Induction of Local Immunosuppression in Allogenic Cell Transplantation by Cell-type-specific Expression of PD-L1 and CTLA4Ig

Immune rejection has long hindered allogenic cell transplantation therapy. Current genetic modification approaches, including direct targeting of major histocompatibility complex or constitutive expression of immune inhibitory molecules, exhibit drawbacks such as severe adverse effects or an elevated risk of tumorigenesis. To overcome these limitations, we propose an innovative approach aimed at inducing cell-type-specific immune tolerance in differentiated cells. By engineering human embryonic stem cells, we enable the exclusive production of immune inhibitory molecules, PD-L1/CTLA4Ig, within differentiated cells following lineage commitment. Leveraging this approach, we have successfully generated hepatocyte-like cells expressing PD-L1 and CTLA4Ig, which effectively induced local immunotolerance. This strategy was evaluated in a humanized mouse model designed to mimic human immune system. Our results demonstrate robust and selective induction of immunotolerance specific to hepatocytes, thereby improving graft survival without tumor formation. This precise immune tolerance strategy holds promise for advancing the development of stem cell-based therapeutics in regenerative medicine. Overall design: wild-type hPSC, ALB-PD-L1/CTLA4Ig hPSC, AAV-PD-L1/CTLA4Ig derived MHLCs were collected, and the total RNA was extracted by Trizol. RNA samples were shipped to the ANOROAD GENOME for analysis as a service. The DNBs were loaded into the patterned nanoarrays and pair-end reads of 150 bp were read through on the MGI platform for the following data analysis study. For this step, the MGI platform combines the DNA nanoball-based nanoarrays and stepwise sequencing using PE150 Method.