A5086767504- HIV Research and Treatment
- Immune Cell Function and Interaction
- HIV/AIDS Research and Interventions
- RNA Interference and Gene Delivery
- T-cell and B-cell Immunology
- Advanced biosensing and bioanalysis techniques
- Neuroinflammation and Neurodegeneration Mechanisms
- HIV/AIDS drug development and treatment
- Epigenetics and DNA Methylation
- HIV-related health complications and treatments
- Cytomegalovirus and herpesvirus research
- Reproductive System and Pregnancy
- RNA Research and Splicing
- MicroRNA in disease regulation
- Systemic Sclerosis and Related Diseases
- Extracellular vesicles in disease
- ATP Synthase and ATPases Research
- Immune cells in cancer
- Eosinophilic Esophagitis
- HIV/AIDS oral health manifestations
- Immune Response and Inflammation
- Transplantation: Methods and Outcomes
- Protein Degradation and Inhibitors
- Monoclonal and Polyclonal Antibodies Research
- RNA regulation and disease
The Wallace H. Coulter Department of Biomedical Engineering
2021-2024
Georgia Institute of Technology
2021-2024
Icahn School of Medicine at Mount Sinai
2023-2024
University School
2013-2022
Case Western Reserve University
2013-2022
University Hospitals of Cleveland
2016
Adding a cationic helper lipid to nanoparticle (LNP) can increase lung delivery and decrease liver delivery. However, it remains unclear whether charge-dependent tropism is universal or, alternatively, depends on the component that charged. Here, we report evidence cholesterol-dependent differ from lipid-dependent tropism. By testing how 196 LNPs delivered mRNA 22 cell types, found charged cholesterols led different lung:liver ratio than lipids. We also combining cholesterol with in heart as...
The major reservoirs for HIV in the CNS are microglia, perivascular macrophages, and to a lesser extent, astrocytes. To study molecular events controlling expression we developed reliable robust method immortalize microglial cells from primary glia fresh tissues commercially available frozen glial cells. Primary human cells, including obtained adult brain tissue, were transformed with lentiviral vectors expressing SV40 T antigen or combination of SVR40 hTERT. immortalized have microglia-like...
Significance The molecular mechanisms leading to the creation and maintenance of latent HIV reservoir remain incompletely understood. Unbiased shRNA screens showed that estrogen receptor acts as a potent repressor proviral reactivation in T cells. Antagonists ESR-1 activate HIV-1 proviruses while agonists, including β-estradiol, potently block reactivation. Using well-matched set male female donors, we found plays an important role regulating transcription both sexes. However, women are much...
We showed previously that the histone lysine methyltransferase (HKMT) H3K27me3 (EZH2) is catalytic subunit of Polycomb repressive complex 2 (PRC2) and required for maintenance HIV-1 latency in Jurkat T cells. Here we show, by using chromatin immunoprecipitation experiments, both PRC2 euchromatic histone-lysine N-methyltransferase (EHMT2), G9a H3K9me2-3 methyltransferase, are highly enriched at proviral 5' long terminal repeat (LTR) rapidly displaced upon reactivation. Clustered regularly...
The role of the negative elongation factor (NELF) in maintaining HIV latency was investigated following small hairpin RNA (shRNA) knockdown NELF-E subunit, a condition that induced high levels proviral transcription latently infected Jurkat T cells. Chromatin immunoprecipitation (ChIP) assays showed latent proviruses accumulate polymerase II (RNAP II) on 5' long terminal repeat (LTR) but not 3' LTR. NELF colocalizes with RNAP II, and its level increases induction. pause sites provirus were...
Multiple toll-like receptors (TLRs) are expressed in cells of the monocytic lineage, including microglia, which constitute major reservoir for human immunodeficiency virus (HIV) infection brain. We hypothesized that TLR receptor mediated responses to inflammatory conditions by microglial central nervous system (CNS) able induce latent HIV proviruses, and contribute etiology HIV-associated neurocognitive disorders. Newly developed cell lines (hµglia), obtained immortalizing primary microglia...
Lipid nanoparticles (LNPs) have delivered RNA to hepatocytes in patients, underscoring the potential impact of nonliver delivery. Scientists can shift LNP tropism lung by adding cationic helper lipids; however, biological response these LNPs remains understudied. To evaluate hypothesis that charged lead differential cellular responses, we quantified how 137 mRNA 19 cell types vivo. Consistent with previous studies, observed lipid-dependent tropism. After identifying and individually...
