Proposal for CARDIA Ancillary Study

Appendix C. Concept Form Title Page

Study Title PI Name Affiliation E-mail Address Mailing Address Co-Investigators’ Names Proposed Start date Proposed End date Funding Source and Internal funds from your own institution Application Date: Neptune Pilot funding NephCure Kidney International-NEPTUNE Ancillary Studies Grant Program Private sector funding (specify source) Extramural/Intramural Funding If already funded, specify agency and grant number:

If planned submission, specify agency and application date:

Abstract (limit to one paragraph):

Revised August 21, 2015 Part 1. Feasibility 1. Define the patient population for your ancillary study, including inclusion and exclusion requirements.

2. How many adult and pediatric NEPTUNE patients meet your study requirements defined in Question #1 above? We encourage you to use tranSMART or work with a DACC member to assess case counts for your study design. If you need access to TranSMART to review NEPTUNE data, please contact [email protected].

Biopsy Cohort: Biopsy Cohort: cNEPTUNE Adult Pediatric Cohort Number of participants:

3. Summarize your request for existing clinical data, including visits. Please refer to tranSMART or the tranSMART ontology map for data collected at baseline and follow up visits. If you need access to tranSMART to review NEPTUNE data, please contact [email protected].

4. In the table below, please indicate the amount of existing sample, at each visit, you are requesting for your proposed study population (#1 & #2 above). Please work with a DACC member to determine current availability in the NEPTUNE biorepositories.

The Ancillary Studies Policy states that you should indicate the absolute minimum amount to complete your proposed study. Attachment 1 shows original sample collections during the longitudinal study period.

If your proposed project will use samples in addition to NEPTUNE samples (e.g., controls from healthy subjects or people with other diseases), list them separately in the table by adding rows, as needed.

Base 4 8 12 18 24 30 36 42 48 54-60 Visit Sequence Biopsy line Mos. Mos. Mos. Mos. Mos. Mos. Mos. Mos. Mos. Mos. Plasma available Serum available 24-hour Urine available Spot Urine available DNA available RNA available Unstained section available

Please specify required sample storage conditions:

Revised August 21, 2015 5. W ill you need a statistician or data analyst from the DACC for you proposed ancillary study?

The DACC will contact you to set up a time to discuss the details of the YES biostatistical support and will generate a cost estimate.

Please provide a description of the biostatistical expertise that will be available from NO your team for this proposed study.

6. Does your proposed study involve additional study visits, procedures or specimen acquisition beyond the core NEPTUNE protocol?

YES NO GO TO Part 2. Study Design, next page.

6a. Describe the rationale for additional procedures or visits.

6b. Use the table below to specify the proposed visit schedule and procedures (questionnaires, intervention, laboratory sampling and tests). Provide a subject level time estimate of additional study procedures (e.g. self-administered questionnaire x 10 minutes).

Procedure Visit (Time) Visit (Time) Visit (Time)

6c. If the ancillary study requires additional blood or urine beyond that collected in the NEPTUNE protocol please describe in table below:

Volume Specimen (Plasma, Serum, Urine, Sampling Times*: Other - specify) Requested

*Specify NEPTUNE Visit (N) Month 0, 4, 8, 12, etc. or specify Additional (A) Visit Time

Revised August 21, 2015 Part 2. Study Design

3. Design and Methods

Describe: the plan for data acquisition; sample collection and handling; specific assays and experiments; planned quality control measures.

If you are proposing use of limited biospecimens, please provide:

a. A justification for the request of limited and irreplaceable biomaterials. The justification should discuss how the proposed project will take advantage of the depth of larger NEPTUNE clinical data set and it should make the case clearly that the proposed study will have significant scientific impact.

b. Preliminary data to support the proposal’s hypothesis and the technical feasibility of the approach proposed. The ancillary studies committee may consider the use of NEPTUNE biosamples to develop preliminary data in an iterative fashion in some cases. In this case, explain why alternative samples cannot be obtained from other sources.

c. Consideration of the source and condition of requested biospecimens, such as how they were collected, prepared and stored if this might affect the proposed analysis.

d. Assay details and methodology, including: amounts and type of samples (applicants should request the minimum volume required for their assay), anticipated assay variability, quality control (assay "robustness") assessment.

Revised August 21, 2015 Attachment 1. Original Sample Collections during Longitudinal Study

The tables below show the samples and quantities requested at each visit in NEPTUNE study. Please note that actual amounts may vary.

Baseli ne Blood

Sampl Seru EDT es by m SST Na EDT A- Follow-up Blood Na EDTA Typea Na Cit Epib Epib Hepb A iced Samples by Typea Serum Epib SST S Hepb EDTA -iced Adult: S Baseli ne Appro x Volum Adult: Follow-up e (mL) 3.2 3.6 3.6 2.4 27.2 4.8 Approx Volume (mL) 1.2 4.8 1 1.2 17.6 4.8 Peds > . 52 lbs: Baseli ne Appro x Volum Peds > 52 lbs: FU e (mL) 3.2 3.6 3.6 2.4 22.4 3.2 Approx Volume (mL) 3.6 4.8 3 2.4 9.6 3.2 Peds . 21-51 lbs: Baslin e Appro x Volum Peds 21-51 lbs: FU e (mL) 3.2 3.6 3.6 2.4 8 0 Approx Volume (mL) 3.6 0 3 2.4 8 0 Peds < . 21 lbs: Baseli ne Appro x Volum Peds < 21 lbs: FU e (mL) 0 1.2 1.2 1.2 4.8 0 Approx Volume (mL) 1.2 0 1 1.2 4.8 0 b Epi indicates samples procured for light sensitive product, stored in light sensitive eppendorf tubes . c Volume variable

5 mL RNA-L PI – Urine 24h & Spot Baseline & 50mL aliquot Whole NaN3 - # PI - # # Follow-up Falcon s: urined # SN: pellet SN pellet:

Revised August 21, 2015 Whole urine samples Per Participant 2 5 Clean catch spot Per Participant 4 4 2 4 2 Total Volume 90 20 6.4 6.4 - 0 - d Whole urine spot samples collected only if >60 mL sample obtained

Columns specified with a specimen type (Na Cit, 50 mL Falcon, etc) indicate the amount accounted for in the Baseline Biospecimens CRF by research coordinators, collectively.

Abbrev Sample Type Na Cit Sodium Citrate Serum Epi Serum, light sensitive SST SST SST Epi SST, light sensitive Na Hep Sodium Heparin EDTA Ethylenediaminetetraacetic Acid 50 mL Falcon 24 hour urine, 1-50mL unprocessed whole urine, 1-50 mL whole urine with protease inhibitor cocktail 5 mL aliquots 24 hour urine, 5-5mL unprocessed whole urine Whole urine Spot sample, 2 mL unprocessed whole urine NaN3 # SN Spot sample, 2 mL spun supernatant with sodium azide RNA-L # Pellet Spot sample, 2 mL spun pellet stored in RNA-Later PI - # SN Spot sample, 2 mL spun supernatant with protease inhibitor cocktail PI - # Pellet Spot sample, 2 mL spun pellet, treated with protease inhibitor cocktail, stored in RNA-Later

Revised August 21, 2015