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 目前阿尔茨海默病 (AD) 的治疗方法不能逆转甚至减缓疾病的进展。 这种情况是可怕的，并且由于针对两种更有希望的抗体的失败而加剧了这种情况 显然，新的治疗方法刻不容缓。 需要。 2017 年，一项大型全基因组研究发现了 PLCG2 的一个自然发生的变体 (P522R)，该基因 编码 PLC-2，可预防迟发性 AD。在后续研究中，这种遗传关联仍然存在。 更令人鼓舞的是，在轻度认知患者的临床研究中，这一结果非常有力且高度可重复。 与非携带者相比，携带 PLCG2 (P522R) 的人认知能力下降的速度较慢。 即使对于 ApoE4（一种与 AD 密切相关的生物标志物）纯合子患者，也观察到了保护作用。 在大脑中，PLC-2 主要在小胶质细胞中表达，控制吞噬细胞和神经炎症 PLC-2 在 AD 患者的病理区域中表达更高。 TREM2（使用 ApoE4 作为配体）和 CSF1R 的下游，这两种跨膜受体 与 AD 密切相关，PLC-2 类似地激活 PKC，这也与 AD 相关。因此，遗传和细胞。 数据有力地支持 PLC-2 作为治疗 AD 的新治疗靶点。 PLC-2 (P522R) 的磷脂酶活性相对于其野生型对应物略有升高，并且 我们建议确定小胶质细胞活性的增加可以预防AD。 并优化选择性激活 PLC-2 的小分子，以重现 PLC-的神经保护作用 2 (P522R) 和治疗 AD 该研究计划依赖于两种互补的高通量检测。 我们将专门为这项研究发明的真核 PLC 荧光底物进行测试。 追求三个目标 在目标 1 中，将对总计约 300,000 种化合物进行内部筛选以寻找活化剂。 将使用 PLC-2 的活性、选择性、组成和纯度经过验证的初级命中； 在目标 2 中，建立与分子动力学相结合的全长 PLC-2 的高质量模型。 在 Aim 3 中，模拟将用于数千万种化合物的计算筛选。 生物化学、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 这些新颖的小分子将成为宝贵的工具。 进一步了解 PLC-2 (P522R) 如何降低 AD 风险 小分子也将用作先导化合物。 开发治疗 AD 的新疗法。