Production of Retrovirus-Based Vectors in Mildly Acidic pH Conditions

Holic, Nathalie; Fenard, David

doi:10.1007/978-1-4939-3753-0_3

Cited by 2 publications

(2 citation statements)

References 21 publications

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“…The scrambled miR‐494 was cloned into the MSCV‐IRES‐GFP vector to use as a control. The plasmid was cotransfected into 293T cells with VSVG and pKAT, using Lipofectamine 2000 (Holic and Fenard, ). Virus supernatant was harvested 48 h after transfection.…”

Section: Methodsmentioning

confidence: 99%

MicroRNA‐494 Activation Suppresses Bone Marrow Stromal Cell‐Mediated Drug Resistance in Acute Myeloid Leukemia Cells

Tian

Zheng

Zhuang

et al. 2016

Journal Cellular Physiology

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Acute myeloid leukemia (AML) is not sensitive to chemotherapy partially because of the protection of AML cells by mesenchymal stromal cells (MSCs). Our previous studies found that MSCs protected AML cells from apoptosis through the c-Myc-dependent pathway. However, the mechanism by which MSCs regulate c-Myc in AML cells is still unknown. To elucidate the mechanism, we performed microRNA array analysis of AML cell lines and validated by TaqMan realtime PCR. The results showed that the expression of microRNA-494 (miR-494) in AML cells after coculture with MSCs was downregulated. Reporter gene analysis confirmed miR-494 as one of the regulators of c-Myc. In the coculture system, activation of miR-494 in AML cells suppressed proliferation and induced apoptosis of AML cells in vitro. After addition of mitoxantrone to the coculture system, the proliferation of AML cells with miR-494 activation was suppressed more than that of control cells. After subcutaneous injection of AML cell lines in combination with MSC, tumor growth was suppressed in mice injected with miR-494-overexpressing AML cells. The rate of tumor formation was even lower after mitoxantrone treatment in the miR-494 overexpressing group. Moreover, miR-494 activation resulted in a decrease of leukemic cell counts in peripheral blood (PB) and bone marrow, and prolonged survival in mice injected with miR-494-overexpressing AML cellls and MSCs compared to the control mice. Our results indicate that miR-494 suppresses drug resistance in AML cells by downregulating c-Myc through interaction with MSCs and that miR-494 therefore is a potential therapeutic target. J. Cell. Physiol. 232: 1387-1395, 2017. © 2016 Wiley Periodicals, Inc.

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Section: Methodsmentioning

confidence: 99%

MicroRNA‐494 Activation Suppresses Bone Marrow Stromal Cell‐Mediated Drug Resistance in Acute Myeloid Leukemia Cells

Tian

Zheng

Zhuang

et al. 2016

Journal Cellular Physiology

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“…LVs were generated as described previously (32). Briefly, HEK293T cells were transiently transfected using calcium phosphate transfection with four plasmids: the gagpol (pKLgagpol) and rev (pKrev) expression plasmids, a transfer plasmid (pCCLsin.cPPT.hPGK.eGFP.WPRE), and a plasmid encoding either the VSV-G (pMDG) envelope glycoprotein (GP), the GALVTR GP (pBA.GALV/Ampho-Kana), or the RD114TR GP (pHCMV-RD114TR).…”

Section: Viral Vector Production and Titeringmentioning

confidence: 99%

Peptides derived from evolutionarily conserved domains in Beclin-1 and Beclin-2 enhance the entry of lentiviral vectors into human cells

Majdoul

Cosette

Seye

et al. 2017

Journal of Biological Chemistry

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Autophagy-related proteins such as Beclin-1 are involved in an array of complex processes, including antiviral responses, and may also modulate the efficiency of gene therapy viral vectors. The Tat-Beclin-1 (TB1) peptide has been reported as an autophagy-inducing factor inhibiting the replication of pathogens such as HIV, type 1 (HIV-1). However, autophagy-related proteins are also essential for the early steps of HIV-1 infection. Therefore, we examined the effects of the Beclin-1 evolutionarily conserved domain in TB1 on viral transduction and autophagy in single-round HIV infection or with nonreplicative HIV-1-derived lentiviral vectors. TB1 enhanced transduction with various pseudotypes but without inducing the autophagy process. TB1 augmented the transduction of human CD34+ hematopoietic stem/progenitor cells while maintaining their capacity to engraft into humanized mice. TB1 was as effective as other transduction additives and functioned by enhancing the adhesion and fusion of viral particles with target cells but not their aggregation. We also found that the N-terminal L1 loop was critical for TB1 transduction-enhancing activity. Interestingly, the Tat-Beclin-2 (TB2) peptide, derived from the human Beclin-2 protein, was even more potent than TB1 in promoting viral transduction and infection. Taken together, our findings suggest that the TB1 and TB2 peptides enhance the viral entry step. Tat-Beclin peptides therefore represent a new family of viral transduction enhancers for potential use in gene therapy.

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Production of Retrovirus-Based Vectors in Mildly Acidic pH Conditions

Cited by 2 publications

References 21 publications

MicroRNA‐494 Activation Suppresses Bone Marrow Stromal Cell‐Mediated Drug Resistance in Acute Myeloid Leukemia Cells

MicroRNA‐494 Activation Suppresses Bone Marrow Stromal Cell‐Mediated Drug Resistance in Acute Myeloid Leukemia Cells

Peptides derived from evolutionarily conserved domains in Beclin-1 and Beclin-2 enhance the entry of lentiviral vectors into human cells

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