Clinical trial NCT02317276
Conditions: Dermatitis, Atopic
Sponsor: University of California, San Francisco
Start date: 2014-12-01; Completion date: 2017-04-06
Enrollment: 11 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
The purpose of this study is to assess the differential expression of AD biomarkers in serum, plasma, and skin biopsies from both lesional and non-lesional skin in moderate to severe AD patients in the presence of TCS and after withdrawal from TCS.

Detailed description:
This exploratory study consists of four visits to the investigator site post screening: at Weeks 0 (baseline), 2, 6 and 8 (Figure 1). At the first visit (baseline), patients who have met the screening eligibility criteria will be started on a stable TCS regimen for a total of two weeks. At the second visit (Week 2) following two weeks of therapy on stable TCS, patients will stop TCS and blood, serum, plasma and two 6mm punch biopsies (one lesional (L) and one non-lesional (NL) skin) will be obtained. At the third visit (Week 6), after four weeks receiving no TCS, the same sample collections will be repeated and the patients will then enter a two week safety follow up. At the end of the safety follow up (last visit, Week 8) and after obtaining written informed consent by the patient, blood, serum and plasma samples mav be collected.

Arms: arm 1: Topical Triamcinolone 0.1% ointment will be provided for twice daily application, during treatment periods.
Interventions: intervention 1: Triamcinolone 0.1%
Intervention descriptions: intervention 1: Topical ointment

Protocol documents:
Prot_SAP_000.pdf: 
 
1
 
Protocol Title: 
 
 
NCT #: 
 
A 
STUDY 
IN 
ATOPIC 
DERMATITIS 
TO 
DETERMINE 
SERUM 
AND 
SKIN 
BIOPSY 
BIOMARKERS 
IN 
PATIENTS 
RECEIVING 
TOPICAL 
CORTICOSTEROIDS 
(TCS) 
AND 
FOLLOWING TCS WITHDRAWAL  
 
NCT02317276 
 
Investigator: 
Sarah Tuttleton Arron, MD, PhD 
Assistant Professor of Dermatology in Residence 
Chief of Mohs Micrographic Surgery, SF VA Medical 
Center  
Director, High Risk Skin Cancer Program 
Director, Dermatology Clinical Research Unit 
UCSF Dermatologic Surgery and Laser Center 
1701 Divisadero Street, Third Floor, Box 316 
San Francisco, CA 94115-0316 
arrons@derm.ucsf.edu 
tel (appointments)415-353-7878 
fax 415-353-7838 
 
Arron Lab 
http://www.dermatology.ucsf.edu/arronlab 
2340 Sutter Street, N431 
San Francisco, CA 94115 
tel 415-514-9769 
 
Study Site: 
UCSF Dermatologic Surgery and Laser Center 
Protocol Version 
Number: 
April 5, 2016 
Sponsor: 
Genentech, Inc. 
Short Title: 
Biosample Collections in Atopic Dermatitis 
 
 
This study will be conducted according to the principles of Good Clinical Practice (GCP), 
including the archiving of essential documents. 


 
2
 
CONTENTS 
INTRODUCTION .................................................................................................. 3 
Study Objective.......................................................................................... 4 
INVESTIGATIONAL PLAN ................................................................................... 4 
Study Design ............................................................................................. 4 
RATIONALE FOR BIOMARKER ASSESSMENTS .............................................. 5 
SAMPLE SIZE ...................................................................................................... 5 
STUDY VISITS ..................................................................................................... 6 
Visit 1 (Baseline – Week 0) ........................................................................ 6 
Visit 2 (Week 2): Patients receiving TCS ................................................... 7 
Visit 3 (Week 6): Patients receiving no TCS .............................................. 7 
Visit 4 (Week 8): End of Study ................................................................... 7 
Study Population...................................................................................... 11 
Inclusion Criteria : ......................................................................... 11 
Exclusion Criteria : ........................................................................ 11 
Study Procedures ............................................................................................... 12 
Patient Treatment Regimen ..................................................................... 12 
Safety  …………………………………………………………………………. 13 
Whole Blood Collection ............................................................................ 14 
Skin Biopsy .............................................................................................. 14 
Length of Study ....................................................................................... 16 
End of Study ............................................................................................ 16 
Ethics 16 
Research Ethics Board ............................................................................ 16 
Ethical Conduct of the Study ................................................................... 16 
Patient Information and Consent ............................................................. 16 
Investigators and Study Administrative Structure ............................................... 17 
References ......................................................................................................... 17 
APPENDIX 1 ...................................................................................................... 18 
 


