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

doi:10.1073/pnas.0404180101

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

^*San Raffaele Telethon Institute for Gene Therapy, ^§Institute for Stem Cell Research, and ^∥Vita Salute San Raffaele University, H. San Raffaele Scientific Institute, Via Olgettina 58, 20132 Milan, Italy; and ^**Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037

Edited by Inder M. Verma, The Salk Institute for Biological Studies, La Jolla, CA, and approved August 23, 2004 (received for review June 16, 2004)

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.

Footnotes

↵ †† To whom correspondence may be addressed. E-mail: vescovi.angelo{at}hsr.it or naldini.luigi{at}hsr.it.
↵ † A.C. and A.G. contributed equally to this work.
↵ ‡ Present address: Laboratory of Genetics, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037.
↵ ¶ Present address: Department of Medicine and Pathology, Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Ullmann 625, 1300 Morris Park Avenue, Bronx, NY 10461.
Author contributions: A.L.V. and L.N. designed research; A.C., A.G., D.D., and A.F. performed research; A.C., A.G., C.B., F.H.G., and L.N. analyzed data; A.C., A.G., A.L.V., and L.N. wrote the paper.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: NSC, neural stem cell; SVZ, subventricular zone; RMS, rostral migratory stream; OB, olfactory bulb; LV, lentiviral vector; FGF2, basic fibroblast growth factor; FACS, fluorescence-activated cell sorter; GFAP, glial fibrillary acidic protein; DAPI, 4,6-diamidine-2-phenylindole dihydrochloride; GCL, granule cell layer; MFI, mean fluorescent intensity.