Amyloid beta peptides and type-2 diabetes sequelae synergistically inhibit insulin signaling and trafficking at the blood brain barrier

淀粉样β肽和2型糖尿病后遗症协同抑制血脑屏障的胰岛素信号传导和运输

• 批准号: 10346468
• 负责人: KARUNYA KUMAR KANDIMALLA
• 金额: $ 54.97万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10346468
• 关键词: APP-PS1; Address; Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid beta-Protein; Automobile Driving; Autophagocytosis; Blood - brain barrier anatomy; Brain; Cerebrovascular Circulation; Clinical; Complex; Coupled; Data; Defect; Development; Diabetes Mellitus; Disease; Down-Regulation; Early Onset Alzheimer Disease; Endocytosis; Endothelium; Epidemiology; Etiology; Exhibits; Exocytosis; Flow Cytometry; Functional disorder; Goals; Health; Hemorrhage; High Fat Diet; Image; Impairment; Insulin; Insulin Receptor; Insulin Resistance; Kinetics; Knowledge; Label; Metabolic; Methods; Mission; Modeling; Molecular; Monitor; Mus; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Pathologic; Pathway interactions; Peptides; Pharmaceutical Preparations; Phase; Physiological; Plasma; Protein Array; Public Health; Research; SNAP receptor; Signal Transduction; Stroke; Techniques; Testing; Total Internal Reflection Fluorescent; United States National Institutes of Health; Western Blotting; Wild Type Mouse; Work; X-Ray Computed Tomography; aging population; base; blood-brain barrier function; brain parenchyma; cerebral microvasculature; cerebrovascular; cognitive change; combat; db/db mouse; imaging modality; inhibitor; innovation; insulin signaling; monolayer; mouse model; novel therapeutic intervention; novel therapeutics; overexpression; receptor expression; single photon emission computed tomography; synergism; targeted treatment; trafficking; transcytosis; uptake; APP-PS1; Address; Alzheimer' s Disease; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid beta-Protein; Automobile Driving; Autophagocytosis; Blood - brain barrier anatomy; Brain; Cerebrovascular Circulation; Clinical; Complex; Coupled; Data; Defect; Development; Diabetes Mellitus; Disease; Down-Regulation; Early Onset Alzheimer Disease; Endocytosis; Endothelium; Epidemiology; Etiology; Exhibits; Exocytosis; Flow Cytometry; Functional disorder; Goals; Health; Hemorrhage; High Fat Diet; Image; Impairment; Insulin; Insulin Receptor; Insulin Resistance; Kinetics; Knowledge; Label; Metabolic; Methods; Mission; Modeling; Molecular; Monitor; Mus; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Pathologic; Pathway interactions; Peptides; Pharmaceutical Preparations; Phase; Physiological; Plasma; Protein Array; Public Health; Research; SNAP receptor; Signal Transduction; Stroke; Techniques; Testing; Total Internal Reflection Fluorescent; United States National Institutes of Health; Western Blotting; Wild Type Mouse; Work; X-Ray Computed Tomography; aging population; base; blood-brain barrier function; brain parenchyma; cerebral microvasculature; cerebrovascular; cognitive change; combat; db/db mouse; imaging modality; inhibitor; innovation; insulin signaling; monolayer; mouse model; novel therapeutic intervention; novel therapeutics; overexpression; receptor expression; single photon emission computed tomography; synergism; targeted treatment; trafficking; transcytosis; uptake

