Cerebral organoid and IPSC derived microglia: Modeling of HIV and methamphetamine co-morbidity

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Principal Investigator: Peng  Jiang
Organization: UNIVERSITY OF NEBRASKA MEDICAL CENTER
Fiscal Year: 2024
Award: $495,815
Funding agency: National Institute on Drug Abuse

Abstract:
An estimated ~1.3 million adults and adolescents are living with human immunodeficiency virus (HIV) infection
in the USA of which 15% of them undiagnosed. The initiation of combination antiretroviral therapy (cART) has
transformed AIDS from a fatal illness into a chronic and manageable disease. The presence of HIV infected
microglia and macrophage reservoirs in the central nervous system (CNS) and variable penetration of anti-
retroviral drugs across the blood brain barrier after cART are likely factors for the persistence of HIV associated
neurocognitive disorder (HAND). Drugs of abuse such as the potent psychostimulant methamphetamine (Meth)
that is abused by an estimated 30 million people in the world further minimize the efficacy of cART and in
conjunction with HIV exacerbate CNS pathology. Thus, significant gap in knowledge in the field is to better
understand the neuropathogenesis and the etiology of clinical outcomes observed in HIV+ patient’s dependent
on Meth abuse. The current grant proposal focuses on developing a human induced pluripotent stem cell based
cerebral organoid model to investigate underlying brain dysfunction during HIV/Meth interaction. Cerebral
organoids are 3-dimensional “mini brains” that can self-organize and recapitulate many milestone events seen
in in vivo brain development. Our recent study on generating a novel organoid model with the feasibility in
controlling the microglia ratio and microenvironment of organoid formation to recapitulate changes in brain
functions under various conditions (e.g. virus and viral infection-induced inflammation and synaptic pruning)
makes this model ideal for our proposed studies. The central hypothesis of this proposal is that Meth in
conjunction with HIV causes significant neuronal damage. The combination of microglia-containing cerebral
organoid model, extracellular vesicle biology, single-cell RNA-sequencing, CRISPR/Cas9-mediated gene editing
will decipher novel intercellular and molecular mechanisms and pathways that underlie abnormalities in neuronal
functions and connectivity caused by HIV infection. We will investigate this hypothesis under three specific aims:
Specific Aim 1 will examine neuronal function, synaptic and mitochondrial perturbations during HIV/meth
interactions in 3D microglia-containing cerebral organoid model; Specific Aim 2 will investigate the role of
extracellular vesicles in microglial organoids treated under HIV/meth conditions; and Specific Aim 3 will
characterize the molecular mechanisms underlying HIV infection-induced neuronal injury and further evaluate
our microglial organoid model as a reliable tool to identify molecular signatures of HAND. Our proposed
experiments will decipher molecular mechanisms, novel signaling events and molecular partners underlying the
neuronal injury in HAND. Through the series of the experiments we aim to present an easily scalable and
reproducible model to study HAND pathogenesis. The data obtained from these studies can be used to design
novel therapeutics to control HIV infection.

