Directed evolution of tissue inhibitor of metalloproteinase 3 (TIMP-3) to develop novel Alzheimer’s disease (AD) therapeutics

金属蛋白酶组织抑制剂 3 (TIMP-3) 的定向进化可开发新型阿尔茨海默病 (AD) 疗法

• 批准号: 10303777
• 负责人: Maryam Raeeszadeh Sarmazdeh
• 金额: $ 30万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10303777
• 关键词: Affect; Affinity; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Alzheimer’s disease biomarker; American; Amyloid beta-Protein; Apoptosis; Binding; Biological Assay; Blood; Blood - brain barrier anatomy; Brain; Cell Culture Techniques; Clinical Research; Competitive Binding; Complex; Degenerative Disorder; Demyelinations; Dendrites; Directed Molecular Evolution; Disease; Disease Progression; Disintegrins; Engineering; Enzyme Inhibitor Drugs; Enzymes; Exposure to; Extracellular Matrix Degradation; Family; Future; Gelatinase B; Goals; Growth Factor; In Vitro; Inhibition of Matrix Metalloproteinases Pathway; Label; Libraries; Ligands; Light; MMP9 gene; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Metalloproteases; Mitochondria; Modeling; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Pathogenicity; Pathologic; Pathologic Processes; Pathway interactions; Peptide Hydrolases; Permeability; Physiological; Physiological Processes; Play; Protein Engineering; Proteins; Recombinants; Research; Research Personnel; Role; Scaffolding Protein; Signal Transduction; Signaling Molecule; Structure; Surface; TIMP3 gene; Testing; Therapeutic; Therapeutic Effect; Tissue Engineering; Tissue Inhibitor of Metalloproteinases; Tissues; Toxic effect; Up-Regulation; Variant; Work; X-Ray Crystallography; Yeasts; base; blood-brain barrier disruption; blood-brain barrier permeabilization; combinatorial; drug development; engineering design; enzyme mechanism; genetic variant; improved; improved outcome; inhibitor; insight; member; molecular targeted therapies; next generation; novel; novel therapeutics; rational design; scaffold; side effect; targeted treatment; tau aggregation; therapeutic target; tool; tumorigenesis

SUMMARY Limited efficient therapeutics are available to control Alzheimer's disease (AD), so there is a strong demand to target new pathogenic biomarkers of AD. The metzincin superfamily, including matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinases (ADAMs), play a multifaceted role in physiological and pathological processes in the central nervous system and therefore are therapeutic targets to limit neurodegeneration in AD. For instance, upregulation of MMP-9 promotes apoptosis, blood-brain barrier disorder, and demyelination and is involved in the amyloid-β processing pathway through proteolytic degradation of extracellular matrix components and tissue remodeling, and increasing tau accumulation which play key roles in AD. But on the other hand, upregulation of ADAM-10 increases soluble amyloid-β formation, which helps control AD. Thus, using highly selective inhibitors to tailor the proteolytic activity of metzincin- induced pathological changes in AD patients’ brains may be a promising therapeutic strategy. Protein inhibitors offer higher selectivity to target MMP-9 compared to smaller synthetic inhibitory molecules with broad-spectrum targets. Tissue inhibitors of metalloproteinases (TIMPs) are the major endogenous inhibitors of MMPs and ADAMs with varying degrees of binding affinity. Among all TIMPs, TIMP-3 has the lowest association with the cell signaling molecules triggering undesired off-target effect such as the ones responsible in tumorigenesis, suggesting that it has a low off-target therapeutic effect in MMP-dependent diseases. This makes TIMP-3 an ideal protein scaffold for engineering improved inhibitors for MMP-9. However, TIMP-3 also binds to other MMPs, ADAMs, and growth factors. Here, we propose to use a combination of rational and random combinatorial approaches to develop TIMP-3-based therapeutics to control neurodegenerative disease progression by targeting MMP-9 with high selectivity. These engineered TIMP-based probes will avoid binding to ADAM-10, which is neuroprotective in AD, to decrease off-target effects. The overarching goal of this project is to engineer and design a new molecularly targeted therapy based on natural MMP inhibitors that target MMP-9 with high selectivity, understand their mechanism of inhibition using structural studies and cell culture models to improve outcomes in neurodegenerative diseases with inadequate treatment options.

总结 有限的有效治疗剂可用于控制阿尔茨海默病（AD），因此强烈需要 靶向AD新的致病性生物标志物。Metzincin超家族，包括基质金属蛋白酶 基质金属蛋白酶（MMPs）和去整合素和金属蛋白酶（亚当斯）在生理和病理过程中发挥多方面的作用。 因此是治疗靶点，以限制 AD中的神经变性。例如，MMP-9的上调促进细胞凋亡，血脑屏障， 疾病和脱髓鞘，并通过蛋白水解参与淀粉样蛋白-β加工途径 细胞外基质成分的降解和组织重塑，以及增加tau积累， 在AD中扮演重要角色。但另一方面，ADAM-10的上调增加了可溶性淀粉样蛋白-β的形成， 有助于控制AD。因此，使用高选择性抑制剂来定制美替辛的蛋白水解活性， 诱导AD患者脑组织病理改变可能是一种有前途的治疗策略。蛋白抑制剂 与具有广谱活性的较小的合成抑制分子相比， 目标的金属蛋白酶组织抑制剂（TIMPs）是MMPs的主要内源性抑制剂， 具有不同程度的结合亲和力的亚当斯。在所有TIMP中，TIMP-3与肿瘤的发生相关性最低。 触发不希望的脱靶效应的细胞信号分子如在肿瘤发生中起作用的细胞信号分子， 表明其在MMP依赖性疾病中具有低脱靶治疗效果。这使得TIMP-3成为一种 理想的蛋白质支架，用于工程化改良MMP-9抑制剂。然而，TIMP-3也结合其他的 MMPs、亚当斯和生长因子。在这里，我们建议使用理性和随机的组合 开发基于TIMP-3的治疗剂以控制神经退行性疾病的组合方法 通过以高选择性靶向MMP-9来进行进展。这些工程化的基于TIMP的探针将避免结合 ADAM-10在AD中具有神经保护作用，以减少脱靶效应。这个项目的首要目标是 是设计和设计一种新的分子靶向疗法， MMP-9具有高选择性，通过结构研究和细胞培养了解其抑制机制 模型，以改善治疗方案不足的神经退行性疾病的结局。