Recombinant AAV2 transduction of primitive human hematopoietic stem cells capable of serial engraftment in immune-deficient mice

doi:10.1073/pnas.0502902102

Recombinant AAV2 transduction of primitive human hematopoietic stem cells capable of serial engraftment in immune-deficient mice

Divisions of ^*Virology and ^†Hematology/Hematopoietic Cell Transplantation, City of Hope National Medical Center, Duarte, CA 91010; and ^§Department of Obstetrics and Gynecology, University of Southern California and Huntington Memorial Hospital, Pasadena, CA 91105

Edited by Kenneth I. Berns, University of Florida College of Medicine, Gainesville, FL, and approved May 24, 2005 (received for review April 7, 2005)

Abstract

A recombinant AAV2 (rAAV2) vector encoding antisense RNA to HIV-1 transactivating region (TAR) was evaluated for transduction of human cord blood CD34⁺CD38^- hematopoietic stem cells (HSC) capable of serial engraftment in nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mice. Results revealed long-term multilineage marking in primary and secondary recipients, and significantly, an enrichment of transduced cells in secondary hosts, indicating efficient transduction of multipotential self-renewing HSC. These results were confirmed by the persistence of rAAV marking of clonogenic progenitors in serial analyses of recipient marrow. Upon HIV-1 challenge, the macrophage progeny of transduced CD34⁺ cells expressed antisense RNA and exhibited sustained and significant inhibition of virus replication as compared with controls in every donor tested, without selective pressure. This study represents a clear in vivo demonstration of efficient rAAV2 transduction of human HSC.

Footnotes

↵ ¶ To whom correspondence should be addressed. E-mail: schatterjee{at}coh.org.
↵ ‡ L.S. and H.P. contributed equally to this work.
Author contributions: L.S., H.P., C.W., L.L., and S.C. performed research; L.S., H.P., C.W., L.L., K.K.W., and S.C. analyzed data; J.M., S.F., K.K.W., and S.C. contributed new reagents/analytic tools; K.K.W. and S.C. designed research; and L.S., H.P., K.K.W., and S.C. wrote the paper.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations: HSC, hematopoietic stem cell; SCID, severe combined immunodeficient; NOD, nonobese diabetic; rAAV, recombinant AAV2; CFU-C, colony-forming unit-culture.