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: 10581056
• 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/10581056
• 关键词: Academia; Adenylate Cyclase; Affect; Agonist; Attenuated; Autopsy; Brain; CNR2 gene; Cathepsins B; Cells; Chronic; Confocal Microscopy; Development; Enzymes; Flow Cytometry; Future; HIV; HIV Infections; HIV Seropositivity; HIV-1; HIV-associated neurocognitive disorder; Human; Immunohistochemistry; In Vitro; Individual; Infection; Infiltration; Inflammation; Inflammatory; Inositol; Knowledge; Label; Laboratories; Learning; Longevity; MAPK3 gene; Mentors; Microglia; Neurodegenerative Disorders; Neurosciences; Nicotinic Receptors; Organoids; Pathway interactions; Patients; Peptide Hydrolases; Persons; Phase; Phospholipase C; Play; 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; base; blood-brain barrier permeabilization; brain tissue; cytokine; effective therapy; macrophage; 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.

项目概要 大约 50% 的 HIV 阳性者会出现与 HIV 相关的神经认知障碍 (HAND)，尽管 正在接受联合抗逆转录病毒治疗（cART）。手部疾病是一种炎症性神经退行性疾病 其特点是受影响者大脑中巨噬细胞浸润增加和艾滋病毒持续复制 个人。目前尚无针对 HAND 的有效疗法。我们的实验室发现增加 组织蛋白酶 B（一种促炎性溶酶体酶）在患有此病的个体死后脑组织中的表达 手。此外，单核细胞源性巨噬细胞（MDM）感染 HIV-1 后，会分泌组织蛋白酶 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，靶向组织蛋白酶 B 是对抗 HAND 的潜在策略。在寻找新的 疗法中，已证明 2 型大麻素受体 (CB2R) 的激活可抑制 HIV-1 复制 在巨噬细胞中，降低血脑屏障 (BBB) 通透性，降低 HIV-1 病毒蛋白的神经毒性， 并减少促炎细胞因子的释放。尽管如此，研究表明， 体外 HIV 感染巨噬细胞和 HAND 患者死后脑组织中的 CB2R。因此，这 受体代表了治疗 HAND 的一个有希望的靶标。然而，CB2R 激活的作用 组织蛋白酶 B 的分泌和神经毒性此前尚未研究过。本提案的总体目标 目的是了解组织蛋白酶 B 分泌中 CB2R 激活的机制以及 HIV 感染者的神经毒性 巨噬细胞。我们的中心假设是 CB2R 激活将阻止组织蛋白酶 B 分泌和神经毒性 通过减弱 MDM 中的细胞内炎症途径并预防 HIV 感染的巨噬细胞 组织蛋白酶 B 与 SAPC 相互作用。我们的假设是根据显示显着的结果制定的 用 CB2R 激动剂治疗的 HIV 感染 MDM 导致组织蛋白酶 B 分泌减少和神经毒性。我们 将测试我们的中心假设，从而通过追求以下目标来实现本提案的目标 具体目标： 1) 确定 CB2R 激活对组织蛋白酶 B 分泌和 HIV 神经毒性的影响 被感染的巨噬细胞。 2) 探索细胞内通路并表征组织蛋白酶 B 相互作用组 CB2R 激活后 HIV 感染巨噬细胞的上清液。 3）了解慢性病的发病机制 手部炎症。这项研究的基本原理是了解 CB2R 激活的作用 组织蛋白酶 B 的分泌和相互作用将允许开发针对 HIV 诱导的组织蛋白酶的策略 B神经毒性。这一贡献意义重大，因为它将提供有关机制的新知识 CB2R 调节组织蛋白酶 B 诱导的 HIV 感染巨噬细胞的神经毒性，并将有助于 制定针对 HAND 的新策略。