 
3
INTRODUCTION 
Atopic Dermatitis (AD) is a chronic relapsing and remitting inflammatory skin disorder 
affecting all age groups.  Clinically AD is characterized by xerosis, erythematous crusting 
rash, lichenification, an impaired skin barrier and intense pruritus (Bieber 2008).  
AD is one of the most common dermatologic diseases and the prevalence has increased 
over the past two to three decades; 15% to 30% of children and 2% to 10% of adults are 
affected (Williams et al. 2006).  Overall, the disease burden is high and quality of life 
(QoL) in AD patients is severely impacted, even when compared to other skin diseases 
such as psoriasis (Keibert et al. 2002).   
 
The therapeutic approach to AD primarily consists of trigger avoidance, skin hydration 
with bathing and use of emollients and anti-inflammatory therapies consisting 
predominantly of topical corticosteroids (TCS). For patients with moderate to severe 
disease, a step up in therapy is required, including an increase in the potency and 
frequency of TCS.  However, long term (months) use of TCS, particularly high potency 
TCS, is not recommended due to the increased incidence of both local and systemic side 
effects (Simpson EL 2010).  In cases where high doses (potency and frequency of use) of 
TCS are unable to control disease, topical calcineurin inhibitors (TCIs) may be used to 
control disease and/or allow a reduction in TCS potency particularly at high-risk sites.  
Recent guidelines provide a number of options for patients not responding to TCS and/or 
TCI depending on the clinical assessment of disease activity (Ring et al. 2012; Schneider 
et al. 2013), that include, phototherapy and immunosuppressive agents such as oral 
corticosteroids, cyclosporine, azathioprine, methotrexate and mycophenolate.  Although 
these medications may be effective, significant toxicities limit their long term use.  The 
continued use of these agents despite the harmful side effects and limitations speaks to 
the high unmet medical need for safer and more effective therapies. 
 
Therapy targeted at central pathologic mediators has promise as a means to maximize 
efficacy and limit toxicity.  The high expression of IL-13 in AD skin (Gittler 2012, Choy 
2012) indicates that lebrikizumab, an anti-IL-13 monoclonal antibody, may represent such 
a targeted therapy.  Genentech plans to perform a Phase II study to assess the efficacy 
and safety of lebrikizumab in patients who have persistent moderate to severe AD that is 
inadequately controlled by TCS.  The design of this study includes a run-in periods on 
daily prescribed TCS for 2 weeks, followed by randomization to placebo and treatment 
arms remaining either on or off background TCS, respectively.  Assessment of pre-
determined biomarker candidates have been incorporated into the planned Phase II study 
design to characterize the mechanism of action (MOA) and treatment effect of 
lebrikizumab on biomarkers of AD disease.  However, the pre-determined biomarkers 
were identified from studies of AD patients who had been withdrawn from TCS weeks 
prior to providing samples, and it remains to be determined how well these markers are 
representative of AD disease on a background of TCS.  Therefore, the purpose of this 
proposed, interventional study will be to collect serum and lesional and non-lesional skin 
from moderate to severe AD patients both on, and after withdrawal from a stable TCS 
regimen in order to provide samples that can answer this outstanding question. 
 


 
4
Study Objective 
To assess the differential expression of AD biomarkers in serum, plasma, and skin 
biopsies from both lesional and non-lesional skin in moderate to severe AD patients in the 
presence of TCS and after withdrawal from TCS.  
 
INVESTIGATIONAL PLAN 
 
Study Design  
This exploratory study consists of four visits to the investigator site post screening: at 
Weeks 0 (baseline), 2, 6 and 8 (Figure 1).  At the first visit (baseline), patients who have 
met the screening eligibility criteria will be started on a stable TCS regimen for a total of 
two weeks.  At the second visit (Week 2) following two weeks of therapy on stable TCS, 
patients will stop TCS and blood, serum, plasma and two 6mm punch biopsies (one 
lesional (L) and one non-lesional (NL) skin) will be obtained.  At the third visit (Week 6), 
after four weeks receiving no TCS, the same sample collections will be repeated and the 
patients will then enter a two week safety follow up. At the end of the safety follow up (last 
visit, Week 8) and after obtaining written informed consent by the patient, blood, serum 
and plasma samples mav be collected. 
 
