Tightly regulated gene expression in human hematopoietic stem cells after transduction with helper-dependent Ad5/35 vectors

Wang, Hongje; Cao, Hua; Wohlfahrt, Martin E.; Kiem, Hans Peter; Lieber, André

doi:10.1016/j.exphem.2008.01.014

Cited by 13 publications

(19 citation statements)

References 33 publications

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“… 13 , 14 Toxicity related to leaky viral gene expression can be circumvented by the use of helper-dependent (HD) Ad5/35 vectors that lack all viral genes. 15–18 Growth of HD-Ad vectors depends on coinfection of the producer cells with helper Ad vector, which provides all necessary Ad proteins in trans . Removal of helper virus from HD vector preparations is based on Cre-recombinase-mediated excision of the packaging signal flanked by loxP sites during coinfection.…”

Section: Introductionmentioning

confidence: 99%

“…We have previously shown that HD-Ad5/35 vectors efficiently transduce human CD34+ cells in vitro without significant signs of cytotoxicity. 17 , 18 , 20 The HD-Ad5/35 vector platform has major advantages for HSC genome engineering. HD-Ad5/35 vectors transduce primitive subsets of HSCs, have a large capacity (~30 kb) that can accommodate large payloads, including several EN expression cassettes and homologous donor template, 18 and can be used for transducing in vivo HSC after intravenous injection.…”

Section: Introductionmentioning

confidence: 99%

“…We produced green fluorescent protein (GFP) expressing HD-Ad5/35 vectors and showed that background expression in 293 cells with Tet induction was suppressed. 17 However, when we replaced that GFP gene with the CCR5 ZFN gene, the resulting HD-Ad genomes isolated from purified particles demonstrated genomic rearrangements and a deletion of parts of the ZFN cassette ( Supplementary Figure S1 ).…”

Section: Introductionmentioning

confidence: 99%

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Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

Saydaminova

Wang

et al. 2015

Molecular Therapy - Methods & Clinical Development

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Genome editing with site-specific endonucleases has implications for basic biomedical research as well as for gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35) vectors for zinc-finger nuclease (ZFN)– or transcription activator-like effector nuclease (TALEN)–mediated genome editing in human CD34+ hematopoietic stem cells (HSCs) from mobilized adult donors. The production of these vectors required that ZFN and TALEN expression in HD-Ad5/35 producer 293-Cre cells was suppressed. To do this, we developed a microRNA (miRNA)-based system for regulation of gene expression based on miRNA expression profiling of 293-Cre and CD34+ cells. Using miR-183-5p and miR-218-5p based regulation of transgene gene expression, we first produced an HD-Ad5/35 vector expressing a ZFN specific to the HIV coreceptor gene ccr5. We demonstrated that HD-Ad5/35.ZFNmiR vector conferred ccr5 knock out in primitive HSC (i.e., long-term culture initiating cells and NOD/SCID repopulating cells). The ccr5 gene disruption frequency achieved in engrafted HSCs found in the bone marrow of transplanted mice is clinically relevant for HIV therapy considering that these cells can give rise to multiple lineages, including all the lineages that represent targets and reservoirs for HIV. We produced a second HD-Ad5/35 vector expressing a TALEN targeting the DNase hypersensitivity region 2 (HS2) within the globin locus control region. This vector has potential for targeted gene correction in hemoglobinopathies. The miRNA regulated HD-Ad5/35 vector platform for expression of site-specific endonucleases has numerous advantages over currently used vectors as a tool for genome engineering of HSCs for therapeutic purposes.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

Saydaminova

Wang

et al. 2015

Molecular Therapy - Methods & Clinical Development

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“…These vectors are devoid of all viral genes and only contain minimal amounts of viral DNA. This completely abolishes the leaky expression of adenoviral genes that had been shown to lead to cytotoxicity in HSCs that have been transduced with earlier Ad vectors that still encoded these genes 59,60 .…”

Section: Examples For In Vivo Hsc Gene Therapy (Fig 1)mentioning

confidence: 99%

In Vivo Hematopoietic Stem Cell Transduction

Richter

Stone

Miao

et al. 2017

Hematology/Oncology Clinics of North America

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Synopsis Current hematopoietic stem cell (HSC) gene therapy approaches rely on the collection of patient HSCs, followed by their culture and expansion in vitro, their modification using γ-retrovirus or lentiviral vectors, and their re-infusion into myelo-conditioned patients. While this approach has been successfully used in numerous clinical trials, its reliance on the extended ex vivo culture of patient HSCs comes with a set of disadvantages. Culturing HSCs in the presence of a cytokine cocktail to facilitate their expansion and transduction is thought to negatively impact the long-term viability of HSCs, and their homing and repopulation capacity. Importantly, the requirement for myeloablative regimens in patients represents a critical risk factor and results in considerable morbidity. In addition to these biological problems, the process of ex vivo HSC manipulation is also challenging from a logistics and regulatory standpoint, as manipulations must be performed in specialized, accredited centers. These complexities lead to a high price of such gene therapeutic regimens, and subsequently severely limited patient access to these treatments. In vivo HSC gene therapy approaches aim to simplify the gene therapy process by eliminating the need for ex vivo handling of patient HSCs. The in vivo approach can be subdivided into three methods. One method is based on the direct modification of HSCs in the bone marrow following intraosseous injection. Another involves intravenous injection of gene delivery vectors that home to HSCs in bone marrow. We have developed an alternative approach in which HSCs from the bone marrow are mobilized into peripheral blood, become gene modified upon intravenous vector administration, and then re-engraft in the bone marrow. We will provide examples of these three approaches and then discuss the advantages and disadvantages of in vivo HSC gene therapy.

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“…It has been proposed that only integrated promoters can be inhibited by the TetR-KRAB system [41]. However, different studies demonstrated efficient regulation of transgene expression in vivo using non-integrating adenoviral vectors [42,43].…”

Section: Krab Teton (Tts Tetr-krab)mentioning

confidence: 99%

In vivo gene regulation using tetracycline-regulatable systems

Stieger

Belbellaa

Guiner

et al. 2009

Advanced Drug Delivery Reviews

110

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Numerous preclinical studies have demonstrated the efficacy of viral gene delivery vectors, and recent clinical trials have shown promising results. However, the tight control of transgene expression is likely to be required for therapeutic applications and in some instances, for safety reasons. For this purpose, several ligand-dependent transcription regulatory systems have been developed. Among these, the tetracycline-regulatable system is by far the most frequently used and the most advanced towards gene therapy trials. This review will focus on this system and will describe the most recent progress in the regulation of transgene expression in various organs, including the muscle, the retina and the brain. Since the development of an immune response to the transactivator was observed following gene transfer in the muscle of nonhuman primate, focus will be therefore, given on the immune response to transgene products of the tetracycline inducible promoter.

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Tightly regulated gene expression in human hematopoietic stem cells after transduction with helper-dependent Ad5/35 vectors

Cited by 13 publications

References 33 publications

Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

Efficient genome editing in hematopoietic stem cells with helper-dependent Ad5/35 vectors expressing site-specific endonucleases under microRNA regulation

In Vivo Hematopoietic Stem Cell Transduction

In vivo gene regulation using tetracycline-regulatable systems

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