................... 12 
4 
Study Objectives / Hypotheses ....................................................................................................................... 13 
4.1 
Primary Hypothesis ................................................................................................................................. 13 
4.2 
Secondary Hypotheses ............................................................................................................................ 13 
5 
Study Design .................................................................................................................................................. 14 
5.1 
Overview of Study Design ...................................................................................................................... 14 
5.2 
Anticipated Duration of the Clinical Investigation ................................................................................. 15 
5.3 
Evaluations and study timeline ................................................................................................................ 15 
5.4 
Study Population ..................................................................................................................................... 17 
5.4.1 
Sample Size ...................................................................................................................................... 17 
5.4.2 
Subject Recruitment ......................................................................................................................... 17 
5.4.3 
Subject Screening ............................................................................................................................. 17 
5.4.4 
Prior and Concomitant Therapy ....................................................................................................... 17 
5.4.5 
Inclusion Criteria .............................................................................................................................. 17 
5.4.6 
Exclusion Criteria ............................................................................................................................. 18 
5.4.7 
Participation of Females ................................................................................................................... 18 
5.4.8 
Study subject Exit/Discontinuation Criteria ..................................................................................... 19 
6 
Study Procedures ............................................................................................................................................ 20 
6.1 
Informed Consent and Assent ................................................................................................................. 20 


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6.2 
Laboratory Testing Procedures ............................................................................................................... 21 
6.2.1 
Specimen Preparation ....................................................................................................................... 21 
6.2.2 
Research Specimen Handling and Storage ....................................................................................... 23 
6.2.3 
Specimen Shipment .......................................................................................................................... 25 
6.2.4 
Serum Sample Procedure for Immunogenicity assays: ........................................................................ 26 
6.3 
Clinical Procedures .................................................................................................................................. 27 
6.3.1 
Medical History ................................................................................................................................ 27 
6.3.2 
Laboratory Evaluations .................................................................................................................... 28 
6.3.3 
Medical examination and Anthropometrics ..................................................................................... 29 
6.3.4 
Radiographic assessments ................................................................................................................ 30 
6.3.4.1 
Dual energy x-ray absorptiometry (DXA)..................................................................................... 30 
6.3.4.2 
Peripheral quantitative computed tomography (pQCT) ................................................................ 30 
6.3.5 
Pain and physical function testing .................................................................................................... 30 
6.3.5.1 
Child Health Questionnaire - Parent Form 50 (CHQ-PF50) ......................................................... 30 
6.3.5.2 
Medical Outcomes Study Short Form-36  (SF-36) ....................................................................... 31 
6.3.5.3 
Patient and parent-reported pain intensity (acute and within the past week).. .............................. 31 
6.3.5.4 
Infectious Outcomes ...................................................................................................................... 31 
6.3.5.5 
In vitro osteoclast function assays ................................................................................................. 31 
6.3.6 
Administration of ACTIMMUNE .................................................................................................... 32 
6.4 
Unscheduled Study Visits ....................................................................................................................... 34 
6.5 
Study Protocol Compliance / Treatment Adherence ............................................................................... 34 
6.6 
Deviations from the Clinical Protocol ..................................................................................................... 34 
6.7 
Protocol Amendments After Study Initiation .......................................................................................... 34 
6.8 
Subject Withdrawal ................................................................................................................................. 34 
6.9 
Subject Compensation ............................................................................................................................. 35 
6.10 
Subject Compensation for Research Related Injury .............................................................................. 35 
6.11 
Sharing Results with Subjects ............................................................................................................... 35 
7 
Data Collection and Analysis ......................................................................................................................... 35 
7.1 
Subject Population(s) for Analysis .......................................................................................................... 35 
7.2 
Statistical Methods .................................................................................................................................. 36 
8 
Safety and Adverse Events ............................................................................................................................. 36 
8.1 
Definitions ............................................................................................................................................... 36 
8.2 
Recording of Adverse Events .................................................................................................................. 37 


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8.3 
Reporting of Serious Adverse Events...................................................................................................... 38 
8.3.1 
Study Sponsor Notification by Investigators .................................................................................... 38 
8.3.2 
IRB Notification by Investigators .................................................................................................... 39 
8.3.3 
UPIRTSO Events ............................................................................................................................. 39 
8.3.4 
Pregnancy ......................................................................................................................................... 39 
8.4 
Safety Monitoring Plan ........................................................................................................................... 39 
8.4.1 
Anticipated Risks / Risk Mitigation ................................................................................................. 40 
8.4.2 
Study Stopping Rules ....................................................................................................................... 41 
8.4.3 
Anticipated / Expected Adverse Events ........................................................................................... 41 
8.5 
Data Safety Monitoring Plan ................................................................................................................... 41 
9 
Data Handling and Record Keeping ............................................................................................................... 41 
9.1 
Confidentiality ......................................................................................................................................... 41 
9.2 
Source Documents ................................................................................................................................... 42 
9.3 
Case Report Forms .................................................................................................................................. 42 
9.4 
Clinical Reports ....................................................................................................................................... 43 
9.5 
Records Retention ................................................................................................................................... 43 
9.6 
Data and Specimen Banking ................................................................................................................... 43 
10 
Study Auditing, and Inspecting .................................................................................................................... 44 
11 
Ethical Considerations .................................................................................................................................. 44 
12 
Study Finances ............................................................................................................................................. 44 
12.1 
Funding Source...................................................................................................................................... 44 
12.2 
Conflicts of Interest ............................................................................................................................... 44 
13 
Publications Plan .......................................................................................................................................... 45 
 


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Study Summary 
Title 
Open-label Phase 1/2 Study with a Single Arm of Interferon Gamma-1b Treatment of 
Osteopetrosis 
Study Sponsor 
Lynda Polgreen, MD, MS 
Assistant Professor, Pediatric Endocrinology 
David Geffen School of Medicine 
Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center 
Principal 
Investigator (PI) 
Lynda Polgreen, MD, MS 
 
Assistant Professor, Pediatric Endocrinology 
David Geffen School of Medicine 
Los Angeles Biomedical Research Institute at Harbor-UCLA Medical Center 
Study Design 
multi-center open-label phase 1/2 clinical trial  
Study Duration 
Subjects will participate in the study for 1 year. It is expected that enrollment will be completed 
in 18 months and the last subject will complete the protocol 2.5 years after study initiation. 
Study Center(s) 
1. LABioMed at Harbor-UCLA 
1124 W. Carson St. 
Torrance, CA 90502 
2. University of Minnesota 
2450 Riverside Avenue 
Minneapolis, MN 55454 
Study Objectives 
1. To determine the feasibility and tolerability of interferon gamma-1b treatment in 
patients with intermediate osteopetrosis. 
2. To determine the safety and efficacy of interferon gamma-1b in patients with 
intermediate osteopetrosis. 
Study Endpoints 
Primary Endpoint: 
The primary objective of this pilot study is to determine the feasibility and tolerability of 
interferon 
gamma-1b 
treatment 
for 
1 
year 
in 
patients 
with 
intermediate 
osteopetrosis.  Specifically, i) the ability to enroll patients, and ii) continued treatment 
throughout the 1-year observational period.    
Secondary Endpoints: 
The secondary objectives of this study will focus on change in immunologic and hematologic 
function, bone mineral density and osteoclast function, physical function and quality of life. 
Number of Subjects 
10 
Main Inclusion 
Criteria 
 
Intermediate Osteopetrosis, defined as: 
o 
Anemia, or 
o 
Neutropenia, or 
o 
≥ 1 serious infection over prior year defined as requiring hospitalization and/or 
IV antibiotics, or 
o 
History of impaired bone healing 
 
 
Age > 1 year 
 
Duration of 
Observation 
1 year 
Statistical Methods 
Descriptive analyses of baseline characteristics and outcomes will include means and 
standard deviations for continuous variables and frequencies for categorical variables. 
Confidence intervals and P-values will be based on a t-distribution. Statistical significance 
will be considered as p<0.05.  The primary endpoint and safety analyses will be primarily 
descriptive reporting the number and percentage of compliance and AEs.  The secondary 
endpoints for hematologic function, immune function, bone mineral density, physical 
function and quality of life will be evaluated by paired t-tests or chi-squared to evaluate 
change from baseline to 12 months.  Secondary endpoints related to in vitro osteoclast assays 
will be descriptive. 
 


Interferon gamma-1b in Osteopetrosis 
 
 
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List of Abbreviations 
 
AE 
 
Adverse Event 
BMI 
 
Body Mass Index 
CAII  
Carbonic Anhydrase II 
CBCD  
Complete Blood Count with Differential 
CFR 
 
Code of Federal Regulations 
CHQ-PF50 
Children’s Health Questionnaire-Parent Form 50 
CLCN7 
Chloride Channel 7 
CNS  
Central Nervous System 
CRF 
 
Case Report Form 
CRP 
 
C-Reactive Protein 
CTSI  
Clinical and Translational Science Institute 
DNA  
Deoxyribonucleic Acid 
DSMB  
Data and Safety Monitoring Board 
EDTA  
Ethylenediaminetetraacetic Acid 
ELISA  
Enzyme Linked Immunosorbent Assay 
FDA  
Food and Drug Administration 
GCP  
Good Clinical Practice 
HCT  
Hematopoietic Cell Transplantation 
HIPAA 
Health Insurance Portability and Accountability Act of 1996 
IATA  
International Air Transport Association 
ICH 
 
International Conference on Harmonization 
IM 
 
Intramuscular 
IV 
 
Intravenous 
IDS 
 
Investigational Drug Services 
Ig 
 
Immunoglobulin 
IL 
 
Interleukin 
IND 
 
Investigational New Drug 
IRB 
 
Institutional Review Board 
ITT 
 
Intent-To-Treat 
LFT 
 
Liver Function Tests 
OSTM1 
Osteopetrosis-associated Transmembrane Protein 1 
PHI 
 
Protected Health Information 
PI 
 
Principal Investigator 
PP 
 
Per Protocol 
pQCT  
Peripheral Quantitative Computed Tomography 
RANKL 
Receptor Activator of Nuclear Factor-κ B Ligand 
REPA  
Report of External Professional Activities 
ROM  
Range of Motion 
SAE 
 
Serious Adverse Event 
SDS 
 
Standard Deviation Score 
SF-36  
Medical Outcomes Study Short Form-36   
SST 
 
Serum Separation Tube 
SC 
 
Subcutaneous 
TBD  
To Be Determined 
TCIRG1 
T-cell Immune Regulator 1 
UMN  
University of Minnesota 
UPIRTSO 
Unanticipated Problems Involving Risk To Subjects or Others 


Interferon gamma-1b in Osteopetrosis 
 
 
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USP 
 
United States Pharmacopeia 
vBMD  
Volumetric Bone Mineral Density 
 
 


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1 
Study Contact Information 
1.1 
Principal Investigator (PI) / Sponsor  
Lynda Polgreen, MD, MS 
Assistant Professor 
Pediatric Endocrinology 
LA BioMed at Harbor-UCLA 
1124 W. Carson St. 
Liu Research Bldg (RB3) 
Torrance, CA 90502 
Telephone: 310-222-1961 
Facsimile: 310-972-2962 
Email: lpolgreen@labiomed.org 
1.2 
UMN Site PI 
Paul Orchard, MD 
Professor 
Pediatric Blood and Marrow 
Transplantation 
University of Minnesota 
420 Delaware St., MMC 366 
Minneapolis, MN 55455 
Telephone: 612-626-2313 
Facsimile: 612-626-4074 
Email: orcha001@umn.edu 
1.3 
Co-Investigators  
 
Kim Mansky, PhD 
Associate Professor 
Develomental/Surgical Sci-Orthodont 
Room 16-146 MoosT 
515 Delaware St SE 
Minneapolis, MN 55455 
Telephone:  612-626-5582  
Facsimile:  612-301-5293 
Email:  kmansky@umn.edu 
 
 
 
 
 
 
 
 
 
 
1.4 
Study Personnel 
Clinical Coordinator (LA BioMed):   
 
Clinical Coordinator (UMN): 
 
  
Allison Barnwell  
 
 
 
 
Ka Xiong 
Los Angeles Biomedical Research Institute at  
University of Minnesota 
Harbor-UCLA Medical Center  
 
 
Masonic Cancer Center 
1124 West Carson Street  
 
 
 
Clinical Trials Office 
Torrance, CA 90502 
 
 
 
 
Minneapolis, MN 55455 
Office: (310) 7803-5623  
 
 
 
Office: (612) 626-5331 
allison.barnwell@labiomed.org   
 
 
herx0088@umn.edu 
 
 
 
 
 
Angel “Jun” Zozobrado  
 
 
 
