Development of Microfluidics for Use in a Novel Way to Characterize Latently HIV-Infected Cells
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2022
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Abstract
Antiretroviral therapy (ART) has been transformative in helping people living with HIV (PLWH) in the last 30 years. Despite the many advances in ART, it cannot eradicate HIV due to its inability to target the HIV latent reservoir. Therapeutic approaches have been considered for eradicating latent HIV such as “shock and kill” and “block and lock”but are still in development and have shown poor results in clinical trials thus far. Laboratory techniques such as STIP-Seq and HIV-Flow have been developed to quantify and characterize the HIV reservoir but do not aim to specifically identify and characterize genuine latently infected cells. Such techniques fall short due to the requirement to activate latently infected cells for identification and analysis, since activation changes cellular gene expression globally. Therefore, we introduce HIV-Seek, a novel approach to characterizing latently HIV infected cells in their pre-activated state. HIV-Seek aims to combine microfluidics, single cell RNA-sequencing, and molecular biology to identify biomarkers for immunological or pharmacological control of the viral reservoir. The focus of this thesis is to establish the development of microfluidics for both its fabrication and function in HIV-Seek alongside the integration of microfluidics and molecular biology. Designs and fabrication conditions were developed for the custom microfluidics for use in HIV-Seek. Encapsulation of beads and cells was performed using microfluidics and a coencapsulation rate of 4.38 ± 0.31% was achieved. Further experimentation with the integration of the molecular biology with the microfluidics is required but shows promise as shown with the dPCR.
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Biomedical engineering, Virology, HIV, HIV Latency, HIV-Seek, microfluidics
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69 pages
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