Modulation of alpha-7 nicotinic acetylcholine receptor in HIV-infected microglia and brain organoids

HIV 感染的小胶质细胞和脑类器官中 α-7 烟碱乙酰胆碱受体的调节

• 批准号: 10615915
• 负责人: Lester José Rosario-Rodríguez
• 金额: $ 8.73万
• 依托单位: UNIVERSITY OF PUERTO RICO MED SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615915
• 关键词: Academia; Adenylate Cyclase; Affect; Agonist; Agreement; Attenuated; Autopsy; Brain; CNR2 gene; Cathepsins B; Cell Reprogramming; Cells; Chronic; Confocal Microscopy; Development; Enzymes; Flow Cytometry; Future; HIV; HIV Infections; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Human; Immunohistochemistry; Immunoprecipitation; In Vitro; Individual; Infection; Infiltration; Inflammation; Inflammatory; Inositol; Knowledge; Label; Laboratories; Learning; Longevity; MAPK3 gene; Macrophage; Mentors; Microglia; Neurodegenerative Disorders; Neurosciences; Nicotinic Receptors; Organoids; Pathway interactions; Patients; Peptide Hydrolases; Persons; Phase; Phospholipase C; Play; Postdoctoral Fellow; Proteins; Proteomics; Research; Research Personnel; Research Project Grants; Role; Serum Amyloid P-Component; Signal Transduction; Techniques; Testing; Training; Validation; Viral Proteins; Western Blotting; Work; antiretroviral therapy; blood-brain barrier permeabilization; brain tissue; cytokine; effective therapy; monocyte; neuron apoptosis; neurotoxic; neurotoxicity; novel therapeutics; prevent; receptor; tandem mass spectrometry; therapy development; treatment strategy; tripolyphosphate

Project Summary Approximately, 50% of HIV-positive people develop HIV-associated neurocognitive disorders (HAND), despite being under combined antiretroviral therapy (cART). HAND is an inflammatory neurodegenerative disease characterized by increased macrophage infiltration and persistent HIV replication in the brain of affected individuals. There are no effective therapies available against HAND. Our laboratory has found increased expression of cathepsin B, a pro-inflammatory lysosomal enzyme, in postmortem brain tissues of individuals with HAND. In addition, after HIV-1 infection of monocyte-derived macrophages (MDM), secreted cathepsin B changes interactions with other proteins such as serum amyloid P component (SAPC) and promotes neurotoxicity. Thus, targeting cathepsin B represents a potential strategy against HAND. In search for new therapies, it was demonstrated that activation of cannabinoid receptor type 2 (CB2R) inhibits HIV-1 replication in macrophages, reduces blood-brain barrier (BBB) permeability, decreases neurotoxicity to HIV-1 viral proteins, and diminishes pro-inflammatory cytokines release. Nonetheless, studies have shown increased expression of CB2R in in vitro HIV-infected macrophages, and in post-mortem brain tissues of HAND patients. Therefore, this receptor represents a promising target for treatment against HAND. However, the role of CB2R activation in cathepsin B secretion and neurotoxicity has not been studied previously. The overall objective of this proposal is to understand the mechanisms of CB2R activation in cathepsin B secretion and neurotoxicity from HIV-infected macrophages. Our central hypothesis is that CB2R activation will prevent cathepsin B secretion and neurotoxicity from HIV-infected macrophages by attenuating intracellular inflammation pathways in MDM and preventing cathepsin B interactions with SAPC. Our hypothesis was formulated based on results that show a significant decrease in cathepsin B secretion and neurotoxicity from HIV-infected MDM treated with a CB2R agonist. We will test our central hypothesis and, thereby, accomplish the objective of this proposal by pursuing the following specific aims: 1) Determine the effect of CB2R activation in cathepsin B secretion and neurotoxicity from HIV- infected macrophages. 2) Explore the intracellular pathways and characterize cathepsin B interactome in supernatants from HIV-infected macrophages after CB2R activation. 3) Understand the mechanisms of chronic inflammation in HAND. The rationale for this proposed research is that understanding the role of CB2R activation in cathepsin B secretion and interactions will permit the development of strategies against HIV-induced cathepsin B neurotoxicity. This contribution is significant because it will provide new knowledge about the mechanisms of CB2R modulation in cathepsin B-induced neurotoxicity from HIV-infected macrophages, and will contribute to the development of new strategies against HAND.

项目摘要 大约有50％ 在抗逆转录病毒疗法（CART）下。手是一种炎症性神经退行性疾病 其特征是巨噬细胞浸润增加和受影响的大脑中的持续性HIV复制 个人。没有有效的疗法可用。我们的实验室发现 expression of cathepsin B, a pro-inflammatory lysosomal enzyme, in postmortem brain tissues of individuals with 手。此外，在HIV-1感染单核细胞衍生的巨噬细胞（MDM）之后，分泌的组织蛋白酶B 改变与其他蛋白质（例如血清淀粉样蛋白P成分（SAPC））的相互作用，并促进 神经毒性。因此，靶向组织蛋白B代表了反对手的潜在策略。在寻找新的 疗法，证明大麻素受体2型（CB2R）的激活抑制了HIV-1复制 在巨噬细胞中，降低了血脑屏障（BBB）的渗透性，可降低对HIV-1病毒蛋白的神经毒性 并减少促炎性细胞因子释放。尽管如此，研究表明 在体外HIV感染的巨噬细胞和手部患者尸体后脑组织中的CB2R。因此，这个 受体代表了对手的有希望的治疗目标。但是，CB2R激活在 先前尚未研究组织蛋白酶B分泌和神经毒性。该提议的总体目标 是要了解组织蛋白酶B分泌中CB2R激活的机制和HIV感染的神经毒性 巨噬细胞。我们的中心假设是CB2R激活将防止组织蛋白酶B分泌和神经毒性 通过减弱MDM中细胞内炎症途径并防止，从HIV感染的巨噬细胞中 组织蛋白酶B与SAPC的相互作用。我们的假设是根据结果表明显着的结果提出的 通过CB2R激动剂治疗的HIV感染的MDM的组织蛋白酶B分泌和神经毒性的降低。我们 将检验我们的中心假设，从而通过追求以下内容来实现该提议的目标 具体目的：1）确定CB2R激活在组织蛋白酶B分泌和HIV-神经毒性中的影响 感染的巨噬细胞。 2）探索细胞内途径并表征组织蛋白酶B相互作用 CB2R激活后，来自HIV感染的巨噬细胞的上清液。 3）了解慢性的机制 炎症。这项拟议研究的理由是了解CB2R激活的作用 在组织蛋白酶B分泌和互动中，将允许制定针对HIV引起的组织蛋白酶的策略 B神经毒性。这项贡献很重要，因为它将提供有关机制的新知识 在组织蛋白酶B诱导HIV感染的巨噬细胞的神经毒性中的CB2R调节，并将有助于 制定反对手的新策略。