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：
10250909
负责人：
Eliezer Masliah
金额：
$ 3.42万
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/10250909
关键词：
Active Immunotherapy Adaptive Immune System Affinity Age-Months Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Amyloid beta-Protein Amyloid beta-Protein Precursor Animal Model Animals Antibodies Area Astrocytes Attenuated Autopsy Axon B-Lymphocytes Behavioral Binding Blood Vessels Brain CD3 Antigens CD4 Positive T Lymphocytes CD8B1 gene Cell model Cells ChIP-seq Clinical Trials Collaborations Combination immunotherapy Corpus striatum structure Cues Data Development Dysautonomias Exhibits FYN gene Functional disorder Future Gene Expression Profile Genetically Engineered Mouse Goals Hippocampus (Brain)Human Immune Immune response Immune system Immunize Immunomodulators Immunotherapeutic agent Immunotherapy Infiltration Inflammation Inflammatory Injections Innate Immune Response LRRK2 gene Laboratories Lewy Body Dementia Link MS4A1 gene Manuscripts Mediating Mediator of activation protein Microglia Modeling Molecular Mouse Strains Mus Myeloid Cells NF-kappa B Natural Immunity Neocortex Nerve Degeneration Neurobiology Neurodegenerative Disorders Neuroimmune Neurons Neurosciences Oligodendroglia Parkinson Disease Parkinson&apos s Dementia Parkinsonian Disorders Pathogenesis Pathology Pathway interactions Patients Peptides Peripheral Pharmacology Phase Phosphotransferases Play Population Preparation Presynaptic Terminals Process Proteins Publishing Reporting Research Role Science Signal Pathway Signal Transduction Susceptibility Gene Synapses System T cell response T-Lymphocyte TLR2 gene TREM2 gene Testing Therapeutic Therapeutic Effect Toll-like receptors Transgenic Mice Tumor-infiltrating immune cells Up-Regulation Wild Type Mouse adaptive immunity aged aging population alpha synuclein astrogliosis base brain tissue cell type cohort combinatorial conditioned fear cytokine experimental study extracellular frontier genetic architecture hippocampal pyramidal neuron immunomodulatory therapies immunoregulation in vitro Model in vivo in vivo Model interest mouse model neurogenetics neuroinflammation neuronal cell body neuropathology neurotoxic novel novel therapeutics overexpression p38 Mitogen Activated Protein Kinase promoter protein TDP-43 protein aggregation response single-cell RNA sequencing synuclein synucleinopathy targeted treatment tau Proteins tau phosphorylation therapy development transcription factor transcriptome sequencing transcriptomics transmission process white matter γδ T cells

Active Immunotherapy Adaptive Immune System Affinity Age-Months Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Amyloid beta-Protein Amyloid beta-Protein Precursor Animal Model Animals Antibodies Area Astrocytes Attenuated Autopsy Axon B-Lymphocytes Behavioral Binding Blood Vessels Brain CD3 Antigens CD4 Positive T Lymphocytes CD8B1 gene Cell model Cells ChIP-seq Clinical Trials Collaborations Combination immunotherapy Corpus striatum structure Cues Data Development Dysautonomias Exhibits FYN gene Functional disorder Future Gene Expression Profile Genetically Engineered Mouse Goals Hippocampus (Brain)Human Immune Immune response Immune system Immunize Immunomodulators Immunotherapeutic agent Immunotherapy Infiltration Inflammation Inflammatory Injections Innate Immune Response LRRK2 gene Laboratories Lewy Body Dementia Link MS4A1 gene Manuscripts Mediating Mediator of activation protein Microglia Modeling Molecular Mouse Strains Mus Myeloid Cells NF-kappa B Natural Immunity Neocortex Nerve Degeneration Neurobiology Neurodegenerative Disorders Neuroimmune Neurons Neurosciences Oligodendroglia Parkinson Disease Parkinson&apos s Dementia Parkinsonian Disorders