 
Los Angeles Biomedical Research Institute at 
Harbor-UCLA Medical Center 
1124 West Carson Street 
Torrance, CA 90502 
Office: (310) 781-3652 
azozobrado@labiomed.org 
 
 
 


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2 
Introduction / Background and Rationale 
2.1 
Description of the disease.   
Osteopetrosis is a rare inherited metabolic bone disease characterized by impaired osteoclast 
function resulting in defective bone resorption and generalized high bone mass and mineral density 
(BMD).  In patients with severe disease, this high bone mass compromises bone marrow space 
leading to marrow failure and frequent infections, along with hepatosplenomegaly from 
extramedullary hematopoiesis (1). Visual defects may result due to impingement of cranial nerves. 
Other sequelae include hypocalcemia and hypocalcemic seizures and “brittle bones” or 
osteopetrotic rickets with an increased risk of fracture (2).  
Most patients with severe, autosomal recessive osteopetrosis have a defect in the osteoclast H+-
ATPase proton pump; mutations affecting the production of carbonic acid and the chloride channel 
have been reported as well (1).  The mutations result in varying degrees of osteoclast dysfunction 
and a wide spectrum of disease.  Osteopetrosis is generally divided into three categories based on 
clinical aspects of disease:  1) infantile or “malignant,” inherited in an autosomal recessive fashion; 
2) “intermediate,” associated with non-lethal recessive mutations, or a more severe phenotype of 
dominant disease; this is the patient population that will be included in this study; and 3) adult 
(“benign”) autosomal dominant osteopetrosis with generally milder clinical course (1).  Currently, 
the only treatment for individuals with severe forms of osteopetrosis is hematopoietic cell 
transplantation (HCT), however survival in patients with autosomal recessive osteopetrosis treated 
with HCT is only around 55% (3).  Therefore, this treatment is only indicated in select individuals 
with life-threatening complications of their disease.  Clearly, additional treatments for osteopetrosis 
are needed both for individuals who are not candidates for HCT and to prolong the time until HCT 
is needed. 
2.2 
Interferon gamma-1b in osteopetrosis.   
Interferon gamma is a naturally occurring cytokine that has been shown to have anti-microbial and 
anti-viral immunomodulatory effects (4), and is a potent stimulator of superoxide anion production 
which in turn promotes the formation and activation of osteoclasts (5). Interferon gamma-1b (IFN-
1b) has been shown to increase osteoclast size, tartrate acid phosphatase staining, and superoxide 
anion production in cell culture from patients with osteopetrosis (6).  In addition, osteopetrotic mice 
had increased marrow space following treatment with IFN-1b (7). Two previous studies of IFN-
1b in a small group of individuals with osteopetrosis found a decrease in trabecular bone area, an 
increase in marrow space, a decrease in the number of severe infections requiring antibiotic therapy, 
and an increase in superoxide generation by granulocyte-macrophage colonies (8,9).    


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Despite these encouraging findings and FDA approval of interferon gamma-1b for the treatment of 
severe, infantile “malignant” osteopetrosis, IFN-1b is not routinely prescribed for the treatment of 
intermediate osteopetrosis because the FDA approval does not include this more milder form of the 
disease; however, many of these patients have disease progression requiring HCT just at a later age 
than that for the severe infantile form. Due to these issues, and the small number of patients 
previously treated (8,9) there is a need to further delineate who will benefit from treatment, as some 
osteoclast defects may be more responsive than others.  
2.3 
Significance of research 
At the conclusion of this study, we will have obtained data on the tolerability and efficacy of IFN-
1b that, if encouraging, will be used to increase awareness about this treatment option among 
clinicians taking care of patients with intermediate osteopetrosis. We expect a decrease in disease 
burden, and improvement in health and quality-of-life for participants in our study.  Finally, the 
proposed project should provide further insight into possible variability in response to treatment 
based on genetic abnormality.  These contributions are significant because there is currently no 
effective treatment for osteopetrosis except HCT, which carries a high mortality rate (3,10). 
3 
Interferon gamma-1b (ACTIMMUNE) Description/Indications 
ACTIMMUNE® (IFN-1b), a biologic response modifier, is a single-chain polypeptide containing 
140 amino acids. ACTIMMUNE is a synthetic version of IFN-1b, a naturally occurring biologic 
response modifier.  
ACTIMMUNE is a sterile, clear, colorless solution filled in a single-use vial for subcutaneous 
injection. Each 0.5 mL of ACTIMMUNE contains: 100 mcg (2 million IU) of Interferon gamma-1b 
formulated in 20 mg mannitol, 0.37 mg disodium succinate hexahydrate, 0.14 mg succinic acid, 
0.05 mg polysorbate 20 and Sterile Water for Injection. Note that the above activity is expressed in 
International Units (1 million IU/50 mcg). This is equivalent to what was previously expressed as 
units (1.5 million U/50 mcg). 
ACTIMMUNE was first approved by the FDA in 1999 (Biologics License Application # 103836) 
and is currently FDA approved for the treatment of chronic granulomatous disease and severe 
malignant osteopetrosis.  FDA has approved ACTIMMUNE to decrease the number and severity of 
infections in patients with chronic granulomatous disease and to delay the progression of severe, 
malignant osteopetrosis. 
The original Phase 1 clinical development of ACTIMMUNE was conducted in oncology/AIDS 
patients and maximum tolerated doses based on intramuscular (IM) or intravenous (IV) 
administration were established for doses ranging from 10-1000 mcg/m2/day.  Subsequently, daily 
doses of 100 mcg/m2 ACTIMMUNE by the subcutaneous (SC) route of administration was chosen 
based on an NCI study comparing immunological activity (including enhanced superoxide/hydrogen 
peroxide production) of different doses and routes of administration in melanoma patients (11). SC 


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administration was compared with IM administration using the 100 mcg/m2 dose. SC administration 
resulted in enhanced hydrogen peroxide production and Fc receptor expression by monocytes. A 
comparison of two schedules, daily and three times weekly, suggested that monocyte activation may 
persist for up to 72 hours after ACTIMMUNE administration.  Of the doses tested, 100 mcg/m2 
administered daily or three times weekly appeared to be the most effective biological response 
modifier regimen, and due to ease of administration, the SC route was preferred. Since then, many 
studies have found similar tolerability and toxicity for 100 mcg/day and 200 mcg/day (12–17).  Thus, 
based on these previous studies of dosing safety and efficacy, we chose a maximum dose of 100 
mcg/m2 SC three times weekly for this Phase 1/2 trial.  
This is an investigator-initiated, investigator-held IND, multi-institutional research study. 
ACTIMMUNE is FDA-approved are described above, but not for the treatment of intermediate 
osteopetrosis. 
4 
Study Objectives / Hypotheses 
4.1 
Primary Hypothesis 
The primary hypothesis to be tested is:  
That treatment with IFN-1b will be feasible and tolerable to patients with intermediate osteopetrosis.  
4.2 
Secondary Hypotheses 
The secondary hypotheses are: 
1. 
Compared to baseline, treatment with IFN-1b will result in: 
a. Decreased tibial volumetric bone mineral density (vBMD) and estimated bone strength 
measured by peripheral quantitative computed tomography (pQCT); 
b. Decreased total body and lumbar spine areal BMD (aBMD) by dual-energy x-ray 
absorptiometry (DXA); 
c. Increased levels of markers of bone remodeling (see Table 1); 
d. Increased osteoclast activity by in vitro testing; 
e. Increased white blood cell (WBC) and platelet counts in patients that have lower than 
expected blood counts at baseline.   
f. Increased height standard deviation score (SDS) for participants <18 years with open growth 
plates; 
g. Decreased inflammation measured by erythrocyte sedimentation rate (ESR); 


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h. Decreased pain score (based on prior 4 weeks); 
i. Increased physical function score (based on prior 4 weeks); and 
2. Compared to the 12 months prior to study entry (based on medical record review or subject 
recollection), treatment with IFN-1b over 12 months will result in: 
a. Decreased number of hospitalizations for infection; 
b. Decreased number of days treated with antibiotics;  
c. Beneficial change in WBC and platelet counts in patients who have lower than expected 
blood counts at baseline;   
d. Beneficial change in ESR; 
e. Increased height SDS for participants <18 years with open growth plates; and 
3. Patients will not develop neutralizing antibodies. 
5 
Study Design 
5.1 
Overview of Study Design 
This is a multi-center, open-label, 12-month clinical trial of ACTIMMUNE treatment of patients with 
intermediate osteopetrosis. Study design is summarized in Figure 1.  Routine clinical safety and 
efficacy data (see Table 1) will be obtained at baseline, weeks 4 and 6, and months 3, 6, 9, and 12 


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following the initiation of therapy.  Additional outcomes (see Table 1) will be obtained at baseline, 
3, 6 and 12 months.  
 
5.2 
Anticipated Duration of the Clinical Investigation 
Each subject will be enrolled in the study for 12 months. It is expected that 15 patients will be screened 
over 18 months to enroll 10 participants, with the final subject completing study participation 2.5 
years after study initiation. The final study report will be completed approximately 3 years after study 
initiation. If the study is terminated prior to completion, a closeout report will be filed with the 
Institutional Review Board (IRB) within 3 months of study termination. 
5.3 
Evaluations and study timeline 
Table 1.  Evaluations and study timeline  
 
Screeninga 
Baseline 
# of months from 1st  
IFN-γ1b treatment  
(± 1 week) 
3 
6 
12 
General 
 
Medical history 
X 
X 
X 
X 
X 
 
Physical exam 
 
X 
X 
X 
X 
 
Genotypeb 
 
X 
 
 
 
 
Anthropometric measurements 
X 
Xf 
X 
X 
X 
 
RNA and DNA assays 
 
X 
X 
X 
X 
Safety Measuresc 
 
Basic metabolic panel (BMP) 
X 
X 
X 
X 
X 
 
Liver function tests (LFT) 
X 
X 
X 
X 
X 
 
Urine pregnancy test (UPT) e 
X 
X 
X 
X 
X 
 
Urinalysis (UA) 
X 
X 
X 
X 
X 
 
ECHO with interpretation 
X 
 
 
 
 
 
EKG with interpretation 
X 
 
 
 
 


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 
Anti-drug antibodies (ADA)g 
 
X 
X 
X 
X 
Bone Biomarker Testingd 
 
Formation Testing (bone specific 
alkaline phosphatase [BAP], type 1 
procollagen [P1NP]) 
- 
X 
X 
X 
X 
 
Resorption testing (C-telopeptide 
[CTX], Tartrate resistant acid 
phosphatase type5b [TRAP5b]) 
- 
X 
X 
X 
X 
 
RANKL, OPG, CK-BB 
- 
X 
X 
X 
X 
Endocrine Assessment 
 
Total 25-OH Vitamin D (25OHD)h 
- 
X 
X 
X 
X 
 
Serum ionized calcium 
- 
X 
X 
X 
X 
 
Urine calcium: creatinine 
- 
X 
X 
X 
X 
 
Intact parathyroid hormone (iPTH) 
- 
X 
X 
X 
X 
Inflammatory/Immunology 
 
CBC with differential and platelets 
- 
Xf 
X 
X 
X 
 
Erythrocyte sedimentation rate (ESR) 
- 
Xf 
X 
X 
X 
 
Immunologic phenotyping (CD3, 4, 8, 
14, 15, 16, 19, 34, 56) 
- 
X 
X 
X 
X 
Neuropsychological testing 
 
Pain assessment 
- 
X 
X 
X 
X 
 
Quality of life survey 
- 
X 
X 
X 
X 
Orthopedic assessment 
 
Dual energy X-ray absorptiometry 
(DXA) lumbar spine, total body, total 
hip and femoral neck (adults ≥18yrs 
only) 
- 
X 
- 
- 
X 
 
Peripheral Quantitative CT (pQCT) left 
tibia 
- 
X 
- 
X 
X 
In vitro osteoclast function assays – Mansky Lab 
 
Gene expression – osteoclast marker 
genes 
- 
X 
X 
X 
X 
 
Osteoclast activity – ability to release 
calcium phos from coated plate 
- 
X 
X 
X 
X 
Infectious outcomes 
 
Number of hospitalizations in prior 3 
months 
- 
Xf 
X 
X 
X 
 
Number of days treated with antibiotics 
in last 3 months 
- 
Xf 
X 
X 
X 
 
Type of infectious organism 
- 
Xf 
X 
X 
X 
RANKL=Receptor activator of nuclear factor kappa-B ligand; OPG=osteoprotegerin; CK-BB=creatinine 
kinase isoenzyme BB. 
a Screening visit evaluations will count for baseline evaluations where applicable. Screening visit will be 
within 3 months of baseline visit 
b Genetic testing for CLCN7, CAII, TCIRG1, OSTM1, and RANKL mutations will be done at the University of 
Minnesota if not already performed clinically 
c Safety labs will also be obtained at weeks 4 and 6 ± 2 days, and month 9 ± 1 week; subjects will be 
contacted on a weekly basis either by email or telephone to monitor safety during the initial 4 week dose 
escalation period. 
d Sample collection will be postponed if needed to ensure collection is more than 2 weeks since last fever 
(>100.4°F [38°C] axillary) or fracture. 
e Post-menarchal females. 
f Height (for subjects <18 years), CBC, and ESR will be obtain from the patient’s medical record when 
available and number of hospitalizations and infectious data from the patient’s medical record or if not 
available then the patient’s recollection at the baseline visit. 
g ADA testing will also be performed at weeks 3 and 6 ± 2 days after the first dose, and 2-3 weeks after last 
dose, and then every 4-6 weeks until values return to baseline. 
h 25OHD level will be checked at 6 weeks as well. 


