PRECLINICAL DEVELOPMENT OF RNA DECOYS

Project: Research project

Description

The goals of the proposed studies are to conduct a thorough preclinical
evaluation of promising HIV resistance genes in primary CD4+ lymphocytes,
and to establish appropriate procedures and necessary logistical support
for clinical studies targeting peripheral T cells. We haven previously
shown that expression of TAR decoy RNA in immortalized CD4+ T-cells
provided significant protection against HIV replication, and initial
studies have indicated that transduction of primary CD4+ PBL obtained from
healthy volunteers with TAR decoy vectors led to a modest though
reproducible inhibition of HIV replication. We have recently identified a
highly potent RRE-decoy comprising of the 13 nt-long minimal Rev binding
domain of the HIV-1 RRE which unlike TAR decoy RNA is not known to bind
cellular factors that could be sequestered in uninfected cells expressing
decoy RNA, and potentially compromise the viability or function of the
genetically modified cells. The specific objectives of the proposed studies are: (a) To develop
increasingly potent inhibition strategies based on RRE-decoys and
combination vectors including TAR decoys, and rev transdominants and
ribozymes. (b) To evaluate inhibition strategies in ex vivo cultured CD4+
PBL obtained from healthy volunteers and HIV infected individuals
including the development of methods to maximize efficiency of retroviral
gene transfer, comparative evaluation of HIV resistance genes against
clinical HIV isolates, and the development of highly accurate and
sensitive PCR-based methods to measure gene transfer and HIV replication
in preclinical models and clinical settings. (c) To establish procedures
and carry out careful and exhaustive safety studies as mandated by the
FDA, in ex vivo cultured CD4+ PBL and in hematochimeric SCID mice
reconstituted with CD4+ PBL obtained from HIV infected individuals. The studies proposed in this application will set the stage for clinical
evaluation of promising HIV resistance genes in a peripheral T-cell-
targeted gene therapy protocol, and will provide the basis for clinical
studies targeting hemopoietic stem cells.
StatusFinished
Effective start/end date5/1/941/31/98

Funding

  • National Institutes of Health: $182,210.00
  • National Institutes of Health: $174,253.00
  • National Institutes of Health
  • National Institutes of Health

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HIV
RNA
Genes
T-Lymphocytes
Healthy Volunteers
Genetic Therapy
HIV-1
Stem Cells
Lymphocytes
Safety
Polymerase Chain Reaction
Catalytic RNA
SCID Mice
Volunteers

ASJC

  • Medicine(all)
  • Immunology and Microbiology(all)