 
Figure 1. Study Design 
 
 
 
 
 
 


 
5
RATIONALE FOR BIOMARKER ASSESSMENTS 
The sensitivity of IL-13/atopic dermatitis–related biomarkers to topical corticosteroid is 
unknown.  This study is intended to increase this understanding and will ultimately inform 
selection of steroid-resistant biomarkers that can be evaluated in subsequent 
interventional trials where Lebrikizumab is provided as an adjunctive therapy to topical 
corticosteroid.   
To that end, blood and skin biomarkers, including but not limited to eosinophils, IgE, IL-
13, CCL-13 and CCL-17, will be evaluated for differential gene expression and protein 
levels in samples obtained from atopic dermatitis patients on and off topical corticosteroid 
treatment.  The skin evaluations will include Hematoxylin & Eosin (H&E) staining to 
evaluate the pathologic skin phenotype, and immunohistochemistry (IHC) for biomarkers 
of dermatopathologies including but not limited to epidermal hyperplasia (eg. KI16, Ki67), 
epidermal differentiation (eg. Filaggrin), and inflammatory cell infiltration (eg. eosinophils). 
Additionally, differential gene expression analysis of lesional and non-lesional skin 
collected from patients on and off topical corticosteroid will be evaluated to measure 
steroid responsive genes, including but not limited to FKBP51.   
Whole blood for genomic analyses of predisposing factors, including, but not limited to, 
the two common filaggrin mutations, R501X and 2282de14 will also be determined. 
SAMPLE SIZE 
Assuming the proportion of non-differentially expressed genes is above 0.997 and that 
the gene expression variances are following an Inverse Gamma(3, 0.3) distribution (as 
observed in Gittler et al, 2012 for non-differentially expressed genes between acute and 
chronic AD), approximately 10 patients would be needed to detect a two-fold change in 
the gene expression levels (corresponding to a difference in log2 expressions of one) 
between the presence of TCS and after withdrawal of TCS, while controlling the false 
discovery rate (FDR) at 5%. 
Figure 1 shows the average power versus sample size for different proportions of non-
differentially expressed genes 


 
6
 
Figure 1: Sample size vs. power for one-sample two-sided t-test with effect size of one 
and various proportions of non-differentially expressed genes with FDR controlled at 5%. 
Variances are assumed to follow an Inverse Gamma(3,0.3) distribution. 
 
Furthermore, the sample size of 10 patients provides over 80% power to detect 
differences between the two regimens (TCS and no TCS) for epidermal thickness and 
epidermal hyperplasia as measured by Ki67.  Differences are either 70 microns in 
epidermal thickness assuming a standard deviation of 50 microns and a two-sided type I 
error rate of 0.05, or 40 Ki67 cells/mm assuming a standard deviation of 30 Ki67 cells/mm 
and a two-sided type I error rate of 0.05, respectively.  
 
STUDY VISITS  
See schedule of assessments for details 
Visit 1 (Baseline – Week 0)  
− Written informed consent will be obtained for those patients meeting eligibility 
criteria. 
− Medical history and concomitant medications will be recorded. 
− Skin severity scores (including EASI and IGA) will be assessed. 


 
7
− Protocol prescribed TCS treatment (Triamcinolone 0.1%, twice daily to lesional 
skin) and daily emollient use will be initiated. 
 
