Discovery of allosteric activators of phospholipase C-gamma2 to treat Alzheimer's disease

发现用于治疗阿尔茨海默病的磷脂酶 C-gamma2 变构激活剂

• 批准号: 10901007
• 负责人: Kenneth Hugh Pearce
• 金额: $ 76.32万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10901007
• 关键词: Affinity; Allosteric Regulation; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease risk; Alzheimer' s disease therapy; American; Amyloid; Amyloid beta-Protein; Antibodies; Area; Biochemical; Biochemistry; Biological; Biological Assay; Biological Markers; Biophysics; Brain; CD8-Positive T-Lymphocytes; CSF1R gene; Cell physiology; Cells; Chemicals; Classification; Clinical Research; Collection; Computing Methodologies; Coupled; Data; Development; Disease; Disease Progression; Docking; Drug Design; Drug usage; Endocytosis; Failure; Fluorogenic Substrate; Follow-Up Studies; Future; Genes; Genetic; Goals; Human; Hydrolysis; Immune response; Impaired cognition; Individual; Informatics; Interleukins; Isoenzymes; Late Onset Alzheimer Disease; Lead; Length; Libraries; Ligands; Link; Lipid Binding; Membrane; Microglia; Modeling; Mutation; PLCG2 gene; PLCgamma2; Pathologic; Patients; Persons; Phagocytes; Phagocytosis; Pharmaceutical Chemistry; Phenocopy; Phosphatidylinositol 4,5-Diphosphate; Phosphatidylinositols; Phospholipase; Phospholipase C; Phosphorylation; Population; Process; Property; Regulation; Reproducibility; Research; Resolution; Risk Reduction; Role; Site; Structural Models; Structure; TREM2 gene; Testing; Variant; Work; apolipoprotein E-4; cheminformatics; drug development; drug discovery; drug-like compound; genetic association; genetic variant; genome wide association study; genome-wide analysis; high throughput screening; invention; mild cognitive impairment; molecular dynamics; nervous system disorder; neuroinflammation; neuroprotection; new therapeutic target; novel; novel therapeutics; pharmacologic; receptor; recruit; reduce symptoms; response; screening; side effect; small molecule; small molecule therapeutics; success; tau-1; tool

ABSTRACT Current therapies for Alzheimer’s disease (AD) do not reverse, or even slow, progression of the disease. This situation is dire and exacerbated by the failure of antibodies directed toward two of the more promising targets—phosphorylated tau and beta-amyloids—to treat the disease. Clearly, new treatments are urgently needed. In 2017, a large genome-wide study associated a naturally-occurring variant (P522R) of PLCG2, the gene encoding PLC-2, with protection from late onset AD. In follow-up studies, this genetic association has remained strong and highly reproducible. Even more encouraging, in clinical studies of patients with mild cognitive impairment, people that carried PLCG2 (P522R) had slower rates of cognitive decline compared to non-carriers. Protection was observed even for patients homozygous for ApoE4, a biomarker strongly linked to AD. In the brain, PLC-2 is primarily expressed in microglial cells where it controls phagocytic and neuroinflammatory processes. It is more highly expressed in pathological areas of patients with AD. In microglia, PLC-2 is activated downstream of both TREM2 (which uses ApoE4 as a ligand) and CSF1R, two transmembrane receptors that are strongly linked to AD. Similarly, PLC-2 activates PKC, which is also linked to AD. Thus, genetic and cellular data strongly support PLC-2 as a novel therapeutic target for treatment of AD. The phospholipase activity of PLC-2 (P522R) is modestly elevated relative to its wild-type counterpart and it is this increased activity in microglia that is generally accepted to protect against AD. We propose to identify and optimize small molecules that selectively activate PLC-2 to reproduce the neuroprotective effects of PLC- 2 (P522R) and treat AD. The research plan relies on complementary high-throughput assays enabled by two fluorogenic substrates for eukaryotic PLCs that we invented explicitly for this research. Consequently, we will pursue three Aims. In Aim 1, in-house collections totaling ~300,000 compounds will be screened for activators of PLC-2 and primary hits verified for activity, selectivity, composition, and purity; cheminformatics will be used to structurally classify hits. In Aim 2, a high-quality model of full-length PLC-2 coupled with molecular dynamics simulations will be used for computational screens of tens of millions of compounds. In Aim 3, a suite of biochemical, biophysical, and cell biological studies will be used to prioritize allosteric activators of PLC-2 with favorable chemical and pharmacological properties. These novel small molecules will be invaluable tools to further understand how PLC-2 (P522R) reduces the risk of AD. The small molecules will also be used as leads for the development of novel therapeutics to treat AD.

抽象的 阿尔茨海默氏病（AD）的当前疗法不会逆转，甚至缓慢的疾病进展。 这种情况因针对两种更有希望的抗体的失败而严重和加剧 靶标 - 磷酸化的tau和β-淀粉样蛋白来治疗该疾病。显然，新的治疗方法是 需要。 2017年，一项全基因组的大型研究与PLCG2的自然存在变体（p522r）相关联基因 编码PLC-2，并防止晚期AD。在后续研究中，这种遗传关联仍然存在 强大，高度可重复。在轻度认知患者的临床研究中，更令人鼓舞 与非携带者相比，携带PLCG2（P522R）的人的认知能力下降速度较慢。 即使对于纯合的APOE4的患者也观察到了保护，APOE4是与AD密切相关的生物标志物。在 大脑，PLC-2主要在小胶质细胞中表达，该细胞控制吞噬和神经炎症 过程。它在AD患者的病理区域中更高表达。在小胶质细胞中，PLC-2被激活 trem2（使用APOE4作为配体）和CSF1R的下游，两个跨膜接收器 与AD密切相关。同样，PLC-2激活PKC，这也链接到AD。那是遗传和细胞 数据强烈支持PLC-2作为AD治疗的新型治疗靶标。 相对于其野生型对应物和 正是小胶质细胞中的活动增加，可以预防AD。我们建议确定 并优化小分子，这些分子有选择地激活PLC-2以再现PLC-的神经保护作用 2（p522r）和治疗AD。该研究计划依赖于两个由两个启用的高通量测定 我们为这项研究明确发明的真核PLC的荧光底物。因此，我们会的 追求三个目标。在AIM 1中，将对激活剂筛选总计约300,000种化合物的内部收藏 PLC-2和主要命中率验证了活性，选择性，组成和纯度；化学信息学将使用 在结构上对命中进行分类。在AIM 2中，全长PLC-2的高质量模型与分子动力学结合 模拟将用于数千万化合物的计算屏幕。在AIM 3中，一套 生化，生物物理和细胞生物学研究将用于优先考虑PLC-2的变构激活剂 有利的化学和药物特性。这些新颖的小分子将是无价的工具 进一步了解PLC-2（P522R）如何降低AD的风险。小分子也将用作铅 用于开发新疗法以治疗AD。