Autoimmune Mechanisms of Diabetic Neuropathy

糖尿病神经病变的自身免疫机制

• 批准号: 7344806
• 负责人: JOHN W WILEY
• 金额: $ 29.32万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7344806
• 关键词: 1-Phosphatidylinositol 3-Kinase; Adenosine Triphosphate; Age; Agonist; Animal Model; Animals; Apoptosis; Apoptotic; Autoantibodies; Autoimmune Process; Autoimmunity; Autophagocytosis; Autophagosome; Binding; Binding Proteins; Biological Markers; Budgets; CD95 Antigens; Cardiovascular system; Caspase; Cell Death; Cell membrane; Cells; Cellular Stress; Cessation of life; Class; Complement; Condition; Data; Death Domain; Development; Diabetes Mellitus; Diabetic Neuropathies; Diagnosis; Diet; Disease; Enrollment; Event; Family; Fatty acid glycerol esters; Female; Functional disorder; Gender; Gene Silencing; Hour; Human; Immunoglobulins; In Situ; In Vitro; Injury; Lead; Link; Longitudinal Studies; Malignant Neoplasms; Mediating; Methods; Mitochondria; Modeling; Monitor; Myenteric Plexus; Nervous system structure; Neuroblastoma; Neuronal Dysfunction; Neuronal Injury; Neurons; Neuropathy; Nodose Ganglion; Non-Insulin-Dependent Diabetes Mellitus; Organelles; Oxidative Stress; Pathway interactions; Patients; Pattern; Peripheral; Peripheral Nervous System Diseases; Play; Process; Proteins; Public Health; Rattus; Reporting; Research; Research Personnel; Role; Sensory; Serum; Spinal Ganglia; Streptozocin; Streptozocin Diabetes; Stress; TNFRSF6 gene; Time; autonomic neuropathy; base; cohort; diabetic; diabetic rat; exhaust; healthy aging; inhibitor/antagonist; injured; insight; member; mitochondrial dysfunction; mitochondrial permeability transition pore; non-diabetic; novel; programs; receptor; release factor; research study; response

The pathophysiology of diabetic neuropathy is poorly understood. We reported previously that sera from patients with type 2 diabetes mellitus with neuropathy induced programmed cell death (PCD) in cultured human neuroblastoma cells via an autoantibody-mediated pathway involving activation of caspase- dependent apoptosis. Recent studies support that a caspase-independent pathway can also contribute to PCD. This process may involve activation of autophagy, a pathway that sequesters proteins and organelles in autophagosomes in response to cellular stress. We propose the novel hypothesis that autoantibodies present in the sera of type 2 diabetic patients with neuropathy sequentially activate autophagy, caspase- dependent and -independent PCD in neurons via a Fas-dependent pathway. Fas (CD95) is a member of the cell membrane-bound death receptor family. Our preliminary data support the hypothesis that agonist autoantibodies bind and activate the Fas receptor. We will monitor the development of autoantibody(-ies) and correlate their presence with established markers of autonomic and peripheral neuropathy in an early and later cohort of patients with type 2 diabetes mellitus. We will dissect the pathway(s) that induce autophagy and PCD. We propose that autophagy is an early cytoprotective response to remove injured mitochondria that progresses sequentially to caspase-dependent and -independent PCD with decrease in ATP levels. We hypothesize that autoantibody-induction of autophagy will involve incorporation of LC3n, a specific marker for autophagosomes, and activation of PI-3 kinase (class III). Cultured SH-SY5Y (human neuroblastoma cells) will be exposed to complement-inactivated sera obtained from: 1. type 2 diabetic patients with documented diabetic neuropathy, 2. Age- and gender- matched type 2 diabetic patients without evidence of neuropathy and 3. Healthy, age- and gender-matched controls. Parallel animal studies will be performed examining whether autophagy is activated in situ in primary neurons (nodose ganglia, dorsal root ganglia and myenteric plexus) obtained from the female Zucker Diabetic Fatty rat, an inducible model of type 2 diabetes and the streptozotocin-induced diabetic rat, a well validated model of diabetic neuropathy. We will also assess whether sera from diabetic rats with neuropathy induce autophagy and caspase-dependent and/or -independent PCD in cultured rat nodose ganglia, DRG and myenteric neurons compared to sera from diabetic rats without neuropathy and lean, non-diabetic controls, and examine the time-course for these events. We hypothesize that mitochondrial dysfunction and decreased levels of ATP will play a pivotal role in sequential activation of caspase-dependent and -independent PCD. Relevance to public health: These studies will lead to novel insights regarding the mechanistic basis of autoimmunity, autophagy and programmed cell death in the pathophysiology of diabetic neuropathy. .

糖尿病神经病的病理生理学知之甚少。我们之前报道了来自 培养的2型糖尿病患者伴有神经病诱导的程序性细胞死亡（PCD）的患者 人类神经母细胞瘤细胞通过自身抗体介导的途径，涉及caspase-激活 依赖性凋亡。最近的研究支持，即caspase独立途径也可以有助于 PCD。此过程可能涉及自噬的激活，自噬是一种隔离蛋白质和细胞器的途径 在响应细胞应激的自噬体中。我们提出了自身抗体的新假设 存在于2型糖尿病患者的血清中，依次激活自噬，caspase- 通过FAS依赖性途径在神经元中的依赖和非依赖性PCD。 FAS（CD95）是 细胞膜结合的死亡受体家族。我们的初步数据支持激动剂的假设 自身抗体结合并激活FAS受体。我们将监视自身抗体的发展（-ies） 并将其存在与早期的自主神经和周围神经病的既定标记相关联 后来的2型糖尿病患者队列。我们将剖析诱导的途径 自噬和PCD。我们建议自噬是一种早期的细胞保护反应，以消除受伤 线粒体顺序依赖于caspase依赖性和非依赖性PCD，并降低 ATP级别。我们假设自噬自动抗体诱导将涉及掺入LC3N，A 自噬体的特定标记和PI-3激酶的激活（III类）。培养的SH-SY5Y（人类 神经母细胞瘤细胞）将暴露于以下补体灭活的血清：1。类型2糖尿病 患有糖尿病神经病的患者，2。年龄和性别匹配的2型糖尿病患者 神经病和3。健康，年龄和性别匹配的对照的证据。平行动物研究将是 进行了检查是否在原发性神经元（nodose Ganglia，背根）的原位激活自噬 神经节和骨髓丛）从雌性Zucker糖尿病脂肪大鼠获得的类型诱导模型 2个糖尿病和链蛋白酶诱导的糖尿病大鼠，这是一个良好的糖尿病神经病模型。我们将 还要评估神经病糖尿病大鼠的血清是否诱导自噬和caspase依赖性 与血清相比 从没有神经病和瘦肉的糖尿病大鼠，非糖尿病控制，并检查这些时间顺序 事件。我们假设线粒体功能障碍和ATP水平降低将起关键作用 在caspase依赖性和非依赖性PCD的顺序激活中。与公共卫生有关：这些 研究将导致有关自身免疫，自噬和自动免疫的机理基础的新见解 糖尿病神经病病理生理学中的程序性细胞死亡。 。