Visit 2 (Week 2): Patients receiving TCS 
− Skin severity scores (including EASI and IGA) will be assessed. 
− Total TCS use will be recorded at each visit by measuring tube weights (g) and 
questioning of the patients to primarily assess treatment compliance. 
− Concomitant medications will be recorded. 
− Blood, serum and plasma samples will be obtained (Table A). 
− UCSF labs will perform CBC and differential. 
− Skin biopsies will be collected from one lesional and one non-lesional site, as 
determined by the site investigator (Table A). 
− Photographs will be taken to record the lesional and non-lesional biopsy sites. 
− Protocol prescribed TCS treatment will be stopped for a subsequent period of 4 
weeks. Patients will continue use of daily emollient 
 
Visit 3 (Week 6): Patients receiving no TCS 
− Skin severity scores (including EASI and IGA) will be assessed. 
− Use of TCS over the prior 4 weeks of the study and other concomitant medications 
will be recorded  
− Blood, serum and plasma samples will be obtained (Table A). 
− UCSF labs will perform CBC and differential. 
− One lesional and one non-lesional skin biopsy will be obtained adjacent to those 
previously collected on Visit 2, as determined by the site investigator (Table A). 
− Photographs will be taken to document the biopsy sites. 
− Patients will enter a two week period of safety follow up during which time protocol 
prescribed TCS treatment will be re-initiated. Patients will use daily emollient. 
 
Visit 4 (Week 8): End of Study   
− Skin severity scores (including EASI and IGA) will be assessed. 
− Use of TCS and other concomitant medications will be recorded  
− Optional blood, serum and plasma samples will be obtained from those patients 
who provide additional consent for these sample collections (Table A). 
 


 
8
Table A – Time and Events Table 
 
 
Mandatory 
Samples 
Visit 2 (week 2) 
Prepare as 
Aliquots 
Labelled as 
Final 
Volume/tube 
Tube Type 
10 ml Whole blood 
EDTA  
1 
CBC 
3.5ml  
Lavendar top 
EDTA  
1 
DNA Collection 
6ml  
Lavendar top 
10 ml Blood  (1 
collection tube) =>      
5 ml Serum 
Serum  
1 
Periostin –  
Split 1 
0.5ml 
Sarstedt FBT 
Serum 
4 
Serum PD – 
Splits 2-5 
1ml 
Polypropylene Tube 
6 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-EDTA 
2 
EDTA Plasma PD –  
Splits 1 through 2 
1ml 
Polypropylene Tube 
4 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-Li 
Heparin 
1 
Li Heparin Plasma 
PD –Split-1 
1ml 
Polypropylene Tube 
6mm punch biopsy 
of lesional skin 
Bisect 
1 
Skin for RNA 
3mm 
RNA Later 
1 
Skin for 
dermatopath 
3mm 
Formalin fixed 
6mm punch biopsy 
of non-lesional skin 
Bisect 
1 
 Skin for RNA 
3mm 
RNA Later 
1 
 Skin for 
dermatopath 
3mm 
Formalin fixed 
 


 
9
 
Mandatory 
Samples 
Visit 3 (week 6) 
Prepare as 
Aliquots 
Labelled as 
Final 
Volume/tube 
Tube Type 
4 ml Whole blood 
EDTA  
1 
CBC 
3.5ml  
Lavendar top 
10 ml Blood  (1 
collection tube) =>      
5 ml Serum 
Serum  
1 
Periostin – 
Split 1 
0.5ml 
Sarstedt FBT 
Serum 
4 
Serum PD – 
Splits 2-5 
1ml 
Polypropylene Tube 
6 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-EDTA 
2 
EDTA Plasma PD –  
Splits 1 through 2 
1ml 
Polypropylene Tube 
4 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-Li 
Heparin 
1 
Li Heparin Plasma 
PD –Split-1 
1ml 
Polypropylene Tube 
6mm punch biopsy 
of lesional skin 
Bisect 
1 
Skin for RNA 
3mm 
RNA later 
1 
Skin for 
dermatopath 
3mm 
Formalin fixed 
6mm punch biopsy 
of non-lesional skin 
Bisect 
1 
 Skin for RNA 
3mm 
RNA later 
1 
 Skin for 
dermatopath 
3mm 
Formalin fixed 
 


 
10 
 
Optional 
Samples 
Visit 4 (week 8) 
Prepare as 
Aliquots 
Labelled as 
Final 
Volume/tube 
Tube Type 
4 ml Whole blood 
EDTA  
1 
CBC 
3.5ml  
Lavendar top 
10 ml Blood  (1 
collection tube) =>      
5 ml Serum 
Serum  
2 
Periostin –  
Splits 1 and 2 
0.5ml 
Sarstedt FBT 
Serum 
4 
Serum PD –  
Splits 3-6 
1ml 
Polypropylene Tube 
6 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-EDTA 
2 
EDTA Plasma PD-
Splits 1 through 2 
1ml 
Polypropylene Tube 
4 ml Blood (1 
collection tube) => 
>2ml Plasma 
Plasma-Li 
Heparin 
1 
Li Heparin Plasma 
PD -Split-1 
1ml 
Polypropylene Tube 
 
