Alzheimers Disease Project: Neuroimmune responses and therapeutics of alpha-synucleinopathies of the aging population

阿尔茨海默病项目：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：
10913166
负责人：
Eliezer Masliah
金额：
$ 10.28万
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/10913166
关键词：
3-Dimensional Aging Alpha-Synuclein transgenic mouse Alzheimer&apos s Disease Alzheimer&apos s disease model Amyloid beta-Protein Animal Model Animals Antibodies Apolipoprotein E Astrocytes Astrocytosis Attenuated B-Lymphocytes Behavior Behavioral Binding Brain Brain Diseases CSF1 gene CSF1R gene CSF2 gene Candidate Disease Gene Cell Aging Cell Nucleus Cell Separation Cell model Cells ChIP-seq Chromatin Chronic Disease Collaborations Combination immunotherapy Communication Cytoplasm DNA Damage Dementia with Lewy Bodies Disease Disease Outcome Disease model Down-Regulation Encephalitis Exhibits Functional disorder Gene Expression Profile Genes Genetic Genetic Risk Genetic Transcription Genetically Engineered Mouse Gliosis Goals Hippocampus Human IL6 gene Immune Immune response Immune system Immunologic Receptors Immunotherapy In Vitro Inflammation Inflammatory Inflammatory Response Injections Innate Immune Response LRRK2 gene Laboratories Lewy Body Disease Link MAP Kinase Gene Manuscripts Maps Mediating Mediator Methods Microglia Modeling Molecular Molecular Profiling Motor Mouse Strains Multiomic Data Mus NF-kappa B Natural Immunity Nature Nerve Degeneration Neurodegenerative Disorders Neuroimmune Neuroimmune system Neurologic Neurons Nuclear Translocation Paper Parkinson Disease Pathogenesis Pathologic Pathology Pathway interactions Patients Peptides Peripheral Phenotype Phosphorylation Phosphotransferases Play Preparation Protein Isoforms Publishing Regulatory Element Research Resolution Role Science Signal Pathway Signal Transduction South Korea Substantia nigra structure Synapses System T cell infiltration T cell response T-Lymphocyte TLR2 gene TNF gene Testing Therapeutic Therapeutic Effect Toll-like receptors Transcriptional Regulation Transgenic Mice Transgenic Model Translational Research Universities Wild Type Mouse adaptive immunity age effect age related aged aging population alpha synuclein brain tissue cell type combinatorial cytokine data integration differential expression epigenome epigenomics experimental study extracellular gene network genetic architecture glial activation immune modulating agents immunomodulatory therapies immunoregulation in vitro Model in vivo in vivo Model interest mouse model multiple omics neurogenetics neuroinflammation neuropathology neurotoxic neurotoxicity novel novel therapeutics nuclear factors of activated T-cells overexpression p38 Mitogen Activated Protein Kinase pharmacologic pre-formed fibril protein TDP-43 protein aggregation proteostasis receptor response risk variant single-cell RNA sequencing small hairpin RNA small molecule synuclein synucleinopathy targeted treatment tau Proteins therapeutic target therapy development transcription factor transcriptome transcriptome sequencing transcriptomics translational medicine γδ T cells

