Targeting the alpha secretase ADAM10 for the treatment of Alzheimer's disease

靶向 α 分泌酶 ADAM10 治疗阿尔茨海默病

• 批准号: 10590899
• 负责人: DIMITAR B NIKOLOV
• 金额: $ 26.55万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10590899
• 关键词: Alzheimer' s Disease; Alzheimer' s disease therapeutic; Amino Acids; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Binding; Biochemical; Biological Assay; Biophysics; Cell Death; Cell Line; Cell surface; Cells; Cognition Disorders; Colon Carcinoma; Complex; Cryoelectron Microscopy; Cysteine; Degenerative Disorder; Dementia; Development; Digestion; Disadvantaged; Disintegrins; Engineering; Ephrins; Epidermal Growth Factor Receptor; Equilibrium; Exhibits; Family; Free Energy; Glioblastoma; Hand; Human; In Vitro; Kinetics; Ligands; Long-Term Potentiation; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Mediating; Memory; Memory Loss; Metalloproteases; Molecular; Molecular Conformation; Molecular Mechanisms of Action; Monoclonal Antibodies; Mus; Names; Neurons; Oncogenic; PC12 Cells; Pathway interactions; Peptide Hydrolases; Peptides; Positioning Attribute; Pre-Clinical Model; Prevalence; Primary Bone Osteosarcoma; Property; Protease Domain; Proteins; Proteolysis; Rattus; Reporting; Research; Resolution; Senile Plaques; Signal Transduction; Spectrum Analysis; Structure; Synapses; Tertiary Protein Structure; Therapeutic; Therapeutic Intervention; Thermodynamics; Toxic effect; Transfection; Up-Regulation; X-Ray Crystallography; Zinc; alpha secretase; amyloid formation; analytical ultracentrifugation; enthalpy; gamma secretase; human monoclonal antibodies; improved; malignant stomach neoplasm; member; microcalorimetry; mouth squamous cell carcinoma; neoplastic cell; neuroblastoma cell; neuroprotection; neurotoxic; notch protein; novel; novel strategies; novel therapeutic intervention; overexpression; protein complex; secretase; sequential proteolysis; side effect; stoichiometry; tumor

Summary/Abstract Alzheimer’s disease (AD), one of the most prevalent degenerative diseases, is characterized by the prevalence of senile plaques that consist of fibrils of amyloid b (Ab). In the amyloidogenic pathway, sequential proteolysis of the b-amyloid precursor protein (APP) by b and g secretases generate Ab (40- 42 amino acid) peptides. Ab multimerizes into neurotoxic oligomers that target the synapse, leading to disruption of long-term potentiation, cell death, and memory loss. Alternatively, in the non-amyloidogenic pathway, APP is cleaved sequentially by a and g secretase to generate sAPPa that exhibits neuroprotective properties and precludes the formation of Ab. Studies have identified ADAM10, a member of the adamlysin family of zinc metalloproteinases, as the constitutive a-secretase in neurons. Activation of ADAM10 is currently being evaluated as a means of therapeutic intervention. However, this strategy has severe disadvantages: ADAM10 is known to cleave multiple substrates and is upregulated in a wide range of malignancies. A non-specific (non-selective) up-regulation/enhancement of ADAM10 as a means of therapeutic intervention in AD would likely have deleterious side effects. The importance of the non-catalytic (disintegrin and cysteine rich) domains of ADAM10 in substrate recognition was shown previously. A conformation specific mAb, 8C7, which recognizes an active conformation of ADAM10 found on tumor cells was developed. Importantly, while 8C7 selectively enhances APP cleavage in cell-based assay, it inhibits Notch-Delta and Eph-ephrin oncogenic signaling in preclinical models, without any discernible toxicity effects in mice, thereby ruling out deleterious side effects that might result from its potential use as AD therapeutic. The proposed studies will characterize the ADAM10-APP interactions in vitro to better understand how the ADAM10 a-secretase interacts with and cleaves APP. The molecular mechanism of action of the 8C7 mAb in promoting this cleavage will also be investigated. A human version of the anti-ADAM10 mAb 8C7, named 1H5, that augments cleavage of peptide substrates and binds to the substrate binding domain of ADAM10, was generated recently. Similar to 8C7, this human mAb (1H5), as well as a single-chain variable fragment (scFv) derived from it, will now be evaluated in cell-based assays in order to determine their efficacies in promoting APP cleavage. The shedding of APP will further be examined in rat neuronal and human glioblastoma cell lines in the presence of the mAbs (8C7 and 1H5) and the scFv. This proposal, to enhance the non- amyloidogenic pathway with anti-ADAM10 mAbs towards the development of novel AD treatments, serves as a proof of principle of therapeutic potential.

摘要/摘要 阿尔茨海默氏病（AD）是最流行的退行性疾病之一，其特点是 在淀粉样蛋白生成途径中，由淀粉样蛋白 b (Ab) 纤维组成的老年斑的患病率， b 和 g 分泌酶对 b-淀粉样前体蛋白 (APP) 进行连续蛋白水解，生成 Ab (40- 42 个氨基酸）肽多聚成针对突触的神经毒性寡聚体，从而导致 或者，在非淀粉样蛋白生成中，长期增强作用被破坏、细胞死亡和记忆丧失。 途径中，APP 被 a 和 g 分泌酶依次裂解，生成 sAPPa，其表现出 ADAM10 具有神经保护特性并阻止抗体的形成。 锌金属蛋白酶 adamlysin 家族的成员，作为神经元中的组成型 α-分泌酶。 目前正在评估 ADAM10 的激活作为一种治疗干预手段。 该策略有严重的缺点：ADAM10 已知可裂解多个底物并且上调 ADAM10 的非特异性（非选择性）上调/增强。 作为 AD 治疗干预的一种手段可能会产生有害的副作用。 ADAM10 在底物识别中的非催化（解整合素和富含半胱氨酸）结构域的 如前所示，构象特异性单克隆抗体 8C7，可识别 的活性构象。 重要的是，开发了在肿瘤细胞上发现的 ADAM10，而 8C7 则选择性增强 APP。 在基于细胞的检测中进行切割，它在临床前抑制 Notch-Delta 和 Eph-ephrin 致癌信号传导 模型，在小鼠中没有任何明显的毒性作用，从而排除了有害的副作用 可能源于其作为 AD 治疗的潜在用途。拟议的研究将描述其特征。 ADAM10-APP 体外相互作用，以更好地了解 ADAM10 α-分泌酶如何与 和 8C7 mAb 促进这种裂解的分子作用机制也将被证实。 人类版本的抗 ADAM10 单克隆抗体 8C7（名为 1H5）可增强裂解。 最近生成了肽底物并与 ADAM10 的底物结合结构域结合。 与 8C7 类似，这种人 mAb (1H5) 以及源自它的单链可变片段 (scFv)， 现在将在基于细胞的测定中进行评估，以确定它们在促进 APP 方面的功效 将在大鼠神经和人胶质母细胞瘤细胞中进一步检查 APP 的脱落。 在存在单克隆抗体（8C7 和 1H5）和 scFv 的情况下，该方案增强了非抗性。 抗 ADAM10 单克隆抗体的淀粉样蛋白生成途径，用于开发新型 AD 治疗方法， 作为治疗潜力原理的证明。