Pathogenesis Pathology Pathway interactions Patients Peptides Peripheral Pharmacology Phase Phosphotransferases Play Population Preparation Presynaptic Terminals Process Proteins Publishing Reporting Research Role Science Signal Pathway Signal Transduction Susceptibility Gene Synapses System T cell response T-Lymphocyte TLR2 gene TREM2 gene Testing Therapeutic Therapeutic Effect Toll-like receptors Transgenic Mice Tumor-infiltrating immune cells Up-Regulation Wild Type Mouse adaptive immunity aged aging population alpha synuclein astrogliosis base brain tissue cell type cohort combinatorial conditioned fear cytokine experimental study extracellular frontier genetic architecture hippocampal pyramidal neuron immunomodulatory therapies immunoregulation in vitro Model in vivo in vivo Model interest mouse model neurogenetics neuroinflammation neuronal cell body neuropathology neurotoxic novel novel therapeutics overexpression p38 Mitogen Activated Protein Kinase promoter protein TDP-43 protein aggregation response single-cell RNA sequencing synuclein synucleinopathy targeted treatment tau Proteins tau phosphorylation therapy development transcription factor transcriptome sequencing transcriptomics transmission process white matter γδ T cells

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项目摘要

The main objective of the Molecular Neuropathology section (MNS) at Laboratory of Neurogenetics (LNG) is to harness the immune system to better understand the mechanisms of neurodegeneration and to develop therapies for a-synucleinopathies of the aging population. We propose 3 Aims, the first investigating the role of innate immune responses and combinatorial immunotherapy targeting LRRK2, Toll-like receptors and the protein aggregates (eg: 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 T cell mediated adaptive immune cell responses in PD/DLB pathogenesis and for developing immunotherapies for a-synucleinopathies. During this period we published over 30 manuscripts mostly focusing understanding the pathogenesis of PD/DLB and developing novel pharmacological and immunotherapeutical approaches. Progress for Aim1. We recently published a review manuscript describing present progress and future directions utilizing immunotherapy in neurodegenerative disorders (Kwon et al Neurotherapeutics 2020) an expanded ou studies targeting the interactions between extracellular -synuclein aggregates and TLR2 in models of synucleinopathy by modulating synuclein transmission and neuroinflammation (Kwon et al Exp Neurobiol 2019). Other important question in the field is if there are specific antibodies targeting synuclein that will block neuron to neuron propagation and ameliorate neurodegeneration In this respect we recently reported the development (Schofield et al Neurobiology of Dis 2019) of a high affinity -synuclein antibody, MEDI1341, that can enter the brain, sequester extracellular -synuclein and attenuate -synuclein spreading in vivo. MEDI1341 binds both monomeric and aggregated forms of -synuclein. Using a novel lentiviral-based in vivo mouse model of -synuclein spreading in the brain, we show that treatment with MEDI1341 significantly reduces -synuclein accumulation and propagation along axons. MEDI1341 is now in Phase 1 human clinical trial testing for PD. Moreover, during FY20 we also investigated the role of combined immunotherapy targeting A and -synuclein in models AD/PD (Mandler et al Alz and Dem 2019). For this purpose Amyloid precursor protein/-syn tg mice were immunized with AFFITOPEs (AFF) peptides specific to A (AD02) or -syn (PD-AFF1) and the combination. Remarkably, we found that AD02 was more effectively reduced A and pTau burden; however, the combination exhibited some additive effects. Both AD02 and PD-AFF1 effectively