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5.4 
Study Population 
This study will include patients diagnosed with osteopetrosis who are ≥1 year of age, have anemia, 
neutropenia, frequent serious infections, or impaired bone healing. 
5.4.1 Sample Size 
This is a pilot study; therefore, the sample size was determined based on clinician expert 
opinion and prevalence of the disease (estimated at 1/100,000-500,000) to estimate the 
number of potential participants who could be enrolled over 18 months. 
5.4.2 Subject Recruitment 
Subjects will be recruited from the Osteopetrosis Society website, clinicaltrials.gov, and 
provider referral. 
5.4.3 Subject Screening 
Due to the rarity of this disease patients will be traveling from all parts of the country to 
participate and therefore the screening may occur prior to the subject traveling to the study 
center to avoid unnecessary travel and therefore consent will be obtained verbally by phone 
when necessary (by the PI or study coordinator). At the Baseline visit, if not signed previously, 
consent and assent forms will be again reviewed in detail and the patient and/or their 
parents/legal guardians will have the opportunity to ask any questions they may have. They 
will also be asked to describe the protocol prior to signing these forms to ensure that all details 
are clearly understood. Medical records will then be reviewed to confirm eligibility. 
5.4.4 Prior and Concomitant Therapy 
Potential participants will not be enrolled if they expect to be treated with HCT within 12 
months of entering the study.  All other therapies are permitted.  
5.4.5 Inclusion Criteria 
Patients will be eligible to participate in the study if all of the following conditions exist: 
1. Diagnosis of osteopetrosis based on radiographic findings and 
2. Anemia (Hemoglobin <12 g/dL) not related to iron deficiency, or 
3. Neutropenia (Neutrophil count <1000 neutrophils/ul unsupported with cytokines), or 
4. Thrombocytopenia (Platelet count <50,000 cells x 109/L), or 
5. History of impaired bone healing, or 
6. ≥ 1 serious infection over prior year defined as requiring hospitalization and/or IV 
antibiotics, and 
7. Age > 1 year; and 
8. Ability to travel to a study center for every 3-6 month study visits; and 


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9. Patient or parent/legal guardian is able and willing to provide informed consent. For 
patients 7 to 17 years of age, assent must also be provided.   
5.4.6 Exclusion Criteria 
Patients will be excluded from participation in the study if any of the following conditions 
exist: 
1. History of HCT; 
2. Pregnancy or breastfeeding; 
3. Known or suspected allergy to interferon gamma-1b or related products;  
4. Participation in simultaneous therapeutic study that involves an investigational study 
drug or agent within 4 weeks of study enrollment;  
5. ALT greater than 3 fold higher than normal;  
6. Thalassemia or other hemoglobinopathy due to potential for inhibition of erythropoiesis 
and/or increased red blood cell destruction (18–21); or 
7. Left ventricular ejection fraction measured by ECHO of <30%; or 
8. Any other social or medical condition that the Investigator believes would pose a 
significant hazard to the subject if the investigational therapy were initiated or be 
detrimental to the study. 
5.4.7 Participation of Females 
The fetal risks associated with IFN-1b are not known, but pre-clinical animal data demonstrate 
some risk (Pregnancy category C).  Females of childbearing potential must agree not to become 
pregnant during the study and must have a pregnancy test at each study visit (baseline, month 3, 
month 6, and month 12).   Because of the risk involved, sexually active subjects and their 
partners must use two methods of birth control.  They must continue to use both methods until 1 
month after stopping study drug.  Two of the birth control methods listed below may be chosen:  
• 
Hormonal contraception  


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• 
Male or female condoms with or without a spermicidal 
• 
Diaphragm or cervical cap with a spermicidal  
• 
Intrauterine device (IUD) 
5.4.8 Study subject Exit/Discontinuation Criteria 
National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events 
(CTCAE), Version 4 will be used to define an Adverse Event (AE).  The CTCAE Grades 1 
through 5 with unique clinical descriptions of severity for each AE will be used and defined 
as follows: 
Grade 1 Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; 
intervention not indicated. 
Grade 2 Moderate; minimal, local or noninvasive intervention indicated; limiting age-
appropriate instrumental Activities of Daily Living (ADL)*. 
Grade 3 Severe or medically significant but not immediately life-threatening; hospitalization 
or prolongation of hospitalization indicated; disabling; limiting self care ADL**. 
Grade 4 Life-threatening consequences; urgent intervention indicated. 
Grade 5 Death related to AE. 
A Semi-colon indicates ‘or’ within the description of the grade.  
*Instrumental ADL refer to preparing meals, shopping for groceries or clothes, using the 
telephone, managing money, etc. 
**Self care ADL refer to bathing, dressing and undressing, feeding self, using the toilet, taking 
medications, and not bedridden. 
Subjects will discontinue participation in study for any unexplained CTCAE Grade 3 or higher 
that is not reasonably related to the underlying disease process or treatments.   In addition, 
subjects will discontinue study participation if any of the following conditions exist: 
1. Subject voluntarily withdraws from the study.  
2. Subject’s death. 
3. Subject completes the protocol.   
4. Pregnancy. 


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5. Subject acquires any of the listed exclusion criteria.  
6. Subject is non-compliant with the protocol.  For example, missing the Month 3 visit (no 
visit by 120 days after beginning therapy). 
7. Subject’s well-being, in the opinion of the Investigator would be compromised by 
continuation in the study. 
8. Subject contracts an intercurrent illness that requires treatment not consistent with the 
protocol requirements. 
 
6 
Study Procedures 
6.1 
Informed Consent and Assent 
Each subject or the subject’s parent/legal guardian will be provided a consent form describing this 
study and providing sufficient information to make an informed decision about their participation in 
this study. The consent form will be submitted with the protocol for review and approval by the IRB 
for the study. The formal consent of an adult subject or minor subject’s parent/legal guardian, using 
the IRB-approved consent form, must be obtained before that subject is submitted to any study 
procedure. This consent form must be signed by the subject and/or parent/legal guardian, and the 
Investigator-designated research professional obtaining the consent. A blank copy of the IRB-
approved form must be kept on-site by the Investigator.  
 
Consent process (in person) will take place in a private room. We will ensure that there is an 
ample waiting period available between informing the prospective subject and obtaining the 
consent. There will be dialogue during each contact with the subject to ensure ongoing 
consent is voluntary. 
 
Non-English Speaking Subjects 
 Spanish speaking subjects will be included. 
 Telephone or interpreter will be used. 
Subjects who are not yet adults (infants, children, teenagers) 
 Subjects in the United States are considered minors if they are 17 years or younger, with 
the exception of Alabama (18 years), Nebraska (18 years or upon marriage), and 
Mississippi (20 years) 
 Parental permission will be obtained from: 
o One parent even if the other parent is alive, known, competent, reasonably available, 
and shares legal responsibility for the care and custody of the child. 


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 Parental or guardian (as defined by DHHS and FDA regulations) consent will be 
obtained.  A guardian must provide written documentation of the legal ability to consent 
to for the child’s participation in research.  A copy of this documentation will be kept with 
the consent document.   
 Assent will be obtained from all minor subjects aged 7 to 17 years.  
 Assent will be documented with their signature on the assent form. 
 
Adults Unable to Consent 
The individuals from whom permission will be obtained are the following in descending 
order of priority: 
 The person's agent pursuant to an advance health care directive. 
 The conservator or guardian of the person having the authority to make health care 
decisions for the person 
 The spouse of the person 
 An individual as defined in Section 297 of the Family Code 
 An adult son or daughter of the person 
 A custodial parent of the person 
 Any adult brother or sister of the person 
 Any adult grandchild of the person 
 An available adult relative with the closest degree of kinship to the person 
The process for assent of the subjects is as follows: 
 Assent will be required of all subjects age 7-17 years [except for Alabama (age 7-18 
years), Nebraska (age 7-18 years or upon marriage), and Mississippi (age 7-20 years)] 
and for cognitively impaired adults. 
 The process for assent will include reviewing the assent document with the subject and 
their parent/guardian in a verbal face-to-face discussion, asking them for questions and 
discussing these, then asking the subject to sign and date the assent document if they 
would like to participate in the study. 
 Assent will be documented in a progress note. 
6.2 
Laboratory Testing Procedures 
6.2.1 Specimen Preparation 
Specimens for analysis will be collected as described in Table 2 at the time points indicated 
in Table 1. Venipuncture will be performed by trained certified nursing assistants, nurses, 
nurse practitioners, or physicians and collection and handling will be per the manufacturer’s 
recommendations. A central venous catheter may be utilized for all blood sampling if the 
patient currently has one. Serum, plasma, and urine will be collected and stored at -80°F.  
 


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Table 2. Laboratory Analytes and Collection Information. 
Analyte 
Specimen 
Tube Type 
Notes 
Batched 
at end of 
study? 
Volume  
High sensitivity erythrocyte 
sedimentation rate (ESR) 
Plasma 
K2EDTA (purple-top tube) 
Quest/Fairview  
no 
8 mL blood 
(2 tubes) 
 
Bone specific alkaline 
phosphatase (BAP) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Type 1 procollagen (P1NP) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Osteoprotegrin (OPG) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
C-telopeptide (CTX) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Tartrate resistant acid 
phosphatase type 5b 
(TRAP5b) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Receptor activator of 
nuclear factor-κ B ligand 
(RANKL) 
Plasma 
K2EDTA (purple-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Complete blood count with 
differential (CBCD) 
Whole blood 
K2EDTA (purple-top tube) 
Quest/Fairview 
no 
4 mL 
(1 tube) 
Immunologic phenotyping 
(CD3, 4, 8, 14, 15, 16, 19, 
34, 56) 
Whole blood 
K2EDTA (purple-top tube) 
Fairview  
no 
Genetic testing for Carbonic 
Anhydrase II (CAII), 
Chloride Channel 7 
(CLCLN7,) Osteopetrosis-
associated Transmembrane 
Protein 1 (OSTM1), 
RANKL, T-cell Immune 
Regulator 1 (TCIRG1) 
gene mutations 
Whole blood 
K2EDTA (purple-top tube) 
Fairview 
no 
4 mL 
(1 tube) 
Liver function tests (LFT) 
Serum 
SST (red-top tube) 
Quest/Fairview 
no 
10 ml blood 
(2 tubes) 
25-hydroxy vitamin D 
(25OHD) 
Serum 
SST (red-top tube) 
Quest/Fairview 
no 
Calcium 
Serum 
SST (red-top tube) 
Quest/Fairview 
no 
Intact parathyroid hormone 
(iPTH) 
Serum 
SST (red-top tube) 
Quest/Fairview 
no 
Basic metabolic panel 
(BMP) 
Serum 
SST (red-top tube) 
Quest/Fairview 
no 
Creatinine kinase -  BB 
isoenzyme (CK-BB) 
Serum 
SST (red-top tube) 
Mortari lab (store 
Orchard lab) 
yes 
Anti-drug antibodies (ADA) 
Serum 
SST (red-top tube) 
Intertek (store 
Polgreen/Orchard labs) 
yes- at 
month 12 
visit 
4 ml blood 
Storage 
Serum 
SST (red-top tube) 
Polgreen/Orchard labs 
NA 
5 ml blood 
Storage 
Plasma 
K2EDTA (purple-top tube) 
Polgreen/Orchard labs 
NA 
4 ml blood 
Storage 
Urine 
sterile container 
Polgreen/Orchard labs 
NA 
8 ml urine 
Urine pregnancy test, 
calcium to creatinine ratio 
(ca:cr) and urinalysis (UA) 
Urine 
sterile container 
Quest/Fairview 
POC UPT 
no 
10 ml urine 
Ex vivo assays (Mansky 
Lab) 
Whole Blood 
K2EDTA (purple-top tube) 
Manksy lab 
no 
10 ml 
(1 tube) 
RNA expression 
Whole Blood 
PAXgene RNA tube 
Chen lab 
yes 
2.5 ml 
DNA 
Buffy coat 
K2EDTA (purple-top tube) 
Chen lab 
NA 
10 ml 