项目摘要

The main focus 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 8 manuscripts mostly focusing understanding the pathogenesis of PD/DLB and developing novel pharmacological and anti-sense 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. We were interested at better characterizing the downstream pathways mediating neurodegeneration by using multiomics approaches. For this purpose we collaboratives with Dr. Jung laboratory in South Korea. First, we establish transcriptomic and epigenomic landscapes of the substantia nigra by profiling 113,207 nuclei obtained from healthy controls and patients with PD. Our multiomics data integration provides cell type annotation of 128,724 cis-regulatory elements (cREs) and uncovers cell type-specific dysregulations in cREs with a strong transcriptional influence on genes implicated in PD. The establishment of high-resolution three-dimensional chromatin contact maps identifies 656 target genes of dysregulated cREs and genetic risk loci, uncovering both potential and known PD risk genes. Notably, these candidate genes exhibit modular gene expression patterns with unique molecular signatures in distinct cell types, highlighting altered molecular mechanisms in immune cells like microglia as well as in oligodendroglial cells (Lee et al, Sci Adv 2023). Together, our single-cell transcriptome and epigenome reveal cell type-specific disruption in transcriptional regulations related to PD that are complementary with studies described for Aim 2. More recently, in collaboration with UCSD we investigated the therapeutical role of downregulating a-synuclein with targeted shRNA delivery into the hippocampus of mouse models of AD and DLB. We showed that downregulation of -syn results in significant reduction in the number of Ab plaques and tau. In addition, mice treated with LV-shRNA -syn had amelioration of abnormal microglial activation and astrocytosis in AD mice. This suggests a link between A and -syn in pathology pointing to a possible therapeutic angle for AD targeting -syn. This paper was published in JAD (Leitao et al JAD 2023). In other studies, in collaboration with the Cookson lab we studied the distribution of p-a-synuclein in control and PD brains using the PLA method (Arlinghaus et al J Park Dis 2023). Progress for Aim 2. We previously showed that once extracellular a-synuclein binds TLR2, (Kim et al Science TM 2020) 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 under revision at Nature Communications. Next, we investigated the role of p38 MAPK as a potential immunomodulatory pathway. We have shown that activation of the p38 MAPK isoform and mislocalization of the p38 MAPK isoform are associated with neuroinflammation and synaptic degeneration in DLB and PD. Therefore, we hypothesized that p38 might be associated with neuronal p38 distribution and synaptic dysfunction in these diseases. To test this hypothesis, we treated in vitro cellular and in vivo mouse models of DLB/PD with SKF-86002, a compound that attenuates inflammation by inhibiting p38/, and then investigated the effects of this compound on p38 and neurodegenerative pathology. We found that inhibition of p38 reduced neuroinflammation and ameliorated synaptic, neurodegenerative, and motor behavioral deficits in transgenic mice overexpressing human -synuclein. Moreover, treatment with SKF-86002 promoted the redistribution of p38 to synapses and reduced the accumulation of -synuclein in mice overexpressing human -synuclein. Treatment with SKF-86002 ameliorated -synuclein-induced neurodegeneration in these neurons only when microglia were pretreated with this compound. These findings provide a mechanistic connection between p38 and p38 as well as a rationale for targeting this pathway in DLB/PD. This study was recently published in Science Translational Medicine (Iba et al Science TM 2023). 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. 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. Similar studies have been performed in collaboration with the Winkler laboratory (Erlangen University) in another model of synucleinopathy, that is a transgenic model of MSA treated with PLX5622, this study showed that deleting microglia delayed onset of neurological alterations (J Neuroscie 2023). 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.

神经遗传学实验室（LNG）的分子神经病理学单元（MNU）的主要重点是利用免疫系统，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8个手稿，主要集中于了解PD/DLB的发病机理，并开发了新型的药理和反义方法。 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。我们有兴趣更好地表征下游途径通过使用多组学方法介导神经退行性的途径。为此，我们与韩国的荣格实验室合作。首先，我们通过分析从健康对照组和PD患者获得的113,207个核来建立黑质的转录组和表观基因组景观。我们的多组学数据集成提供了128,724个顺式调节元件（CRE）的细胞类型注释，并发现对与PD中有关基因有强大影响的CRE中的细胞类型特异性失调。高分辨率的三维染色质接触图的建立确定了656个CRE和遗传风险基因座的目标基因，从而发现了潜在的和已知的PD风险基因。值得注意的是，这些候选基因在不同的细胞类型中具有独特的分子特征表现出模块化基因表达模式，突出了免疫细胞（如小胶质细胞以及寡头胶质细胞）中的分子机制改变（Lee等人，SCI ADV 2023）。总之，我们的单细胞转录组和表观基因组揭示了与PD相关的转录法规中细胞类型特异性的破坏，这些破坏与AIM 2所述的研究相辅相成。最近，与UCSD合作，我们研究了下调A-突触核蛋白与靶向shrna与AD和DLB小鼠模型的Hippocampus递送的A-突触核蛋白的治疗作用。我们表明-SYN的下调导致AB斑块和TAU的数量显着减少。此外，用LV -sHRNA -SYN治疗的小鼠在AD小鼠中对异常小胶质细胞活化和星形细胞增多症具有改善。这表明在病理学中，A和-SYN之间的联系指向AD靶向-Syn的可能的治疗角度。本文发表在Jad（Leitao等人Jad 2023）中。在其他研究中，我们与库克森实验室合作研究了使用PLA方法（Arlinghaus et al J Park dis 2023）在对照和PD大脑中的P-A-核蛋白分布。 AIM 2的进展。我们先前表明，曾经的细胞外A-核蛋白结合TLR2（Kim等人TM 2020）LRRK2 LRK2通过选择性地磷酸化并诱导活化T细胞的免疫转录因子核因子的核转移来促进神经炎的级联反应。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。本文正在自然通讯中进行修订。接下来，我们研究了p38 MAPK作为潜在的免疫调节途径的作用。我们已经表明，p38 MAPK同工型的激活和p38 MAPK同工型的错误定位与DLB和PD中的神经炎症和突触退化有关。因此，我们假设p38可能与这些疾病中的神经元p38分布和突触功能障碍有关。为了检验这一假设，我们用SKF-86002处理了DLB/PD的体外细胞和体内小鼠模型，该化合物通过抑制p38/来减弱炎症，然后研究了该化合物对p38和神经退化性病理学的影响。我们发现，抑制p38可减少神经炎症，并改善过表达人 - 鼻核蛋白的转基因小鼠的突触，神经退行性和运动行为缺陷。此外，用SKF -86002的治疗促进了p38的重新分布，从而突触并减少了过表达人-synuclein的小鼠中-Synclein的积累。 SKF-86002的处理仅在用这种化合物预处理小胶质细胞时，可以改善这些神经元的突触核蛋白诱导的神经变性。这些发现提供了p38和p38之间的机械连接，以及针对DLB/PD的途径的理由。这项研究最近发表在科学转化医学上（IBA等人TM 2023）。 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。其他实验包括研究使用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。与Winkler实验室（Erlangen University）合作进行了类似的研究，这是另一种突触核力疗法模型，该模型是用PLX5622处理的MSA的转基因模型，这项研究表明，删除小胶质细胞延迟神经系统变化的发作（J Neuroscie 2023）。我们还正在研究删除T细胞和小胶质细胞或CSF1和CSF2的模型中衰老和炎症的影响，所有这些研究的手稿都在准备中。 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。

