Augmented transgene expression in transformed cells using a parvoviral hybrid vector

Krüger, Lara Katharina; Eskerski, Helmut; Dinsart, Christiane; Cornelis, Jan J.; Rommelaere, Jean; Haberkorn, Uwe; Kleinschmidt, Jürgen A.

doi:10.1038/sj.cgt.7701113

Cited by 6 publications

(5 citation statements)

References 95 publications

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“…These isolates are ideally suited for development into human gene therapy vectors due to their varied tissue tropisms. Advances in the study of mutated capsids, chimeric capsids and modified capsids (such as the addition of single chain antibodies) may further contribute to differences in viral tropism (Bowles et al, 2003; Kruger et al, 2008; Rabinowitz and Samulski, 2000; Wu et al, 2006a; Xiao et al, 1999). Through such modifications of the rAAV combined with cell specific promoters, it may be possible to uniquely tailor a vector allowing transduction of specific cell/tissue types, and increased potential for use as a successful gene therapy technique.…”

Section: Discussionmentioning

confidence: 99%

Convection-enhanced delivery and systemic mannitol increase gene product distribution of AAV vectors 5, 8, and 9 and increase gene product in the adult mouse brain

Carty

Lee

Dickey

et al. 2010

Journal of Neuroscience Methods

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The use of recombinant adeno-associated viral (rAAV) vectors as a means of gene delivery to the central nervous system has emerged as a potentially viable method for the treatment of several types of degenerative brain diseases. However, a limitation of typical intracranial injections into the adult brain parenchyma is the relatively restricted distribution of the delivered gene to large brain regions such as the cortex, presumably due to confined dispersion of the injected particles. Optimizing the administration techniques to maximize gene distribution and gene expression is an important step in developing gene therapy studies. Here, we have found additive increases in distribution when 3 methods to increase brain distribution of rAAV were combined. The convection enhanced delivery (CED) method with the step-design cannula was used to deliver rAAV vector serotypes 5, 8 and 9 encoding GFP into the hippocampus of the mouse brain. While the CED method improved distribution of all 3 serotypes, the combination of rAAV9 and CED was particularly effective. Systemic mannitol administration, which reduces intracranial pressure, also further expanded distribution of GFP expression, in particular, increased expression on the contralateral hippocampi. These data suggest that combining advanced injection techniques with newer rAAV serotypes greatly improves viral vector distribution, which could have significant benefits for implementation of gene therapy strategies.

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Section: Discussionmentioning

confidence: 99%

Convection-enhanced delivery and systemic mannitol increase gene product distribution of AAV vectors 5, 8, and 9 and increase gene product in the adult mouse brain

Carty

Lee

Dickey

et al. 2010

Journal of Neuroscience Methods

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“…Potential promoter activity through the left-end ITR was blocked by insertion of the MVM poly(A) sequence at H1-NS1 position nt1493. It was constructed as follows: pTRH1-Gfp [65]was first cleaved with PmeI/Bst1107I, ligated. The PmeI/Not cleaved GFP cassette was then inserted into the StuI/NotI cleaved P4-less pTRH1-Gfp.…”

Section: Site-directed Mutagenesis and Cloning Proceduresmentioning

confidence: 99%

Vesicular Transport of Progeny Parvovirus Particles through ER and Golgi Regulates Maturation and Cytolysis

2013

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Progeny particles of non-enveloped lytic parvoviruses were previously shown to be actively transported to the cell periphery through vesicles in a gelsolin-dependent manner. This process involves rearrangement and destruction of actin filaments, while microtubules become protected throughout the infection. Here the focus is on the intracellular egress pathway, as well as its impact on the properties and release of progeny virions. By colocalization with cellular marker proteins and specific modulation of the pathways through over-expression of variant effector genes transduced by recombinant adeno-associated virus vectors, we show that progeny PV particles become engulfed into COPII-vesicles in the endoplasmic reticulum (ER) and are transported through the Golgi to the plasma membrane. Besides known factors like sar1, sec24, rab1, the ERM family proteins, radixin and moesin play (an) essential role(s) in the formation/loading and targeting of virus-containing COPII-vesicles. These proteins also contribute to the transport through ER and Golgi of the well described analogue of cellular proteins, the secreted Gaussia luciferase in absence of virus infection. It is therefore likely that radixin and moesin also serve for a more general function in cellular exocytosis. Finally, parvovirus egress via ER and Golgi appears to be necessary for virions to gain full infectivity through post-assembly modifications (e.g. phosphorylation). While not being absolutely required for cytolysis and progeny virus release, vesicular transport of parvoviruses through ER and Golgi significantly accelerates these processes pointing to a regulatory role of this transport pathway.

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“…packaged rAAV genomes into the B19 capsid, which allowed efficient and specific targeting of erythroid cells. Additionally, a chimeric H1/AAV vector, consisting of the tumor-specific replication gene and promoters of the autonomous parvovirus H1, was packaged into an AAV2 capsid, and it was designed to selectively kill tumor cells in vitro 16 . Finally, the most recent report of a chimeric parvovirus vector was published in 2013 17 .…”

Section: Introductionmentioning

confidence: 99%

Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses

Fakhiri

Schneider

Puschhof

et al. 2019

Molecular Therapy - Methods & Clinical Development

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Parvoviruses are highly attractive templates for the engineering of safe, efficient, and specific gene therapy vectors, as best exemplified by adeno-associated virus (AAV). Another candidate that currently garners increasing attention is human bocavirus 1 (HBoV1). Notably, HBoV1 capsids can cross-package recombinant (r)AAV2 genomes, yielding rAAV2/HBoV1 chimeras that specifically transduce polarized human airway epithelia (pHAEs). Here, we largely expanded the repertoire of rAAV/BoV chimeras, by assembling packaging plasmids encoding the capsid genes of four additional primate bocaviruses, HBoV2–4 and GBoV (Gorilla BoV). Capsid protein expression and efficient rAAV cross-packaging were validated by immunoblotting and qPCR, respectively. Interestingly, not only HBoV1 but also HBoV4 and GBoV transduced pHAEs as well as primary human lung organoids. Flow cytometry analysis of pHAEs revealed distinct cellular specificities between the BoV isolates, with HBoV1 targeting ciliated, club, and KRT5+ basal cells, whereas HBoV4 showed a preference for KRT5+ basal cells. Surprisingly, primary human hepatocytes, skeletal muscle cells, and T cells were also highly amenable to rAAV/BoV transduction. Finally, we adapted our pipeline for AAV capsid gene shuffling to all five BoV isolates. Collectively, our chimeric rAAV/BoV vectors and bocaviral capsid library represent valuable new resources to dissect BoV biology and to breed unique gene therapy vectors.

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Augmented transgene expression in transformed cells using a parvoviral hybrid vector

Cited by 6 publications

References 95 publications

Convection-enhanced delivery and systemic mannitol increase gene product distribution of AAV vectors 5, 8, and 9 and increase gene product in the adult mouse brain

Convection-enhanced delivery and systemic mannitol increase gene product distribution of AAV vectors 5, 8, and 9 and increase gene product in the adult mouse brain

Vesicular Transport of Progeny Parvovirus Particles through ER and Golgi Regulates Maturation and Cytolysis

Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses

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