Doctor of Philosophy, The Ohio State University, 2021, Speech and Hearing Science

More than 70% of adults living with human immunodeficiency virus (HIV) or acquired immunodeficiency syndrome (AIDS) are taking lifelong highly active antiretroviral therapy (HAART) for the management of their disease. This includes HIV-positive (HIV+) women who are pregnant and nursing, for whom the World Health Organization (WHO) recommends taking HAART in order to avoid transmitting HIV to their children. To date, studies of HAART exposure in utero have indicated that, though these children are often born HIV-negative (HIV-), they are at increased risk for neurological abnormalities, including auditory effects, when compared to unexposed peers. Previous studies have suggested that non-auditory damage resulting from exposure to HAART is caused by mitochondrial damage and endoplasmic reticulum stress, though it has been noted that these toxicities vary across different cells, organs, and tissues. Little has been done to date to explore the specific manifestations of the auditory differences or their etiology. This is likely due to a high loss-to-follow-up in this population, as many children exposed to HAART in utero leave the healthcare system after being born HIV-, and more than 50% of parents in resource-constrained countries where these treatments are most prevalent report not believing that children can be born with hearing loss. Mining is the largest industry in many resource-constrained countries where HIV is most common, and the current literature suggests that there is significant risk of noise-induced hearing loss associated with work in this area. Additionally, the use of aminoglycoside antibiotics is far more common in these areas for the treatment of diseases like tuberculosis (TB), with more than 1.3 million cases of TB in WHO's African Region alone in 2017. Both of these are potential causes of hearing loss for which those exposed to HAART in utero may be at increased risk. As such, this study also evaluated how exposure to HAART during gestation (open full item for complete abstract)

Committee: Eric Bielefeld (Advisor); Christina Roup (Committee Member); Eric Healy (Committee Member); Ruili Xie (Committee Member) Subjects: Audiology; Pharmacology; Toxicology

Doctor of Philosophy, Case Western Reserve University, 2017, Molecular Biology and Microbiology

Human Immunodeficiency Virus (HIV) infection, while now treatable with antiretroviral therapy (ART), remains a problem for millions of patients worldwide. Patients on ART continue to have an increased risk of Acquired Immunodeficiency Syndrome (AIDS)-related and non-AIDS-related diseases such as cardiovascular disease, cancer, and neurocognitive defects. One potential reason for the increased morbidity and mortality seen in HIV (+) patients is the intestinal barrier defect known to be present in these patients. Decreased barrier function allows for microbial translocation, localized inflammation, partially due to Toll-Like receptor (TLR) ligation, and eventually systemic immune activation. We analyzed the phenotype of monocytes, known players in cardiovascular disease which travel to sites of inflammation, by flow cytometry, as well as TLR phenotype in the colon by immunofluorescence. Our results showed an increase in monocyte activation (HLA-DR, CD86) and decreased chemokine receptor expression (CCR2, CX3CR1) in viremic HIV (+) patients when compared to uninfected controls, which was only partially normalized with ART. Additionally, CD40 is further disrupted on monocytes with ART. Similarly, we find increased TLR3, TLR4, and TLR9 expression in the colons of Viremic HIV (+) patients, whose abundance and location do not fully normalize (TLR4, TLR9), or was further disrupted by ART (TLR3). Together our data suggest that ART, which is successful at controlling viral replication in tissue and HIV RNA levels found in the blood, does not allow for normalization of the immune activation seen in the HIV (+) patients, and in fact could be contributing to the continued pathology.

Committee: Alan Levine (Advisor); John Tilton (Committee Chair); David McDonald (Committee Member); Calvin Cotton (Committee Member) Subjects: Immunology; Pathology; Virology

PHD, Kent State University, 2007, College of Biomedical Sciences

HIV/AIDS pandemic presents a huge challenge to public health with over 40 million people living with HIV/AIDS worldwide. Current treatment is the highly active antiretroviral therapy (HAART). HAART is a combination of two nucleoside reverse transcriptase inhibitors (NRTI), a protease inhibitor (PI) or a non-nucleoside reverse transcriptase inhibitor (NNRTI), with the fusion inhibitor added as required. HAART has increased patient life expectancy by an average of 8 years. However; these drugs are still expensive, unavailable to over 90% of eligible individuals and associated with widespread drug resistance culminating into therapy failure. Thus, there is an urgent need to identify and characterize novel antiretroviral drugs. In this dissertation, small molecule libraries were screened with either a GFP-tagged lentiviral vector or replicative-competent, fluorescent protein-tagged HIV-1 isolates. Rationally designed compounds were also analyzed for their antiretroviral activity. Characterization of the mechanism(s) of action of these compounds included assessment of compound cellular toxicity (CC50) by trypan blue exclusion and tetrazolium-based colorimetric assays. Viral susceptibility was determined from the IC50 of these compounds against primary and drug-resistant isolates. Their mechanism(s) of antiretroviral activity was confirmed by specific in vitro biochemical assays, time of addition assays and serial passages to select resistant HIV-1 strains. The relevance of drug resistant mutations was assessed by antiviral assays, viral growth kinetics and site-directed mutagenesis. Nine potential candidates for further antiretroviral drug development were identified in this dissertation. Three compounds, CBL 4.0, CBL 4.1 and CBL 4.3 were structurally similar to INDOPY-1, a first-in-class novel nucleoside-competing reverse transcriptase inhibitor (NcRTI) recently described. CBL 17, CBL 21 and CBL 26 were novel non-nucleoside reverse transcriptase inhibitors. However, CBL (open full item for complete abstract)