项目成果

期刊论文数量（42）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

α-Synuclein oligomers induce early axonal dysfunction in human iPSC-based models of synucleinopathies.

DOI：
10.1073/pnas.1713129115
发表时间：
2018-07-24
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Prots I;Grosch J;Brazdis RM;Simmnacher K;Veber V;Havlicek S;Hannappel C;Krach F;Krumbiegel M;Schütz O;Reis A;Wrasidlo W;Galasko DR;Groemer TW;Masliah E;Schlötzer-Schrehardt U;Xiang W;Winkler J;Winner B
通讯作者：
Winner B

Correction to: A candidate gene study of intermediate histopathological phenotypes in HIV-associated neurocognitive disorders.

更正：HIV 相关神经认知障碍中间组织病理学表型的候选基因研究。

DOI：
10.1007/s13365-020-00871-y
发表时间：
2020
期刊：
Journal of neurovirology
影响因子：
3.2
作者：
Levine,AndrewJ;Soontornniyomkij,Virawudh;Masliah,Eliezer;Sinsheimer,JanetS;Ji,SarahS;Horvath,Steve;Singer,ElyseJ;Kallianpur,Asha;Moore,DavidJ
通讯作者：
Moore,DavidJ

Evolvability and Neurodegenerative Disease: Antagonistic Pleiotropy Phenomena Derived from Amyloid Aggregates.

DOI：
10.3233/jpd-181365
发表时间：
2018
期刊：
Journal of Parkinson's disease
影响因子：
0
作者：
Hashimoto M;Ho G;Takamatsu Y;Shimizu Y;Sugama S;Takenouchi T;Waragai M;Masliah E
通讯作者：
Masliah E

The Leukotriene Receptor Antagonist Montelukast Reduces Alpha-Synuclein Load and Restores Memory in an Animal Model of Dementia with Lewy Bodies.

DOI：
10.1007/s13311-020-00836-3
发表时间：
2020-07
期刊：
Neurotherapeutics : the journal of the American Society for Experimental NeuroTherapeutics
影响因子：
0
作者：
Marschallinger J;Altendorfer B;Rockenstein E;Holztrattner M;Garnweidner-Raith J;Pillichshammer N;Leister I;Hutter-Paier B;Strempfl K;Unger MS;Chishty M;Felder T;Johnson M;Attems J;Masliah E;Aigner L
通讯作者：
Aigner L

Therapeutic Potential of α-Synuclein Evolvability for Autosomal Recessive Parkinson's Disease.