Efficient generation of CD4 & CD8 T cell derived iPSC for potential use in allogeneic cellular therapies

Background & Aim The clinical success of approved chimeric antigen receptor T-Cell (CAR-T) therapy to treat B-cell leukemias has generated interest to apply similar approach to other forms of cancer. However, processing patient T cells that are often exhausted and dysfunctional poses a challenge. This and the steep cost associated with autologous therapies has led to exploration of allogeneic approaches using T cells from healthy donors. An appealing alternate source are induced pluripotent stem cells (iPSCs) since they can be expanded indefinitely and differentiated into healthy T cells. Reprogramming T cells to generate T-iPSC is especially beneficial when working with antigen specific T cells, such as tumor infiltrating lymphocytes or viral specific T cells, where preserving antigen recognition is of importance. Additionally, T-cell derived iPSC (T-iPSC) with an unique TCR serves as an intrinsic barcode enabling in vivo tracking that can be a beneficial for any iPSC-based therapy. Methods, Results & Conclusion We had previously reported a Sendai-virus based reprogramming specifically designed for translational and clinical applications. Here, we studied the effect of diverse cellular phenotypes in different donor T cells on reprogramming. A robust method of reprogramming was established for CD3+ cells isolated using different methods and enriched for CD4+ and CD8+ T cells. Resulting iPSC were analyzed using NGS-based immune repertoire to generate high-throughput sequencing data to examine the diversity of TCR. Comprehensively characterized T-iPSC cell banks offer a renewable cell source that can be differentiated into cell types of choice for the development of translational therapies. The clinical success of approved chimeric antigen receptor T-Cell (CAR-T) therapy to treat B-cell leukemias has generated interest to apply similar approach to other forms of cancer. However, processing patient T cells that are often exhausted and dysfunctional poses a challenge. This and the steep cost associated with autologous therapies has led to exploration of allogeneic approaches using T cells from healthy donors. An appealing alternate source are induced pluripotent stem cells (iPSCs) since they can be expanded indefinitely and differentiated into healthy T cells. Reprogramming T cells to generate T-iPSC is especially beneficial when working with antigen specific T cells, such as tumor infiltrating lymphocytes or viral specific T cells, where preserving antigen recognition is of importance. Additionally, T-cell derived iPSC (T-iPSC) with an unique TCR serves as an intrinsic barcode enabling in vivo tracking that can be a beneficial for any iPSC-based therapy. We had previously reported a Sendai-virus based reprogramming specifically designed for translational and clinical applications. Here, we studied the effect of diverse cellular phenotypes in different donor T cells on reprogramming. A robust method of reprogramming was established for CD3+ cells isolated using different methods and enriched for CD4+ and CD8+ T cells. Resulting iPSC were analyzed using NGS-based immune repertoire to generate high-throughput sequencing data to examine the diversity of TCR. Comprehensively characterized T-iPSC cell banks offer a renewable cell source that can be differentiated into cell types of choice for the development of translational therapies.