The identification of human immunodeficiency virus type 1 (HIV-1) as the etiological agent of AIDS has led to the proposal of novel intervention strategies to block HIV-1 infection and viral replication. Because HIV-1 infection can be detected early, when the vast majority of the target cells are uninfected, gene therapy is an attractive approach. Our long-ter strategy is to stably transduce pluripotent hematopoietic stem cells with anti-HIV-1 agents that will confer """"""""intracellular immunity"""""""" to progeny CD4+ lymphocytes and macrophages, athe permissive host for HIV-1. This approach necessitates; (1) the ability to functionally transduce pluripotent hematopoietic stem cells; and (2) an agent that confers resistance to HIV- 1. We will systematically test the ability of established oncoretroviral and AAV-based gene transfer vectors to functionally transduce human and murine hematopoietic stem cells. We will also examine various well- established methods for gene transfer for their ability to introduce and express transgenes with minimal disruption to cell viability and renewal potential. We will also actively explore the use of HIV-1 pseudotyped vectors. As described in the proposal, these vectors may be advantageous for targetting non-dividing cells., such as pluripotent stem cells. The efficacy of these HIV-based vectors will be assessed in parallel with oncoretroviral and AAV-vectors. Secondly, we will continue to develop the inhibitory potential of two novel peptide inhibitors of the essential HIV-1 regulatory protein, Rev. We have previously identified two novel protein inhibitors of Rev; a new class of dominant-negative Rev mutants and a selected peptide that binds to Rev's effector domain. The potency of these new Rev inhibitors suggest they may be efficacious anti-viral agents. We will perform a series of experiments to directly test their ability to block HIV-1 replication. We will continue to analyze their mechanism of action using biochemical structure- function experiments. These studies will facilitate the development of other potent peptide derivatives.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL056920-05
Application #
6358985
Study Section
Project Start
2000-09-29
Project End
2002-08-31
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
5
Fiscal Year
2000
Total Cost
$153,773
Indirect Cost
Name
University of Massachusetts Medical School Worcester
Department
Type
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
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Colvin, Gerald A; Lambert, Jean-Francois; Dooner, Mark S et al. (2007) Murine allogeneic in vivo stem cell homing(,). J Cell Physiol 211:386-91
Kieusseian, Aurelie; Chagraoui, Jalila; Kerdudo, Cecile et al. (2006) Expression of Pitx2 in stromal cells is required for normal hematopoiesis. Blood 107:492-500
Aliotta, Jason M; Keaney, Patrick; Passero, Michael et al. (2006) Bone marrow production of lung cells: the impact of G-CSF, cardiotoxin, graded doses of irradiation, and subpopulation phenotype. Exp Hematol 34:230-41
Zhang, Hui Z; Degar, Barbara A; Rogoulina, Svetlana et al. (2006) Hematopoiesis following disruption of the Pitx2 homeodomain gene. Exp Hematol 34:167-78
Abedi, Mehrdad; Greer, Deborah A; Foster, Bethany M et al. (2005) Critical variables in the conversion of marrow cells to skeletal muscle. Blood 106:1488-94
Quesenberry, Peter J; Colvin, Gerald; Abedi, Mehrdad (2005) Perspective: fundamental and clinical concepts on stem cell homing and engraftment: a journey to niches and beyond. Exp Hematol 33:9-19
Quesenberry, Peter J; Colvin, Gerald A; Abedi, Mehrdad et al. (2005) The stem cell continuum. Ann N Y Acad Sci 1044:228-35
Colvin, Gerald A; Lambert, Jean-Francois; Abedi, M et al. (2004) Differentiation hotspots: the deterioration of hierarchy and stochasm. Blood Cells Mol Dis 32:34-41
Nowakowski, Grzegorz S; Dooner, Mark S; Valinski, Helen M et al. (2004) A specific heptapeptide from a phage display peptide library homes to bone marrow and binds to primitive hematopoietic stem cells. Stem Cells 22:1030-8

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