Preserving cellular aspects of aging in patient-specific models of ALS

在 ALS 患者特异性模型中保留衰老的细胞方面

基本信息

批准号：
9467166
负责人：
Samuel V Alworth
金额：
$ 22.49万
依托单位：
ACURASTEM, INC.
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9467166
关键词：
Age Aging Amyotrophic Lateral Sclerosis Autopsy Big Data Biological Assay Biological Preservation Brain C9ORF72 Cell model Cells Cellular biology Characteristics Clinical Confocal Microscopy Data Defect Development Disease Disease model Embryo Evaluation Feasibility Studies Fibroblasts Frontotemporal Dementia Gene Expression Gene Expression Profile Genes Genomics Government Heart Human Immunofluorescence Immunologic In Vitro Licensing Measures Mediating Methods Modeling Motor Neurons Mutation Nature Neurodegenerative Disorders Neurons Nuclear Pathogenicity Patients Persons Phenotype Preclinical Drug Evaluation Process Property Protocols documentation Research Proposals Rest Risk Factors Sampling System Testing Therapeutic Tissues Validation age related blastomere structure computerized data processing disease phenotype genetic makeup in vitro testing individual patient induced pluripotent stem cell innovation neurodegenerative phenotype nucleocytoplasmic transport pre-clinical therapy outcome transcription factor transcriptome virtual

Age Aging Amyotrophic Lateral Sclerosis Autopsy Big Data Biological Assay Biological Preservation Brain C9ORF72 Cell model Cells Cellular biology Characteristics Clinical Confocal Microscopy Data Defect Development Disease Disease model Embryo Evaluation Feasibility Studies Fibroblasts Frontotemporal Dementia Gene Expression Gene Expression Profile Genes Genomics Government Heart Human Immunofluorescence Immunologic In Vitro Licensing Measures Mediating Methods Modeling Motor Neurons Mutation Nature Neurodegenerative Disorders Neurons Nuclear Pathogenicity Patients Persons Phenotype Preclinical Drug Evaluation Process Property Protocols documentation Research Proposals Rest Risk Factors Sampling System Testing Therapeutic Tissues Validation age related blastomere structure computerized data processing disease phenotype genetic makeup in vitro testing individual patient induced pluripotent stem cell innovation neurodegenerative phenotype nucleocytoplasmic transport pre-clinical therapy outcome transcription factor transcriptome virtual

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项目摘要

Preserving cellular aspects of aging in patient-specific models of ALS Project Summary Induced pluripotent stem cell (iPSC) biology holds great promise for human in vitro neurodegenerative disease modeling because these cells can give rise to any cell in the human brain, a living “virtual brain” amenable to experimental manipulation, having the exact same genetic makeup as individual patients and displaying neurodegenerative phenotypes previously identified in postmortem and clinical samples. These “patient-specific” in vitro testing systems enable target discovery, drug screening and therapeutic proof of concept studies in patient cells much earlier in the translational process than is currently possible. Despite these unique advantages, the preservation of age as a key pathogenic risk factor is presently a major limitation of these systems. This is in part due to 1) the loss of age-related characteristics in cells that are rejuvenated to an embryonic state, and 2) to deficiencies in the differentiation protocols that are unable to produce mature neurons from embryonic cells. The direct, transcription factor mediated reprogramming, also known as “lineage conversion”, of patient fibroblasts into induced motor neurons represents an alternative approach for generating human neurons in vitro. New data from our lab, our collaborators’ and others’ show that neurons generated through lineage conversion better retain age-related and disease-associated deficits. In this 1-year feasibility study we will compare motor neurons generated by lineage conversion from fibroblasts (fib-MNs), with those generated by directed differentiation from iPSCs reprogrammed from the same fibroblast samples (iPSC-MNs). The fibroblast samples are from patients having the GGGGCC hexanucleotide repeat expansion mutation in the C9ORF72 gene, which is known to cause a form of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia, and matched controls. Our 1st hypothesis is that fib-MN transcriptomes will be significantly more similar to those of post-mortem tissue and those from ALS patients. Our collaborator Verge Genomics has created an innovative, big-data-driven ALS gene expression signature using public and proprietary gene expression data from 39 ALS-relevant studies that we will use in the project. Nucleocytoplasmic transport defects have emerged as one phenotype where both age and C9-ALS related differences have been identified. Therefore, our 2nd hypothesis is that fib-MNs will have significantly more pronounced age-related nucleocytoplasmic transport defects than iPSC-MNs as measured by immunofluorescence confocal microscopy. Taken together, these two studies would prove the principle that fib-MNs create superior ALS in-vitro testing systems retaining important, disease-relevant aspects of aging having tremendous impact on the iPSC banking and disease modeling fields. AcuraStem Inc. develops human cell models and assays for preclinical human validation of its own CNS therapeutics, as well as those of its development partners.

在ALS的患者特异性模型中保留衰老的细胞方面项目摘要诱导多能干细胞（IPSC）生物学对人类体外神经退行性疾病具有巨大的希望建模是因为这些细胞会引起人脑中的任何细胞，这是一个活着的“虚拟大脑” 实验性操纵，具有与单个患者完全相同的基因组成并显示先前在死后和临床样品中鉴定出的神经退行性表型。这些 “特定于患者”的体外测试系统可以发现目标发现，药物筛查和治疗证明在翻译过程中，患者细胞中的概念研究比目前可能早得多。尽管存在这些独特的优势，但将年龄作为关键的致病风险因素保存是主要的这些系统的限制。这部分是由于1）细胞中与年龄相关的特征的丧失恢复到胚胎状态，以及2）在差异协议中的缺陷从胚胎细胞产生成熟的神经元。直接的转录因子介导的重编程，也被称为“谱系转化”，患者成纤维细胞诱导的运动神经元代表了一种替代在体外产生人类神经元的方法。来自我们实验室，合作者和其他人节目的新数据通过谱系转化产生的神经元可以更好地保留与年龄相关和疾病相关的缺陷。在这项为期1年的可行性研究中，我们将比较谱系转化产生的运动神经元成纤维细胞（FIB-MNS），由与IPSC的定向分化产生的成纤维细胞。相同的成纤维细胞样品（IPSC-MNS）。成纤维细胞样品来自患有GGGGCC的患者六核苷酸重复膨胀突变在C9orf72基因中，已知会导致一种形式肌萎缩性侧索硬化症（ALS）和额颞痴呆，以及匹配的对照。我们的第一个假设是 Fib-MN的转录组将与验尸组织和来自 ALS患者。我们的合作者Verge Genomics创造了创新的，大数据驱动的ALS基因使用公共和专有基因表达数据的表达签名来自39个与ALS相关的研究我们将在项目中使用。核细胞质转运缺陷已成为一种表型，其中年龄和C9-ALS相关已经确定了差异。因此，我们的第二个假设是纤维MN将具有更多与IPSC-MN相比，明显的与年龄相关的核细胞质运输缺陷免疫荧光共聚焦显微镜。综上所述，这两项研究将证明 FIB-MN创建了优越的ALS室内测试系统，这些系统保留了重要的，与疾病相关的衰老方面对IPSC银行和疾病建模领域产生巨大影响。 Acurastem Inc.发展人类细胞模型和临床前人类验证其中枢神经系统疗法的测定及其的测定发展伙伴。