Neuroimmune responses and therapeutics of alpha-synucleinopathies of the aging population

老年人群α-突触核蛋白病的神经免疫反应和治疗

基本信息

批准号：
10688878
负责人：
Eliezer Masliah
金额：
$ 8.36万
依托单位：
NATIONAL INSTITUTE ON AGING
依托单位国家：
美国
项目类别：
财政年份：
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/10688878
关键词：
2019-nCoV Academia Address Affect Aging Alzheimer&apos s Disease Alzheimer&apos s disease related dementia Amygdaloid structure Amyloid beta-Protein Animal Model Animals Antibodies Antibody titer measurement Apolipoprotein E Area Astrocytes Axon B-Lymphocytes Behavior Behavioral Binding Blood - brain barrier anatomy Brain COVID-19 COVID-19 pandemic effects COVID-19 patient CSF1 gene CSF1R gene CSF2 gene Cell Aging Cell model Cells ChIP-seq Chronic Disease Collaborations Combination immunotherapy DNA Damage DNA Vaccines Dementia with Lewy Bodies Disease Outcome Disease model Elderly Epitopes Gene Expression Profile Genes Genetic Genetically Engineered Mouse Goals Human IL6 gene Immune Immune response Immune system Immunologic Receptors Immunomodulators Immunotherapeutic agent Immunotherapy In Vitro Inflammation Inflammatory Inflammatory Response Injections Innate Immune Response Interneurons LRRK2 gene Laboratories MAP Kinase Gene MAPK12 gene Manuscripts Mediating Mediator of activation protein Methods Microglia Modeling Molecular Mouse Strains Mus NF-kappa B National Institute of Neurological Disorders and Stroke Natural Immunity Nerve Degeneration Neurodegenerative Disorders Neuroglia Neuroimmune Neuroimmune system Neurologic Neurons Nuclear Translocation Paper Parkinson Disease Parkinson&apos s Dementia Pathogenesis Pathogenicity Pathologic Pathology Pathway interactions Peptides Peripheral Pharmacology Phenotype Phosphotransferases Play Preparation Publications Publishing Research Role Science Signal Pathway Signal Transduction Synapses System T cell response T-Lymphocyte TLR2 gene TNF gene Technology Testing Therapeutic Therapeutic Effect Toll-like receptors Transgenic Mice Transgenic Model Tumor-infiltrating immune cells Vaccinated Vaccines Viral Vector Wild Type Mouse adaptive immunity age effect age related aged aging population alpha synuclein base brain tissue cell type combinatorial cytokine differential expression env Gene Products experimental study extracellular gene network genetic architecture immunomodulatory therapies immunoregulation immunosenescence in vitro Model in vivo in vivo Model inhibitor neurogenetics neuroinflammation neuropathology neurotoxic neurotoxicity novel novel therapeutics nuclear factors of activated T-cells p38 Mitogen Activated Protein Kinase protein TDP-43 protein aggregation proteostasis receptor response sex single-cell RNA sequencing small molecule synaptic function synucleinopathy targeted treatment tau Proteins therapeutic target therapy development trafficking transcription factor transcriptome sequencing transcriptomics transmission process γδ T cells

项目摘要