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6.2.2 Research Specimen Handling and Storage (See Manual of Operations for instructions 
for non-LA BioMed sites)  
Samples for Bone Biomarkers and Storage 
Plasma. Collect blood in 3 purple-top (ethylenediaminetetraacetic acid [EDTA]) tubes (4 mL 
each). After collection, thoroughly mix the tube by gentle inversion 8 times. Label the tube 
with the Subject’s Study ID number, study visit, date of visit, and “plasma”. Do not write the 
subject’s name or any other identifying information on the blood tube. Place the tube 
upright in a rack in the refrigerator (2 to 8°C) until processed and stored at -80°C.  
If need to transport to the lab for processing, wrap the tube in a paper towel and place it in a 
plastic bag with an absorbent pad. Place the plastic bag containing the tube in a Styrofoam 
mailer with a frozen gel pack or on ice. The gel pack will ensure refrigerator temperature (2 
to 8°C) during transport to the laboratory. 
2 tubes: Centrifuge for 15 minutes at 4000 x g within 30 minutes of collection. Aliquot 0.5 ml 
into each 2 ml cryovial (8 cryovials) and store samples at -80°C.   
1 tube: Invert 8-10 times immediately after drawing. Keep tube cooled on ice, or with a cold 
pack, or in a 4C refrigerator. Transport to Translational Genomics & Population Sciences 
laboratory on ice for processing and storage of buffy coat. 
Serum. Collect blood in 2 red-top (serum separator tube [SST]) tubes (5 mL each). After 
collection, allow blood to clot for 30 minutes. Label the tube with the Subject’s Study ID 
number, study visit, date of visit, and “serum”. Do not write the subject’s name or any other 
identifying information on the blood tube. Place the tube upright in a rack in the refrigerator 
(2 to 8°C) until processed and stored at -80°C.  
If need to transport to the lab for processing, wrap the tube in a paper towel and place it in a 
plastic bag with an absorbent pad. Place the plastic bag containing the tube in a Styrofoam 
mailer with a frozen gel pack or on ice. The gel pack will ensure refrigerator temperature (2 
to 8°C) during transport to the laboratory. 
Centrifuge for 15 minutes at 4000 x g within 30 minutes of collection. Aliquot 0.5 ml into 
each 2 ml cryovial (10 cryovials) and store samples at -80°C.   
Urine   
Collect urine in a sterile container (5 mL each). Label the tube with the Subject’s Study ID 
number, study visit, date of visit, and “urine”. Do not write the subject’s name or any other 


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identifying information on the urine tube. Place the tube upright in a rack in the refrigerator 
(2 to 8°C).  
If need to transport to the lab for storage prior to shipping, wrap the tube in a paper towel and 
place it in a plastic bag with an absorbent pad. Place the plastic bag containing the tube in 
Styrofoam mailer with a frozen gel pack or on ice. The gel pack will ensure refrigerator 
temperature (2 to 8°C) during transport to the laboratory. 
 
Whole Blood 
Ex vivo assays: 
Collect 1 purple-top (EDTA) tube (10 mL).  Label the tube with the Subject’s Study ID 
number, study visit, date of visit, and “whole blood”. Do not write the subject’s name or 
any other identifying information on the blood tube.  Send to Dr. Kim Mansky per 
instructions below for arrival at Mansky lab within 36 hours of sample collection. 
Genetic Testing 
Collect 1 purple-top (EDTA) tube (5 mL).  Label the tube with the Subject’s Study ID number, 
study visit, date of visit, and “whole blood”. Do not write the subject’s name or any other 
identifying information on the blood tube. Send to Troy Lund/Kelly Miettunen per 
instructions below. 
Immunologic phenotyping 
Collect 1 purple-top (EDTA) tube (5 mL).  Label the tube with the Subject’s Study ID number, 
study visit, date of visit, and “whole blood”. Do not write the subject’s name or any other 
identifying information on the blood tube. Send to Troy Lund/Kelly Miettunen per 
instructions below. 
RNA expression 
Collect 1 PAXgene RNA tube (2.5 ml blood in a 10 ml tube).  Invert 8-10 times immediately 
after drawing.  Maintain tubes in upright position at room temperature for a minimum of 2  
hours.  Store PAXgene tubes in a wireframe rack (Styrofoam racks may cause the tubes to 
crack).  Transport to TGPS laboratory on ice. 
 


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***For EDTA (1) and PAXgene RNA tube (1) please email the following people the day 
prior to a subject’s scheduled visit to expect samples from the visit: Dr. Ida Chen, 
ichen@labiomed.org, Vicki Liu, yehengliu@labiomed.org, Ubaydah Nasri 
unasri@labiomed.org, and Kaye Roll, kroll@labiomed.org. 
 
 
 
All specimens must be labeled with a code. Unlabeled specimens will be destroyed.   
All specimens must be accompanied by a completed lab sheet.   
6.2.3 Specimen Shipment 
Whole blood for immunologic phenotyping, genetic testing, and ex vivo assays by Dr. 
Mansky collected at Harbor-UCLA will be shipped at ambient temperature by Fed Express 
overnight, following International Air Transport Association (IATA) regulations (double 
package or bag all specimens; include an absorbent square in each bag; affix "Exempt Human 
Specimens" label to box).  Ship for arrival within 36 hours of collection to: 
 
Troy Lund/Kelly Miettunen  
425 East River Parkway   
MCRB 660  
Minneapolis, MN (612) 626-5768 
 
Contact Kelly Miettunen 48 hours in advance by Phone (above) or Email: landb018@umn.edu 
prior to shipping. Deliver before 10:30 am on Tuesday – Thursday.   
 
Contact Dr. Kim Mansky by Phone: 612-626-5582 or email: kmansky@umn.edu 48 hours 
prior to shipping. 
 
 
Bone Biomarkers 
Samples for bone biomarkers will be stored at each site at -80°C until the end of the study at 
which time all samples will be shipped/transported on an adequate amount of dry ice to Dr. 
Angela Mortari, Cytokine Reference Lab.  For shipping samples to Dr. Mortari, samples will 
be shipped on an adequate amount of dry ice to the study PI following International Air 
Transport Association (IATA) regulations (double package or bag all specimens; include an 
absorbent square in each bag; affix "Exempt Human Specimens" label to box).  Ship for 
arrival within 36 hours: 
 
Angela Mortari 


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Cytokine lab 
13-127 PWB 
516 Delaware St. SE 
Minneapolis, MN 55455 
Contact Dr. Mortari 48 hours in advance by Phone:  (612) 626-7057 or email: 
panos001@umn.edu. Deliver before 10:30 on Tuesday – Thursday. 
 
Storage 
Samples for storage will be kept at each study site at -80°C until the end of the study at which 
time all samples will be shipped on an adequate amount of dry ice to the study PI following 
International Air Transport Association (IATA) regulations (double package or bag all 
specimens; include an absorbent square in each bag; affix "Exempt Human Specimens" label 
to box).  Ship for arrival within 36 hours: 
 
Lynda Polgreen 
1124 W. Carson St. 
RB-3, Rm 221 
Torrance, CA 90502 
Contact Dr. Polgreen 48 hours in advance by Phone:  (310) 222-1961 or email: 
lpolgreen@labiomed.org prior to shipping. Deliver before 10:30 on Tuesday – Thursday. 
 
6.2.4 Serum Sample Procedure for Immunogenicity assays: 
Collect blood samples to analyze Anti-Drug Antibodies (ADA) to IFN-γ and Neutralizing 
antibodies (Nab) to IFN-γ levels using 4.0 mL blood collection tube tubes (BD Vacutainer® SST™ 
Serum Separation Tubes).   
Collect a minimum of 2mL serum per time point described in Table 1. 
After obtaining the blood sample, inverting the collection tube 5 times allow the blood to clot for 30 
minutes at ambient temperature (19-24˚C).   
Centrifuge sample at approximately 1000-1300 RCF for 10 minutes at room temperature in a 
swinging bucket centrifuge.   
Transfer duplicate serum aliquots of approximately equal volume (minimum of 2 x 100 µL for 
ADA and 500 µl for Nab with backup of 500 µl per aliquot), using standard laboratory technique, 
into 2 polypropylene cryovials. 
Secure a label to each storage tube.  Labels should contain the following information:  
Sponsor: Dr. Lynda Polgreen 


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Sample Type: Human serum 
Sample date: DD-MMM-YYYY 
Sample time: XX:XX 
Protocol: 21549-01 
Subject Number: TBD 
Time point: TBD 
Serum Aliquot: TBD 
Within 90 minutes of collection, store both aliquot samples upright at -20°C +/-5°C or -70°C ± 
10˚C. 
Store at -70°C ± 10˚C until collection of month 12 sample, then ship all study subject samples as 
follows. If samples are collected after month 12, then ship immediately to Intertek as follows. 
Ship as separate shipments Serum Aliquot 1 and Serum Aliquot 2 samples for potential analysis of 
ADA and Nab with an adequate supply of dry ice via Fedex Monday through Wednesday to: 
 
Attn: Beth Cecil (ADA samples) or Lynn Jiang (Nab samples)  
Intertek Pharmaceutical Services 
10420 Wateridge Circle 
San Diego, CA  92121 
 
Provide an inventory log listing of all samples shipped with each shipment.  Include a paper copy 
with the shipment and an electronic version via email to Beth Cecil (beth.cecil@interteck.com) and 
Lynn Jian (lynn.jiang@intertek.com). 
Send a shipment notification including a confirmation of the shipment date, courier name, and    
tracking information via email to Beth Cecil (beth.cecil@interteck.com) and Lynn Jian 
(lynn.jiang@intertek.com). 
 
6.3 
Clinical Procedures 
6.3.1 Medical History 
A study physician and/or study coordinator will review the following with the subject/parent 
or legal guardian at the screening baseline visit: 
 Current and past 1 year prescription medications and doses,  
 current and past medical history,  
 review of systems,   
 family history, 
 number of hospitalizations over the last 12 months, and 


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 number of days treated with antibiotics over the last 12 months.  
For patients <18 years of age, height data will be collected from 6 and/or 12 months prior to 
the baseline visit. 
Changes in the medical history and a complete review of systems will be reviewed at each 
subsequent visit.    
If a subject is unable to return to the clinical site for the Month 3 visit they will be contacted 
via telephone to discuss any changes in health since their last study visit and then withdrawn 
from the study.  Three attempts will be made to contact the subject by phone before 
terminating the patient on the study.  
6.3.2 Laboratory Evaluations 
Blood and urine will be collected at the time points described in Table 1.   
CBCD, LFT, ESR, 25OHD, Calcium, iPTH, UA, urine ca:cr will be performed in real time 
by the Fairview Diagnostics Laboratory at UMN and at Quest for subjects seen at Harbor-
UCLA at baseline, 3, 6, and 12 months.  BMP, LFT, and UA will be performed at UMN, 
Quest, or the subject’s local clinical lab at weeks 4 and 6, and month 9.  Anti-drug antibody 
testing will be performed by Intertek Pharmaceutical Services at Baseline, at weeks 3 and 6, 
and months 3, 6 and 12, after the first dose, and 2-3 weeks after last dose, and then every 4-6 
weeks until values return to baseline.  Genetic testing for mutations in TCIRG1, CLCN7, 
CAII, OSTM1 and RANKL genes will be performed at the Baseline visit at the University of 
Minnesota for patients who have not had an osteopetrosis related mutation previously 
identified. 
25OHD will be measured by liquid chromatography tandem mass spectrometry, and an 
attempt will be made to maintain levels between 30 ng/ml and 75 ng/ml. Treatment of vitamin 
D insufficiency/deficiency will be as follows: 
 
Vitamin D insufficiency:  total 25-OH vitamin D 20-29 ng/ml 
Age 
Vitamin D 
Calcium 
(elemental)* 
0-1 years 
1,000 IU/d x 8 weeks 
NA 
1-3 years 
1,000 IU/d x 8 weeks 
500 mg/d 
4-8 years 
1,000 IU/d x 8 weeks 
800 mg/d 
>8 years 
2,000 IU/d x 8 weeks 
1,300 mg/d 
*Add calcium supplements if oral intake is inadequate or serum calcium is low. 
 
 
 
Vitamin D deficiency: total 25-OH vitamin D 10-20 ng/ml: obtain PTH and serum calcium 
levels. 


