Autoimmune mechanisms of gastrointestinal dysmotility in multiple sclerosis

项目摘要/摘要 多发性硬化症(MS)患者除了自主神经功能障碍外 神经肌肉疾病代表他们的标志性症状，也是最常见和最虚弱的 他们的自主神经问题之一是便秘。我们的理论是便秘是一种附属物。 在此期间，MS患者的中枢神经系统(CNS)炎症反应造成的损害。 炎症反应，患者的免疫系统产生针对细胞内蛋白质的抗体 受影响区域死亡的神经元和神经胶质细胞的碎片，这些自身抗体可能在遥远的 网站。肠道神经系统(Ens)的独特之处在于它包含调节 肠道的运动、分泌和血管张力，不依赖于中枢神经系统的支配。ENS含有神经胶质细胞。 具有少突胶质细胞和星形胶质细胞的转录组特征，包括蛋白脂蛋白和髓鞘 碱性蛋白质。这是多发性硬化症中的两个已知的抗原靶向，可以偶然地在它们所在的地方被靶向 也在ENS中表达。这项拨款提案旨在检验自身抗体 多发性硬化症患者产生的蛋白质可以靶向神经元膜和神经胶质膜上的蛋白质，导致改变 肠神经反射活动导致便秘。以支持我们的假设和我们的 建议的研究，我们已经证明了实验性自身免疫小鼠的胃肠道功能发生了改变。 脑脊髓炎(EAE)，MS的主要小鼠模型，其方式与 便秘的发展。此外，我们还发现MS患者和EAE小鼠的血液样本 含有与胃肠道中的神经元和神经胶质细胞结合的抗体。在我们建议的研究中，我们将研究 EAE小鼠胃肠功能障碍的特点，证实肠动力改变与循环有关 自身抗体。我们将确定MS自身抗体结合的ENS细胞类型，以及这些类型的影响 抗体对幼稚小鼠的胃肠道功能有影响。我们还将直接检测神经胶质细胞的生理活动。 细胞和神经元，我们将阐明这些活动是如何在EAE小鼠和在 多发性硬化症患者自身抗体治疗的反应。在这些研究中，我们还将使用来自 通过肠道神经胶质细胞或特定肠道群体表达钙指示蛋白的小鼠 神经元，我们将使用细胞内记录来评估兴奋和抑制的强度 神经肌肉传递。最后，我们将测试潜在的治疗方法在 减轻MS小鼠模型中的便秘。了解便秘的发病机制 多发性硬化症是一个临床相关的目标，因为它是开发有效的治疗方案的关键一步 受影响的MS患者和其他自身免疫介导的运动障碍患者的胃肠道疾病。

Project Summary/Abstract Individuals with multiple sclerosis (MS) frequently suffer from autonomic dysfunction in addition to the neuromuscular ailments that represent their hallmark symptoms, and one of the most common and debilitating of their autonomic problems is constipation. Our theory is that constipation develops as a form of collateral damage from the central nervous system (CNS) inflammatory response that occurs in MS. During this inflammatory response, the patient's immune system generates antibodies that target proteins in the cellular debris of dying neurons and glial cells in affected areas, and these autoantibodies could have actions at distant sites. The enteric nervous system (ENS) is unique in that it contains intrinsic reflex circuitry that regulates motility, secretion and vascular tone in the gut, independently of CNS innervation. The ENS contains glial cells that share transcriptome features of oligodendrocytes and astrocytes, including proteolipid protein and myelin basic protein. These are two known antigenic targets in MS that could be incidentally targeted where they are also expressed in the ENS. This grant proposal is designed to test the hypothesis that autoantibodies generated in MS patients can target proteins on neuronal and glial membranes in the ENS, resulting in altered enteric neural reflex activity leading to constipation. In support of our hypothesis and the feasibility of our proposed studies, we have demonstrated that GI function is altered in mice with experimental autoimmune encephalomyelitis (EAE), the predominant mouse model of MS, in ways that are consistent with the development of constipation. Furthermore, we have found that blood samples from MS patients and EAE mice contain antibodies that bind to neurons and glial cells in the GI tract. In our proposed studies, we will examine the features of GI dysfunction in EAE mice, and confirm that the changes in gut motility involve circulating autoantibodies. We will determine the ENS cell types that MS autoantibodies bind to, and what effects these antibodies have on GI function in naïve mice. We will also directly examine the physiological activities of glial cells and neurons in the GI tract, and we will elucidate how these activities are altered in EAE mice, and in response to treatment with autoantibodies from MS patients. In these studies, we will also use colons from mice in which a calcium indicator protein is expressed by enteric glial cells or select populations of enteric neurons, and we will use intracellular recording to evaluate the strength of excitatory and inhibitory neuromuscular transmission. Finally, we will test potential therapeutic approaches for their effectiveness in alleviating constipation in the MS mouse model. Understanding the mechanisms responsible for constipation in MS is a clinically relevant goal since it is a critical step toward developing an effective therapeutic solution for the GI ailments of affected MS patients and others with autoimmune-mediated dysmotility.