The main objective of the Molecular Neuropathology unit (MNU) at Laboratory of Neurogenetics (LNG) is to harness the immune system to better understand the mechanisms of neurodegeneration and to develop therapies for synucleinopathies of the aging population such as DLB, PD and AD. We propose 3 Aims, the first investigating the role of innate immune responses and combinatorial immunotherapy targeting LRRK2, Toll-like receptors, p38, NFAT and the protein aggregates (eg: a-synuclein, Abeta, tau) in PD/DLB; the second assessing downstream pro-inflammatory signaling pathways including MAPK-p38, NFAT and NFkB and the third evaluating the role of aging in T cell mediated adaptive immune cell responses in PD/DLB pathogenesis and for developing immunotherapies for synucleinopathies. During this period, we published 9 manuscripts mostly focusing understanding the pathogenesis of PD/DLB and developing novel pharmacological and immunotherapeutical approaches. Progress for Aim1. In previous studies we identified several immune receptors that mediate neuroinflammation in synucleinopathies, including Toll-like receptor 2 (TLR2) and have shown which species of a-synuclein bind TLR2 and mediates neuroinflammation. These results suggest that cell type-specific modulation of TLR2 may be a multifaceted and promising therapeutic strategy for synucleinopathies; inhibition of neuronal and astroglial TLR2 decreases pathogenic a-syn transmission, but activation of microglial TLR2 enhances microglial extracellular a-syn clearance. Along these lines we recently published a manuscript (Torres et al Neurobiol of Dis 2022) characterizing in vivo in our a-synuclein transgenic model the relationship between degeneration of interneurons and neuroinflammation in the amygdala. As an extension and given the impact of the COVID19 pandemic and potential role of SARS-CoV2 in AD/ADRD we investigated (in collaboration with the Nath laboratory) inflammation in the brains of COVID19 patients with evidence of neurological involvement, we found increased a-synuclein accumulation in axons and glial cells as well as astrocytic activation and T cell trafficking in affected CNS areas. Moreover, preliminary studies indicate that the SARS-CoV2 envelope protein might interact with TLR2 and play a role in inflammation and a-synuclein driven pathology, we published a manuscript addressing this possibility (Szabo et al Exp Mol Med 2022). Finally, this year in collaborative projects with academia we made considerable progress developing novel ADRD immunotherapies that either target 3RTau (Spencer et al Methods Mol Biol 2022) or simultaneously target multiple epitopes of a-synuclein. For Tau we developed a novel brain penetrating single chain antibody packed in a viral vector that can be applied peripherally. For a-synuclein, we used the MultiTEP based DNA vaccine technology which is an approach especially useful for the elderly with immunosenescence. All vaccines induced high titers of antibodies specific to a-synuclein that significantly reduced PD/DLB-like pathology in mice. The most significant effects were observed in mice vaccinated with PV-1949D and PV-1950D in a sex-dependent manner. We published two recent studies (Kim et al Vaccines 2022 and Zagorski et al Int J Mol Scie 2022). Progress for Aim 2. In collaboration with the Cookson group (Kim et al Science TM 2020) showed that LRRK2 plays a critical role in the activation of microglia by extracellular a-synuclein. We determined that LRRK2 promoted a neuroinflammatory cascade by selectively phosphorylating and inducing nuclear translocation of the immune