Model for SorCS1-mediated Diabetes with Dementia

SorCS1 介导的糖尿病伴痴呆模型

• 批准号: 8599496
• 负责人: SAMUEL E. GANDY
• 金额: $ 36.71万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-15 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8599496
• 关键词: Aging; Alzheimer' s Disease; Alzheimer' s disease risk; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Brain; Characteristics; Complex; Cultured Cells; Data; Dementia; Diabetes Mellitus; Disease; Family; Female; Functional disorder; Gender; Gene Transfer; Generations; Genetic; Genetic Polymorphism; Human; Insulin; Insulin Resistance; Link; Mediating; Metabolic; Metabolism; Modeling; Molecular; Mus; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Patients; Phenotype; Phosphorylation; Protein Precursors; Proteins; Risk; Role; Serine; Structure; Tertiary Protein Structure; Testing; Transgenic Mice; Vacuolar Protein Sorting; Vertebral column; Viral Genes; Work; amyloid peptide; genetic association; genetic linkage; glucose metabolism; glucose tolerance; hippocampal morphometry; hypercholesterolemia; insulin tolerance; learned behavior; overexpression; protein complex; protein metabolism; protein transport; sexual dimorphism; sortilin; trafficking

SUMMARY Type 2 diabetes mellitus (T2D or T2DM) increases the risk for Alzheimer's disease (AD), and SORCS1 is genetically linked to both T2D and AD. We have undertaken a study of the possible role(s) for SorCS1 in metabolism of the Alzheimer's amyloid-¿ (¿¿) precursor protein (APP), in order to define the molecular mechanisms underlying this coordinate genetic linkage to both diseases. Overexpression of SorCS1c¿-myc in cultured cells caused a reduction (p=0.002) in ¿¿ generation (Lane et al., 2010). Endogenous murine A¿40 and ¿¿42 levels were increased (A¿40, p=0.044; A¿42, p=0.007) in the brains of female Sorcs1 hypomorphic mice, possibly paralleling the sexual dimorphism that is characteristic of the genetic associations of SORCS1 with AD and DM. Since SorL1, another AD-linked Vps10-domain protein, directly interacts with Vps35 to modulate APP metabolism, we investigated the possibility that SorCS1c¿-myc might interact with APP, SorL1, and/or Vps35. We readily recovered SorCS1:APP, SorCS1:SorL1, and SorCS1:Vps35 complexes from nontransgenic mouse brain. Notably, total Vps35 protein levels were decreased by 49% (p=0.009) and total SorL1 protein levels were decreased by 29% (p=0.003) in the brains of female Sorcs1-/- mice. We hypothesize that dysfunction of SorCS1 may contribute to both the APP/¿¿ disturbance underlying AD and the insulin/glucose metabolism disturbance underlying DM. In order to test this hypothesis further, we propose the following specific aims: Specific Aim 1. To evaluate the importance of SorCS1 protein interaction motifs and SorCS1/SorL1/APP complex formation on APP metabolism by: (a) Characterizing APP metabolism in cultured cells overexpressing SorCS1; (b) Testing the effects of mutations of protein-protein interacting motifs in the cytoplasmic and ectodomains of SorCS1 on both the formation of tripartite SorCS1/SorL1/APP complexes and APP metabolism; (c) Testing the effect of a putative pathogenic SorCS1 polymorphism on both the formation of tripartite SorCS1/SorL1/APP complexes and APP metabolism; (d) Confirming the importance of functional domains identified in Aim 1aii and 1aiii by viral gene transfer into primary cultures. Specific Aim 2. To employ Sorcs1 hypomorphic and plaque- forming human Swedish APP/PS bigenic mice crossed with Sorcs1 hypomorphic mice for characterization of: (i) endogenous APP metabolism; (ii) hippocampal morphometry, dendritic arborization, and spine structure; (c) learning behavior. Aging (3 mo, 6 mo, 12 mo) effects will also be studied. Specific Aim 3. To perform standard glucose and insulin tolerance tests and metabolic profile phenotyping of Sorcs1-/- mice and plaque-forming" human Swedish APP/PS co-transgenic mice crossed with Sorcs1 -/- mice.

概括 2型糖尿病（T2D或T2DM）增加了阿尔茨海默氏病（AD）的风险，而Sorcs1为 遗传与T2D和AD相关。我们已经研究了Sorcs1可能作用的研究 为了定义分子，阿尔茨海默氏症淀粉样蛋白（APP）的代谢 将这种坐标为两种疾病的遗传联系的机制。 sorcs1c¿-myc的过表达 在培养的细胞中，引起了»生成（Lane等，2010）的降低（P = 0.002）。内源鼠 a¿40和€€增加了42个级别（a¿40，p = 0.044;a¿42，p = 0.007）女性sorcs1 型型小鼠，可能与遗传特征的性二态性平行 Sorcs1与AD和DM的关联。自SORL1（另一种广告连接的VPS10域蛋白）以来，直接 与VPS35相互作用以调节APP代谢，我们研究了Sorcs1c¿-myc可能的可能性 与应用程序，SORL1和/或VPS35进行交互。我们很容易恢复sorcs1：app，sorcs1：sorl1和 SORCS1：来自非转基因小鼠大脑的VPS35复合物。值得注意的是，总VPS35蛋白水平为 在大脑中，降低49％（P = 0.009），总Sorl1蛋白水平降低29％（P = 0.003） 雌性sorcs1 - / - 小鼠的。我们假设SORCS1的功能障碍可能有助于应用程序/�` DM的胰岛素/葡萄糖代谢障碍的障碍和胰岛素/葡萄糖代谢障碍。为了测试 该假设进一步提出了以下特定目的：特定目的1。评估重要性 Sorcs1蛋白相互作用基序和Sorcs1/Sorl1/App复合形成在APP代谢上作者：（a） 表征过表达Sorcs1的培养细胞中的应用代谢； （b）测试 蛋白质 - 蛋白质相互作用基序的突变在两种质质和sorcs1的胞外域的突变 三方Sorcs1/sorl1/应用复合物和应用代谢的形成； （c）测试a的效果 在三方Sorcs1/Sorl1/App Complexs的形成上，假定的致病性Sorcs1多态性 和应用代谢； （d）确认在AIM 1AII和1AIII中确定的功能域的重要性 病毒基因转移到原发性培养物中。特定目的2。对员工Sorcs1 hydormforic和Plaque- 形成与Sorcs1型型型肌big鼠的人类瑞典应用/PS临界小鼠进行表征 的：（i）内源性应用代谢； （ii）海马形态计量学，树突状树博和脊柱 结构; （c）学习行为。衰老（3 mo，6 mo，12个月）的效果也将研究。特定目标3。 执行标准的葡萄糖和胰岛素耐受性测试以及Sorcs1 - / - 小鼠的代谢表型 和形成斑块的“人类瑞典应用程序/PS的共转基因小鼠与Sorcs1 - / - 小鼠交叉。