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Age 
Vitamin D 
Adequate 
Intake 
Calcium 
(elemental)* 
If PTH>65 pg/ml or serum calcium 
is low, add additional calcium as 
follows: 
Calcium (elemental) max dose 
2000 mg/day 
Start at higher dose and taper over 
4 weeks to AI. 
0-1 years 
2,000 IU/d x 8 weeks 
NA 
30-75 mg/kg/day divided 2-4 
times daily 
1-3 years 
2,000 IU/d x 8 weeks 
500 mg/d 
30-75 mg/kg/day divided 2-4 
times daily 
4-8 years 
4,000 IU/d x 8 weeks 
800 mg/d 
30-75 mg/kg/day divided 2-4 
times daily 
>8 years 
50,000 IU/wk x 8 weeks (Rx) 
1,300 mg/d 
30-75 mg/kg/day divided 2-4 
times daily 
*Ensure oral intake of calcium is adequate. 
 
 
Severe Vitamin D deficiency: total 25-OH vitamin D <10 ng/ml: obtain PTH and serum 
calcium levels. 
Age 
Vitamin D 
For all patients with severe vitamin D deficiency: 
Calcium (elemental) max dose 2000 mg/day 
Start at higher dose and taper over 4 weeks to AI. 
0-1 years 
2,000 IU/d x 8 weeks 
30-75 mg/kg/day divided 2-4 times daily 
1-3 years 
2,000 IU/d x 8 weeks 
30-75 mg/kg/day divided 2-4 times daily 
4-8 years 
4,000 IU/d x 8 weeks 
30-75 mg/kg/day divided 2-4 times daily 
>8 years 
50,000 IU/wk x 8 weeks (Rx) 
30-75 mg/kg/day divided 2-4 times daily 
 
 
For post-menarchal female patients, a urine pregnancy test will be performed at Screening 
and before the Baseline, Month 3, and Month 6 visits. Plasma and serum will be collected as 
described in Table 1. Immunologic phenotyping will be performed by University of 
Minnesota.  BAP, P1NP, OPG, CTX, TRAP5b, RANKL, and CK-BB will be performed by 
the Fairview Cytokine Reference Laboratory at the University of Minnesota.  Expression, 
methylation, and genotype data will be generated at LABioMed in the laboratories of Drs. Ida 
Chen and Kent Taylor. 
6.3.3 Medical examination and Anthropometrics 
A trained pediatric provider will perform a physical examination at each visit. Tanner staging 
of pubertal development, including breast and pubic hair development in girls, and testicular 
size and pubic hair development in boys will be performed at baseline, 6, and 12 months for 
participants < 18 years of age or < Tanner stage 5. Height will be measured 3 times standing 
without shoes. The average of 3 measurements by the same observer using identical technique 
with a wall-mounted stadiometer will be recorded. Each subject will be repositioned between 
each measurement. Height standard deviation score (SDS) will be calculated based on the 


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National Center for Health Statistics 2000 data as provided by the Center for Disease Control. 
Weight to be measured standing, once with subject in light clothing or dressing gown with 
shoes removed. These measurements will be used to calculate body mass index (BMI) as 
weight (kg) divided by height (m2). 
6.3.4 Radiographic assessments 
6.3.4.1 Dual energy x-ray absorptiometry (DXA) 
DXA (Hologic [Harbor] and GE [Minnesota]) measurements will be obtained for total body 
less head, L1-L4 lumbar spine, and for subjects ≥18 years of age femoral neck and total hip as 
well.  Standardized procedures for participant positioning and data processing will be used for 
all scans. Standardization of data from different machines is done by standardized phantom 
scanning protocols.   
6.3.4.2 Peripheral quantitative computed tomography (pQCT) 
Detailed measures of bone mass and strength will be assessed using pQCT (XCT 2000L 
[Harbor-UCLA] and XCT-3000 [UMN], Orthometrix Inc., White Plains, NY). This is a 
non-invasive technique that allows for 3-dimensional assessment of bone volumetric 
density, bone geometry and estimates of bone strength. Images will be taken at the 3%, 
38%, and 66% sites of the left tibia and the 3% and 38% sites of the non-dominant radius. 
The reference line will be the proximal end of the distal growth plate. Standardized 
procedures for participant positioning and data analysis will be used for all scans. Image 
processing and calculation of bone parameters will be completed using the manufacturer’s 
software (version 5.5) and thresholds established in the literature.  
6.3.5 Pain and physical function testing 
6.3.5.1 Child Health Questionnaire - Parent Form 50 (CHQ-PF50) 
The CHQ-PF50 is a 50-item, parent-completed questionnaire designed to measure the 
physical and psychosocial well-being of children between the ages of 5 and 18. The CHQ-
PF50 has been widely used in studies of chronic illness in childhood. Twelve components of 
quality of life are measured including: physical functioning, role/social limitations due to 
physical health, bodily pain/discomfort, general health, mental health, roe/social limitations 
due to emotional-behavioral difficulty, behavior, and self-esteem.  The impact of the child’s 
disorder on parents and family functioning (i.e., time impact, emotional impact, family 
activities, family cohesion) are also measured.  Items are measured on a likert scale, 
summed for each subscale and linearly transformed to a 0 to 100 scale, where higher scores 
reflect better functioning. Composite scores for physical functioning (Physical Summary 
Score) and psychosocial functioning (Psychosocial Summary Score) are calculated based on 
population norms. This questionnaire will be administered to all participants between the 
ages of 5 and 17 years. 


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6.3.5.2 Medical Outcomes Study Short Form-36  (SF-36) 
The SF-36 is a widely used measure of patient-reported quality of life for participants 18 
years of age and older.  The SF-36 measures 8 health domains, including physical 
functioning, role limitations due to physical health, bodily pain, general health perceptions, 
vitality, role limitations due to emotional problems, and mental health. Items are rated on a 
likert scale.  Summed scores of the scales are then transformed linearly to values from 0 
(worst health) to 100 (best health). Two summary measures (Physical and Mental Component 
Summary scores) are derived from population-based normative data. The SF-36 has been used 
in studies of adult chronic illness, and as an important component of treatment outcome 
studies. This questionnaire will be administered to all participants aged 18 years and above. 
6.3.5.3 Patient and parent-reported pain intensity (acute and within the past week). 
Visual analogue scales from the Pediatric Pain Questionnaire (PPQ) (22) will be used 
to assess intensity of the child’s pain from the child’s and parent’s perspective (for 
children ages 5-17). Visual analogue scales are widely used in both pediatric and adult 
pain studies due to their good reliability and validity, and they do not tend to result in 
the clustering of scores as can be seen with Likert-type scales (23,24). The visual 
analogue scales included in the PPQ consist of two 100 mm horizontal lines that 
measure both present pain and pain in the past week.  Each line is anchored with 
developmentally appropriate pain descriptions (i.e., “No pain, Not hurting” to 
“Hurting a whole lot, Severe pain”) and accompanying happy and sad faces. Scores on 
the PPQ range from 0 to 100, with higher scores representing more pain. For adults, 
the Visual Analogue Scale (VAS) for Pain (25,26), a 100 mm horizontal line with 
anchors appropriate for adults, ranging from “no pain” to “pain as bad as it could be; 
worst possible pain,” will be used to assess pain intensity in the present moment and 
over the past week.  As in the PPQ visual analogue scales, the VAS score is 
determined by measuring the distance (mm) on the 100 mm horizontal line between 
“no pain” and the patient’s mark, providing a range of scores from 0-100, with higher 
scores representing more pain intensity.  
For participants aged 18 years and older, pain intensity and its impact on daily 
functioning will be captured as part of the SF-36 questionnaire. 
6.3.5.4 Infectious Outcomes 
The frequency of infectious outcomes will be quantified by asking participants for the number 
of hospitalizations in the last 3 months and the number of days treated with antibiotics over 
the last 3 months (IV and PO will be counted separately).  The type of infectious organism 
will be documented as well. 
6.3.5.5 In vitro osteoclast function assays 
Osteoclasts will be isolated from whole blood by Ficoll gradient and CD14+ selection. The 
CD14+ cells will be cultured in macrophage colony stimulating factor (M-CSF) and 


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RANKL until multinuclear, TRAP positive cells are present in the culture.    Assays will be 
performed to determine the direct and indirect effects of IFNγ-1b as follows: 
- 
normal osteoclasts 
- 
normal osteoclasts + IFN-1b (0.25 ng/ml, 0.5 ng/ml, and 1.0 ng/ml),  
- 
diseased osteoclasts (baseline) 
- 
diseased osteoclasts (baseline)+ IFN-1b (0.25 ng/ml, 0.5 ng/ml, and 1.0 ng/ml)  
- 
diseased osteoclasts (month 6)   
- 
diseased osteoclasts (month 6) + IFN-1b (0.25 ng/ml, 0.5 ng/ml, and 1.0 ng/ml) 
- 
normal osteoclasts + 10% serum from treated patients 
- 
diseased osteoclasts (baseline) + 10% serum from treated patients  
- 
diseased osteoclasts (month 6) + 10% serum from treated patients  
 
 
 
Outcome measures will be as follows: 
For phenotypic analysis, gene expression (osteoclast marker genes such as but not limited to 
c-fos, Nfatc1, DC-STAMP, cathepsin K) will be measured by real time RT-PCR.  Osteoclast 
cultures will be stained for TRAP, photographed and osteoclast size and number will be 
measured.  Osteoclast cultures will also be plated on calcium phosphate coated dishes and 
activity of the osteoclasts (ability to release calcium phosphate from coated plate) will be 
evaluated.  Calcium phosphate dishes will be treated, photographed and percent area of the 
well absorbed, number of resorption pits and average size of the pit will be measured.   All 
of the data collected will be used to determine if changes in differentiation and/or activity 
are occurring in the osteoclasts in response to treatment. 
6.3.6 Administration of ACTIMMUNE 
The first SC injection of ACTIMMUNE will be administered by the patient or their 
parent/guardian under the guidance of a medical assistant, nurse or physician.  Subsequent 
injections will be administered independently by the patients or their parent/guardian. 
Titration dosing will be used to reduce the severity of ACTIMMUNE related flu-like 
symptoms as described by Devane et al (27).  Body surface area (BSA) will be calculated by 
the Mosteller method (28). 
For subjects with BSA >0.5 m2: 
- 
10 mcg/m2 SC three times weekly during week 1 
- 
20 mcg/m2 SC three times weekly during week 2 


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- 
30 mcg/m2 SC three times weekly during week 3  
- 
40 mcg/m2 SC three times weekly during week 4  
- 
50 mcg/m2 SC three times weekly during week 5  
- 
60 mcg/m2 SC three times weekly during week 6  
- 
70 mcg/m2 SC three times weekly during week 7  
- 
80 mcg/m2 SC three times weekly during week 8  
- 
90 mcg/m2 SC three times weekly during week 9  
- 
100 mcg/m2 SC three times weekly during week 10 and the remainder of the study if 
tolerated. ACTIMMUNE may be reduced on a case-by-case basis to manage 
subsequent drug-related AEs (e.g. flu-like reactions, elevated liver function tests or 
any drug-related Grade 2 or higher AE). 
For subjects with BSA ≤ 0.5 m2: 
- 
0.3 mcg/kg SC three times weekly during week 1 
- 
0.6 mcg/kg SC three times weekly during week 2 
- 
0.9 mcg/kg SC three times weekly during week 3  
- 
1.2 mcg/kg SC three times weekly during week 4  
- 
1.5 mcg/kg SC three times weekly during week 5  
- 
1.8 mcg/kg SC three times weekly during week 6  
- 
2.1 mcg/kg SC three times weekly during week 7  
- 
2.4 mcg/kg SC three times weekly during week 8  
- 
2.7 mcg/kg SC three times weekly during week 9  
- 
3.0 mcg/kg SC three times weekly during week 10 and the remainder of the study if 
tolerated. ACTIMMUNE may be reduced on a case-by-case basis to manage 
subsequent drug-related AEs (e.g. flu-like reactions, elevated liver function tests or 
any drug-related Grade 2 or higher AE). 
Over-the-counter and prescription medications can be used to treat flu-like reaction side effects 
per the study physician’s discretion. Possible treatment options for the flu-like reaction include 


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administering ACTIMMUNE 1-2 hours prior to bedtime, acetaminophen administration 30 
minutes prior to injection, and/or prednisone 5 mg po in morning on day of injection (titrate up to 
a maximum of 60 mg po day of injection – based on dosing for inflammatory disorder of 
musculoskeletal system; adjunct). 
6.4 
Unscheduled Study Visits 
Additional visits will be scheduled with study physicians or local physicians as needed throughout 
the study period to address concerning signs or symptoms reported by subjects. 
Subjects may withdraw voluntarily from the study at any time. If a subject opts to discontinue, 
attempts will be made to schedule and conduct an Early Termination visit for medical assessment.  
If a subject withdraws after an AE, they will be followed until resolution of the issue or until their 
condition is stable. In the event that a subject becomes pregnant, then discontinuation, reporting, and 
follow-up procedures outlined in Section 8.3.4 will be followed. 
 