transcription factor nuclear factor of activated T cells, cytoplasmic 2 (NFATc2). Our results suggest that modulation of LRRK2 and its downstream signaling mediator NFATc2 might be therapeutic targets for treating synucleinopathies. Next, we investigated the hypothesis that blocking NFATc2 translocation might ameliorate a-synuclein-mediated microglial neurotoxicity. Preliminary studies identified a novel peptide compound (11r VIVIT) that when administered peripherally into -syn tg mice reduced neuroinflammation, restores NFATc2 cytoplasmic localization and ameliorates behavioral deficits. We are currently expanding these studies to include detailed characterization of microglial transcriptomic, phenotypic profiling (M1 vs M2), neuropathological and functional examination. Our results suggest that modulation of LRRK2 and its downstream signaling mediator NFATc2 might be therapeutic targets for treating synucleinopathies. This paper is currently under preparation for submission. The next step is to investigate the immunomodulatory role of p38 inhibitors in models of synucleinopathy. We have previously shown that the synaptic MAPK p38-gamma plays a role synaptotoxicity in DLB/PD, while p38-alpha plays a role in neuroinflammation. We treated our animal models of DLB/PD with a p38 inhibitor that crosses the blood brain barrier and evaluated the effects on synaptic function and neuroinflammation. The results of this study were presented at the intramural NIA retreat and at SfN and a manuscript was recently submitted to Science TM. Progress for Aim 3. In collaboration with the Sen laboratory we showed that T cell infiltration with potential participation of NKT cells play an important role in DLB/PD. Next, we explored the role of aging in the pathogenesis of PD/DLB by analyzing, behavior, neuropathology and transcriptomics and immune responses in young and aged wildtype mice that received intrastriatal injections with a-synuclein preformed fibrils (pff). We found that aged mice showed more extensive accumulation of a-synuclein and behavioral deficits that was associated with greater infiltration of T cells and microgliosis. Distinct inflammatory patterns of gene expression in microglia showed that a-synuclein pff-induced genes networks in young mice (eg: CSF2, TNF, IL1b, IL6) that overlaps with genes differentially expressed in microglia in the aged mice. These results indicate that the aged and more inflamed brain micro-environment directly influences the disease outcome of pathological accumulation of a-syn in age-related chronic diseases such as DLB and PD. This manuscript was recently re-submitted to Molecular Neurodegeneration. Other experiments underway includes investigating the effects of aging and deleting microglia with the PLX3387 (a compound that targets CSF1R) on synucleinopathies in DLB/PD models.We are also investigating the effects of aging and inflammation in models where T cells and microglia are deleted or where CSF1 and CSF2 are modulated, manuscripts for all these studies are under preparation. Other collaborations at LNG are with Drs. Cookson, Singleton, Traynor and Scholz on the genetic architecture of FTD, DLB and MSA by providing expert neuropathological assessment, animal models and human postmorten brain tissues. In addition to the collaborations with LNG, we have been collaborating with the laboratory of Dr. Avi Nath at NINDS investigating the neuropathology of COVID19 that resulted on a recent publication on the microvascular alterations in NeuroCOVID (Nath et al NEJM 2021) and with Dr. Robert Tycho on modeling mechanisms of a-syn protein aggregation.

