Engineering chimeric antigen receptors for durable control over HIV-1 replication / Rachel Shira Leibman.

Format:

Book

Manuscript

Thesis/Dissertation

Author/Creator:

Leibman, Rachel Shira, author.

Contributor:

Riley, James L., degree supervisor.

Hoxie, James A., degree committee member.

Kohli, Rahul M., degree committee member.

Vonderheide, Robert H., degree committee member.

Wherry, E. John, degree committee member.

University of Pennsylvania. Department of Cell and Molecular Biology, degree granting institution.

Language:

English

Subjects (All):

Penn dissertations--Cell and molecular biology.

Cell and molecular biology--Penn dissertations.

Local Subjects:

Penn dissertations--Cell and molecular biology.

Cell and molecular biology--Penn dissertations.

Physical Description:

x, 176 leaves : illustrations ; 29 cm

Production:

[Philadelphia, Pennsylvania] : University of Pennsylvania, 2017.

Summary:

This thesis project aimed to develop chimeric antigen receptors (CARs) capable of durably suppressing the Human Immunodeficiency Virus Type 1 (HIV) replication, by building upon a previous CD4-based CAR that was employed in several clinical trials. We applied lessons learned from cancer-targeting CARs to optimize the CAR vector backbone, promoter, HIV targeting moiety, and transmembrane and signaling domains, in an effort to determine which components augmented the ability of CD8 T cells to control HIV replication. CD8 T cells expressing the optimized CARs were at least 50-fold more potent in vitro at controlling HIV replication than the original CD4 CAR or TCR-based approaches and substantially better than broadly neutralizing antibody-based CARs. We then utilized a humanized mouse model of HIV infection to demonstrate superior control over HIV replication, better protection of CD4 T cells, and greater CAR T cell expansion with the optimized vectors compared to the original clinical trial vector. Compared to optimized CD4 CARs containing the CD28 costimulatory domain, CARs containing 4-1BB expanded better in vivo in the absence of antigen and resulted in greater control over HIV replication.We found that the CD4 CAR promoted infection of transduced CD8 T cells and employed CCR5 zinc finger nucleases (ZFNs) or a GP41-based fusion inhibitor to protect the CAR T cells. We employed ZFN-pretreated, CAR-transduced CD8 T cells in our mouse models and saw an enrichment of the disrupted alleles in HIV-infected mice compared to mock controls.In humans, a functional cure will require CAR T cells to prevent the spread of HIV following virus reactivation from the latent reservoir. We modeled this scenario in vitro using ART patient T cells and latency reversing agents (LRAs). Preliminary data suggest that CD4 CAR T cells can respond to low levels of antigen produced by resting ART patient cells in the presence of LRAs. Together, these data indicate that potent HIV-specific T cells can be generated using improved CAR design and provide optimism that CAR T cells could help achieve a functional cure.

Notes:

Ph. D. University of Pennsylvania 2017.

Department: Cell and Molecular Biology.

Supervisor: James L. Riley.

Includes bibliographical references.

OCLC:

1231652201