 
Blood volumes collected per patient per visit:  
Visit 2 (week 2) ~30ml 
Visit 3 (week 6) ~24ml 
Visit 4 (week 8) ~24ml (optional) 
 
A patient may donate ~80ml blood during the trial. 
 


 
11 
 Study Population 
15 subjects with moderate to severe AD are planned to be enrolled in this study which, 
based on a predicted 30% drop-out rate, is considered to be adequate so that at least 10 
patients complete the study.  
 
Study Eligibility 
 
Subjects who meet all of the inclusion criteria and none of the exclusion criteria at the 
time of screening will be eligible for enrollment in the study.  
 
 
Inclusion Criteria : 
The following inclusion criteria must be met for study entry: 
 
1. Male or female patients 18 − 75 years of age  
2. AD diagnosed by the Rajka/Hanifin criteria at the time of screening and that has 
been present for at least 1 year  
3. At least one area of moderate to severe active AD amenable to study biopsy and 
TCS treatment. 
4. A washout period prior to screening for those patients who have previously received 
the following medications: 
a) Cyclosporine/Oral Steroids/Imuran/Mycophenolate Mofetil/Other systemic 
immunosuppressants: 4 weeks 
b) Phototherapy: 4 weeks 
c) Biologics: 5 half lives of the drug 
 
Exclusion Criteria : 
Patients who meet any of the following criteria will be excluded from study entry: 
 
1. Evidence of other concomitant skin conditions (e.g., psoriasis or contact 
dermatitis) 
2. Use of topical calcineurin inhibitors within 4 weeks of screening 
3. Hypersensitivity to TCS or to any other ingredients contained by the emollient or 
TCS product used during the study  
4. Evidence of active skin infection at screening or baseline visit 
5. Evidence or history of active or latent infections such as tuberculosis or hepatitis C 
6. Patient clinical condition is not appropriate for treatment with protocol prescribed 
TCS 
7. Use of an investigational agent within 4 weeks prior to screening or within 5 half-
lives of the investigational agent, whichever is longer  
8. Use of a tanning booth/parlor within 4 weeks before the baseline visit 
9.  Use of any anti-histamine medication within 4 weeks before the baseline visit. 


 
12 
10. History of any condition (e.g. bleeding diathesis) that may predispose the patient 
to complications associated with the planned skin biopsy procedures  
11. Known current malignancy or current evaluation for a potential malignancy, 
including basal or squamous cell carcinoma of the skin or carcinoma in situ 
12. Other clinically significant medical disease that is uncontrolled despite treatment 
that is likely, in the opinion of the investigator, to impact the patient's ability to 
participate in the study or to impact the study pharmacodynamic (PD), or safety 
assessments 
13. Unwillingness or inability to comply with the study protocol for any other reason. 
 
 
Rules for withdrawal from the Study in Individual Participants: 
• The investigator feels study withdrawal is in the patient’s best interest 
• Withdrawal of consent by patient (or representative) 
• The need for prohibited therapies, including: 
• Phototherapy  
• Systemic agents including oral corticosteroids, cyclosporine or other systemic 
immunosuppressants 
• TCS with higher potency than protocol-prescribed, for more than one day in any 
given week of the study (baseline to Week4) 
• Skin infection 
• Any other serious infections 
• Inadequate compliance to the study protocol 
Patients withdrawn from the study will no longer receive study drug (Triamcinolone 
0.1%) but will continue in the study for safety assessments until completion. 
 