6.5 
Study Protocol Compliance / Treatment Adherence 
Adherence to treatment will be evaluated by querying subjects every month either at scheduled study 
visits or by phone.  
6.6 
Deviations from the Clinical Protocol 
When a deviation from the protocol is necessary for an individual subject, the Investigator must 
contact the study PI prior to the deviation (unless the deviation is safety related). The subject may 
continue in the study by mutual agreement of the Sponsor and the Investigator. A description of the 
deviation from the protocol and justification must be recorded on the Protocol Deviation Form. 
6.7 
Protocol Amendments After Study Initiation 
Should changes in the study plan or protocol become necessary in the course of the study, those 
specific changes will be discussed and agreed upon by the Investigators and appropriate IRB approval 
obtained before the changes are implemented. All changes must be documented as protocol 
amendments. 
6.8 
Subject Withdrawal 
Subjects will be encouraged to complete the study; however, they may voluntarily withdraw at any 
time. If a subject is withdrawn prior to study completion, attempts will be made to schedule and 
conduct an Early Termination visit for medical assessment. If a subject cannot be contacted after 5 
telephone calls and 3 letters, they will be considered lost to follow-up. 
Subjects who discontinue study medication (e.g., due to non-compliance, side effects, etc.) will be 
asked to remain in the study for follow-up per the study schedule to preserve the ability to perform a 


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safety analysis. If a subject or their legal guardian (as applicable) withdraws consent to participate in 
the study, attempts will be made to obtain permission to record survival and adverse event 
(malignancy, hospitalization for infection) data at the 12-month study point. 
As long as the link between the sample ID code and the subject name is maintained by the clinical 
study site, it will be possible for subjects to ask that their samples be withdrawn.  However, we 
anticipate this link will be destroyed at some point in the future, after which withdrawal of samples 
will not be possible.  In addition, once DNA samples have been provided to collaborators, we cannot 
guarantee that all samples will be able to be tracked down. 
 
6.9 
Subject Compensation 
Travel reimbursement up to $1,500 will be provided for each of the Screening, Baseline, Month 3, 
Month 6, and Month 12 study visits.  This reimbursement can be applied to flights for subject or 
subject and 1 parent/guardian, gas, hotel (to be chosen and reserved by study coordinator for the 
number of nights required to complete study visit[s]), and/or food (up to $150/visit).  Receipts will 
need to be provided to study coordinator for reimbursement. Subjects will also receive $50 at 
Screening, Baseline, Month 3, Month 6, and Month 12 study visits as compensation for their time and 
inconvenience.  
6.10 Subject Compensation for Research Related Injury  
If a subject is injured as a result of taking part in this study they will receive medical 
treatment, however this care will be billed to their insurance in the usual manner and 
will not necessarily be free of charge to the subject. 
6.11  Sharing Results with Subjects 
All results obtained in this study are for research only.  No results will be shared 
with the subjects other than those needed to care for the subject during study 
participation (e.g. abnormal lab test).  When results are shared with the subject, they 
will be discussed in person or by telephone. 
 
7 
Data Collection and Analysis 
7.1 
Subject Population(s) for Analysis 
 Treatment-compliant population: Any subject who received at least 85% of prescribed doses and 
attended all scheduled study visits. 
 Safety population: All subjects who receive any amount of treatment.   


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7.2 
Statistical Methods 
7.2.1 Sample size Determination 
This is a pilot study to obtain data for a larger, definitive clinical trial.  The sample size of 10 
was determined based on feasibility estimates including potential screening failures  
7.2.2 Statistical Analysis 
Descriptive analyses of baseline characteristics and outcomes will include means and standard 
deviations for continuous variables and frequencies for categorical variables. Confidence 
intervals and P-values will be based on a t-distribution. Statistical significance will be 
considered as p<0.05.  The primary endpoint and safety analyses will be primarily descriptive 
reporting the number and percentage of compliance and AEs.  The secondary endpoints for 
hematologic function, immune function, and bone mineral density will be evaluated by paired 
t-tests or chi-squared to evaluate change from baseline to 12 months.  Secondary endpoints 
related to in vitro osteoclast assays will be descriptive. 
8 
Safety and Adverse Events 
8.1 
Definitions 
Adverse Event  
An adverse event (AE) is any symptom, sign, illness or experience that develops or worsens in 
severity during the course of the study. Intercurrent illnesses or injuries will be regarded as AEs. 
Abnormal results of laboratory or diagnostic procedures are considered to be AEs if the 
abnormality: 
 Results in study withdrawal 
 Is associated with a serious adverse event (SAE) 
 Is associated with clinical signs or symptoms 
 Leads to additional treatment or to further diagnostic tests 
 Is considered by the Investigator to be of clinical significance. 
Serious Adverse Event (SAE) 
An SAE is any AE that is: 
 Fatal 


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 Life-threatening 
 Requires or prolongs a hospital stay 
 Results in persistent or significant disability or incapacity 
 A congenital anomaly or birth defect 
Important medical events are events that may not be immediately life-threatening, but are 
clearly of major clinical significance and may be SAEs. They may jeopardize the subject, and 
may require intervention to prevent one or the other serious outcomes noted above.  
Hospitalization  
Hospitalization shall include any initial admission (even if less than 24 hours) to a healthcare 
facility as a result of a precipitating clinical adverse effect; to include transfer within the hospital 
to an intensive care unit. Hospitalization or prolongation of hospitalization in the absence of a 
precipitating, clinical adverse effect (e.g., for a preexisting condition not associated with a new 
adverse effect or with a worsening of the preexisting condition; admission for a protocol-
specified procedure) is not, in itself, a serious adverse effect. 
Expected Adverse Event 
Expected AEs are those that are known to be associated with or have the potential to arise as a 
consequence of participation in the study. This will include any AE or SAE that are expected 
consequences of having osteopetrosis or receiving treatment with IFN-1b described in Section 
8.4.4. 
Unexpected Adverse Event 
An AE or suspected adverse reaction is considered unexpected if it is not listed in the drug 
package insert or Protocol at the specificity or severity that has been observed. 
Unanticipated Problems Involving Risk To Subjects or Others (UPIRTSO) 
An AE that in the opinion of the PI is unexpected, related to the study procedures, and serious. 
8.2 
Recording of Adverse Events 
At each contact with the subject, the Investigator/study coordinator must seek information on AEs by 
specific questioning and, as appropriate, by examination. Information on all AEs should be recorded 
immediately in the source document, and also in the appropriate AE module of the case report form 
(CRF). All clearly related signs, symptoms, and abnormal diagnostic procedures resulting from a 
single clinical condition should be recorded individually in the source document and listed under the 


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single unifying diagnosis. (For example, fever, rigors, sore throat, and high neutrophil count might 
all be listed under a diagnosis of pharyngitis.) 
 
All AEs occurring during the study period must be recorded. The clinical course of each event should 
be followed until resolution, stabilization, or until it has been determined that the study treatment or 
participation is not the cause. Serious adverse events that are still ongoing at the end of the study 
period must be followed up to determine the final outcome. Any SAE that occurs after the study 
period and is considered to be possibly related to study participation should be recorded and reported 
immediately. 
 
 
AEs will be classified according to the National Cancer Institute (NCI) Common Toxicity Criteria 
(CTC) V 4.1 and causality will be classified as: 
Unrelated - The AE is clearly NOT related to study procedures 
Unlikely - The AE is doubtfully related to study procedures 
Possible - The AE may be related to study procedures 
Probable - The AE is likely related to study procedures 
Definite - The AE is clearly related to study procedures 
8.3 
Reporting of Serious Adverse Events 
8.3.1 Study Sponsor Notification by Investigators 
An SAE must be reported to the study PI by telephone, fax, or email (preferred) within 
24 hours of the event. An SAE Form must be completed by the Investigator and faxed or 
emailed to the study Sponsor within 24 hours. The Investigator will keep a copy of this SAE 
form on file at the study site. Report SAEs by phone, fax or email to: 
 
Lynda Polgreen, MD, MS 
310-222-1961 (telephone) 
 
 
 
 
 
310-222-3887 (facsimile) 
lpolgreen@labiomed.org 
At the time of the initial report the following information should be provided: 
 Study Identifier 
 Study Center 
 


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 Subject Number 
 A description of the event  
 The reason why the event is 
classified as serious  
 Date of onset 
 Current Status 
 Investigator assessment of the 
association between the event 
and study procedures 
Within the following 48 hours, the Investigator must provide further information on the SAE 
in the form of a written narrative. This should include a copy of the completed SAE Form, 
and any other diagnostic information that will assist the understanding of the event. 
Significant new information on ongoing SAEs should be provided promptly to the study PI. 
8.3.2 IRB Notification by Investigators 
Reports of all SAEs (including follow-up information) must be submitted to the IRB within 
10 working days if it falls under the UPIRTSO guidelines (unexpected and related to study 
drug). All deaths, regardless of expectedness or causality, must be reported to the IRB within 
10 working days. Copies of each report and documentation of IRB notification and receipt 
will be kept in the Clinical Investigator’s binder. 
8.3.3 UPIRTSO Events 
Investigators are required to submit a report of UPIRTSO events to the IRB within 10 working 
days of first learning of the event.  
8.3.4 Pregnancy 
If a female subject becomes pregnant during the study, she will be withdrawn from the study 
immediately. Though pregnancy is not by definition an SAE, it will be subject to SAE 
reporting requirements. 
Though the subject will be withdrawn from the study, permission to record survival and 
adverse event (malignancy, hospitalization for infection) data for the mother to the end of the 
pregnancy and of survival data for the baby at birth will be requested. 
8.4 
Safety Monitoring Plan 
The plan for ensuring subject safety will include the following elements: 
1. Clinical procedures will only be performed by properly trained personnel who are qualified by 
training and licensure to perform the procedures. 
2. Investigator and all study personnel have completed required training regarding Human Subject 
Protections. All personnel will comply with all related regulations and laws. 


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3. Patients will be rigorously screened against inclusion/exclusion criteria to ensure that their 
participation is safe. 
4. Adverse events and SAEs will be assessed and followed throughout study. Subjects will have 
contact information to enable them to contact study personnel easily and quickly. 
5. Study data and information will be kept confidential and managed in accordance with requirements 
of Health Insurance Portability and Accountability Act of 1996 (HIPAA). All data will be stored 
in locked offices and not released without subject permission.  
6. Subject may discontinue participation at any time, for any reason. Any subject who fails to 
complete the research procedures within acceptable parameters, or is unable to safely tolerate 
participation in the study will be withdrawn. 
7. Institutional Review Board approval will be obtained.  
8. Monitoring will be conducted by an external Data and Safety Monitor as described in the Data and 
Safety Monitoring Plan (appendix A).  
8.4.1 Anticipated Risks / Risk Mitigation 
8.4.1.1 Venipuncture 
The risks of venipuncture are transient pain, bleeding, lightheadedness, bruising, 
possible vasovagal reaction, and infection. Only trained certified nursing assistants, 
nurses, nurse practitioners, or physicians will be allowed to perform venipuncture to 
minimize the risk of complication. 
8.4.1.2 pQCT and DXA 
The pQCT and DXA scan will expose the subject to ionizing radiation. The level of 
exposure for this study will be approximately 3% of that received from natural sources 
of radiation by a northern US resident in 1 year. Trained staff will perform the scans to 
minimize the risk of additional exposure. 
8.4.1.3 Provisions to Monitor the Data to Ensure the Safety of Subjects 
Genetic studies have raised concern as to whether the studies would place research 
subjects at risk for discrimination based on genetics.  The federal Genetic Information 
Nondiscrimination Act (GINA) was passed to address this concern.  GINA makes it 
illegal for medical insurance companies and most employers to discriminate based on 
genetic information.  The protections of GINA do not apply to life, disability, or long-
term-care insurance.    


