Innovative therapeutic approaches to address excitotoxic CNS/neuronal damage in opioid-neuroHIV comorbidity

解决阿片类药物-神经艾滋病毒合并症中的兴奋性中枢神经系统/神经元损伤的创新治疗方法

• 批准号: 10684110
• 负责人: Kurt F Hauser
• 金额: $ 67.87万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684110
• 关键词: AMPA Receptors; Acceleration; Acute; Address; Adult; Affect; Agonist; Animals; Architecture; Behavior; Behavioral; Biochemical; Brain; Brain-Derived Neurotrophic Factor; Breeding; CLC Gene; Cell membrane; Cells; Chronic; Clinical Pathology; Contracts; Corpus striatum structure; Data; Disinhibition; Dopamine Receptor; Electrophysiology (science); Enhancers; Epilepsy; Equilibrium; Event; Exposure to; Functional disorder; Glutamates; Gramicidin; HIV; HIV Envelope Protein gp120; HIV-1; Heroin; Homeostasis; Human; Immunosuppression; Impaired cognition; In Vitro; Incidence; Infection; Inflammation; Injury; Intervention; Intractable Epilepsy; Link; Membrane Potentials; Minor; Morphine; Mus; N-Methyl-D-Aspartate Receptors; Neuroglia; Neurologic; Neuronal Injury; Neurons; Opiate Addiction; Opioid; Opioid Receptor; Optics; Outcome; Output; Pattern; Perforation; Phosphorylation; Physiological; Physiology; Population; Probability; Prodrugs; Property; Proteins; Ramp; Regulation; Reporter; Rest; Role; Severities; Signal Transduction; Spinal cord injury; System; Temporal Lobe; Testing; Therapeutic; Therapeutic Intervention; Transgenic Mice; Transgenic Model; autism spectrum disorder; awake; behavior measurement; central nervous system injury; cohort; comorbidity; cytokine; deafness; driving force; excitotoxicity; experimental study; genetic regulatory protein; in vitro Model; in vivo; induced pluripotent stem cell; inhibitor; injection drug use; innovation; motor behavior; motor deficit; nervous system disorder; neuroAIDS; neuronal excitability; neuronal survival; neurotoxic; opioid abuse; opioid exposure; opioid use disorder; optical imaging; painful neuropathy; patch clamp; sex; sodium-potassium-chloride cotransporter 1 protein; spinal cord and brain injury; symporter; theories; therapy design; trafficking; transmission process; uptake; voltage

Injection drug use increases the probability of contracting HIV, and opioid use disorder (OUD) accelerates HIV- 1 infection through immune suppression and direct CNS actions. Glutamatergic excitotoxicity is a major factor in HIV-dependent CNS injury, but emerging evidence also suggests a loss of inhibitory GABAergic function in neuroHIV. The parallel loss of Cl− homeostasis and GABAergic tone will worsen excitatory outcomes since ‘disinhibition’ results in net excitation. If this is true, then interventions that protect inhibitory systems are predicted to at least partially negate ‘excitotoxic’ effects of HIV. KCC2 is the main transporter responsible for maintaining [Cl−]i homeostasis and GABAergic function in the adult CNS and is the focus of proposed studies. Although the idea of disinhibition as a driving force is embraced for other neurological disorders (e.g., autism, certain epilepsies, opioid dependence, traumatic brain/spinal cord injury), it represents a conceptual shift about mechanisms underlying synaptodendritic dysfunction in neuroHIV. The Cl− concentration inside neurons ([Cl−]i) is small and the Cl− reversal potential (ECl) is close to the resting membrane potential. Thus, minor changes in [Cl−]i can greatly affect the strength and polarity of inhibitory (e.g., GABAA) transmission. NKCC1 (Cl− uptake) and KCC2 (Cl− efflux) co-transporters are key regulators of [Cl−]i, and KCC2 expression/function is essential for adult neuron survival. Their balance can be regulated by cytokines/trophic factors (e.g., BDNF) from glia, therapeutically (CLP290), genetically, and by opioids. Importantly, we find that increasing KCC2 levels/function strongly protects against exposure to Tat, gp120, infectious HIV, and opioids in human and mouse neurons. In vivo, maintaining KCC2 phosphorylation can normalize KCC2 localization in Drd2-expressing striatal medium spiny neuron (MSN) cell membranes and reverse motor deficits due to HIV-1 Tat. Aim 1 uses in vitro models including iPSC MSNs, infective HIV, and optical electrophysiology to identify mechanisms by which HIV and/or opioids alter [Cl−]i homeostasis, dysregulate D1/D2 MSN excitability, how this triggers synaptodendritic injury, and protective strategies. Despite its fundamental importance, the role of KCC2 in excitatory/inhibitory (E-I) imbalances and altered ECl and EGABA have never been explored in the context of HIV/OUD. Aim 2 extends the studies in vivo/ex vivo. 2 transgenic models (Tat+/-, HIVTg26) that both mimic clinical pathology with considerable fidelity are crossed with transgenic mice expressing Drd1a-tdTomato (D1) and Drd2-eGFP (D2) to identify both striatal MSN populations. Acute (2 wk) and chronic (8 wk) HIV/Tat exposure times are examined; separate cohorts of mice (both sexes) receive concurrent, ramping exposure to morphine (s.c.). Comprehensive studies of [Cl−]i regulation in MSNs (gramicidin-perforated patch physiology), synaptodendritic injury, and behavior related to striatal function are performed ± the KCC2 enhancer prodrug CLP290 as an intervention strategy. Alternative [Cl−]i regulation through NKCC1, TMEM16A, and CLC-1 are also tested. GCaMP8f expression in D1/D2 MSNs in awake, behaving mice links striatal output activity to behavioral change (Inscopix miniscopes).

注射吸毒增加了感染艾滋病毒的可能性，阿片类药物使用障碍（OUD）加速了艾滋病毒的传播