Insertional Gene Activation by Lentiviral and Gammaretroviral Vectors

Bokhoven, Marieke; Stephen, Sam L.; Knight, Sean; Gevers, Evelien; Robinson, I. C. A. F.; Takeuchi, Yasuhiro; Collins, Mary

doi:10.1128/jvi.01865-08

Cited by 119 publications

(102 citation statements)

References 39 publications

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“…The resulting vector backbone not only contains the shortest leader sequence among all retroviral constructs we are aware of (Fig. 5) but also lacks the retroviral major splice donor (in contrast to HIV-1, spumavirus, and MLV vectors), which may prevent some forms of readthrough-dependent insertional gene activation (3,24). Besides minimizing the risk of RCR formation, the split-packaging design also avoids the potentially immunogenic expression of viral proteins from transduced cells and increases the theoretical packaging capacity (at least 8.4 kb, considering the size of deleted alpharetroviral coding sequences) of the alpharetroviral vector.…”

Section: Discussionmentioning

confidence: 99%

“…Murine leukemia virus (MLV) and its derived gammaretroviral vectors have a tendency to integrate close to the transcriptional start sites (TSS), into promoterproximal and CpG-rich regions (43), increasing the risk of activating cellular proto-oncogenes by enhancer-or promotermediated mechanisms. In contrast, human immunodeficiency virus type 1 (HIV-1) and related lentiviral vectors preferentially integrate within active transcription units (34), which may activate growth-promoting genes by both enhancer-and truncation-mediated mechanisms (3,23,24). Vectors based on RSV, belonging to the genus Alpharetrovirus, have been shown to have the most neutral, and therefore the most favorable, integration pattern (21).…”

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confidence: 99%

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Self-Inactivating Alpharetroviral Vectors with a Split-Packaging Design

Suerth

Maetzig

Galla

et al. 2010

J Virol

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Accidental insertional activation of proto-oncogenes and potential vector mobilization pose serious challenges for human gene therapy using retroviral vectors. Comparative analyses of integration sites of different retroviral vectors have elucidated distinct target site preferences, highlighting vectors based on the alpharetrovirus Rous sarcoma virus (RSV) as those with the most neutral integration spectrum. To date, alpharetroviral vector systems are based mainly on single constructs containing viral coding sequences and intact long terminal repeats (LTR). Even though they are considered to be replication incompetent in mammalian cells, the transfer of intact viral genomes is unacceptable for clinical applications, due to the risk of vector mobilization and the potentially immunogenic expression of viral proteins, which we minimized by setting up a split-packaging system expressing the necessary viral proteins in trans. Moreover, intact LTRs containing transcriptional elements are capable of activating cellular genes. By removing most of these transcriptional elements, we were able to generate a self-inactivating (SIN) alpharetroviral vector, whose LTR transcriptional activity is strongly reduced and whose transgene expression can be driven by an internal promoter of choice. Codon optimization of the alpharetroviral Gag/Pol expression construct and further optimization steps allowed the production of high-titer self-inactivating vector particles in human cells. We demonstrate proof of principle for the versatility of alpharetroviral SIN vectors for the genetic modification of murine and human hematopoietic cells at a low multiplicity of infection.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Self-Inactivating Alpharetroviral Vectors with a Split-Packaging Design

Suerth

Maetzig

Galla

et al. 2010

J Virol

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“…[1][2][3] However, recent studies have also identified this risk using lentiviruses. 4 Non-integrating lentiviruses (NILVs) have been developed by introducing class 1 mutations into the integrase gene, and may obviate some of these difficulties. The vector genome exists episomally as functional double-stranded linear templates or circular forms produced by host nuclear enzymes.…”

Section: Introductionmentioning

confidence: 99%

Efficient gene delivery to the adult and fetal CNS using pseudotyped non-integrating lentiviral vectors

et al. 2009

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Non-integrating lentiviral vectors show considerable promise for gene therapy applications as they persist as long-term episomes in non-dividing cells and diminish risks of insertional mutagenesis. In this study, non-integrating lentiviral vectors were evaluated for their use in the adult and fetal central nervous system of rodents. Vectors differentially pseudotyped with vesicular stomatitis virus, rabies and baculoviral envelope proteins allowed targeting of varied cell populations. Efficient gene delivery to discrete areas of the brain and spinal cord was observed following stereotactic administration. Furthermore, after direct in utero administration (E14), sustained and strong expression was observed 4 months into adulthood. Quantification of transduction and viral copy number was comparable when using nonintegrating lentivirus and conventional integrating vector. These data support the use of non-integrating lentiviral vectors as an effective alternative to their integrating counterparts in gene therapy applications, and highlight their potential for treatment of inherited and acquired neurological disorders.

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“…However, genome integration by lentiviral vectors may lead to insertional mutations. So, human immunodeficiency virus (HIV)-based vectors may integrate into the growth hormone receptor locus [28]. Also, a potentially oncogenic insertion of HIV integration has been reported in rare T-cell lymphomas [29].…”

Section: Periostin and Tumorigenesismentioning

confidence: 99%

Periostin Secreted by Mesenchymal Stem Cells Supports Tendon Formation in an Ectopic Mouse Model

Noack

Seiffart

Willbold

et al. 2014

Stem Cells and Development

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True tendon regeneration in human patients remains a vision of musculoskeletal therapies. In comparison to other mesenchymal lineages the biology of tenogenic differentiation is barely understood. Specifically, easy and efficient protocols are lacking that might enable tendon cell and tissue differentiation based on adult (stem) cell sources. In the murine mesenchymal progenitor cell line C3H10T½, overexpression of the growth factor bone morphogenetic protein 2 (BMP2) and a constitutively active transcription factor, Smad8 L + MH2, mediates tendon cell differentiation in vitro and the formation of tendon-like tissue in vivo. We hypothesized that during this differentiation secreted factors involved in extracellular matrix formation exert a major impact on tendon development. Gene expression analyses revealed four genes encoding secreted factors that are notably upregulated: periostin, C-type lectin domain family 3 (member b), RNase A4, and follistatin-like 1. These factors have not previously been implicated in tendon biology. Among these, periostin showed a specific expression in tenocytes of adult mouse Achilles tendon and in chondrocytes within the nonmineralized fibrocartilage zone of the enthesis with the calcaneus. Overexpression of periostin alone or in combination with constitutively active BMP receptor type in human mesenchymal stem cells and subsequent implantation into ectopic sites in mice demonstrated a reproducible moderate tenogenic capacity that has not been described before. Therefore, periostin may belong to the factors contributing to the development of tenogenic tissue.

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Insertional Gene Activation by Lentiviral and Gammaretroviral Vectors

Cited by 119 publications

References 39 publications

Self-Inactivating Alpharetroviral Vectors with a Split-Packaging Design

Self-Inactivating Alpharetroviral Vectors with a Split-Packaging Design

Efficient gene delivery to the adult and fetal CNS using pseudotyped non-integrating lentiviral vectors

Periostin Secreted by Mesenchymal Stem Cells Supports Tendon Formation in an Ectopic Mouse Model

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