STUDY PROCEDURES 
Patient Treatment Regimen 
1) Baseline (Week 0) until 2nd visit (Week 2):  All patients will be treated with a stable 
topical treatment regimen. Patients must demonstrate compliance with this baseline 
therapy, as determined by the investigator’s interview of the patient and measuring TCS 
tube weights at Visit 2 (Week 2).  The regimen will consist of: 
i) Emollients:  
• Applied to all xerotic skin daily  
• Only basic bland emollient which may include 
o 
Vaseline (Unilever, London, UK); moisturizers such as 
o 
Aquaphor (Beiersdorf AG, Hamburg, Germany), 
o 
Plastibase (E.R. Squibb & Sons Inc., New Brunswick, NJ, USA) 
o 
Cetaphil cream (Galderma Laboratories, SA, Lausanne, Switzerland)  
o 
CeraVe cream (Coria Laboratories Ltd, Fort Worth, TX, USA) 


 
13 
 
ii) Triamcinolone 0.1%:  
•  To be applied twice daily to all lesional skin; for 2 weeks (Week 0 to Week 2)  
 
2) Visit 2 (Week 2): Triamcinolone will be stopped and for the following 4 weeks patients 
will only apply emollient to xerotic skin daily. 
 
3) Visit 3 (Week 6): Patients will resume the protocol prescribed TCS (Triamcinolone 
0.1%, applied twice daily to lesional skin) treatment regimen for the duration of the two 
week safety follow up i.e. end of the study. 
 
4) Visit 4 (End of Study, Week 8): Final safety assessments will be performed and 
optional blood, serum and plasma samples will be obtained from those patients who 
provide additional consent for these sample collections. 
 
Unscheduled Visits 
Patients who experience significant worsening of AD between visits 2 and 3 will return to 
the study site for evaluation of their clinical condition. At this time the site Investigator will 
determine if it is safe and in the patient’s best interests to continue off TCS.  If not, it is at 
the Investigator's discretion to perform Visit 3 assessments at the unscheduled visit and 
restart TCS.  In this case the End of Study visit 4 would occur two weeks after the 
unscheduled visit. 
 
Safety 
Safety will be assessed in all patients who are entered into the study and until study 
completion (Week 8) or early termination. 
Safety assessment will be performed at each study visit (Baseline, Wk2, Wk6 & Wk8) and 
patients will be provided a study site telephone number to contact the Investigator at any 
time to report any adverse events or to discuss any questions or concerns.  
Safety will be assessed by non-directive questioning and symptom directed physical 
examination and all identified adverse events (AEs) will be recorded by the study site. 
Other evaluations, including clinical laboratory tests may be performed at the discretion of 
the study site investigator. 
After informed consent and initiation of TCS, all AEs will be reported until study 
completion (Week 8) or early termination.   Verbatim descriptions of adverse events will 
be coded using appropriate MedDRA terms. If collected, clinical laboratory data, which 
may include serum chemistry, hematology evaluations, or skin cultures will be 
summarized and analyzed by descriptive statistics. If collected, descriptive summaries of 
ECG findings will be summarized. 
During the study the following specific safety monitoring for skin infection will be 
performed: 
 


 
14 
• If, in the opinion of the site investigator, a skin infection is present, a culture of the 
affected area(s) should be performed to identify the microbe and exclude fungi or 
viruses, including eczema herpeticum. Treatment of infections with antimicrobial 
medications will be at the Investigator’s discretion and as per local Institutional 
practice.  Therapy will be modified as appropriate based on the results of microbial 
culture results.  
• If the patient’s infection does not demonstrate improvement following antibiotic 
therapy, the patient will stop study drug but continue to participate in the study until 
completion of the study.  
 
The safety outcome measures for this study are frequency and severity of treatment-
emergent adverse events  
 
Whole Blood Collection 
Blood collection timepoints will be at week 2 and week 6 with an additional blood 
collection at week 8 from patients who give specific consent to participate in this optional 
collection.  
 
At visits where blood will be collected, venipuncture will be performed using aseptic 
techniques and universal precautions, and drawn into vacutainers.  
 
Whole blood will be collected at baseline for genotyping that includes but is not restricted 
to filaggrin status as described in Table A. 
 
Whole blood will be collected for complete blood counts (CBC) as described in Table A. 
 
Whole blood will be collected for serum as described in Table A. 
 
Additionally, plasma will be prepared from a final blood collection as follows.   
• 6mL of whole blood will be collected for EDTA plasma.   
• 4mL of whole blood will be collected for Li plasma as described in Table A. 
 