神经遗传学实验室（LNG）的分子神经病理学单元（MNU）的主要目标是利用免疫系统，以更好地了解神经变性的机制，并开发诸如DLB，PD和AD的衰老人群的核核核酸核酸甘蓝核酸的疗法。我们提出了3个目标，这是第一个研究先天免疫反应和靶向LRRK2，Toll-like受体，p38，NFAT和蛋白质聚集体的作用（例如：a- synuclein，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abeta，abet abeta，abeta，abeta，abeta，abeta，abeta，abeta。第二次评估包括MAPK-P38，NFAT和NFKB在内的下游促炎信号通路以及第三个评估衰老在PD/DLB发病机理中衰老的作用，以及为突触核肾上腺病的免疫疗法而产生免疫疗法的作用。在此期间，我们发表了9条手稿，主要集中于了解PD/DLB的发病机理，并开发了新型的药理和免疫治疗方法。 AIM1的进度。在先前的研究中，我们鉴定了几种免疫受体，这些免疫受体介导突触核蛋白病中的神经炎症，包括Toll样受体2（TLR2），并显示了哪种A-突触核蛋白的物种结合TLR2并介导神经毒素。这些结果表明，TLR2的细胞类型特异性调节可能是突触性核疾病的多方面且有希望的治疗策略。神经元和星形胶质细胞TLR2的抑制可降低致病性A-Syn传播，但小胶质细胞TLR2的激活增强了小胶质细胞外A-Syn清除率。沿着这些线路，我们最近发表了一个手稿（DIS 2022的Torres等人的神经bioliol），该手稿在我们的A-突触核蛋白转基因模型中表征了体内的手稿，杏仁核中神经元的变性与神经发炎之间的关系。 As an extension and given the impact of the COVID19 pandemic and potential role of SARS-CoV2 in AD/ADRD we investigated (in collaboration with the Nath laboratory) inflammation in the brains of COVID19 patients with evidence of neurological involvement, we found increased a-synuclein accumulation in axons and glial cells as well as astrocytic activation and T cell trafficking in affected CNS areas.此外，初步研究表明，SARS-COV2包膜蛋白可能与TLR2相互作用，并在炎症和A-核蛋白驱动的病理学中发挥作用，我们发表了一种针对这种可能性的手稿（Szabo等人Exp Mol Med Med 2022）。最后，今年在与学术界的合作项目中，我们取得了相当大的进步，开发了针对3RTAU的新型ADRD免疫疗法（Spencer等人方法MOL BIOL 2022），或者同时靶向A-核蛋白的多个表位。对于tau，我们开发了一种新型的大脑穿透性单链抗体，该抗体堆积在病毒载体中，可以外周应用。对于A-突触核蛋白，我们使用了基于MultiTEP的DNA疫苗技术，该方法对具有免疫衰老的老年人特别有用。所有疫苗均诱导针对A-核蛋白特异的抗体的高滴度，可显着降低小鼠的PD/DLB样病理。在以性别依赖性方式接种PV-1949D和PV-1950D接种的小鼠中观察到了最重要的作用。我们发表了两项最近的研究（Kim等人疫苗2022和Zagorski等人Int J Mol Scie 2022）。 AIM 2的进展。与库克森组合作（Kim等人TM 2020）表明，LRRK2在细胞外A-突触核蛋白的小胶质细胞激活中起着至关重要的作用。我们确定LRRK2通过选择性地磷酸化和诱导活化T细胞的免疫转录因子核因子的核转运来促进神经炎症级联反应，即细胞质2（NFATC2）。我们的结果表明，LRRK2及其下游信号介体NFATC2的调节可能是治疗突触核苷的治疗靶标。接下来，我们研究了以下假设：阻断NFATC2易位可能会改善A-核蛋白介导的小胶质细胞神经毒性。初步研究确定了一种新型的肽化合物（11R活体），该化合物将外周施用到-Syn TG小鼠中会减少神经炎症，恢复NFATC2细胞质定位并减轻行为缺陷。我们目前正在扩大这些研究，以包括小胶质转录组，表型分析（M1 vs M2），神经病理学和功能检查的详细表征。我们的结果表明，LRRK2及其下游信号介体NFATC2的调节可能是治疗突触核苷的治疗靶标。本文目前正在准备提交。下一步是研究p38抑制剂在突触核苷模型中的免疫调节作用。我们先前已经表明，突触MAPK p38-gamma在DLB/PD中起角色突触毒性，而p38-alpha在神经炎症中起作用。我们用p38抑制剂处理了DLB/PD的动物模型，该模型越过血脑屏障，并评估了对突触功能和神经炎症的影响。这项研究的结果是在壁内NIA静修处和SFN呈现的，最近将手稿提交给Science TM。 AIM 3的进展。与SEN实验室合作，我们表明，NKT细胞潜在参与的T细胞浸润在DLB/PD中起重要作用。接下来，我们通过分析，行为，神经病理学和转录组学以及在接受A-突触核蛋白预科蛋白预先形成的原纤维（PFF）的年轻和老年野生型小鼠中探索了衰老在PD/DLB发病机理中的作用。我们发现，老年小鼠显示出与T细胞更大的浸润和小胶质细胞增多有关的A-突触核蛋白和行为缺陷的积累。小胶质细胞中基因表达的明显炎症模式表明，年轻小鼠中的A-核蛋白PFF诱导的基因网络（例如：CSF2，TNF，IL1B，IL6）与年龄小鼠中小胶质细胞中差异表达的基因重叠。这些结果表明，老化和更具发炎的脑微环境直接影响与年龄相关的慢性疾病（如DLB和PD）中A-Syn病理积累的疾病结果。该手稿最近被重新提交给分子神经退行性。正在进行的其他实验包括调查使用PLX3387（一种靶向CSF1R的化合物）对DLB/PD模型中突触核苷的衰老和删除小胶质细胞的影响。我们还研究了T细胞和小胶质细胞在所有csf1和CSF1中都进行了模型中的模型中衰老和炎症的影响。 LNG的其他合作是与Drs。 Cookson，Singleton，Traynor和Scholz通过为FTD，DLB和MSA的遗传结构提供了专家神经病理学评估，动物模型和人类后乳腺癌脑组织的遗传结构。除了与LNG的合作外，我们还与Ninds的Avi Nath博士实验室合作，调查了Covid19的神经病理学，该研究导致了最近关于Neurocovid（Nath et al Nejm 2021）微血管变化的出版物，并与Robert Tycho博士对Robert Tycho博士进行了建模机械的启示剂A a agenigrign of a-Synnectiencation of a-Synn of a a-Synn蛋白质。