Immunomodulation targeting abnormal conformation and the influence of apoE

针对异常构象的免疫调节及apoE的影响

• 批准号: 10428586
• 负责人: Fernando goni
• 金额: $ 46.39万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10428586
• 关键词: 3xTg-AD mouse; APP-PS1; Active Immunization; Affect; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid; Amyloid beta-Protein; Animal Model; Animals; Antibodies; Antibody Binding Sites; Antibody Response; Apolipoprotein E; Autoimmune; Behavioral; Binding; Binding Sites; Biochemical; Biological Markers; Blood - brain barrier anatomy; Blood Vessels; Brain; Cerebral Amyloid Angiopathy; Cerebrovascular system; Characteristics; Clinical Trials; Cognitive; Complication; Elderly; Female; Future; Hemorrhage; Human; Image; Immune Targeting; Immunization; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Immunotherapy; Injections; Intraperitoneal Injections; Kinetics; Knock-in; Legal patent; Mass Spectrum Analysis; Measures; Mediating; Modeling; Molecular Conformation; Monoclonal Antibodies; Mus; Neurodegenerative Disorders; Neurofibrillary Tangles; Passive Immunization; Passive Immunotherapy; Pathogenesis; Pathologic; Pathology; Peptoids; Peripheral; Permeability; Proteome; Proteomics; Reporting; Safety; Senile Plaques; Sterility; Structure; Testing; Therapeutic Effect; Toxic effect; Vaccination; Western Blotting; abeta oligomer; amyloid structure; apolipoprotein E-4; autoimmune toxicity; beta pleated sheet; blood-brain barrier penetration; cognitive testing; conformer; extracellular; human tissue; hyperphosphorylated tau; immunological intervention; immunoregulation; innovation; insight; male; misfolded protein; mouse model; nanomolar; neuropathology; novel strategies; oAβ; prion-like; side effect; small molecule; tau Proteins; tau aggregation; vascular abnormality

SUMMARY The neuropathology of AD includes fibrillary amyloid β (Aβ) in plaques, cerebral amyloid angiopathy (CAA), and hyperphosphorylated tau fibrils in neurofibrillary tangles (NFT). However, the most toxic species are Aβ and tau oligomers characterized by generic structural β-sheet secondary structure. These misfolded Aβ and tau conformers are suitable targets for immunological intervention, although numerous clinical trials thus far have failed due to: 1) autoimmune toxicity; 2) lack of specific concomitant targeting of oligomeric Aβ and tau species; and 3) Amyloid Related Imaging Abnormalities (ARIA), particularly among apolipoprotein (apo)E4 carriers, thought to be associated with fibrillar vessel amyloid clearance. To overcome these limitations, we developed innovative antibody combining sites (aβComAb paratopes) that only recognize the dominant β- sheet secondary structure of misfolded proteins, a generic characteristic of all pathologic oligomers found in neurodegenerative diseases. Our preliminary results show that some aβComAb combining sites on either IgM or IgG class can, without side effects, penetrate the BBB of an AD Tg mouse model with pre-existing Aβ and tau pathology, achieve significant cognitive rescue, reduce levels of Aβ and tau pathological oligomers, and in a CAA AD Tg mouse model (TgSwDI) act without ARIA-like toxicity (both models on murine apoE background). In this project, we will test the hypothesis that aβComAbs (IgM or IgG), unlike mAbs directed against Aβ sequences (6E10), can be infused safely and produce modifying therapeutic effects without vascular ARIA-like toxicity in 3xTg and the vascular TgSwDI or APP/PS1dE9 mouse models cross-bred to human apoE2, E3, or E4 backgrounds. Furthermore, we anticipate that the vascular amyloid proteomes of these mice will resemble those that Projects 1 and 2 associate with different apoE backgrounds, treatments, and presence or absence of microhemorrhages. This information is critically needed to elucidate the mechanism associated with ARIA and select aβComAbs paratopes for future clinical trials. The specific aims are to: 1) Produce and characterize IgM and IgG forms of combining sites (AβComAb paratopes) that specifically recognize the β-sheet secondary structures of toxic oligomers. 2) Determine the biochemical and histochemical interaction of the four aβComAbs from Aim 1 with the brain vasculature of peripherally infused TgSwDI mice crossed on KI human ApoE2, E3, and E4 backgrounds. 3) Determine behavioral, histochemical, and biochemical changes after passive immunotherapy with two AβComAb paratopes on an IgM and an IgG selected from Aim 2, in 3xTg, Tg APP/PS1dE9, and TgSwDI mice crossed on KI human ApoE2, E3, and E4 backgrounds.

总结 AD的神经病理学包括斑块中的β淀粉样蛋白（Aβ）、脑淀粉样血管病（CAA）、 和神经纤维缠结（NFT）中的过度磷酸化tau纤维。然而，毒性最大的种类是Aβ 和特征在于通用结构β-折叠二级结构的tau寡聚体。这些错误折叠的Aβ和 tau构象异构体是免疫干预的合适靶标，尽管迄今为止的许多临床试验 失败的原因是：1）自身免疫毒性; 2）缺乏寡聚体Aβ和tau的特异性伴随靶向 种;和3）淀粉样蛋白相关成像抗体（ARIA），特别是载脂蛋白（apo）E4 携带者，被认为与纤维状血管淀粉样蛋白清除有关。为了克服这些限制，我们 开发了创新的抗体结合位点（βComAb互补位），仅识别显性β- 错误折叠的蛋白质的折叠二级结构，这是所有病理性寡聚体的一般特征， 神经退行性疾病我们的初步研究结果表明，在IgM和IgM上的一些α βComAb结合位点， 或IgG类可以无副作用地穿透预先存在Aβ的AD Tg小鼠模型的BBB， tau病理学，实现显著的认知拯救，降低Aβ和tau病理性寡聚体的水平， CAA AD Tg小鼠模型（TgSwDI）没有ARIA样毒性（两种模型均对鼠apoE 背景）。在这个项目中，我们将测试一个假设，即β ComAb（IgM或IgG），不像单克隆抗体直接 针对Aβ序列（6 E10），可以安全输注，并产生修饰性治疗效果， 3xTg和血管TgSwDI或APP/PS1 dE 9杂交小鼠模型中的血管ARIA样毒性 人类apoE 2、E3或E4背景。此外，我们预计血管淀粉样蛋白 这些小鼠的蛋白质组将类似于项目1和2与不同apoE相关的蛋白质组 背景、治疗和是否存在微血管造影。这一信息至关重要， 需要阐明ARIA的相关机制，并为将来的临床应用选择aβComAbs的互补位 审判具体目标是： 1)产生并表征特异性结合位点的IgM和IgG形式（AβComAb互补位）， 识别有毒低聚物的β折叠二级结构。 2)确定来自Aim 1的4种aβ ComAb与脑的生物化学和组织化学相互作用 外周输注TgSwDI小鼠的血管系统在KI人ApoE 2、E3和E4背景上杂交。 3)确定被动免疫治疗后的行为，组织化学和生化变化， 在3xTg、Tg APP/PS1 dE 9和TgSwDI小鼠中，AβComAb与选自Aim 2的IgM和IgG互补位 在KI人ApoE 2、E3和E4背景上杂交。