Statement_of_Work.pdf

Statement of Work

A. BACKGROUND INFORMATION:

There is increasing evidence that the endogenous microbiome affects many aspects of human health. Gut microbiota affect the development and functioning of the immune system, and can promote immune responses to attack self-components in autoimmune diseases. Autoimmune uveitis is estimated to underlie 10–15% of blindness. Many patients exhibit immune responses to retinal components that are thought to be involved in progression of the disease, but the primary cause of these disorders is unknown. Microbial triggers have been suspected in uveitis, but never been proven. We have developed a transgenic (Tg) mouse model of spontaneous autoimmune uveitis in NEI. We hypothesized and proved that gut microflora provide a trigger in this model: depletion of commensal microbiota with broad-spectrum antibiotics treatment strongly attenuated disease. Our collaboration with Dr. Kenya Honda (Keio University and RIKEN, Japan) confirmed that our Tg mice under the germ-free condition also show significantly attenuated disease. These findings suggest that endogenous gut microbiota of healthy individuals promotes development of autoimmunity to the eye.

In the current project of “Germ-Free Work on Transgenic Mice at Sankyo Labo Service Corporation, Japan”, we are collaborating with Dr. Katsuko Sudo who is an advisor of Sankyo Labo Service, and Dr. Kikuji Itoh at Japan SLC (a parental vendor of Sankyo Labo Service), to examine germ-free and gnotobiotic TCR Tg mice and to isolate the commensal bacteria that are responsible for promoting uveitis. We will cover the costs associated with germ-free breeding, maintenance and gnotobiotic analysis that will be performed at Sankyo Labo Service Corporation, Japan.

B. PURPOSE AND OBJECTIVES OF THE PROCUREMENT:

The overall project objective is to continue our research in the germ-free condition to identify environmental causes of uveitis. To accomplish this NEI will cover the costs for germ-free breeding, maintenance and gnotobiotic analysis. The contract personnel and team at Sankyo Labo Service will: 1) breed and maintain our germ-free Tg mice in a germ-free isolator, 2) perform ear tagging, tail clipping and genotyping of these mice, 3) perform the gnotobiotic reconstitution and follow-up study in another germ-free isolator, and 4) ship the requested tissue samples (such as eyes and intestines) in fixatives to NEI. NEI will pay a part of the cost, $20,000 USD, for this project and transfer the fund to Sankyo Labo Service.

C. CONTRACT REQUIREMENTS:

1. The employees at Sankyo Labo Service shall provide the service in maintaining germ-free status of our Tg mice and in performing requested experiments in the germ-free facility. Duties include, but are not limited to 1) receive and respond to requests for data/samples from the germ-free Tg mouse colony, 2) ship samples out to requesting researchers, 3) verify all the forms/documents are correctly completed and 4) access sample data in the database.

2. All should possess:

• Laboratory skills: Animal handling and molecular biology

• Administrative skills: Effectively and efficiently communicates with the collaborators involved in this work

• Proficient in Microsoft Word, Excel, PowerPoint, Email and other Internet applications. Proficient with database entry, analysis and report running.

Demonstrated skills in the development and performance of laboratory standard operating procedures.

Period of Performance 9/01/2016 – 8/31/2017

D. GOVERNMENT RESPONSIBILITIES: NEI will provide the laboratory space, equipment, supplies, and consumables necessary to accomplish this project. Shipping of samples for this project will be covered by NEI.

E. REPORTING REQUIREMENTS: NEI and Sankyo Labo Service shall share the inventory of germ-free animals and summarize the research results timely.

F. INSPECTION AND ACCEPTANCE CRITERIA: The final manuscript(s) shall be inspected and approved by the Chief of Immunoregulation Section.

G. ATTACHMENTS: N/A

H. CLEARANCES: None required.

8/15/2016