Comprehensive multimodal analysis of patients with neuroimmunological diseases

神经免疫疾病患者的综合多模态分析

• 批准号: 8158238
• 负责人: Bibiana Bielekova
• 金额: $ 58.56万
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8158238
• 关键词: Disease; Patients

Neuroimmunological diseases of the central nervous system (CNS) represent a broad spectrum of very diverse diagnoses, most of which are considered rare disorders. With the exception of multiple sclerosis (MS), acute demyelinating encephalomyelitis (ADEM), transverse myelitis (TM) and CNS lupus, reported cohorts in the literature rarely exceed 10-20 patients, and it takes years to collect these numbers. Additionally, with the exception of MS, virtually all reports focus on clinical findings and there is a great paucity of data characterizing intrathecal or systemic immune responses in these patients. As a result, the pathophysiology of these diseases is poorly understood and effective therapies are very rare. Emerging data indicate that the immune response is shaped not only by pathogens, but also by the tissue where the inflammatory process develops. From this standpoint, CNS tissue is rather unique. Elegant animal data indicate that foreign grafts survive indefinitely if transplanted into CNS tissue, bypassing systemic presentation of its antigens, whereas they are readily rejected when transplanted into other organs. Indeed, interactions of T cells with neurons can shape T cell effector phenotype, from pathogenic to more regulatory. There is little doubt that these complex immunoregulatory mechanisms emerged as an assurance that CNS tissue, which is vital for the function and survival of the host, will be protected from inadvertent damage by the immune system. Therefore, the apparent failure of the immune system that presents clinically as CNS autoimmunity may originate as a breakdown of natural immunoregulatory mechanisms that govern CNS-immune system interactions. This project studies intrathecal and systemic immune responses in patients referred to NIB for diagnostic work-ups of neuroimmunological CNS disorders. The goal of this study is to define the pathophysiological mechanisms underlying the development of disability in immune-mediated disorders of the CNS and to distinguish these from physiological (and often beneficial) responses of the human immune system to CNS injury. We have established natural history protocol (09-N-0032) under which all untreated patients with suspected immune-mediated disorders of the CNS undergo detailed evaluation at NIB, consisting of the collection of clinical and paraclinical quantitative measures of disease activity and disability, standard and novel quantitative neuroimaging markers and immunological and molecular biomarkers originating from cerebrospinal fluid (CSF), serum and immune cells collected both from peripheral blood and CSF. Additionally, patients may undergo skin biopsy for collection of fibroblasts for their transformation into induced pluripotent stem (iPS) cells in order to develop autologous source of neural cells to study physiological neural-immune interactions in humans, which has not been possible thus far. All patients are coded and analysis of paraclinical, neuroimaging and molecular biomarkers are performed in an unbiased (i.e. blinded) fashion in order to define which biomarkers are associated with specific neuroimmunological disease or phenotype. Our results obtained so far are summarized below: 1. We have developed the methodology for expansion of CD4+ and CD8+ T cells from the limited sample of cerebrospinal fluid (CSF) 2. We have developed methodology for expansion and immortalization of B cells from the limited sample of CSF. We have unblinded our data and summarized the efficacy of EBV CSF B cell transformation for the past 2 years: we observed significantly higher (5 fold increased) efficacy of CSF B cell immortalization by EBV in MS patients, as compared to both inflammatory and non-inflammatory neurological diseases controls. Additionally, we studied phenotype of EBV-immortalized B cells derived from MS patients versus controls in blinded fashion, by combination of surface and intracellular cytokine staining by flow cytometry applied to over 100 immortalized CSF B cell lines. We observed significant differences in the phenotype of CSF B cells derived from MS versus control cohorts. The paper describing our findings is currently in preparation. 3. We analyzed CSF antibody (Ab) reactivity to human myelin and found that intrathecally-produced antibodies of MS patients do not recognize proteins and lipids in their native conformation that are present in human myelin derived from MS patients or control diseased subjects. The paper describing our findings is currently in preparation. 4. We are developing and applying unbiased novel methodologies that aim to determine the target(s) of the intrathecal Ab production in MS patients (collaborative project with Mike Lenardo) 5. We are developing and validating multiple biomarkers of intrathecal oxidative stress 6. We are investigating CSF and blood biomarkers of IL-2 and IL-7 signaling pathways as our studies on MOA of daclizumab indicate that these 2 cytokine pathways are related and mutually regulated and both of these pathways have been genetically linked to MS (collaborative project with Hyun Park) The long-term objective of the study is to acquire knowledge that would allow us to therapeutically inhibit the pathogenic mechanisms and enhance repair mechanisms in immune-mediated CNS diseases, thereby minimizing the extent of CNS tissue damage and promoting recovery. Additionally, we expect that these studies will lead to the development of biomarkers (imaging, immunological and molecular) reflecting concurrent immune-mediated and neurodegenerative pathophysiological mechanisms and those that would able to distinguish among different diagnostic entities. This will lead to rational development and faster screening of process-specific therapies, and will permit the identification of patients with prevailing disease mechanisms, which is a requirement for an individualized approach to medicine. Ultimately, understanding the mechanisms of disease processes will impact the management of immune-mediated diseases of the CNS as a whole.

