SOW_Sankyo_JPN_2017.pdf

Statement of Work

A. BACKGROUND INFORMATION:

In recent years, it has been recognized that the commensal microbiome has a huge impact on all aspects of our biology, including immune function and disease development. Autoimmune uveitis is estimated to be responsible for 10–15% of blindness in the US. 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 an IRBP T cell receptor transgenic (TCR Tg) mouse model of spontaneous autoimmune uveitis in NEI. We demonstrated that gut microbiota can provide a trigger in this model: depletion of commensal microbiota with broad-spectrum antibiotics treatment strongly attenuated disease. Our collaboration with Japanese colleagues from Sankyo Labo Service Corporation confirmed that these Tg mice reared under germ-free conditions also attenuated disease significantly. These findings suggest that endogenous gut microbiota of healthy individuals may promote development of autoimmune disease in the eye.

In the current project of “Germ-Free Work on Transgenic Mice at Sankyo Labo

Service Corporation”, we are collaborating with Dr. Katsuko Sudo who is an advisor 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 re-derivation, breeding and maintenance of the colony in the germ-free isolators, and gnotobiotic analysis using the isolators that will be performed at Sankyo Labo Service Corporation, Japan.

B. PURPOSE AND OBJECTIVES OF THE PROCUREMENT:

The overall project objective is to continue and massively expand our research using increased numbers of germ-free isolators to identify environmental causes of uveitis.

We also obtained the bench to bedside award this year on the related projects to test the effects human gut microbiota in the development of uveitis using our germ-free

TCR Tg animal model, which is already kept in Sankyo Labo Service. To accomplish this, NEI will cover the costs for germ-free re-derivation, breeding, maintenance of germ-free Tg mouse colony and gnotobiotic studies. The contract personnel and team at Sankyo Labo Service will: 1) re-derive, breed and maintain our germ-free TCR Tg mice in germ-free isolators, 2) perform ear tagging, tail clipping and genotyping of these mice, 3) perform gnotobiotic reconstitutions with human flora or isolated microorganisms and maintain them in separate germ-free isolators, and 4) collect and ship requested tissue samples (such as eyes, intestines, fecal matters and intestinal contents) in fixatives or as frozen samples to NEI. NEI will transfer the fund to

Sankyo Labo Service.

C. CONTRACT REQUIREMENTS:

1. The contrctor shall provide the service in re-driving and maintaining germ-free status of our TCR Tg mice and in performing requested collections and 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 TCR 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 9/01/2017 – 8/31/2018

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

E. REPORTING REQUIREMENTS: NEI and contractor shall share the inventory of germ-free animals on a monthly basis and summarize the research results as needed.

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

G. ATTACHMENTS: N/A

H. CLEARANCES: None required.

5/26/2017