reduced -syn, ameliorated degeneration of pyramidal neurons, and reduced neuroinflammation. We concluded that combined active immunotherapy targeting A and/or -syn may be of potential interest for the treatment of dementia with Lewy bodies. Progress for Aim 2. Alterations in kinase pathways including mitogen activated protein kinase (MAPK) p38 has been proposed to play a in PD/DLB and AD. We recently published a new study (Iba et al Frontiers in Neuroscience 2020) showing that while activation of p38 has been linked to neuro-inflammation, alterations in p38 has been associated with Tau phosphorylation in AD. Although p38 has been studied in AD less is known as to the role of MAPK p38 in DLB/PD and other synucleinopathies. While in healthy control human and Non-Tg mice p38 was associated with neurons and astroglial cells, p38 localized to pre-synaptic terminals and in the brains of DLB and -syn Tg mice immunostaining in the synaptic terminals was reduced and p38 was redistributed to the neuronal cell bodies. Further, double immunolabeling showed that p38 co-localized with -syn aggregates in DLB patient and -syn Tg mice. In contrast, in the brains of DLB cases and -syn Tg mice p38 was increased in astroglial cells. These results suggest that -syn might interfere with the p38 pathway and might play a role in the mechanisms of synaptic dysfunction in DLB/PD. Next in collaboration with the Cookson group we are investigating the cross talk between TLR2 and LRRK2 and downstream signaling involving NFATC leading to neurodegeneration and inflammation in PD/DLB, the manuscript is in press in Science TM. Progress for Aim 3. In collaboration with the Sen laboratory we recently published (Iba et al J Neuroinflammation 2020) one of the first studies in the field demonstrating that neuroinflammation in synucleinopathies involves considerable T cell infiltration with potential participation of NKT cells. To understand the involvement of the peripheral immune system in PD/DLB, we evaluated T cell populations in the brains of -syn transgenic (tg) mice (e.g., Thy1 promoter line 61) and DLB patients. Our analysis showed perivascular and parenchymal infiltration by CD3+/CD4+ helper T cells, but not cytotoxic T cells (CD3+/CD8+) or B cells (CD20+), in the neocortex, hippocampus, and striatum of -syn tg mice. CD3+ cells were found in close proximity to the processes of activated astroglia, particularly in areas of the brain with significant astrogliosis, microgliosis, and expression of pro-inflammatory cytokines. A subset of CD3+ showed cytometric data consistent with NKT cells. In post-mortem DLB brains, we similarly detected increased numbers of infiltrating CD3+/CD4+ T cells in close proximity with blood vessels. These results suggest that infiltrating adaptive immune cells play an important role in neuroinflammation and neurodegeneration in synucleinopathies and that modulating peripheral T cells may be a viable therapeutic strategy for PD/DLB. Similarly we have investigated neuroinflammation in other models of synucleinopathy including MSA a neurodegenerative disorder with parkinsonism, dysautonomia and accumulation of -syn in oligodendroglial cells. This study was recently published (Hoffman et al Brain Pathol 2019) and showed increased neuroinflammation in regions with a high -syn load, A distinct gene expression profile including upregulation of cytokines important for myeloid cell attraction and proliferation was detected in -syn overexpressing oligodendrocytes. Collectively, these results imply an early crosstalk between neuroinflammation and oligodendrocytes containing -syn inclusions leading to an immune response locally restricted to white matter regions in MSA. More recently we investigated in collaboration with the Sen