Abstract
Human immunodeficiency virus continues to persist in millions of people worldwide. While antiretroviral drug therapies have improved life for many, a cure remains elusive. Long-term antiretroviral drug therapy can also lead to toxicity, the development of drug resistant strains and the persistence and maintenance of latent reservoirs in resting cluster of differentiation 4 positive T-lymphocytes. Gene therapy offers a promising alternative strategy for eradicating human immunodeficiency virus infection through the development of lentiviral vectors comprised of highly potent anti-HIV transgenes capable of inhibiting human immunodeficiency virus infection during the preintegration phases. Cellular reprogramming allows for the generation of induced pluripotent stem cells (iPSC) that are similar to embryonic stem cell-like cells with the potential to differentiate into any cell type in the body. In this study, hematopoietic stem cells were isolated from umbilical cord blood, transduced with a triple combination lentiviral vector containing three potent anti-HIV transgenes encoding a chemokine-chemokine receptor 5 short hairpin RNA, a chimeric tripartite motif-containing protein 5, and a viral transactivation response element decoy. These cells were then de-differentiated back into an undifferentiated pluripotential state by transduction with specific transcription factors octamer-binding transcription factor 4 (OCT4), sex determining region Y-box 2 (SOX2), cytoplasmic c-MYC, and Krüppel-like factor 4 (KLF4). The resulting colonies were analyzed and found to express the anti-HIV genes, by enhanced green fluorescent protein detection. Additionally, the pluripotency markers were also determined to be expressed indicating successful transduction and reprogramming. Reprogramming patient-specific cells to derive personalized iPSCs is an excellent source for potentially transplantable tissue that will not cause an immune response and subsequent rejection. This could be used to treat a myriad of human blood and degenerative diseases without the ethical concerns attached to the acquisition and use of ES cells. In addition, this iPSC approach could provide a highly beneficial and personalized patient specific platform for future treatments while diminishing the need to rely on potentially toxic drug therapies and resistant strains.