Generation of HIV-1 Resistant and Functional Macrophages From Hematopoietic Stem Cell–derived Induced Pluripotent Stem Cells

Abstract

Induced pluripotent stem cells (iPSCs) have radically advanced the field of regenerative medicine by making possible the production of patient-specific pluripotent stem cells from adult individuals. By developing iPSCs to treat HIV, there is the potential for generating a continuous supply of therapeutic cells for transplantation into HIV-infected patients. In this study, we have used human hematopoietic stem cells (HSCs) to generate anti-HIV gene expressing iPSCs for HIV gene therapy. HSCs were dedifferentiated into continuously growing iPSC lines with four reprogramming factors and a combination anti-HIV lentiviral vector containing a CCR5 short hairpin RNA (shRNA) and a human/rhesus chimeric TRIM5α gene. Upon directed differentiation of the anti-HIV iPSCs toward the hematopoietic lineage, a robust quantity of colony-forming CD133(+) HSCs were obtained. These cells were further differentiated into functional end-stage macrophages which displayed a normal phenotypic profile. Upon viral challenge, the anti-HIV iPSC-derived macrophages exhibited strong protection from HIV-1 infection. Here, we demonstrate the ability of iPSCs to develop into HIV-1 resistant immune cells and highlight the potential use of iPSCs for HIV gene and cellular therapies.

Generation of an HIV-1-Resistant Immune System with CD34 + Hematopoietic Stem Cells Transduced with a Triple-Combination Anti-HIV Lentiviral Vector

“…One promising method to achieve this scenario is to use clonal induced pluripotent stem cells (iPSCs) which express anti-HIV genes. This was demonstrated in a recent study where complete protection from HIV-1 infection was observed due to the expression of a combination of anti-HIV genes in every macrophage derived from the iPSC line (17).…”

“…In this regard, by combining multiple preintegration anti-HIV genes into a single vector, potent preintegration protection from HIV infection could be conferred (5,17). In a previous report by our group, strong preintegration protection from HIV-1 infection, in vitro, was established by a triple-combination anti-HIV lentiviral vector containing a human/rhesus macaque TRIM5␣ isoform, a CCR5 short hairpin RNA (shRNA), and a TAR decoy (5).…”

Seamless modification of wild-type induced pluripotent stem cells to the natural CCR5Δ32 mutation confers resistance to HIV infection

“…Using iPSCs as target cell sources for potential HIV gene and cellular therapy has been explored (28). In that study, the CCR5 gene was modified by knockdown using a lentiviral vector that delivered short hairpin RNA targeting CCR5.…”

Searching for the Key to Expand Hematopoietic Stem Cells

“…Raya et al reprogrammed dermal fibroblasts and/or epidermal keratinocytes of Fanconi anemia patients to generate iPSC, which were genetically corrected with lentiviral vectors encoding FANCA or FANCD2, to obtain hematopoietic progenitors of the myeloid and erythroid lineages that are phenotypically normal, that is, disease-free (Raya et al, 2009). Similar strategies were performed to correct the Hurler syndrome (Tolar et al, 2011) and for the production of macrophages from iPSCs which were resistant to HIV infection (Kambal et al, 2011).…”