Date of Award

3-2020

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Program

Pharmaceutical Sciences

Track

Pharmacometrics

Research Advisor

Santosh Kumar, Ph.D.

Committee

Sarka Beranova, Ph.D.; Theodore J. Cory, Ph.D., Pharm.D.; Francesca-Fang Liao, Ph.D.; Murali M. Yallapu, Ph.D.

Keywords

Blood-brain barrier, Elvitegravir, HIV-associated neurocognitive disorder, Macrophages, Microglia, Nanoparticle

Abstract

Over the last two decades, the use of antiretroviral therapy (ART) has remarkably decreased the morbidity associated with HIV-1 infection. However, the prevalence of HIV-1-associated neurocognitive disorders (HAND) is still increasing. The appearance and persistence of HAND are partially due to the entry of HIV-1-infected monocytes into the brain. Within the central nervous system (CNS), HIV-1-infected macrophages and microglia serve as the major viral reservoirs and provide active viral replication even when systemic viral suppression has been achieved by ART. Efficient viral suppression of CNS macrophages and microglia is important for an effective HIV-1 treatment in the brain. The primary hindrance to the treatment of HAND is mainly contributed by the inability of antiretrovirals (ARVs) to cross the blood-brain barrier (BBB) after the systemic administration. Conventional regimens of ARVs are not sufficient to penetrate the BBB or improve outcomes in HAND. Thus, there is a need for novel treatment regimens that cross the BBB and deliver therapeutic ARVs into the CNS to suppress the HIV-1 replication in these viral reservoirs. Our objective in this study was to improve the efficacy of ART in CNS HIV-1 reservoirs, specifically in macrophages and microglia. Our central hypothesis was that the selected ARV, elvitegravir (EVG), in nanoformulation would have increased transmigration across the BBB, as well as increased drug uptake in macrophages and microglia after crossing the BBB, leading to increased antiviral activity in macrophages and microglia, relative to the native drug. This study was expected to provide an optimized treatment strategy that has potential for therapeutic interventions in reducing HAND.In this study, poly (lactic-co-glycolic acid) (PLGA)-based elvitegravir nanoparticles (PLGA-EVG NPs) were prepared by nano-precipitation. Firstly, we analyzed the physicochemical properties of PLGA-EVG NPs using transmission electron microscopy, dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FTIR). We measured cellular uptake of PLGA NPs by fluorescence microscopy and flow cytometry. We also measured the intracellular drug concentration and viral replication in HIV-1-infected macrophages by using LC-MS/MS and p24 ELISA, respectively. The PLGA-E

ORCID

https://orcid.org/0000-0001-6373-5440

DOI

10.21007/etd.cghs.2020.0497

Additional Files
2020-003-Gong-DOA.pdf (423 kB)
Declaration of Authorship
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