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8.4.2 Study Stopping Rules  
This study may be terminated by the PI at any time. Reasons for terminating the study may 
include the following: 
 The incidence or severity of AEs in this or other studies indicates a potential health hazard 
to subjects. 
 Subject enrollment is unsatisfactory. 
 Data recording is inaccurate or incomplete. 
 1 patient develops a CTCAE Grade 4 or higher. 
8.4.3 Anticipated / Expected Adverse Events 
The following AEs have been reported in individuals treated with ACTIMMUNE:  fever 
(52%), headache (33%), rash (17%), chills (14%), injection-site erythema or tenderness 
(14%), fatigue (14%), diarrhea (14%), vomiting (13%), and nausea (10%). Less common 
adverse events observed more frequently in the IFN-1b group compared with placebo 
(occurring in <10% of patients) included myalgia, arthralgia, back pain, abdominal pain, and 
depression. 
Adverse events related to osteopetrosis are expected. These expected events include but are 
not 
limited 
to, 
pain, 
fracture, 
infection, 
osteonecrosis, 
anemia, 
neutropenia, 
thrombocytopenia, hepatosplenomegaly, cranial nerve impingement, and hypocalcemia. 
These events will be considered expected for the purposes of expedited SAE reporting. 
8.5 
Data Safety Monitoring Plan 
Data safety monitoring plan is described in detail in Appendix A. 
9 
Data Handling and Record Keeping 
9.1 
Confidentiality 
Study data and information will be kept confidential and managed in accordance with requirements 
of Health Insurance Portability and Accountability Act of 1996 (HIPAA). We will follow institutional 
policies regarding HIPAA, we will answer any questions/concerns the subjects and their parents may 
have in regards to their privacy, address items, and document. If at any time subject/parent/family/ or 
child appear uneasy during exams, questions, etc. the team member will stop the visit to refer to PI or 
doctor performing visit to determine if the study visit can continue. Research team is permitted to 
access sources of information about the subjects for research purposes only, document all requests if 
not in protocol.  


Interferon gamma-1b in Osteopetrosis 
 
 
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HIPAA regulations require a signed subject authorization informing the subject of the following:  
 What protected health information (PHI) will be collected from subjects in this study 
 Who will have access to that information and why 
 Who will use or disclose that information 
 The rights of a research subject to revoke their authorization for use of their PHI.  
In the event that a subject revokes authorization to collect or use PHI, the Investigator, by regulation, 
retains the ability to use all information collected prior to the revocation of subject authorization. For 
subjects that have revoked authorization to collect or use PHI, attempts should be made to obtain 
permission to collect at least vital status (i.e., that the subject is alive) at the end of their scheduled 
study period. 
The samples sent to laboratories will include the subject’s ID number and not their name. Those who 
have access to data and specimens are the research team who all have HIPAA training; these same 
people are response for receipt and/or transmission of data and specimens. 
All interactions with the subject and his/her parents will take place in a private setting, e.g., a private 
exam room. 
The genetic information obtained in this study is kept in strict confidence. The laboratories in which 
genotyping and assays are performed use only study ID codes and no other identifiers. The 
information is identified in the database by a unique code. The genotyping results are merged with 
the phenotype data for statistical genetic analysis using only study ID codes to identify individuals. 
We will secure hard copy information with keys and computer databases with password protection 
so that only research personnel related to this project will have access to information.    
 
9.2 
Source Documents 
Source Data are the clinical findings and observations, laboratory and test data, and other information 
contained in Source Documents. Source Documents are the original records (and certified copies of 
original records); including, but not limited to, hospital medical records, physician or office charts, 
physician or nursing notes, subject diaries or evaluation checklists, pharmacy dispensing records, 
recorded data from automated instruments, x-rays, etc. When applicable, information recorded on the 
CRF shall match the Source Data recorded on the Source Documents.  
9.3 
Case Report Forms 
The study CRF is the primary data collection instrument for the study. All data requested on the CRF 
must be recorded. All missing data must be explained. If a space on the CRF is left blank because the 


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procedure was not done or the question was not asked, “N/D” will be entered. If the item is not 
applicable to the individual case, “N/A” will be entered. All entries should be printed legibly in black 
ink. If any entry error has been made, to correct such an error, a single straight line will be drawn 
through the incorrect entry and the correct data entered above it. All such changes must be initialed 
and dated. DO NOT ERASE OR WHITE OUT ERRORS. For clarification of illegible or uncertain 
entries, the clarification will be printed above the item, then initialed and dated. 
A Case Report Form will be completed for each subject enrolled into the clinical study. The 
Investigator-Sponsor 
will 
review, 
approve 
and 
sign/date 
each 
completed 
CRF; 
the 
Investigator-Sponsor’s signature serving as attestation of the Investigator-Sponsor’s responsibility for 
ensuring that all clinical and laboratory data entered on the CRF are complete, accurate and authentic.  
All data including source documents and CRFs will be maintained in a locked filing cabinet in a 
locked office with limited access to only those involved in the research. Individual subject files will 
be labeled with the subjects unique study identification number and not their name.  
 
9.4 
Clinical Reports 
An annual progress report will be submitted to the funding source and the IRB. Investigators will 
submit a final report of the clinical study to the reviewing IRB within 3 months of termination or 
completion of the clinical study or the Investigator’s part of the clinical study. 
9.5 
Records Retention 
The Investigator-Sponsor will maintain a file of all documents concerning the use of human subjects 
in research for 3 years from completion of IRB related work and 6 years for HIPAA. 
9.6 
Data and Specimen Banking 
Stored samples will be identified by an indirect identifier (study ID) indefinitely.  The study PI and 
study coordinator will have access to the link between subject information and study ID. 
 
If stored samples are used in the future to look at new markers the subject or parent/guardian will 
not be notified nor will they receive the results from the analysis.  If the samples are used by other 
research groups for analysis, the subject and guardian will not be notified.  However, any samples 
that are sent to another researcher will be identified only with a number and will not be able to be 
traced back to the subject.  The subject will not be identified in any publication or report from this 
study.  The subject or guardian can request that blood, urine, RNA, and DNA samples be destroyed 
at any time by contacting the study PI. 
 


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10 
Study Auditing, and Inspecting 
The Investigator will permit study-related monitoring, audits, and inspections by the IRB, government 
regulatory bodies, and University compliance and quality assurance groups of all study related 
documents (e.g., source documents, regulatory documents, data collection instruments, study data 
etc.). The Investigator will ensure the capability for inspections of applicable study-related facilities 
(e.g., pharmacy, diagnostic laboratory, etc.). 
Participation as an Investigator in this study implies acceptance of potential inspection by government 
regulatory authorities and applicable University compliance and quality assurance offices. 
11 
Ethical Considerations 
This study is to be conducted according to US and international standards of GCP (Code of Federal 
Regulations [CFR] 21 CFR 312 and International Conference on Harmonization [ICH] guidelines), 
applicable government regulations and Institutional research policies and procedures. 
This protocol and any amendments will be submitted to a properly constituted independent IRB, in 
agreement with local legal prescriptions, for formal approval of the study conduct. The decision of the IRB 
concerning the conduct of the study will be made in writing to the Investigator and a copy of this decision 
will be provided to the Sponsor before commencement of this study.  
12 
Study Finances 
12.1 Funding Source 
Research related costs will be paid for through an investigator initiated grant from Horizon Pharma.  
ACTIMMUNE will be provided by Horizon Pharma. 
12.2 Conflicts of Interest 
Any Investigator who has a conflict of interest with this study (patent ownership, royalties, or 
financial gain greater than the minimum allowable by their institution, etc.) must refer to the Regents 
Policies on Individual Conflict of Interest Policy or Institutional Conflict of Interest Policy. These 
policies require University Faculty and staff to report external professional activities and business 
and significant financial interests related to his or her University activities by submitting a REPA 
(Report of External Professional Activities) at least once per year. Faculty and staff should also file 
a REPA when substantial changes in business or financial interests occur, when an activity that 
presents a potential conflict of interest is anticipated, or when submitting an application for research 
support or technology transfer, submitting research protocols to the IRB, or receiving financial 
contributions. All investigators will follow their University’s conflict of interest policy. 


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13 
Publications Plan 
Neither the complete nor any part of the results of the study carried out under this protocol, nor any of the 
information provided by the Sponsor for the purposes of performing the study, will be published or passed 
on to any third party without the consent of the study Sponsor. Any Investigator involved with this study is 
obligated to provide the Sponsor with complete test results and all data derived from the study. 
 
 


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References 
1.  
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2.  
Gonen KA, Yazici Z, Gokalp G, Ucar AK. Infantile osteopetrosis with superimposed rickets. Pediatr 
Radiol. 2013 Jan;43(2):189–95.  
3.  
Tolar J, Bonfim C, Grewal S, Orchard P. Engraftment and survival following hematopoietic stem cell 
transplantation for osteopetrosis using a reduced intensity conditioning regimen. Bone Marrow Transplant. 
2006 Dec;38(12):783–7.  
4.  
Schroder K, Hertzog PJ, Ravasi T, Hume DA. Interferon-gamma: an overview of signals, mechanisms and 
functions. J Leukoc Biol. 2004 Feb;75(2):163–89.  
5.  
Garrett IR, Boyce BF, Oreffo RO, Bonewald L, Poser J, Mundy GR. Oxygen-derived free radicals 
stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. J Clin Invest. 1990 
Mar;85(3):632–9.  
6.  
Madyastha PR, Yang S, Ries WL, Key LLJ. IFN-gamma enhances osteoclast generation in cultures of 
peripheral blood from osteopetrotic patients and normalizes superoxide production. J Interferon Cytokine 
Res. 2000 Jul;20(7):645–52.  
7.  
Rodriguiz RM, Key LL, Ries WL. Combination macrophage-colony stimulating factor and interferon-
gamma administration ameliorates the osteopetrotic condition in microphthalmic (mi/mi) mice. Pediatr 
Res. 1993 Apr;33(4 Pt 1):384–9.  
8.  
Key LL, Ries WL, Rodriguiz RM, Hatcher HC. Recombinant human interferon gamma therapy for 
osteopetrosis. J Pediatr. 1992 Jul;121(1):119–24.  
9.  
Key LLJ, Rodriguiz RM, Willi SM, Wright NM, Hatcher HC, Eyre DR, et al. Long-term treatment of 
osteopetrosis with recombinant human interferon gamma. N Engl J Med. 1995 Jun 15;332(24):1594–9.  
10.  Driessen GJ, Gerritsen EJ, Fischer A, Fasth A, Hop WC, Veys P, et al. Long-term outcome of 
haematopoietic stem cell transplantation in autosomal recessive osteopetrosis: an EBMT report. Bone 
Marrow Transplant. 2003 Oct;32(7):657–63.  
11.  Maluish AE, Urba WJ, Longo DL, Overton WR, Coggin D, Crisp ER, et al. The determination of an 
immunologically active dose of interferon-gamma in patients with melanoma. J Clin Oncol. 1988 
Mar;6(3):434–45.  
12.  Meyskens FL, Kopecky KJ, Taylor CW, Noyes RD, Tuthill RJ, Hersh EM, et al. Randomized trial of 
adjuvant human interferon gamma versus observation in high-risk cutaneous melanoma: a Southwest 
Oncology Group study. J Natl Cancer Inst. 1995 Nov 15;87(22):1710–3.  
13.  Dawson R, Condos R, Tse D, Huie ML, Ress S, Tseng C-H, et al. Immunomodulation with recombinant 
interferon-gamma1b in pulmonary tuberculosis. PLoS ONE. 2009;4(9):e6984.  
14.  Muir AJ, Sylvestre PB, Rockey DC. Interferon gamma-1b for the treatment of fibrosis in chronic hepatitis 
C infection. J Viral Hepat. 2006 May;13(5):322–8.  