Terms: <21+ years old><3-D><3-D modeling><3-Dimensional><3D><3D modeling><AIDS><AIDS Virus><Abuse Reporting><Acquired Immune Deficiency><Acquired Immune Deficiency Syndrome><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome><Acquired Immunodeficiency Syndrome Virus><Address><Adolescent><Adolescent Youth><Adult><Adult Human><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-Retroviral Agents><Anti-inflammatory><Applications Grants><Assay><Astrocytes><Astrocytus><Astroglia><Autopsy><Bioassay><Biogenesis><Biological Assay><Biology><Blood - brain barrier anatomy><Blood-Brain Barrier><Brain><Brain Nervous System><CNS Nervous System><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Causality><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Central Nervous System><Cerebrum><Chronic><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Co-culture><Cocultivation><Coculture><Coculture Techniques><Crystal Meth><Crystal methamphetamine><Data><Dendritic Spines><Deoxyephedrine><Desoxyephedrine><Development><Diagnosis><Disease><Disorder><Drug abuse><Drugs><Encephalon><End Point Assay><Endpoint Assays><Ensure><Erythro><Etiology><Event><Genes><Glia><Glial Cells><Goals><Grant Proposals><GvHD><HIV><HIV 1 associated neurocognitive disorder><HIV Genome><HIV Infections><HIV associated neurocognitive deficit><HIV associated neurocognitive impairment><HIV induced neurocognitive deficit><HIV induced neurocognitive impairment><HIV neurocognitive impairment><HIV-1 associated neurocognitive deficit><HIV-1 associated neurocognitive disorder><HIV-1 associated neurocognitive impairment><HIV-1 genome><HIV-associated neurocognitive disorder><HIV1 genome><HTLV-III Infections><HTLV-III-LAV Infections><Hemato-Encephalic Barrier><High Prevalence><Homologous Wasting Disease><Hortega cell><Human><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Infections><Individual><Induced Neurons><Inflammation><Inflammatory><Intracellular Communication and Signaling><Knowledge><Kolliker's reticulum><LAV-HTLV-III><Lymphadenopathy-Associated Virus><Macrophage><Measures><Mediating><Medication><Methamphetamine><Methylamphetamine><Mice><Mice Mammals><Micro RNA><MicroRNAs><Microglia><Mitochondria><Modeling><Modern Man><Molecular><Molecular Fingerprinting><Molecular Profiling><Monkeys><Murine><Mus><Myeloid Progenitor><Myeloid Progenitor Cells><Myeloid Stem Cells><Mφ><N-Methylamphetamine><Nerve Cells><Nerve Degeneration><Nerve Unit><Neural Cell><Neuraxis><Neurocognitive Impairment in HIV><Neurocognitive Impairment in HIV-1><Neurocyte><Neuroglia><Neuroglial Cells><Neuroimmune><Neuron Degeneration><Neuronal Injury><Neurons><Neuropathogenesis><Non-neuronal cell><Nonneuronal cell><Organoids><Origin of Life><Outcome><PK/PD><Pathogenesis><Pathology><Pathway interactions><Patients><Pattern><Penetration><Persons><Pharmaceutical Preparations><Property><Proteins><Receptor Protein><Regimen><Reproducibility><Role><Runt Disease><SIV><Sampling><Series><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Simian Immunodeficiency Viruses><Societies><Synapses><Synaptic><Technology><Toxic effect><Toxicities><Treatment Efficacy><Viral><Viral Diseases><Virus Diseases><Virus-HIV><Yolk Sac><abnormal brain function><abuse of drugs><abused drug><abused drugs><abuses drugs><adulthood><anti-retroviral><antiretroviral therapy><antiretroviral treatment><astrocytic glia><biological signal transduction><bloodbrain barrier><brain dysfunction><brain impairment><brain tissue><causation><cell type><cerebral><co-morbid><co-morbidity><comorbidity><dendrite spine><design><designing><developmental><disease causation><drug abused><drug of abuse><drug/agent><drugs abused><drugs of abuse><dysfunctional brain><experiment><experimental research><experimental study><experiments><extracellular vesicles><genome editing><genomic editing><gitter cell><graft versus host disease><graft vs host disease><graft vs. host disease><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><human pluripotent stem cell><humanized mice><humanized mouse><iNeuron><iPS><iPSC><iPSCs><in vivo><induced human pluripotent stem cells><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><intervention efficacy><juvenile><juvenile human><mesoglia><metabolic rate><meth><meth abuse><methamphetamine abuse><miRNA><miRNAs><microglial cell><microgliocyte><mitochondrial><molecular profile><molecular signature><myeloid stem and progenitor cell><necropsy><nerve cement><neural degeneration><neural inflammation><neurodegeneration><neurodegenerative><neuroinflammation><neuroinflammatory><neurological degeneration><neuron injury><neuron toxicity><neuronal><neuronal degeneration><neuronal toxicity><neurotoxicity><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><non-invasive imaging><noninvasive imaging><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><pathway><perivascular glial cell><pharmacokinetics and pharmacodynamics><postmortem><postsynaptic><presynaptic><progenitor><progenitor cell population><progenitor population><psychostimulant><receptor><response><scRNA-seq><self organization><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single-cell RNA sequencing><social role><stem and progenitor cell population><stem cell population><success><synapse><synaptic pruning><therapeutic efficacy><therapeutic evaluation><therapeutic testing><therapy efficacy><three dimensional><three-dimensional modeling><tool><viral infection><virus infection><virus-induced disease><vitelline sac>