Humanized Mouse Avatars for T1D

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ABSTRACT Our ignorance of the key molecules and cells mediating the effector phase of human T1D may well underlie the paucity of promising new therapeutic interventions. This supplemental funding request is for June 1, 2023 through May 31, 2024 for our HIRN project 5U01DK104218, “Humanized Mouse Avatars for T1D”. This proposed time period is predicated on the 2-4 months needed to perform each of many in vivo experiments.The budget request will support only the critical experiments that we were not able to perform due to COVID-related delays. These issues have been resolved and we are poised to complete the experiments that will establish models for investigation of human SC-islet/autoreactive T cell interaction in vivo. This funding will permit us: 1) to retain only essential staff needed to perform these experiments, 2) to maintain and carry out experiments with the novel stocks of immunodeficient mice required, 3) to preserve unique iPSC lines we developed for this project, 4) to generate the SC-islets that are essential for all of the proposed studies, and 5) to perform the essential experiments needed to establish in vivo models for human autoreactive T cell-mediated destruction of SC-islets. Our group currently consists of faculty from 4 institutions, Dr. Melton at Harvard, Dr. Daley at Boston Children's Hospital, Dr. Shultz at The Jackson Laboratory, and Drs. Greiner, Brehm, Maehr, Kent and Harlan at UMass Chan Medical School. Dr. Melton' laboratory is closing on April 30, 2023 and the production of human SC-islets from iPSCs required for this project has been seamlessly assumed by Dr. Harlan at UMass Chan. Although there are two aims in the original proposal, we will focus solely on the critical experiments needed to accomplish Aim 1, the effector phase model of human T1D. To accomplish this aim we will use autoreactive T cells from the blood of T1D individuals (Drs. Brehm, Greiner), T cell lines (generated by Dr. Kent) and iPSC-derived CAR T cells (generated by Dr. Daley) engrafted together with autologous SC-islet cells (generated by Dr. Harlan) into OPTI-MICE (generated by Dr. Shultz). We will identify the TCR specificity of the infiltrating effector T cells and the in vivo beta cell response to immune attack using 10X Genomic sequencing (Dr. Maehr). We request support only for the essential reagents, mouse strains, and personnel that are required to perform these critical studies. Validation of these models will provide much needed in vivo model systems for the investigation of the T cells that mediate beta cell destruction and their in vivo interaction with autologous SC-islets. We anticipate that these models will be widely used by the scientific community to identify which of the many identified T1D autoreactive T cell specificities are “causal” and which are “non-pathogenic”, allow interrogation of the effector response of autoreactive T cells and the response of beta cells to autoimmune attack, and to provide a platform for testing T cell specific immunotherapeutics by the scientific community.