Topical analgesia will be offered for all venous puncture procedures, according to local 
guidelines and practices. 
 
Skin Biopsy  
The skin biopsies will be performed as described in Appendix 1. 
Each 6mm punch biopsy tissue will be divided in two with one half placed into RNALater 
for PCR (as described in Table A) and the other half formalin fixed and paraffin 
embedded for subsequent dermatopathology evaluation. 
 
 
Responsibilities for collected samples: 


 
15 
 
UCSF Responsibilities: 
 
1. Dr. Arron will be responsible for: 
• Conducting the study according to the final study protocol as summarized in 
Figure 1 and according to the principles of Good Clinical Practice (GCP). 
• Providing the Research Ethics Board (REB) with all appropriate material, 
including the informed consent document.   
• Obtaining REB approval for the proposed studies.   
• Providing patient clinical data associated with these samples and a current 
sample inventory log within the clinical manifest. 
• Providing the Biologic Material, defined as follows: blood, serum, plasma 
and skin samples from study participants as outlined in Table A. 
• Perform CBC and differential through UCSF clinical labs (Table A). 
• Storing Biological Material in alarm-monitored <-70C freezer within 1 hour of 
collection and until shipment to sponsor. 
• Shipping Biological Material with each patient visit according to Sponsor’s 
specifications. 
2. Unless specifically outlined in the study-specific guidelines and plans, Dr. Arron will 
perform all tasks and activities in accordance with UCSF standard operating 
procedures (SOPs).  
• Dr. Arron will provide Genentech with copies of all UCSF SOPs utilized in 
the execution of this project.  
 
 
Genentech Responsibilities: 
1. Provide labels, collection tubes/containers and materials described in Table A and 
Appendix 1. 
2. Receive and process all shipped biosamples in accordance with all relevant 
Genentech SOPs. 
3. Genentech will receive requested biosamples for evaluation of prospective disease 
biomarkers intended for future interventional trials in atopic dermatitis patients on a 
background of TCS.   
4. The samples may be evaluated by a third party selected by Genentech and may 
be used for further exploratory analyses as deemed necessary by Genentech. 
5. Any leftover blood samples may be used by Genentech and its representatives, 
affiliates, and collaborators for additional biomarker research and/or the 
development of diagnostic tests that help with the detection or understanding of 
medical conditions. 
 
 
 


 
16 
Length of Study 
This study will be for a total of 8 weeks, composed of 3 time periods: TCS treatment (2 
weeks), no TCS treatment (4 weeks), and safety follow-up with protocol prescribed TCS 
treatment ( 2 weeks). 
End of Study 
The end of this study is defined as the date when the last patient, last visit (LPLV) occurs 
or safety follow-up is received from the last patient, whichever occurs later.  
 
ETHICS 
Research Ethics Board 
The Principal Investigator (PI) will provide the Research Ethics Board (REB) with all 
appropriate material, including the informed consent document.  The study will not be 
started until appropriate REB approval of the protocol, the informed consent (IC) 
document, and all recruiting materials is obtained in writing by the investigator and 
Sponsor.  Appropriate reports on the progress of the study will be made to the REB and 
Sponsor by the Principal Investigator in accordance with applicable governmental 
regulations and in agreement with the policy established by Sponsor. 
 
Ethical Conduct of the Study 
This study will be conducted in accordance with the protocol, Good Clinical Practice 
(GCP) according to the International Conference on Harmonization (ICH) guidelines, and 
regulatory requirements for the participating institution.  The study will be conducted 
under a protocol reviewed by an REB; the study will be conducted by scientifically and 
medically qualified persons; the benefits of the study are in proportion to the risks; the 
rights and welfare of the patients will be respected; the physicians conducting the study 
do not find the hazards outweigh the potential benefits; and each patient will give his or 
her informed consent without duress. 
 
Patient Information and Consent 
A properly executed, written, informed consent form, in compliance with GCP according 
to ICH guidelines will be submitted by the investigator to the REB for review and approval 
prior to the start of the study.  Before entering patients into the study, a copy of the REB-
approved IC will be reviewed with the potential participant, and signed and dated.  The 
investigator will provide a copy of the signed informed consent form to each patient and 
will maintain a copy in the patient’s study file.   
 
