Efficiency of adeno-associated virus type-2 vectors in non-human primate Schwann cells

J. Neurochem. (2010) 115, 1137–1149. Abstract For biotechnological applications, insect cell lines are primarily known as hosts for the baculovirus expression system that is capable to direct synthesis of high levels of recombinant proteins through use of powerful viral promoters. Here, we demonstrate the implementation of two alternative approaches based on the baculovirus system for production of a mammalian recombinant glycoprotein, comprising the extracellular part of the cell adhesion molecule L1, with potential important therapeutic applications in nervous system repair. In the first approach, the extracellular part of L1 bearing a myc tag is produced in permanently transformed insect cell lines and purified by affinity chromatography. In the second approach, recombinant baculoviruses that express L1‐Fc chimeric protein, derived from fusion of the extracellular part of L1 with the Fc part of human IgG1, under the control of a mammalian promoter are used to infect mammalian HEK293 and primary Schwann cells. Both the extracellular part of L1 bearing a myc tag accumulating in the supernatants of insect cultures as well as L1‐Fc secreted by transduced HEK293 or Schwann cells are capable of increasing the motility of Schwann cells with similar efficiency in a gap bridging bioassay. In addition, baculovirus‐transduced Schwann cells show enhanced motility when grafted on organotypic cultures of neonatal brain slices while they retain their ability to myelinate CNS axons. This proof‐of‐concept that the migratory properties of myelin‐forming cells can be modulated by recombinant protein produced in insect culture as well as by means of baculovirus‐mediated adhesion molecule expression in mammalian cells may have beneficial applications in the field of CNS therapies.

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“…While mammalian viral vectors derived from retroviruses, lentiviruses, adenovirus or adeno‐associated virus (Girard et al. 2005; Watanabe et al.…”

Section: Discussionmentioning

confidence: 99%

Soluble forms of the cell adhesion molecule L1 produced by insect and baculovirus‐transduced mammalian cells enhance Schwann cell motility

Lavdas

Efrose

Douris

et al. 2010

Journal of Neurochemistry

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“…Adeno-associated viral vectors are increasingly regarded as safe and are well-tolerated following application to the human brain. AAV2 and 8 transduce primate and rat Schwann cells ( Girard et al, 2005 ; Homs et al, 2011 ). As a first step toward optimizing AAV-mediated gene transfer to Schwann cells we performed a screen of all 9 common serotypes and showed that AAV2 vectors outperform other serotypes in transducing Schwann cells in human peripheral nerve explants, whereas several AAV serotypes efficiently transduced rat Schwann cells ( Hoyng et al, 2015 ).…”

Section: Perspective On Future Steps To Accomplish Recovery Of Functimentioning

confidence: 99%

GDNF Gene Therapy to Repair the Injured Peripheral Nerve

Eggers

Winter

Tannemaat

et al. 2020

Front. Bioeng. Biotechnol.

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A spinal root avulsion is the most severe proximal peripheral nerve lesion possible. Avulsion of ventral root filaments disconnects spinal motoneurons from their target muscles, resulting in complete paralysis. In patients that undergo brachial plexus nerve repair, axonal regeneration is a slow process. It takes months or even years to bridge the distance from the lesion site to the distal targets located in the forearm. Following ventral root avulsion, without additional pharmacological or surgical treatments, progressive death of motoneurons occurs within 2 weeks ( Koliatsos et al., 1994 ). Reimplantation of the avulsed ventral root or peripheral nerve graft can act as a conduit for regenerating axons and increases motoneuron survival ( Chai et al., 2000 ). However, this beneficial effect is transient. Combined with protracted and poor long-distance axonal regeneration, this results in permanent function loss. To overcome motoneuron death and improve functional recovery, several promising intervention strategies are being developed. Here, we focus on GDNF gene-therapy. We first introduce the experimental ventral root avulsion model and discuss its value as a proxy to study clinical neurotmetic nerve lesions. Second, we discuss our recent studies showing that GDNF gene-therapy is a powerful strategy to promote long-term motoneuron survival and improve function when target muscle reinnervation occurs within a critical post-lesion period. Based upon these observations, we discuss the influence of timing of the intervention, and of the duration, concentration and location of GDNF delivery on functional outcome. Finally, we provide a perspective on future research directions to realize functional recovery using gene therapy.

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Gene delivery to rat and human Schwann cells and nerve segments: a comparison of AAV 1–9 and lentiviral vectors

et al. 2015

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Schwann cells (SCs) in an injured peripheral nerve form pathways for regenerating axons. Although these cells initially support regeneration, SCs lose their pro-regenerative properties following a prolonged period of denervation. Gene transfer to SC can enhance their therapeutic potential. In this article, we compared adeno-associated viral (AAV) vectors based on serotypes 1-9 for their capability to transduce cultured primary rat and human SCs and nerve segments. AAV1 is the best serotype to transduce rat SCs, whereas AAV2 and AAV6 performed equally well in human SCs. Transduction of monolayers of cultured rat and human SCs did not accurately predict the transduction efficiency in nerve segments. Rat nerve segments could be genetically modified equally well by a set of four AAV vectors (AAV1, AAV5, AAV7, AAV9), whereas AAV2 was superior in human nerve segments. The current experiments were undertaken as a first step towards future clinical implementation of ex vivo AAV-based gene therapy in surgical nerve repair. The transduction of rat and human SCs and nerve segments by entirely different AAV serotypes, as documented here, highlights one of the challenges of translating gene therapy from experimental animals to human patients.

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Efficiency of adeno-associated virus type-2 vectors in non-human primate Schwann cells

Cited by 3 publications

References 25 publications

Soluble forms of the cell adhesion molecule L1 produced by insect and baculovirus‐transduced mammalian cells enhance Schwann cell motility

Soluble forms of the cell adhesion molecule L1 produced by insect and baculovirus‐transduced mammalian cells enhance Schwann cell motility

GDNF Gene Therapy to Repair the Injured Peripheral Nerve

Gene delivery to rat and human Schwann cells and nerve segments: a comparison of AAV 1–9 and lentiviral vectors

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