Committee: Miguel Quiñones-Mateu (Advisor) Subjects:

Master of Sciences, Case Western Reserve University, 2011, Clinical Research

Background: Lipoatrophy, subcutaneous fat wasting, is a stigmatizing complication of antiretroviral therapy (ART). Thiazolidenediones are potent PPARγ agonists and expected to reverse lipoatrophy. Methods: We evaluated rosiglitazone's effect on several facets of lipoatrophy: 1) limb fat, 2) surrogate markers cardiovascular disease, and 3) mechanistic measures. We conducted a 48-week randomized, double-blind placebo-controlled trial of rosiglitazone in HIV-infected subjects with lipoatrophy on thymidine nucleoside reverse transcriptase inhibitors (tNRTI)-sparing regimens. Results: Rosiglitazone significantly (p=0.02) improved peripheral lipoatrophy in the absence of tNRTIs. Carotid intima media thickness (IMT), oxidative, and some inflammatory markers worsened over time in these subjects on stable ART, regardless of the rosiglitazone assignment. Conclusion: In the absence of tNRTIs, rosiglitazone improved peripheral lipoatrophy. Observed limb fat improvement after rosiglitazone was not associated with changes in surrogate markers of cardiovascular disease or mitochondrial indices. Lipoatrophy can in part be overcome by the PPAR-γ pathway independent of mitochondrial toxicity.

Committee: James Spilsbury PhD (Committee Chair); Grace McComsey MD (Committee Member); Wilson Tang MD (Committee Member) Subjects: Medicine

Master of Sciences, Case Western Reserve University, 2009, Epidemiology and Biostatistics

Understanding sexual behaviors of persons on Antiretroviral therapy(ART) is critical designing and implementation of positive prevention programs. 559 HIV infected adults were enrolled in clinic based prospective observational study and sexual behaviors ascertained at ART initiation and semi-annually thereafter. Using Logistic regression with generalized estimating equations, factors associated with sexual activity and unprotected sex were examined. Over the first three years of ART, the proportion sexually active did not change at ~52% (χ2 Trend, p=0.94) while the proportion reporting unprotected sex decreased (χ2 Trend, p<0.0001). Men reported unprotected sex less often than women (p<0.0001). In all analyses, having no children and female gender (controlling for any other factors) was associated with the practice of unprotected sex. The interventional effect of comprehensive HIV care resulted in marked reductions in unprotected sex particularly among men. Strengthening of positive prevention interventions, especially among females are needed in ART programs in this setting.

Committee: Ajay Sethi Phd,MHS (Committee Chair); Christopher Whalen MD, MS (Committee Member); Jeffrey Albert Phd (Committee Member) Subjects: Biostatistics; Epidemiology; Health; Health Care; Virology

Master of Sciences, Case Western Reserve University, 0, Biology

The single nucleotide polymorphisms (SNPs) in CCR5 (-2459 G>A [promoter], ¿32 deletion polymorphism [open reading frame, ORF]) influences the expression of CCR5 chemokine receptor on human T lymphocytes. CCR5 is a major coreceptor for HIV to bind to the T cell prior to fusion. Individuals have various frequencies of these polymorphisms, which influence their susceptible to HIV infection and disease progression. This thesis is focused on determining the association between these polymorphisms and CCR5 expression on T cells, as well as between these polymorphisms and in vitro HIV infection. The CCR5 -2459 genotype results that were obtained showed clear associations with CCR5 expression on the central memory T cell (TCM) subpopulation. That is, CCR5 -2459 G/G had lower levels of CCR5 expression as compared to A/A and G/A on the TCM cells. While with the CCR5 ORF genotypes, wildtype (Wt)/¿32 had very low levels of CCR5 expression as compared to Wt/Wt. Furthermore, results obtained for CCR5 -2459 genotype and HIV infectivity of the TCM cells also clearly showed significant associations. These were observed when comparing CCR5 -2459 genotypes G/G and A/A (p=0.029), and also between G/A and G/G (p=0.049). That is, G/G had lower HIV infectivity of its TCM cells in comparison to those of both A/A and G/A genotypes. These findings, especially those of CCR5 -2459 G allele demonstrates its protective effect in the in vitro assay, thus, providing insights into expression and infection at the T cell subpopulation level.

Committee: Peter Zimmerman (Advisor) Subjects: Biology; Biomedical Research; Cellular Biology; Genetics; Health Sciences; Immunology