Attachment III NCC Task 2 SOW FINAL.pdf

TASK ORDER 2 STATEMENT OF WORK

NIEHS DIVISION OF THE NATIONAL TOXICOLOGY PROGRAM

HUMAN NEURAL CREST CELL MIGRATION ASSAY DATA ANALYSIS AND REPORTING (TASK 2)

1.0. INTRODUCTION: Under this task order, the Contractor shall analyze data generated in Task 1 from the human neural crest cell (NCC) migration assay . All services, labor, equipment, and supplies provided under this task order shall be performed under accordance with the terms and conditions of the contract Statement of Work (SOW) and task order SOW.

2.0. TASK: The contractor shall evaluate the neurodevelopmental effects of 115 environmental compounds using two (2) neurodevelopmental endpoints in the NCC: 1) NCC migration; 2) Viability of migrating NCCs. In Task 2 the Contractor shall analyze and report the data generated from Task 1.

2.1. NCC migration assay data analysis and reporting: NCC that have migrated into the region of interest (ROI) shall be determined using both an automated imaging algorithm and the RingAssay software tool (http://invitrotox.uni-konstanz.de/) to generate concentration-response curves from at least three separate experiments. Migration results shall be normalized to untreated controls combining the migration and viability data into a prediction model to classify test compounds as either inactive or specific inhibitors of NCC migration, or non-specific inhibitors of NCC migration using both an automated imaging algorithm and software tool to generate concentration-response curves from at least three separate experiments. Migration results shall be normalized to untreated controls combining the migration and viability data into a prediction model. The prediction model shall be based on two endpoints, migration

(M) and viability (V). The EC90V/EC75M ratio shall be used to separate specific and unspecific compounds as well as the extent of migration inhibition at the EC90V. EC90V shall be defined as the effective concentration bringing viability down to 90% and EC75M shall be defined as the effective concentration bringing migration down to 75%. The threshold value for the prediction model using migration at the EC90V shall be set at 25%. Migration shall be measured at EC90V, and a hit shall be classified as a compound that inhibits migration ≥ 25% at this concentration.If cytotoxicity is noted in greater than three (>3) concentrations then the dilution series shall be repeated at a lower range.

Experiments shall be performed at least but not limited to three times. Test compounds identified as specific inhibitors of NCC migration shall be repeated in a second test of concentration response experiments. Test concentrations shall be prepared in a 1:3 dilution series from a starting concentration of 100 µm.

3.0. DATA RIGHTS: See IDIQ base contract

4.0. DELIVERABLES AND REPORTING REQUIREMENTS: See IDIQ base contract

4.1. Required Reports: A draft and final report for this effort shall provide a summary of the methods used and the results obtained in sufficient detail and quality to meet the requirements for peer reviewed literature. The draft report shall be delivered in both hard and electronic formats (MS Word) for review and comment by the Contracting Officers Representative (COR); the final report shall be delivered in hard and electronic (PDF) formats. All final electronic documents shall be section 508 compliant. All raw data shall be attached as an appendix to the report and submitted electronically to the COR as an Excel spreadsheet. The final report shall include but not be limited to:

4.1.1. A detailed description of all experimental procedures

4.1.2. A full set of all raw data, summarized/normalized data

4.1.3. Guidance on the interpretation of the DNT in vitro NCC testing results.

4.1.4. Concentration response curves f providing the EC90V/EC75M ratio for each test compound

4.1.5. Identification of compounds as "specific DNT compounds," unspecific DNT compounds," or "negative compounds" using an NCC-specific classification model

4.2. Contract Management Reports: See IDIQ base contract.

http://invitrotox.uni-konstanz.de/

4.3. Periodic Status Reports: See IDIQ base contract.

4.4. Draft Final Report: See IDIQ base contract.

4.5. General Reporting Requirements: See IDIQ base contract.

4.6. Electronic Deliverables: See IDIQ base contract

5.0. PLACE OF PERFORMANCE: See IDIQ base contract.

6.0. PERIOD OF PERFORMANCE: 12 months

7.0. Section 508 Requirements for Deliverables: See IDIQ base contract

8.0. CONTRACTOR FURNISHED MATERIALS: See IDIQ base contract.