In vivo validation of Universal Donor Cells

DESCRIPTION provided by applicant Pluripotent stem cells have the potential to treat diseases affecting almost every organ system However their clinical use is limited by host rejection due to differences in the diverse Major Histocompatibility Complex MHC genes In the past this problem has been addressed by using MHC matched donors and or dangerous immunosuppressive drug regimens Here we propose to demonstrate that MHC negative pluripotent stem cells can avoid recognition as allogeneic in clinical transplantation applications and therefore be used as universal donor stem cells Prior experiments using MHC class I negative human embryonic stem cells ESCs derived by targeted disruption of the Beta Microglobulin gene B M showed that these cells are not recognized by immune cells in vitro We will now perform in vivo mouse experiments in order to show that MHC class I negative cells are not rejected and can engraft in a normal mouse after allogeneic transplantation We will first derive induced Pluripotent stem cells iPSCs from the well characterized B m model which are MHC class I deficient Then we will induce teratoma formation using these cells in order to analyze the allogeneic immune response against B m iPSC derived teratomas We will derive cardiac progenitor cells from B m iPSCs and transplant them into allogeneic recipients We hypothesize that these cells will engraft and survive after transplantation contrary to B m allogeneic iPSCs bringing in vivo proof of concept that the MHC class I negative pluripotent cells can act as universal donors The long term goal of this proposal is to use MHC engineering to create stem cell lines that can be used for regenerative medicine applications in multiple recipients without rejection thereby facilitating the translation of stem cell based therapies into the clinic PUBLIC HEALTH RELEVANCE A major limitation in the clinical use of stem cells is their rejection after transplantation Here we will validate the use of pluripotent stem cells engineere to act as universal donor cells that can be used for various cell therapy applications without rejection These cells will be ideally suited for clinical use since only one cell line needs to b prepared and approved by regulatory agencies