Harnessing the mesenchymal stromal cell secretome to target alpha-synuclein associated dysfunction in Parkinson's disease and related disorders.

利用间充质基质细胞分泌组来靶向帕金森病和相关疾病中与α-突触核蛋白相关的功能障碍。

• 批准号: 10045791
• 负责人: Jeremy Daniel Burgess
• 金额: $ 4.7万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10045791
• 关键词: Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease risk; Biochemical; Biological Process; Blood Vessels; Brain; Caregivers; Cell model; Cell physiology; Cells; Chemicals; Clinical Trials; Complement; Data; Deterioration; Development; Disease; Disease Progression; Disease model; Environment; Failure; Functional disorder; Gene-Modified; Genes; Genetic; Genetic Risk; Goals; Health; Health Care Costs; Human; Lead; Link; Mediating; Mesenchymal Stem Cells; Modeling; Multipotent Stem Cells; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Organoids; Parkinson Disease; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Physiological; Property; Proteins; Proteomics; Quality of life; Research; Research Project Grants; Rodent; Rotenone; Stains; Stimulus; Techniques; Testing; Therapeutic Intervention; Time; Tissues; Toxic effect; Translating; Variant; Western Blotting; alpha synuclein; base; cell type; combat; comparative; conditioning; cost; differential expression; genetic association; genetic risk factor; genome wide association study; human subject; improved; induced pluripotent stem cell; interest; mesenchymal stromal cell; multipotent cell; neuron loss; paracrine; post-doctoral training; prevent; regenerative; relating to nervous system; risk variant; symptom treatment; synucleinopathy; therapeutic target; transcriptome; transcriptome sequencing; transcriptomics

Parkinson’s disease (PD) is the second most common neurodegenerative condition, with serious negative impacts on quality of life, as well as financial and personal burden on sufferers and caregivers. There remains no disease modifying therapy. PD is strongly linked to aberrant oligomerization and accumulation of alpha-synuclein (αsyn) protein. Mesenchymal stem cells (MSCs) are multipotent cells that can be isolated from vascularized tissue throughout the body and exert regenerative effects through their dynamic secretome, which is influenced by multiple factors including their microenvironment and any surrounding pathology. In this proposal, we hypothesize that by conditioning MSCs with PD-simulating stimuli, both chemical and patient-derived, we will enrich their secretome for factors which are particularly beneficial in combating αsyn-associated pathology. The potency of conditioned MSCs in targeting αsyn pathology will be assessed using biochemical and staining methods, using protein complementation, semi-native western blot and immunohistochemical analysis of oligomeric αsyn in cellular models of synucleinopathy, as well as assessing toxicity. By comparative proteomic analysis of the MSC secretome in the presence and absence of PD-simulating stimuli we will identify these beneficial secreted molecules and validate them for efficacy in alleviating αsyn-associated pathology in cellular models of synucleinopathy. For the K00 portion of this proposal, focus shifts to Alzheimer’s disease (AD), an even more common neurodegenerative condition with no meaningful therapeutic interventions. Clinical trials based on rodent studies fail to translate efficacy to human subjects, highlighting the need for physiologically relevant translatable models. Further, genetic association studies have highlighted more than 20 risk loci, usually relatively common but with moderate effect sizes. In order to mechanistically probe these different genetic risk factors in a relevant, pathology-developing model we propose to generate human brain organoids from iPSCs, gene edited to reflect various distinct genetic risk profiles. We will perform longitudinal comparative transcriptomics (RNAseq) to identify common and distinct pathways associated with the progression of pathology in these models and determine therapeutic targets which may be generalizable to AD or specific to certain genetic backgrounds.

帕金森氏病（PD）是第二大最常见的神经退行性疾病，对生活质量以及对患者和看护者的经济和个人烧伤产生了严重的负面影响。没有修改疗法的疾病。 PD与异常的寡聚和α-Syn（αSyn）蛋白的积累密切相关。间充质干细胞（MSC）是多能细胞，可以从整个体内的血管化组织中分离出来，并通过其动态分泌组来锻炼再生效应，这受到多种因素的影响，包括其微环境和任何周围的病理学。在此提案中，我们假设通过对化学和患者衍生的PD减少刺激进行调节MSC，我们将使他们的分泌组丰富了他们的分泌组，这些因素对于打击α-Syn相关病理特别有益。将使用生化和染色方法评估条件MSC在靶向α综合病理学中的效力，使用蛋白质的完成，半本地蛋白质印迹和免疫组织化学分析在突触核蛋白酶病的细胞模型中的寡聚体α综合分析，以及评估毒性。通过在存在和不存在PD刺激刺激的情况下对MSC分泌组的比较蛋白质组学分析，我们将鉴定出这些有益的分泌分子，并验证它们以减轻突触核酸的细胞模型中的αSyn相关病理的效率。对于本提案的K00部分，重点转移到了阿尔茨海默氏病（AD），这是一种更常见的神经退行性疾病，没有任何有意义的治疗干预措施。基于啮齿动物研究的临床试验无法将有效性转化为人类受试者，从而强调了对物理相关的可翻译模型的需求。此外，遗传关联研究突出了20多个风险基因座，通常是为了在相关的，病理发展的模型中机械探测这些不同的遗传风险因素，我们建议从IPSC产生人类脑器官，从IPSC中产生人类脑器官，以反映各种不同的遗传风险谱。我们将执行纵向比较转录组学（RNASEQ），以识别这些模型中与病理发展相关的常见和不同的途径，并确定可以推广到AD或特定于某些遗传背景的治疗靶标。