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15.  Li JT, Yunginger JW, Reed CE, Jaffe HS, Nelson DR, Gleich GJ. Lack of suppression of IgE production 
by recombinant interferon gamma: a controlled trial in patients with allergic rhinitis. J Allergy Clin 
Immunol. 1990 May;85(5):934–40.  
16.  Bitran JD, Green M, Perry M, Hollis DR, Herndon JE. A phase II study of recombinant interferon-gamma 
following combination chemotherapy for patients with extensive small cell lung cancer. CALGB. Am J 
Clin Oncol. 1995 Feb;18(1):67–70.  
17.  Ghanei M, Panahi Y, Mojtahedzadeh M, Khalili ARH, Aslani J. Effect of gamma interferon on lung 
function of mustard gas exposed patients, after 15 years. Pulm Pharmacol Ther. 2006;19(2):148–53.  
18.  Maciejewski J, Selleri C, Anderson S, Young NS. Fas antigen expression on CD34+ human marrow cells 
is induced by interferon gamma and tumor necrosis factor alpha and potentiates cytokine-mediated 
hematopoietic suppression in vitro. Blood. 1995 Jun 1;85(11):3183–90.  
19.  Selleri C, Maciejewski JP, Sato T, Young NS. Interferon-gamma constitutively expressed in the stromal 
microenvironment of human marrow cultures mediates potent hematopoietic inhibition. Blood. 1996 May 
15;87(10):4149–57.  
20.  Wanachiwanawin W, Wiener E, Siripanyaphinyo U, Chinprasertsuk S, Mawas F, Fucharoen S, et al. 
Serum Levels of Tumor Necrosis Factor-alpha, Interleukin-1, and Interferon-gamma in beta-
Thalassemia/HbE and Their Clinical Significance. Journal of Interferon & Cytokine Research. 1999 Feb 
1;19(2):105–11.  
21.  Butthep P, Wisedpanichkij R, Jindadamrongwech S, Fucharoen S. Elevated erythropoietin and cytokines 
levels are related to impaired reticulocyte maturation in thalassemic patients. Blood Cells, Molecules, and 
Diseases. 2015 Feb;54(2):170–6.  
22.  Varni JW, Thompson KL, Hanson V. The Varni/Thompson Pediatric Pain Questionnaire. I. Chronic 
musculoskeletal pain in juvenile rheumatoid arthritis. Pain. 1987 Jan;28(1):27–38.  
23.  Cohen LL, Lemanek K, Blount RL, Dahlquist LM, Lim CS, Palermo TM, et al. Evidence-based 
assessment of pediatric pain. J Pediatr Psychol. 2008 Oct;33(9):939-955-957.  
24.  Thompson KL, Varni JW, Hanson V. Comprehensive assessment of pain in juvenile rheumatoid arthritis: 
an empirical model. J Pediatr Psychol. 1987 Jun;12(2):241–55.  
25.  Burckhardt CS, Jones KD. Adult measures of pain: The McGill Pain Questionnaire (MPQ), Rheumatoid 
Arthritis Pain Scale (RAPS), Short‐Form McGill Pain Questionnaire (SF‐MPQ), Verbal Descriptive Scale 
(VDS), Visual Analog Scale (VAS), and West Haven‐Yale Multidisciplinary Pain Inventory (WHYMPI). 
Arthritis Care & Research. 2003;49.S5:S96–104.  
26.  Hawker GA, Mian S, Kendzerska T, French M. Measures of adult pain: Visual Analog Scale for Pain 
(VAS Pain), Numeric Rating Scale for Pain (NRS Pain), McGill Pain Questionnaire (MPQ), Short-Form 
McGill Pain Questionnaire (SF-MPQ), Chronic Pain Grade Scale (CPGS), Short Form-36 Bodily Pain 
Scale (SF-36 BPS), and Measure of Intermittent and Constant Osteoarthritis Pain (ICOAP). Arthritis Care 
Res (Hoboken). 2011 Nov;63 Suppl 11:S240-252.  


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27.  Devane JG, Martin ML, Matson MA. A short 2 week dose titration regimen reduces the severity of flu-like 
symptoms with initial interferon gamma-1b treatment. Curr Med Res Opin. 2014 Jun;30(6):1179–87.  
28.  Mosteller RD. Simplified calculation of body-surface area. N Engl J Med. 1987 Oct 22;317(17):1098.  
 
 
 


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APPENDIX A:  
Data and Safety Monitoring Plan (Multi-Centered, No DSMB) 
Project No. :  21549-01 
  
Protocol Title:  Open-label Phase 1/2 Study with a Single Arm of Interferon Gamma-
1b Treatment of Osteopetrosis 
 
Principal Investigator:  Lynda Polgreen, MD, MS 
 
Monitor: 
Peter Liu, MD 
 
UCLA CTSI: 
  UCLA-CHS 
  LA Biomed/ Harbor-
UCLA 
  
Cedar-
Sinai 
Medical 
Center 
  Charles 
Drew University 
Version:  1.0 
Version 
Date: 
June 23, 2015 
 
 
1. Categorization of risk level (low, medium, or high) and a discussion of the relevant specific 
considerations. 
 
Risk Level: 
  Low 
  Moderate 
  High 
Justification of Risk Level: 
 
Potential for direct subject benefit.  
 
2. Specific information to be provided at the time of each periodic review and the bases for 
selecting this data set. 
 
Enrollment Data: 
The investigator will prepare a periodic report to include Local target, interval and cumulative data in a 
tabular format as follows:  
 
 
Number of subjects screened 
 
 
Number of subjects enrolled 
 
 
Number of screen failures with reason for screen failure 
 
 
Dropout number with reason for dropout and stage of study at dropout 
 
 
Completion number 
 
 
Other study specific enrollment data such as subset enrolled in each group, 
site, etc.  Specify below: enrollment at each site 
 
For Entire Study: 
 
 
Number of subjects enrolled 
 
 
Completion number 
 
 


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Local Adverse Event Reporting:   
 
 
All study-related adverse events (including loss of confidentiality and 
moderate-severe discomfort with study sample collection) with severity and 
attribution (possibly, probably or definitely related).  
 
 
The rates of routine illnesses (URI, headaches, muscle aches, etc.) and 
traumatic injuries are not expected to be different than usual and will not be 
reported or tracked as unexpected AEs but will be tracked and reported as 
AEs for the DSM Reports. 
Serious Adverse Event and Unexpected Adverse Events: 
 
 
For all study-related serious adverse events (SAEs) and study-related 
unexpected adverse events (UAEs), the clinical summary will also be 
provided in a cumulative fashion. 
 
 
Due to the large number of disease-related SAEs such as  
 
 
fracture, bone pain, osteonecrosis, anemia, neutropenia, thrombocytopenia, 
cranial nerve impingement, and hypocalcemia, 
 
 
these serious adverse events will be tracked and evaluated by treatment 
group at each DSM periodic reporting but will not be reported by expedited 
reporting to the IRB.   
 
 
 
 
Entire Study Serious Adverse Event Reporting:   
 
All study-related serious adverse events (SAEs) and study-related unexpected 
adverse events (UAEs) 
 
 
For all study-related serious adverse events (SAEs) and study-related 
unexpected adverse events (UAEs), the clinical summary will also be 
provided in a cumulative fashion. 
 
The investigator will comply with the Policies for Reporting of Adverse Events to the local 
IRB that requires reporting of adverse events that in the opinion of the investigator may 
represent unanticipated problems involving risks to the other subjects in the research. 
 
 
The investigator will provide a tabulation of the unanticipated problems that 
meet the IRB reporting requirement with their evaluation of each events in the 
DSM Report 
 
 
Monitor’s Review: 
 
 
 
The investigator will provide the report to the Monitor at least 30 day before 
the CTRC due date for DSM Report submission. 
 
 
 
 
 
 


Interferon gamma-1b in Osteopetrosis 
 
 
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The Monitor will review the report and may request additional information 
from the investigator as needed to evaluate the safety and progress of the 
study.  The monitor will submit the periodic report to the PI with the following 
by the due date. 
 
 
 
Narrative summary of information presented. 
 
 
 
Any concerns regarding adverse event rates, recruitment rates, drop-
out rates, study integrity, etc 
 
 
 
Any concerns regarding adverse event rates, recruitment rates, drop-
out rates, study integrity, etc. with statement of how these concerns 
were resolved with the PI. 
 
 
 
Recommendations for future study conduct (continuation, modification 
or termination). 
 
 
The PI will submit the DSM Report with the Monitor’s review to the CTRC by 
the due date via iMedris. 
 
 
 
 
 
3. Frequency of review, and rationale for recommended frequency. 
 
Frequency of Review: 
Every 6 months 
. 
Rationale for Review Frequency: Limited number of subjects in the study (10 will be 
enrolled over 18 months) and the level of moderate 
risk. 
 
 
4. Designation of individual(s) as independent monitor(s) reviewing the information.  An 
explicit review of any conflict of interest issues should be presented. 
 
 
Dr. Jennifer Yee will act as the Monitor for this clinical study.  Dr. Yee is a faculty member at Harbor-
UCLA with an appointment in the Department of Pediatrics, division of Endocrinology.  She has 
expertise in the area of clinical trials and metabolic bone disease.  She has no conflict of interest with 
the local PI or the study sponsor. 
 
 
 
 
 
 


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APPENDIX B 
Schedule of Events (next page) 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Interferon gamma-1b in Osteopetrosis 
 
 
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Schedule of Events 
If ADA Positive 
 
Screeninga 
Baseline 
time from 1st 
IFN-γ1b dose 
time from last  
IFN-γ1b dose 
3 weeks 
(± 2 days) 
4 weeks 
(±2 days) 
6 weeks 
(±2 days) 
3 months 
(± 1 week) 
6 months 
(± 1 week) 
9 months 
(± 1 week) 
12 months 
(± 1 week) 
2-3 
weeks 
every 4-6 
weeks 
General 
Medical history 
X 
X 
 
 
 
X 
X 
 
X 
 
 
Physical exam 
 
X 
 
 
 
X 
X 
 
X 
 
 
Genotypeb 
 
X 
 
 
 
 
 
 
 
 
 
RNA and DNA samples 
 
Xf 
 
 
 
X 
X 
 
X 
 
 
Anthropometric measurements 
X 
Xf 
 
 
 
X 
X 
 
X 
 
 
Safety Measuresc 
Basic metabolic panel (BMP) 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
 
Liver function tests (LFT) 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
 
Urine pregnancy test (UPT) e 
X 
X 
 
 
 
X 
X 
X 
X 
 
 
Urinalysis (UA) 
X 
X 
 
X 
X 
X 
X 
X 
X 
 
 
ECHO with interpretation 
X 
 
 
 
 
 
 
 
 
 
 
EKG with interpretation 
X 
 
 
 
 
 
 
 
 
 
 
Anti-drug antibodies (ADA) 
 
X 
X 
 
X 
X 
X 
 
X 
X 
X 
Bone Biomarker Testingd,g 
Formation Testing (bone specific alkaline 
phosphatase [BAP], type 1 procollagen 
[P1NP]) 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Resorption testing (C-telopeptide [CTX], 
Tartrate resistant acid phosphatase type5b 
[TRAP5b]) 
- 
X 
 
 
 
X 
X 
 
X 
 
 
RANKL, OPG, CK-BB 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Endocrine Assessment 
Total 25-OH Vitamin D (25OHD) 
- 
X 
 
 
X 
X 
X 
 
X 
 
 
Serum ionized calcium 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Urine calcium: creatinine 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Intact parathyroid hormone (iPTH) 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Inflammatory/Immunology 
CBC with differential and platelets 
- 
Xf 
 
 
 
X 
X 
 
X 
 
 
Erythrocyte sedimentation rate (ESR) 
- 
Xf 
 
 
 
X 
X 
 
X 
 
 
Immunologic phenotyping (CD3, 4, 8, 14, 15, 
16, 19, 34, 56)g 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Neuropsychological testing 
Pain assessment 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Quality of life survey 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Orthopedic assessment 
Dual energy X-ray absorptiometry (DXA) 
lumbar spine, total body, total hip and 
femoral neck (adults ≥18yrs only) 
- 
X 
 
 
 
- 
- 
 
X 
 
 
Peripheral Quantitative CT (pQCT) left tibia 
- 
X 
 
 
 
- 
X 
 
X 
 
 
In vitro osteoclast function assays – Mansky Labh 
Gene expression – osteoclast marker genes 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Osteoclast activity – ability to release 
calcium phos from coated plate 
- 
X 
 
 
 
X 
X 
 
X 
 
 
Infectious outcomes 
Number of hospitalizations in prior 3 months 
- 
Xf 
 
 
 
X 
X 
 
X 
 
 
Number of days treated with antibiotics in 
last 3 months 
- 
Xf 
 
 
 
X 
X 
 
X 
 
 
Type of infectious organism 
- 
Xf 
 
 
 
X 
X 
 
X 
 
 
Summary of samples collected per visit 
Serum (red top) 
1 
4 
1 
 
1 
4 
4 
 
4 
1 
1 
Plasma (purple top) 
- 
5 
- 
 
- 
5 
5 
 
5 
- 
- 
Urine (non-sterile) 
1 
1 
- 
 
- 
1 
1 
 
1 
- 
- 
Whole blood  
 
1 
 
 
 
1 
1 
 
1 
 
 
RANKL=Receptor activator of nuclear factor kappa-B ligand; OPG=osteoprotegerin; CK-BB=creatinine kinase isoenzyme BB. 
a Screening visit evaluations will count for baseline evaluations where applicable. 
b Genetic testing for CLCN7, CAII, TCIRG1, OSTM1, and RANKL mutations will be done at the University of Minnesota if not already performed clinically 
c Subjects will be contacted on a weekly basis either by email or telephone to monitor safety during the initial 4 week dose escalation period. 
d Sample collection will be postponed if needed to ensure collection is more than 2 weeks since last fever (>100.4°F [38°C] axillary) or fracture. 
e Post-menarchal females. 
f Height (for subjects <18 years), CBC, and ESR will be obtain from the patient’s medical record when available and number of hospitalizations and infectious data from the patient’s medical record or if not available then the 
patient’s recollection at the baseline visit. 
g Samples batch shipped every 3 months to Troy Lund/Kelly Miettunen at UMN 
h Whole blood shipped overnight to Kim Mansky at UMN