‘Advanced’ generation lentiviruses as efficient vectors for cardiomyocyte gene transduction in vitro and in vivo

Bonci, Desirée; Cittadini, Antonio; Latronico, Michael V.G.; Borello, Ugo; Aycock, Joyce; Drusco, Alessandra; Innocenzi, A; Follenzi, Antonia; Lavitrano, Marialuisa; Monti, Maria Gaia; Ross, John; Naldini, Luigi; Peschle, Cesare; Cossu, Giulio; Condorelli, Gianluigi

doi:10.1038/sj.gt.3301936

Cited by 107 publications

(75 citation statements)

References 15 publications

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“…3,[13][14][15] Transplantation of freshly transduced cardiomyocytes into the adult rat brain and muscle resulted in long-term transgene expression for at least 12 weeks (data not shown 16 ).…”

Section: Discussionmentioning

confidence: 92%

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Shuttle of lentiviral vectors via transplanted cells in vivo

et al. 2004

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Section: Discussionmentioning

confidence: 92%

“…3,[13][14][15] Cardiomyocytes survived transduction and long-term culture for at least 14 days, although progressive lentivirus-independent de-differentiation became visible during the culture.…”

Section: Resultsmentioning

confidence: 99%

Shuttle of lentiviral vectors via transplanted cells in vivo

et al. 2004

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“…[12][13][14] Immunohistochemistry analysis of cross-sectional myocardial slices sampled each millimeter with anti-hemoagglutinin (HA) monoclonal antibody showed that approximately 70% of cardiomyocytes were positive ( Figure 1, panels a and b). Moreover, tissue histology analysis demonstrated a uniform distribution of the adenoviral infection with adenovirus with wild-type Akt (Ad.Akt) throughout the myocardium.…”

Section: Cardiac Gene Transfermentioning

confidence: 99%

“…31 The viral delivery procedure has been described previously in detail. 12 Briefly, rats were anesthetized with a mixture of ketamine hydrochloride (Sigma, 50 mg/kg BW) and xylazine (Sigma, 10 mg/kg BW) and then orally intubated and ventilated. The animals were subsequently cooled with ice bags and their temperature monitored with a thermistor catheter.…”

Section: Experimental Protocolmentioning

confidence: 99%

Adenoviral gene transfer of Akt enhances myocardial contractility and intracellular calcium handling

et al. 2005

Self Cite

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The serine-threonine kinase Akt/PKB mediates stimuli from different classes of cardiomyocyte receptors, including the growth hormone/insulin like growth factor and the b-adrenergic receptors. Whereas the growth-promoting and antiapoptotic properties of Akt activation are well established, little is known about the effects of Akt on myocardial contractility, intracellular calcium (Ca 2+ ) handling, oxygen consumption, and b-adrenergic pathway. To this aim, Sprague-Dawley rats were subjected to a wild-type Akt in vivo adenoviral gene transfer using a catheter-based technique combined with aortopulmonary crossclamping. Left ventricular (LV) contractility and intracellular Ca 2+ handling were evaluated in an isolated isovolumic bufferperfused, aequorin-loaded whole heart preparations 10 days after the surgery. The Ca 2+ -force relationship was obtained under steady-state conditions in tetanized muscles. No significant hypertrophy was detected in adenovirus with wild-type Akt (Ad.Akt) versus controls rats (LV-to-body weight ratio 2.670.2 versus 2.770.1 mg/g, controls versus Ad.Akt, P, NS). LV contractility, measured as developed pressure, increased by 41% in Ad.Akt. This was accounted for by both more systolic Ca 2+ available to the contractile machinery (+19% versus controls) and by enhanced myofilament Ca 2+ responsiveness, documented by an increased maximal Ca 2+ -activated pressure (+19% versus controls) and a shift to the left of the Ca 2+ -force relationship. Such increased contractility was paralleled by a slight increase of myocardial oxygen consumption (14%), while titrated dose of dobutamine providing similar inotropic effect augmented oxygen consumption by 39% (Po0.01). Phospholamban, calsequestrin, and ryanodine receptor LV mRNA and protein content were not different among the study groups, while sarcoplasmic reticulum Ca 2+ ATPase protein levels were significantly increased in Ad.Akt rats. b-Adrenergic receptor density, affinity, kinase-1 levels, and adenylyl cyclase activity were similar in the three animal groups. In conclusion, our results support an important role for Akt/PKB in the regulation of myocardial contractility and mechanoenergetics.

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“…30 While the relative ability of lentiviral vectors to transduce postmitotic cells versus dividing cells has been controversial, several modifications to the vector backbone may have resulted in an increased ability to transduce nondividing cells. 13,31,32 One such modified, third-generation VSV-Gpseudotyped lentiviral vector has been used to transfer an EPO cDNA into rat skeletal muscles. 13 MacKenzie et al 14 reported that VSV-G-pseudotyped lentiviral vectors 14 we explored the potential of VSV-G vectors to transduce a variety of mouse skeletal muscle cells both in vitro and in vivo.…”

Section: Discussionmentioning

confidence: 99%

Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin

Kimura

Fall

et al. 2005

Gene Ther

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Gene therapy for Duchenne muscular dystrophy (DMD) will require sustained expression of therapeutic dystrophins in striated muscles. Lentiviral vectors have a relatively large transgene carrying capacity and can integrate into nondividing cells. We therefore explored the use of lentiviral vectors for transferring genes into mouse skeletal muscle cells. These vectors successfully transferred a minidystrophin expression cassette into mdx muscles, and minidystrophin expression persisted and prevented subsequent muscle fiber degeneration for at least 6 months. However, only low to moderate levels of skeletal muscle transduction could be obtained by intramuscular injection of the highest currently available lentiviral doses. Using cultured cells, the lentiviral vectors effectively transduced proliferating and terminally differentiated muscle cells, indicating that cell cycling is not essential for transduction of myogenic cells. We further showed that lentiviral vectors efficiently transduced both primary myoblasts and multipotent adult progenitor cells (MAPCs) in vitro, and the cells persistently expressed transgenes without any obvious toxicity. When mdx primary myoblasts were genetically modified with minidystrophin vectors and transplanted into mdx skeletal muscles, significant numbers of dystrophin-expressing myofibers formed. Finally, we showed that a short, highly active CK6 regulatory cassette directed muscle-specific activity in the context of the lentiviral vectors. The ability of lentiviral vectors to transduce myogenic progenitors using a minidystrophin cassette regulated by a muscle-specific promoter suggests that this system could be useful for ex vivo gene therapy of muscular dystrophy.

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‘Advanced’ generation lentiviruses as efficient vectors for cardiomyocyte gene transduction in vitro and in vivo

Cited by 107 publications

References 15 publications

Shuttle of lentiviral vectors via transplanted cells in vivo

Shuttle of lentiviral vectors via transplanted cells in vivo

Adenoviral gene transfer of Akt enhances myocardial contractility and intracellular calcium handling

Stable transduction of myogenic cells with lentiviral vectors expressing a minidystrophin

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