Robust in vivo gene transfer into adult mammalian neural stem cells by lentiviral vectors

Abstract

Stable genetic modification of adult stem cells is fundamental for both developmental studies and therapeutic purposes. Using in vivo marking studies, we showed that injection of lentiviral vectors (LVs) into the subventricular zone of the adult mouse brain enables efficient gene transfer into long-term self-renewing neural precursors and steady, robust vector expression in their neuronal progeny throughout the subventricular zone and its rostral extension, up to the olfactory bulb. By clonal and population analysis in culture, we proved that in vivo-marked neural precursors display self-renewal and multipotency, two essential characteristics of neural stem cells (NSCs). Thus, LVs efficiently target long-term repopulating adult NSCs, and the effect of the initial transduction is amplified by the continuous generation of NSC-derived, transduced progeny. LVs may thus allow novel studies on NSCs' physiology in vivo, and introduction of therapeutic genes into NSCs may allow the development of novel approaches for untreatable CNS diseases.

Adeno-associated virus type 4 (AAV4) targets ependyma and astrocytes in the subventricular zone and RMS

“…6 NPCs have also been successfully labeled with recombinant retroviruses. 7,8 Adeno-associated virus (AAV) vectors mediate longterm stable expression and induce minimal immune responses, which make them attractive vehicles for in vivo gene transfer to NPCs. While prior work with AAV type 2 (AAV2) vectors showed poor NPC transduction following direct SVZ injection, AAV2 did successfully transduce human NPCs in vitro.…”

Adult neurogenesis in non‐mammalian vertebrates

“…(54) Lentiviruses, in contrast, can incorporate into quiescent cells in vivo, and therefore have been used to trace the progeny of SEZ and SGZ stem cells. (55,56) Using this approach, the multipotency of single infected cells has to date mostly been followed using the neurosphere method, upon dissociation and culture of a lentivirus-infected progenitor zone. (55) In teleosts, neurospheres from the brown ghost telencephalon and cerebellum can differentiate into neurons and GFAP-or vimentin-positive glial cells, (47) but the limitations involved in the interpretation of neurosphere studies, clearly apply here as well.…”

“…(55,56) Using this approach, the multipotency of single infected cells has to date mostly been followed using the neurosphere method, upon dissociation and culture of a lentivirus-infected progenitor zone. (55) In teleosts, neurospheres from the brown ghost telencephalon and cerebellum can differentiate into neurons and GFAP-or vimentin-positive glial cells, (47) but the limitations involved in the interpretation of neurosphere studies, clearly apply here as well. In order to address the multipotentiality of progenitors in vivo, it is crucial to follow these cells without passaging in culture.…”

Evaluation of the specificity and sensitivity of ferritin as an MRI reporter gene in the mouse brain using lentiviral and adeno-associated viral vectors

“…As LVs efficiently transduce non-dividing post-mitotic and quiescent cells, they are ideal vehicles for gene transfer into the central nervous system (CNS), including endogenous neural stem and progenitor cells. [20][21][22] LVs are integrating vectors that stably mark cells and their progeny. After transduction of cells with LVs, the labeling will expand stoichiometrically as cells proliferate, providing a non-invasive means to follow cell progeny longitudinally.…”