 


 
17 
INVESTIGATORS AND STUDY ADMINISTRATIVE STRUCTURE 
The study will be conducted by Dr. Sarah Tuttleton Arron, MD, PhD.  The medical 
procedures will be conducted or supervised by Dr Sarah Tuttleton Arron, MD, PhD. 
 
 
REFERENCES 
1. Bieber 2008 
2. Williams et al. 2006 
3. Simpson EL 2010 
4. Ring et al. 2012 
5. Schneider et al. 2013 
6. Gittler 2012  
7. Choy 2012 
 
 
 


 
18 
APPENDIX 1 
Skin Biopsy Procedures 
 
 
At intervals stated in the protocol 6 mm skin punch biopsies will be obtained from lesional 
and non­lesional skin. It is important to carefully follow the steps outlined below.  
 
A. MATERIALS - Provided in bulk by Genentech 
 
Methanol 
Scalpel 
Suture kit 
6 mm punch 
Labels 
- 
50 ml conical tube 
Formalin filled collection jars 
- 
2x2 Gauze Pads 
RNAlater filled collection tubes 
- 
 
 
 
B. BIOPSY SITE SELECTION 
 
Biopsies should be performed by the Investigator as per the site’s standard practices and 
following the instructions described below.   
 
1.   First biopsies – week 4 (visit 2): 
a.  Lesional skin 
i.  The 6 mm punch biopsy should be taken from the most involved chronic 
active erythematous (scaly) lesions. 
 
ii.  Biopsies from the folds of the upper extremities, followed by lower 
extremities are preferred, followed by the trunk, in that order 
(although investigators should have flexibility). 
iii.  Each biopsy should be divided in two using a scalpel, with one half placed 
into labeled collection jars containing formalin, and the other half prepared 
in RNAlater as described below. 
 
 
b.  Non-lesional skin 
i.  The 6 mm non-lesional sample should be collected from the most normal 
appearing skin in a relative proximity to the lesional skin biopsy site, at 
least 5 cm away from the lesion (at least 1 cm away, if 5 cm is not 
possible). 
ii.  Each biopsy should be divided in two using a scalpel, with one half placed 
into labeled collection jars containing formalin, and the other half prepared 
in RNAlater as described below. 
 
 


 
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c.  Photograph the site from which the biopsies will be collected to document the 
site of biopsy, identify biopsy sites at the time of subsequent visits and as reference 
to assess any biopsy related complications. 
 
2.  Second biopsies - week 6 (visit 3): 
a.   Lesional skin 
i.  The week 8 "lesional" biopsy should be taken from the same area location as 
the week 4 lesional sample.  If the lesion has cleared, collect the sample 
close to the previous biopsy scar (>5mm but not further than 1cm from the 
biopsy scar). Do not biopsy the previous biopsy scar. 
ii.  Each biopsy should be divided in two using a scalpel, with one half placed into 
labeled collection jars containing formalin, and the other half prepared in 
RNAlater as described below. 
 
b.   Non-lesional skin 
i.  The week 8 non-lesional sample should be taken close to the area of the 
previous biopsy scar of the healthy appearing nonlesional skin biopsy (>5mm 
but not further than 1cm from the biopsy scar).  Do not biopsy the previous 
biopsy scar. 
ii.  Each biopsy should be divided in two using a scalpel, with one half placed into 
labeled collection jars containing formalin, and the other half prepared in 
RNAlater as described below. 
 
c.  Photograph the site from which the biopsies will be collected to document the 
site of biopsy, identify biopsy sites at the time of subsequent visits and as reference 
to assess any biopsy related complications. 
 
 
C. FREEZING THE BIOPSY SPECIMEN IN RNALATER 
 
 
1)  DO NOT leave the tissue at room temperature or on wet ice for an extended period of time. 
 
2)   Place the remaining half of the biopsy specimen in the tube of RNALater solution at 4°C. 
It is important that the sample be completely immersed in the solution. 
 
3}  Store overnight at 4°C, then move samples to  -70°/-80°C until shipment.  Samples must be 
frozen before shipment. 
 
 
D. SHIPMENT OF BIOPSY SPECIMENS 
 
 
Ship samples according to sponsor’s specifications based on sample type.