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Principal Investigator: German Gustavo Gornalusse
Organization: UNIVERSITY OF WASHINGTON
Fiscal Year: 2024
Award: $829,396
Funding agency: National Institute of Allergy and Infectious Diseases
ABSTRACT
Despite the effectiveness of antiretroviral treatment (ART) at treating HIV, it fails to completely eradicate the
viral reservoir. HIV reactivation occurs naturally, as evidenced by occasional low-level viremia (“blips”)
observed in patients on ART. Studying the anatomical sites of HIV reactivation and the molecular mechanisms
driving these blips can provide essential information for the development of HIV cure strategies. Non-human
primate studies, autopsies, and clinical trials indicate that the gastrointestinal (GI) tract contains up to 98%
of the HIV reservoir and that its immunophysiology could be especially conducive for HIV reactivation. In our
previous work, we found that the epithelium of the intestinal mucosa contains a population of cells producing
extremely high levels of type I/III interferon-stimulated proteins, among them, IFN-stimulated gene 15 (ISG15).
These specialized enterocytes in the gut express glycoprotein 2, suggesting that some of these epithelial cells
with heightened immunity are microfold cells (M cells). Type I IFN stimulation drives bystander T cell
proliferation in vivo, which likely include latently infected cells, thus contributing to reservoir
persistence. There is also evidence that type I IFN efficiently reactivates HIV-1 in vitro and ex vivo.
Furthermore, mouse studies indicate that blocking type I interferon signaling diminishes immune activation,
restores T cell function, and reduces the size of the HIV reservoir. Taken together, the data suggest that the GI
tract, and particularly M cells, may contribute to HIV persistence and reactivation.
Our overarching hypothesis is that immunologically hyperactive microfold (“M”) cells in the gut create
a microenvironment that fosters proliferation of latently infected bystander CD4+ T cells, sporadic HIV
reactivation from these cells, or both. We will investigate our hypothesis by ex vivo studies of mucosal GI
tissues, including from people living with HIV (PLH), and in vitro M cell cultures to model and manipulate their
effect on HIV latency. In Aim 1, we will investigate whether the interaction with M cells (ISG15high enterocytes)
increases immune activation of bystander CD4+ T cells and macrophages using a single-cell spatial
transcriptomics/proteomics approach. In Aim 2, we will test the hypothesis that T cells/macrophages
containing HIV-1 proviral DNA/mRNA are found in high number in the vicinity of M cells using duodenal and
rectal biopsies from a clinical study of 40 PLH on stable ART. We will quantify HIV-1 DNA and mRNA copies
by digital PCR, determine T cell clonality by spatial transcriptomics and analyze the spatial relationship of HIV-
1 DNA+ and mRNA+ cells by DNA/RNAscope. In Aim 3, we will use in vitro models to test the hypothesis that
M cells drive T cell proliferation and HIV reactivation via their increased activity of type I/III IFN pathways. This
study will provide key information about the intestinal microarchitecture of the HIV reservoir that is necessary
for the development of novel latency reversal agents or strategies to target latently infected cells via M cells.
Terms: <(IFN) α><(IFN)-α><(IFN)α><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Alferon><Alimentary Canal><Anatomic Sites><Anatomic structures><Anatomy><Anti-Retroviral Agents><Automobile Driving><Autopsy><Binding><Biopsy><Body Tissues><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><Cell Body><Cell Communication and Signaling><Cell Culture Techniques><Cell Function><Cell Physiology><Cell Process><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Clinical><Clinical Research><Clinical Study><Clinical Trials><Clonality><Co-culture><Cocultivation><Coculture><Coculture Techniques><DNA><Data><Deoxyribonucleic Acid><Development><Digestive Tract><Drugs><Duodenum><Effectiveness><Enterocytes><Environment><Epithelial Cells><Exhibits><Expression Signature><Fostering><GI Tract><Gastrointestinal Tract><Gastrointestinal tract structure><Gene Activation><Gene Expression><Gene Expression Profile><Genes><Genome><Glycoproteins><Goals><Grant><Gut Mucosa><HIV><HIV Infections><HIV-1><HIV-I><HIV1><HTLV-III Infections><HTLV-III-LAV Infections><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Infections><Human immunodeficiency virus 1><Hyperactivity><IFN><IFN Alpha><IFN α><IFN-α><IFNa><IFNα><Immune><Immune Cell Activation><Immunes><Immunity><Immunobiology><Immunochemical Immunologic><Immunologic><Immunological><Immunologically><Immunologics><Immunophysiology><In Situ><In Vitro><Individual><Inflammation><Interferon Alfa-n3><Interferon Type I><Interferon alpha><Interferon-α><Interferons><Intestinal><Intestinal Mucosa><Intestines><Intracellular Communication and Signaling><Investigators><LAV-HTLV-III><Length of Life><Leukocyte Interferon><Longevity><Lymphadenopathy-Associated Virus><Lymphoblast Interferon><Lymphoblastoid Interferon><Macrophage><Maintenance><Medication><Messenger RNA><Mice><Mice Mammals><Modeling><Molecular><Molecular Interaction><Mucosa><Mucosal Tissue><Mucous Membrane><Murine><Mus><Mφ><NIAID><NIH Program Announcements><National Institute of Allergy and Infectious Disease><Operative Procedures><Operative Surgical Procedures><Organoids><Pathway interactions><Patients><Persons><Phagocytosis><Pharmaceutical Preparations><Physiology><Population><Program Announcement><Proliferating><Proteins><Proteomics><Proviruses><RNA Seq><RNA sequencing><RNAseq><Rectum><Research><Research Personnel><Researchers><Risk><Role><Signal Transduction><Signal Transduction Systems><Signaling><Source><Subcellular Process><Surgical><Surgical Interventions><Surgical Procedure><System><T cell clonality><T-Cell Proliferation><T-Cells><T-Lymphocyte><T-cell receptor clonality><T4 Cells><T4 Lymphocytes><TCR clonality><Testing><Tissues><Transcript><Viral><Viral reservoir><Viremia><Virus><Virus reservoir><Virus-HIV><Work><alimentary tract><anti-retroviral><antiretroviral therapy><antiretroviral treatment><biological signal transduction><bowel><cell culture><cell cultures><cell imaging><cellular imaging><developmental><digestive canal><digital><driving><drug/agent><gastrointestinal><gene expression pattern><gene expression signature><global gene expression><global transcription profile><humanized mice><humanized mouse><immune activation><in vitro Model><in vivo><innovate><innovation><innovative><intestinal epithelium><latent HIV reservoir><latent HIV-1 reservoir><latent HIV1 reservoir><mRNA><multiomics><multiple omics><nano-string><nanostring><necropsy><non-human primate><nonhuman primate><novel><panomics><pathogen><pathway><postmortem><prevent><preventing><social role><spatial relationship><surgery><thymus derived lymphocyte><tissue culture><transcriptional profile><transcriptional signature><transcriptome><transcriptome sequencing><transcriptomic sequencing><transcriptomics><transcytosis><viraemia><viral rebound><viral sepsis><virus rebound><virusemia>