中枢神经系统（CNS）的神经免疫性疾病代表了非常多样化的诊断的广泛范围，其中大多数被认为是少数疾病。除多发性硬化症（MS）外，急性脱髓鞘脑脊髓炎（ADEM），横向骨髓炎（TM）和CNS狼疮报告在文献中报道了同类文献很少超过10-20名患者，并且需要数年的时间才能收集这些数字。此外，除了MS外，几乎所有报告都集中在临床发现上，并且这些患者中表征鞘内或全身免疫反应的数据很少。结果，这些疾病的病理生理学知之甚少，有效疗法非常罕见。 新兴数据表明，免疫反应不仅由病原体形成，而且由炎症过程发展的组织形成。从这个角度来看，CNS组织是相当独特的。优雅的动物数据表明，如果移植到中枢神经系统组织中，外国移植物将无限期地生存，绕过了其抗原的全身性表现，而将它们移植到其他器官中时很容易被拒绝。实际上，T细胞与神经元的相互作用可以塑造从致病性到更调节性的T细胞效应表型。毫无疑问，这些复杂的免疫调节机制出现是一种保证，即CNS组织对宿主的功能和存活至关重要，将受到免疫系统的无意损害。因此，由于CNS自身免疫性，在临床上呈现的免疫系统的明显失败可能起源于控制CNS-免疫系统相互作用的天然免疫调节机制。 该项目研究了转介给NIB的患者的鞘内和全身免疫反应，用于神经免疫性中枢神经系统疾病的诊断检查。这项研究的目的是确定中枢神经系统免疫介导的疾病中残疾发展的基础的病理生理机制，并将其与人类免疫系统对CNS损伤的生理（且经常有益）反应区分开。我们已经建立了自然历史方案（09-N-0032），在该方案下，所有未经治疗的中枢神经系统的未经治疗的患者在NIB处进行了详细的评估，包括收集临床和旁层次的疾病活性和残疾的疾病活性，标准和新颖的神经性神经疾病和免疫学的熟人（CERENALICAL ECORALICAL ECORPROCTALICALICAL INFIRALICAL ECORTIAL ECORPORTAILIAL ECORPROCTARICAL EREF）的CN，该测量值包括从外周血和CSF收集的免疫细胞。此外，患者可能会进行皮肤活检以收集成纤维细胞的转化为诱导多能干（IPS）细胞，以开发神经细胞的自体源以研究人类的生理神经免疫相互作用，到目前为止尚未实现。 所有患者均进行编码，并以无偏见的（即盲人）方式进行层链，神经影像学和分子生物标志物，以定义哪些生物标志物与特定的神经免疫学疾病或表型相关。 到目前为止，我们获得的结果总结了下面： 1。我们开发了从有限的脑脊液样品（CSF）样品中扩展CD4+和CD8+ T细胞的方法。 2。我们开发了从有限的CSF样品中B细胞扩展和永生化的方法。在过去的两年中，我们没有盲目的数据，并总结了EBV CSF B细胞转化的疗效：与炎症性和非炎性神经系统疾病的对照相比，EBV在MS患者中的CSF B细胞永生化的疗效显着更高（增加了5倍）。此外，我们研究了通过表面和细胞内细胞因子染色，通过将超过100多名永生化的CSF B细胞系的流式细胞术结合结合，以盲目的方式研究了EBV剥离的B细胞的表型。我们观察到来自MS与对照组的CSF B细胞的表型显着差异。描述我们发现的论文目前正在准备。 3。我们分析了对人髓磷脂的CSF抗体（AB）反应性，发现MS患者的鞘内生产的抗体不识别来自MS患者或对照疾病受试者的人髓磷脂中存在的本机构象中的蛋白质和脂质。描述我们发现的论文目前正在准备。 4。我们正在开发和应用公正的新方法，旨在确定MS患者鞘内AB产生的靶标（与Mike Lenardo的协作项目） 5。我们正在开发和验证鞘内氧化应激的多个生物标志物 6。我们正在研究IL-2和IL-7信号传导途径的CSF和血液生物标志物，因为我们对Daclizumab MOA的研究表明，这两种细胞因子途径是相关的，并且相互调节，并且这两种途径均与MS（与Hyun Park的协作性项目）相关。 该研究的长期目标是获取知识，使我们能够治疗抑制病原机制并增强免疫介导的中枢神经系统疾病中的修复机制，从而最大程度地减少中枢神经系统组织损害的程度并促进康复。此外，我们预计这些研究将导致生物标志物（成像，免疫学和分子）的发展，反映出并发的免疫介导的和神经退行性的病理生理机制以及能够区分不同诊断实体的生物标志物。 这将导致合理发展和更快的过程特定疗法筛查，并允许鉴定具有流行疾病机制的患者，这是对医学方法的要求。最终，了解疾病过程的机制将影响整个中枢神经系统的免疫介导的疾病的管理。