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) 的神经免疫疾病代表了一系列非常多样化的诊断，其中大多数被认为是罕见疾病。除多发性硬化症 (MS)、急性脱髓鞘性脑脊髓炎 (ADEM)、横贯性脊髓炎 (TM) 和中枢神经系统狼疮外，文献中报道的队列很少超过 10-20 名患者，并且需要数年时间才能收集这些数字。此外，除了多发性硬化症外，几乎所有报告都集中在临床发现上，并且非常缺乏表征这些患者的鞘内或全身免疫反应的数据。因此，人们对这些疾病的病理生理学知之甚少，有效的治疗方法也非常罕见。 新数据表明，免疫反应不仅受到病原体的影响，还受到炎症过程发生的组织的影响。从这个角度来看，中枢神经系统组织相当独特。优雅的动物数据表明，如果将外来移植物移植到中枢神经系统组织中，则可以无限期地存活，绕过其抗原的全身呈递，而当移植到其他器官中时，它们很容易被排斥。事实上，T 细胞与神经元的相互作用可以塑造 T 细胞效应表型，从致病性到更具调节性。毫无疑问，这些复杂的免疫调节机制的出现保证了对宿主的功能和生存至关重要的中枢神经系统组织将受到保护，免受免疫系统的无意损伤。因此，临床上表现为中枢神经系统自身免疫的免疫系统明显故障可能源于控制中枢神经系统-免疫系统相互作用的自然免疫调节机制的崩溃。 该项目研究转诊至 NIB 的患者的鞘内和全身免疫反应，以进行神经免疫学中枢神经系统疾病的诊断检查。本研究的目的是确定免疫介导的中枢神经系统疾病导致残疾的病理生理机制，并将其与人类免疫系统对中枢神经系统损伤的生理（通常是有益的）反应区分开来。我们建立了自然史方案（09-N-0032），根据该方案，所有未经治疗的疑似中枢神经系统免疫介导疾病的患者在NIB接受详细评估，包括收集疾病活动性和残疾的临床和临床旁定量测量、标准和新型定量神经影像标记以及源自脑脊液（CSF）、血清的免疫学和分子生物标记。 以及从外周血和脑脊液中收集的免疫细胞。此外，患者可能会接受皮肤活检，收集成纤维细胞，将其转化为诱导多能干（iPS）细胞，从而开发自体神经细胞来源，以研究人类的生理神经免疫相互作用，而这迄今为止是不可能的。 对所有患者进行编码，并以无偏见（即盲法）的方式对临床旁、神经影像和分子生物标志物进行分析，以确定哪些生物标志物与特定的神经免疫疾病或表型相关。 迄今为止我们获得的结果总结如下： 1. 我们开发了从有限的脑脊液 (CSF) 样本中扩增 CD4+ 和 CD8+ T 细胞的方法 2. 我们开发了从有限的脑脊液样本中扩增和永生化 B 细胞的方法。我们公开了数据并总结了过去 2 年 EBV CSF B 细胞转化的功效：我们观察到，与炎症性和非炎症性神经系统疾病对照相比，多发性硬化症患者中 EBV 的 CSF B 细胞永生化功效显着更高（增加了 5 倍）。此外，我们通过流式细胞术结合表面和细胞内细胞因子染色，对 100 多种永生化 CSF B 细胞系进行了盲法研究，以盲法方式研究了来自 MS 患者与对照的 EBV 永生化 B 细胞的表型。我们观察到来自 MS 的 CSF B 细胞与对照群体的表型存在显着差异。描述我们发现的论文目前正在准备中。 3.我们分析了脑脊液抗体（Ab）对人髓磷脂的反应性，发现多发性硬化症患者鞘内产生的抗体不能识别来自多发性硬化症患者或对照患病受试者的人髓磷脂中存在的天然构象的蛋白质和脂质。描述我们发现的论文目前正在准备中。 4. 我们正在开发和应用公正的新方法，旨在确定多发性硬化症患者鞘内抗体生产的目标（与 Mike Lenardo 的合作项目） 5.我们正在开发和验证鞘内氧化应激的多种生物标志物 6. 我们正在研究 IL-2 和 IL-7 信号通路的 CSF 和血液生物标志物，因为我们对 daclizumab MOA 的研究表明这 2 条细胞因子通路是相关且相互调节的，并且这两条通路都与 MS 有遗传关联（与 Hyun Park 的合作项目） 该研究的长期目标是获得知识，使我们能够在治疗上抑制免疫介导的中枢神经系统疾病的致病机制并增强修复机制，从而最大限度地减少中枢神经系统组织损伤的程度并促进康复。此外，我们预计这些研究将导致生物标志物（影像学、免疫学和分子）的发展，反映同时存在的免疫介导和神经退行性病理生理机制以及能够区分不同诊断实体的生物标志物。 这将导致特定过程疗法的合理开发和更快筛选，并将允许识别具有流行疾病机制的患者，这是个体化医学方法的要求。最终，了解疾病过程的机制将影响整个中枢神经系统免疫介导疾病的管理。