Ocular Gene Therapy

National Eye Institute

Investigators

Abstract

The Core has strong expertise in the design and cloning of AAV constructs, and packaging and purification of AAV vectors. We provide a broad selection of capsid serotypes to meet diverse research needs. AAV vectors are aomng the most effective means to transduce a gene construct into all retinal cell types in vivo via subretinal or intravitreal injections, and as such are being used widely in developing gene therapies and in fundamental research. Outside of the eye, AAV is also the gene delivery vector of choice for the brain, liver, skeletal muscle, heart, cells of hematopoietic lineages, stem cells, organoids and primary cultures. It became apparent to us that intramural investigators from multiple NIH ICs have needs for custom AAV vector production service that are not being met elsewhere. As the NEI OGTC has become the only standing AAV packaging Core on the entire NIH Bethesda campus and as inquiries pour in from non-NEI researchers, we opened up our service to non-NEI laboratories, first to NINDS and now to all NIH ICs. This is encouraged by both NEI and NIH leadership as it leads to more efficient utilization of resources. While traditionally animal models have typically been used to test AAV-vector mediated gene transduction, advances in retinal organoids are increasingly making them a viable alternative. The Core encourages investigators to employ organoid based platforms to address many of their research questions, where appropriate, before resorting to the use of animal models. Questions appropriate for the organoid models may include validation of vector design, vector transduction efficiency, effectiveness of a chosen promoter, biochemical and functional analyses of expressed transgenes, and phenotype modification in the targeted retinal cells following gene delivery. Research conducted at the Core in the past few years support support the expanded use of human retinal organoids as a valuable tool for both basic research and translational studies. Testing of candidate therapeutic compounds in organoids is also a viable option. Such an approach aligns with NIH’s goal of reducing animal use in research. The recently announced NIH new initiative (https://www.nih.gov/news-events/news-releases/nih-prioritize-human-based-research-technologies ) adds a renewed sense of urgency to this transition. Below is a partial list of AAV vectors we have produced for NEI and NIH investigators. 05/04/24 Anand Swaroop NEI 10RB 7m8 7m8 CMV hNRL-AS321 P51S X 06/11/24 Wei Li NEI 5RB 7m8 7m8 CMV rDA3m X 06/18/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV hNRL_Frog 06/18/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV hNRL_Shark 06/18/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV hNRL_Tunicate 06/25/24 Gu Yi NINDS 5RB AAV1 AAV1 EF1 Alpha 06/25/24 Gu Yi NINDS 5RB AAV1 AAV1 CamKIIP 07/15/24 Mark Wagner NINDS 5RB AAV8 AAV8 fD10 07/15/24 Mark Wagner NINDS 5RB AAV8 AAV8 R-CamP2 07/25/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV EGFP X 07/15/24 Susan Wray NINDS 5RB 7m8 7m8 GnRH NPYpHluorin X 07/23/24 Gu Yi NINDS 5RB AAV5 AAV5 RAM X 07/23/24 Gu Yi NINDS 5RB AAV9 AAV9 RAM X 11/04/24 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 HTRA1 X 11/04/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV Amphioxus 11/04/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV Lamphry 11/05/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV Cartilaginous fish 11/05/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV hNRL L160fs X 11/05/24 Martha Garcia NINDS 5RB AAV8 AAV8 FRT FS_REv 11/13/24 Anand Swaroop NEI 5RB 7m8 7m8 CMV hNRLmH125Q X 11/13/24 Anand Swaroop NEI 10RB 7m8 7m8 CMV hNRLAS321 X 01/29/25 Narendra Derek NINDS 5RB AAV9 AAV9 Su9 EGFPmCherry 01/29/25 Narendra Derek NINDS 5RB AAV9 AAV9 EGFPmCherry_FISI 01/29/25 Gu Yi NINDS 5RB AAV5 AAV5 CAG Flex mRuby2GSG X 02/04/25 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 EmGFPmiR62 X 02/04/25 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 EmGFPmiR LacZ X 02/11/25 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 SOCS3 X 02/11/25 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 PPARGCIA X 02/11/25 Kapil Barti NEI 5RB AAV5 AAV5 hVMD2 MPU17L X 03/11/25 Mark Wagner NINDS 5RB PHP.eB PHP.eB syn DOHBjRGE Cola X 03/24/25 Tiansen Li NEI 1RB PHP.eB PHP.eB CMV EGFP for general testing X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 04/20/25 Anand Swaroop NEI 1RB PHP.eB PHP.eB CMV NRL derivative series X 4/28/25 Claire Le Pichon NIDDK planning multiple x 4/28/25 Martyna Grochowska NIA planning multiple x 4/28/25 Stefano Antonucci NICHD planning multiple xx 5/23/25 Narendra Derek NINDS 1 RB myoAAV-4 myoAAV-4-EJS1096 Dual-sgRNA.Design 1 guides modified 5/23/25 Mark Wagner NINDS 1 RB AAV8 AAV8-DIO-ATLASsnFlp In addition to AAV vectors, the Core has also provided human retinal organoids for several groups.