Laboratory the role of aging in T cell infiltration and neuroinflammation in synucleinopathies, for this purpose we utilized a novel model, for this -syn pff was injected into the striatum of wild type mice at 4 and 18 months of age and analyzed for behavioral deficits, neuropathology and immune responses at 1- and 3-month post injection periods. Following intracranial injection of -syn pff we found that in the aged mice cohort there was a significant increase in -syn pathology, infiltration of T cells, microgliosis and astrogliosis. Behavioral analysis showed increase in activity, decreased cued fear conditioning in old -syn pff mice. In conclusion, the results points to the possibility that targeting neuro-immune responses might be important in developing treatments for DLB/PD. This manuscript is under preparation for submission. 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）的分子神经病理学科（MNS）的主要目标是利用免疫系统，以更好地了解神经变性的机理，并开发衰老人群的A-鼻核的疗法。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样受体和蛋白质聚集体（例如：Synuctulin，abeta，abeta，tau）的作用的作用；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T细胞介导的适应性免疫细胞反应在PD/DLB发病机理中的作用以及为A-突触核肾上腺病的免疫疗法的作用。在此期间，我们发表了30多种手稿，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。我们最近发表了一份评论手稿，描述了利用免疫疗法在神经退行性疾病中的当前进展和未来方向（Kwon等人神经治疗学2020年）一项扩展的OU研究，针对细胞外 - 舌 - 舌 - 舌核聚集体与TLR2之间相互作用的相互作用，在模含量的促舌链蛋白促进蛋白和Neuroin neuroin neuroin neuroin neuroin neuroin neuroin（Kw）中， 2019）。 Other important question in the field is if there are specific antibodies targeting synuclein that will block neuron to neuron propagation and ameliorate neurodegeneration In this respect we recently reported the development (Schofield et al Neurobiology of Dis 2019) of a high affinity -synuclein antibody, MEDI1341, that can enter the brain, sequester extracellular -synuclein and attenuate -synuclein在体内传播。 MEDI1341结合了 - 突触核蛋白的单体和聚集形式。使用一种新型的基于慢病毒的在大脑中 - 突触核蛋白扩散的体内小鼠模型，我们表明使用MEDI1341的治疗显着降低了 - 突触核蛋白的积累和沿轴突的传播。 MEDI1341现在正在对PD的第1阶段人类临床试验测试中。此外，在20财年期间，我们还研究了靶向A和 - 核蛋白在AD/PD中的合并免疫疗法的作用（Mandler等人Alz和Dem，2019年）。为此，用特有的（AD02）或-Syn（PD-AFF1）的染色（AFF）肽和组合对淀粉样蛋白/-SYN TG小鼠进行免疫。值得注意的是，我们发现AD02更有效地减轻了A和PTAU负担。但是，该组合表现出一些添加效应。 AD02和PD -AFF1都有效地降低了-SHN，改善了锥体神经元的变性，并减少了神经炎症。我们得出的结论是，靶向A和/或SYN的主动免疫疗法可能具有潜在的利用Lewy身体治疗痴呆症。 AIM 2的进展。激酶途径的改变，包括有丝分裂原活化蛋白激酶（MAPK）p38在PD/DLB和AD中播放p38。我们最近发表了一项新的研究（IBA等人的神经科学2020年的IBA等人），表明，虽然p38的激活与神经炎症有关，但p38的变化与AD中的Tau磷酸化有关。尽管已在AD中研究了p38，但对于MAPK p38在DLB/PD和其他突触核管中的作用而言，p38已知。虽然在健康对照中，人类和非TG小鼠p38与神经元和星形胶质细胞有关，但p38位于突触前末端以及在突触末端中免疫染色的DLB和-SYN TG小鼠的大脑中，p38降低了P38，并将p38降低为Neuronal细胞体。此外，双重免疫标记表明，p38与DLB患者和-Syn TG小鼠中的-SYN聚集体共定位。相反，在星形胶质细胞中，在DLB病例和-Syn TG小鼠的大脑中，P38增加了。这些结果表明-syn可能会干扰p38途径，并且可能在DLB/PD中突触功能障碍的机理中发挥作用。接下来，在与库克森集团（Cookson Group）合作，我们正在研究TLR2和LRRK2之间的串扰，以及涉及NFATC的下游信号传导，导致PD/DLB的神经变性和炎症，该手稿在Science TM中发表。 AIM 3的进展。与SEN实验室合作，我们最近发表了（IBA等人神经炎症2020年），该领域的第一批研究表明，突触核苷的神经炎症涉及NKT细胞潜在参与的T细胞浸润。为了了解周围免疫系统在PD/DLB中的参与，我们评估了-SYN转基因（TG）小鼠（例如THY1启动子61）和DLB患者的T细胞群体。我们的分析表明，在新皮层，海马室和-Syn TG小鼠的新皮层，海马和纹状体中，CD3+/CD4+辅助T细胞CD3+/CD4+辅助T细胞（CD3+/CD4+辅助T细胞）的周围和实质性浸润。发现CD3+细胞与活化的星形胶质细胞的过程非常接近，尤其是在大脑区域，具有明显的星形胶质细胞增多症，小胶质细胞增多症和促炎性细胞因子的表达。 CD3+的子集显示了与NKT细胞一致的细胞计量数据。在验尸后DLB大脑中，我们类似地检测到与血管紧密相邻的浸润CD3+/CD4+ T细胞的数量增加。这些结果表明，浸润的适应性免疫细胞在突触核断病中的神经炎症和神经退行性中起重要作用，调节外周T细胞可能是PD/DLB的可行治疗策略。同样，我们还研究了其他突触性疾病模型中的神经炎症，包括MSA具有帕金森氏症，功能障碍和-syn在寡头细胞中的神经退行性疾病。这项研究最近发表（Hoffman等人Brain Pathol，2019年），显示出较高的SYN负载的区域的神经炎症增加，在-syn中检测到 - 在-syn中检测到了对髓样细胞吸引力重要的细胞因子的独特基因表达谱，以表达寡糖细胞的过表达。总的来说，这些结果意味着神经炎症与含有-SYN夹杂物的少突胶质细胞之间的早期串扰，导致免疫反应在局部限于MSA的白质区域。最近，我们与SEN实验室合作调查了衰老在突触核断病中的T细胞浸润和神经炎症中的作用，为此，我们利用了一种新型模型，因为这种-Syn PFF被注入4和18个月的野生型小鼠的纹状体，并在4和18个月中分析了行为缺乏症，并进行了神经疗法，并进行了神经疗法，并进行了神经疗法和3个年龄。颅内注射-Syn PFF后，我们发现在老年小鼠队列中，-SYN病理学，T细胞的浸润，小胶质细胞增多和星形胶质细胞增多显着增加。行为分析显示活性增加，旧-Syn PFF小鼠的提示恐惧调节减少。总之，结果表明，靶向神经免疫反应对于开发DLB/PD治疗可能很重要。此手稿正在准备提交。 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。