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

Stem Cell Program, Department of Internal Medicine, University of California, Davis, Sacramento, California 95817, USA.
Molecular Therapy (Impact Factor: 6.23). 11/2010; 19(3):584-93. DOI: 10.1038/mt.2010.269
Source: PubMed


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.

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Available from: Jan A Nolta, Mar 24, 2015
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    • "HSCs were derived from fresh human umbilical cord blood as described in the Supporting Information) and seeded in one well of a six-well tissue-culture-treated plate and cultured in 4 ml CD34 1 culture medium (Kambal et al., 2011): "
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    • "Several groups have published methods for producing monocytes/and macrophages from PSC [3], [4], [5], [6], [7]. However, these methods are technically complicated, because they involve coculture on mouse stromal cells (e.g. "
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    • "some of them inter-dependent: (i) virus entrapment into TRIM5 cytoplasmic bodies 47 (Campbell et al., 2008), (ii) decreased stability of the virus core (Black and Aiken, 2010; 48 Perron et al., 2007; Stremlau et al., 2006; Zhao et al., 2011) (iii) targeting to a 49 proteasome-dependent degradation pathway (Anderson et al., 2006; Diaz-Griffero et al., 50 2006; Lienlaf et al., 2011; Rold and Aiken, 2008); , and (iv) inhibition of nuclear import 51 (Anderson et al., 2006; Berthoux et al., 2004; Wu et al., 2006). The linear structure of 52 TRIM5 reveals a partition between effector domains at the N-terminus and a target 53 recognition domain called PRYSPRY (also known as SPRY or B30.2) found at the C- 54 terminus. "
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