Shuttle of lentiviral vectors via transplanted cells in vivo

Blömer, Ulrike; Gruh, Ina; Witschel, H; Haverich, Axel; Martin, Ulrich

doi:10.1038/sj.gt.3302384

Cited by 29 publications

(25 citation statements)

References 26 publications

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“…These studies, along with PCR results in transplantation studies, indicate that vector particle transfer results in proviral integration and that mere protein transfer (i.e., pseudotransduction) accounts for only a minor portion of GFP expression in recipient fibroblasts. We also found that particle transfer diminishes with additional posttransduction washes of primary cells but is not completely abolished, consistent with studies by Blomer and coworkers (5). Importantly, particles shed from primary targets (i.e., hematopoietic cells) remain fully infectious, and the transfer efficiency to secondary targets is a function of vector particle loading (i.e., MOI).…”

Section: Discussionsupporting

confidence: 76%

“…Indeed, in our own studies we recovered only ϳ10% infectious particles in the non-cell-associated fraction after a 1-h exposure of vector to primary hematopoietic cells, consistent with results by Cole and colleagues (10). Moreover, retention of particles on the cell surface can result in localized secondary transduction in recipients after direct injection of lentivectorexposed cardiomyocytes into brain and muscle tissue (5). However, direct extrapolation of those findings to protocols involving ex vivo gene transfer to and subsequent intravenous injection of washed hematopoietic cells is difficult, and the quality and quantity of any potential inadvertent transfer and secondary transduction in vivo under these circumstances is unclear.…”

Section: Discussionsupporting

confidence: 75%

“…In studies using recombinant retrovirus particles bearing alternate pseudotypes, others have previously demonstrated prolonged binding after ex vivo exposure of cardiomyocytes that was resistant to washing procedures, with resultant tissue transduction from attached particles after local injection in the brain (5). We reasoned that the mechanisms underlying this observation might be more widely applicable and undertook the studies reported here.…”

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confidence: 99%

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Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Pan¹,

Scarlett²,

Luoh³

et al. 2007

J Virol

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Human immunodeficiency virus type 1-derived lentivirus vectors bearing the vesicular stomatitis virus G (VSV-G) envelope glycoprotein demonstrate a wide host range and can stably transduce quiescent hematopoietic stem cells. In light of concerns about biosafety and potential germ line transmission, they have been used predominantly for ex vivo strategies, thought to ensure the removal of excess surface-bound particles and prevent in vivo dissemination. Studies presented here instead reveal prolonged particle adherence after ex vivo exposure, despite serial wash procedures, with subsequent transduction of secondary target cells in direct and transwell cocultures. We explored the critical parameters affecting particle retention and transfer and show that attachment to the cell surface selectively protects virus particles from serum complement-mediated inactivation. Moreover, studies with nonmyeloablated murine recipients show that transplantation of vectorexposed, washed hematopoietic cells results in systemic dissemination of functional VSV-G/lentivector particles. We demonstrate genetic marking by inadvertent transfer of vector particles and prolonged expression of transgene product in recipient tissues. Our findings have implications for biosafety, vector design, and cell biology research.

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Section: Discussionsupporting

confidence: 76%

Section: Discussionsupporting

confidence: 75%

mentioning

confidence: 99%

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Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Pan¹,

Scarlett²,

Luoh³

et al. 2007

J Virol

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“…S4). Transduced host cells were not observed in the culture medium injected controls (n ¼ 6 eyes), and the virally transfected cells were washed extensively and FAC-sorted prior to transplantation, but we cannot rule out the possibility that a few viral particles have been shuttled with the cells, as has been shown previously for lentiviral vectors [24].…”

Section: Esc-derived Retinal Cells Transplanted To the Adult Retinamentioning

confidence: 99%

Defining the Integration Capacity of Embryonic Stem Cell-Derived Photoreceptor Precursors

West¹,

Gonzalez‐Cordero²,

Hippert³

et al. 2012

Stem Cells

119

116

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Retinal degeneration is a leading cause of irreversible blindness in the developed world. Differentiation of retinal cells, including photoreceptors, from both mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), potentially provide a renewable source of cells for retinal transplantation. Previously, we have shown both the functional integration of transplanted rod photoreceptor precursors, isolated from the postnatal retina, in the adult murine retina, and photoreceptor cell generation by stepwise treatment of ESCs with defined factors. In this study, we assessed the extent to which this protocol recapitulates retinal development and also evaluated differentiation and integration of ESC-derived retinal cells following transplantation using our established procedures. Optimized retinal differentiation via isolation of Rax.GFP retinal progenitors recreated a retinal niche and increased the yield of Crx 1 and Rhodopsin 1 photoreceptors. Rod birth peaked at day 20 of culture and expression of the early photoreceptor markers Crx and Nrl increased until day 28. Nrl levels were low in ESC-derived populations compared with developing retinae. Transplantation of early stage retinal cultures produced large tumors, which were avoided by prolonged retinal differentiation (up to day 28) prior to transplantation. Integrated mature photoreceptors were not observed in the adult retina, even when more than 60% of transplanted ESCderived cells expressed Crx. We conclude that exclusion of proliferative cells from ESC-derived cultures is essential for effective transplantation. Despite showing expression profiles characteristic of immature photoreceptors, the ESC-derived precursors generated using this protocol did not display transplantation competence equivalent to precursors from the postnatal retina. STEM CELLS

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“…A recent study by Blomer et al 18 reports the presence of infectious VSV-G pseudotype lentiviral particles on cells injected into animals after a 5-h ex vivo transduction (at MOI 7), and persisting at low levels even after extensive washing. We have not evaluated to what extent injected cells in our model serve as a 'shuttle' for unincorporated viral particles, but it is worthwhile to emphasize the differences between their protocol and the one proposed here.…”

mentioning

confidence: 99%

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

2005

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Efficient gene transfer to hematopoietic stem cells by Moloney murine leukemia virus-derived retroviral vectors benefits from ex vivo culture and cytokine support. Both also increase the risks of apoptosis and differentiation among cells targeted for transduction. In an effort to maximize the retention of stem cell properties in target cells, we developed a transduction protocol with a focus on minimizing graft manipulation, cytokine stimulation, and ex vivo exposure duration. Based on their wide host range and ability to transduce quiescent cells, human immunodeficiency virus (HIV)-derived lentivirus vectors are ideally suited for this purpose. Our present studies in a murine model show that whole bone marrow cells are readily transduced after a 1-hour vector exposure in the presence of stem cell factor and CH296 fibronectin fragment. Using this rapid transduction protocol, we achieved long-term, multilineage reconstitution of murine recipients with up to 25% GFP-expressing cells in primary and secondary recipients. Our results demonstrate the unique ability of HIV-derived vectors to transduce hematopoietic stem cells in the absence of enrichment, under minimal cytokine stimulation, and following brief exposures.

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Shuttle of lentiviral vectors via transplanted cells in vivo

Cited by 29 publications

References 26 publications

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Prolonged Adherence of Human Immunodeficiency Virus-Derived Vector Particles to Hematopoietic Target Cells Leads to Secondary Transduction In Vitro and In Vivo

Defining the Integration Capacity of Embryonic Stem Cell-Derived Photoreceptor Precursors

Rapid 1-hour transduction of whole bone marrow leads to long-term repopulation of murine recipients with lentivirus-modified hematopoietic stem cells

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