The HIV transactivator protein, Tat, enhances transcription by recruiting P-TEFb from the inactive 7SK snRNP complex and directing it to proviral elongation complexes. To test hypothesis that T-cell receptor (TCR) signaling induces critical post-translational modifications leading enhanced interactions between we employed affinity purification–tandem mass spectrometry analyze P-TEFb. TCR or phorbal ester (PMA) strongly induced phosphorylation of CDK9 kinase at Ser175. Molecular modeling...
Abstract Plasma human immunodeficiency virus type 1 (HIV-1) RNA levels in women are lower early untreated HIV-1 infection compared with those men, but have higher T-cell activation and faster disease progression when adjusted for viral load. It is not known whether these sex differences persist during effective antiretroviral therapy (ART), or they would be relevant the evaluation implementation of cure strategies. We prospectively enrolled a cohort reproductive-aged matched men on...
To predict whether preclinical lipid nanoparticle (LNP) delivery will translate in humans, it is necessary to understand the mechanism used by LNPs enter cells conserved across species. In mice, non-human primates, and deliver RNA hepatocytes adsorbing apolipoprotein E (ApoE), which binds low-density lipoprotein receptor (LDLR). A growing number of can nonhepatocytes, suggesting that ApoE- LDLR-independent interactions could affect LNP tropism. evaluate this hypothesis, we developed a...
Latently infected cells remain a primary barrier to eradication of HIV-1. Over the past decade, better understanding molecular mechanisms by which latency is established and maintained has led discovery number compounds that selectively reactivate latent proviruses without inducing polyclonal T cell activation. Recently, histone deacetylase (HDAC) inhibitor vorinostat been demonstrated induce HIV transcription from latently when administered patients. While will be given in context...
Current primary cell models for HIV latency correlate poorly with the reactivation behavior of patient cells. We have developed a new model, called QUECEL, which generates large and homogenous population latently infected CD4 + memory By purifying HIV-infected cells inducing quiescence defined cocktail cytokines, we eliminated largest problems previous latency: variable infection levels, ill-defined polarization states, inefficient shutdown cellular transcription. Latency reversal in QUECEL...
Romidepsin (RMD) is a histone deacetylase inhibitor reported to reverse HIV-1 latency. We sought identify doses of RMD that were safe and induced expression.Enrollees had RNA <40 copies/ml on ART. Measurements included levels, plasma viremia by single copy assay, DNA, cell-associated unspliced (CA-RNA), acetylation H3-lysine-9 (H3K9ac+) phosphorylation transcription factor P-TEFb. Wilcoxon tests used for comparison.43 participants enrolled in the dose Cohorts 1-3 0.5, 2, 5 mg/m 2 (36 RMD; 7...
Abstract Background Sex differences in human immunodeficiency virus (HIV) reservoir dynamics remain underexplored. Methods Longitudinal samples from virally suppressed midlife women (n = 59, median age 45 years) and age-matched men 31) were analyzed retrospectively. At each time point, we measured sex hormones (by means of enzyme-linked immunosorbent assay) cellular HIV DNA RNA digital droplet polymerase chain reaction). Number inducible RNA+ cells, which provides an upper estimate the...
We have developed models of HIV latency using microglia derived from adult human patient brain cortex and transformed with the SV40 T large hTERT antigens. Latent clones infected by reporter viruses display high levels spontaneous reactivation in culture. BrainPhys, a medium highly representative CNS extracellular environment, containing low glucose 1% FBS, reduced, but did not prevent, reactivation. hypothesized that culture was due to expression pro-inflammatory genes, such as TNF-α,...
One strategy for a functional cure of HIV-1 is “block and lock”, which seeks to permanently suppress the rebound quiescent by epigenetic silencing. For bivalent promoter in HIV LTR, both histone 3 lysine 27 tri-methylation (H3K27me3) DNA methylation are associated with viral suppression, while H3K4 (H3K4me3) correlated expression. However, H3K27me3 readily reversed upon activation T-cells through T-cell receptor. In an attempt latent stable fashion, we knocked down expression or inhibited...
Anti-retroviral therapy (ART) generally suppresses HIV replication to undetectable levels in peripheral blood, but immune activation associated with increased morbidity and mortality is sustained during ART, infection rebounds when treatment interrupted. To identify drivers of potential sources viral rebound, we modified RNAscope situ hybridization visualize HIV-producing cells as a standard against which compare the following assays virus rebound interruption: (i) envelope detection by...