PROJECT SUMMARY Type-2 diabetes mellitus (T2DM) sequelae damage the cerebral microvasculature and augment Alzheimer's pathology by inducing brain insulin resistance characterized by sub-physiological insulin levels and impaired insulin-signaling in the brain. Conversely, soluble amyloid beta (sAβ) peptides that accumulate in the plasma and brain during Alzheimer's progression exacerbate the impact of T2DM and aggravate brain insulin resistance. A critical need exists to identify how T2DM sequelae and sAβ exposure inhibit insulin delivery to the brain and intensify brain insulin resistance. The long-term goal is to elucidate cerebrovascular and metabolic contributions to Alzheimer's disease and facilitate the development of novel therapeutic interventions. The overall objective in this application is to determine the combined effects of T2DM sequelae and sAβ on insulin delivery to the brain and to identify the underlying cellular and molecular mechanisms. The central hypothesis is that T2DM sequelae and sAβ peptides perturb insulin signaling/trafficking at the cerebrovascular endothelium [referred to as the blood brain barrier (BBB)] and reduce insulin delivery to the brain. It is also hypothesized that these effects are further aggravated by the pathological synergism between T2DM sequelae and sAβ. The rationale for the proposed research is that a mechanistic understanding of how sAβ exposure and T2DM sequelae disrupt brain insulin delivery will allow us to develop novel therapeutic strategies to address brain insulin resistance in Alzheimer's disease and T2DM. Guided by preliminary data, the following three specific aims are proposed: 1) Determine the effect of T2DM sequelae on insulin trafficking/signaling at the BBB; 2) Determine the effects of sAβ alone and in conjunction with T2DM sequelae on insulin trafficking/signaling at the BBB; and 3) Identify insulin trafficking pathways at the BBB, vulnerable to sAβ exposure and impaired insulin signaling. Under the first and second aims, dynamic SPECT/CT imaging will be used to characterize insulin uptake kinetics at the BBB in mouse models that exhibit T2DM and Alzheimer's sequelae. Moreover, the dysregulation in insulin signaling at the BBB will be captured by reverse phase protein arrays. For the third aim, flow cytometry and TIRF microscopy will be used to determine the effects of sAβ ± insulin signaling inhibitors on insulin transcytosis in BBB monolayers. The proposed research is potentially innovative because it employs dynamic imaging methods coupled with quantitative modeling techniques to capture changes in insulin trafficking kinetics at the BBB in T2DM and Alzheimer's mouse models. The proposed research is significant because the contribution it is expected to have broad translational importance in repurposing existing drugs to treat brain insulin resistance and in identifying candidate targets to discover novel drugs. Upon completion of the work, the new knowledge generated is expected to have an important positive impact by facilitating the identification of novel therapeutic strategies to combat brain insulin resistance in Alzheimer's patients with T2DM.

项目摘要 2型糖尿病（T2DM）后遗症损害了脑微脉和增强阿尔茨海默氏症 病理学通过诱导以亚物理学胰岛素水平为特征的脑胰岛素抵抗和受损 大脑中的胰岛素信号。相反，积聚在血浆中的可溶性淀粉样β（SAβ）肽 阿尔茨海默氏症过程中的大脑加剧了T2DM的影响并加重了大脑胰岛素 反抗。存在关键需求，以确定T2DM后遗症和SAβ暴露如何抑制胰岛素的递送到 大脑并增强大脑胰岛素抵抗。长期目标是阐明脑血管和 对阿尔茨海默氏病的代谢贡献，并促进新疗法的发展 干预措施。该应用程序的总体目的是确定T2DM后遗症的综合作用 SAβ在胰岛素传递到大脑方面，并确定潜在的细胞和分子机制。这 中心假设是T2DM后遗症和SaβPeperides扰动胰岛素信号传导/运输 脑血管内皮[称为血脑屏障（BBB）]，并减少胰岛素的递送 脑。还假设这些效应是通过在 T2DM后遗症和SAβ。拟议研究的理由是，机械理解 SAβ暴露和T2DM后遗症破坏脑胰岛素的递送将使我们能够发展新的治疗 解决阿尔茨海默氏病和T2DM中脑胰岛素抵抗的策略。在初步数据的指导下， 提出了以下三个特定目的：1）确定T2DM后遗素对胰岛素的影响 BBB的贩运/信号； 2）确定单独的SAβ和与T2DM后遗症的影响 关于BBB的胰岛素运输/信号传导； 3）识别BBB的胰岛素运输途径，脆弱 SAβ暴露和胰岛素信号受损。在第一个和第二个目标下，动态SPECT/CT成像 将使用在暴露T2DM和 阿尔茨海默氏症的后遗症。此外，BBB处胰岛素信号传导的失调将通过反向捕获 相蛋白阵列。对于第三个目标，流式细胞仪和TIRF显微镜将用于确定 SAβ±胰岛素信号抑制剂对BBB单层胰岛素转介异增生的影响。拟议的研究 具有创新性，因为它采用动态成像方法以及定量建模 捕获T2DM和阿尔茨海默氏症的BBB胰岛素运输动力学变化的技术 型号。拟议的研究很重要，因为预计其贡献将具有广泛的翻译 重新利用现有药物以治疗大脑胰岛素抵抗并确定候选靶标的重要性 发现新型药物。完成工作后，预计产生的新知识将具有 通过促进鉴定新的治疗策略来对抗大脑的重要积极影响 阿尔茨海默氏症患者T2DM患者的胰岛素抵抗。