Use of IPSC to define role of astrocytes in specifying risk for onset of cerebral adrenoleukodystrophy

使用 IPSC 来定义星形胶质细胞在确定脑肾上腺脑白质营养不良发作风险中的作用

基本信息

批准号：
10435433
负责人：
Jaspreet Singh
金额：
$ 35.37万
依托单位：
HENRY FORD HEALTH SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10435433
关键词：
Accounting Address Adrenoleukodystrophy Adrenomyeloneuropathy Allogenic Amino Acids Area Astrocytes Autologous Automobile Driving Autopsy Benign Biochemical Body Fluids Brain Brain Diseases CRISPR/Cas technology Cells Cerebrum Citric Acid Cycle Clinical Clinical Course of Disease DNA Sequence Alteration Data Defect Deletion Mutation Demyelinations Development Diagnosis Disease Disease Progression Distant Drug Design Energy Metabolism Exposure to Eye Female Fibroblasts Genes Genetic Diseases Genotype Glucose Glycolysis Goals Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human Inborn Errors of Metabolism Inflammatory Response Inherited Investigation Knock-in Knowledge Laboratories Lead Life Link Membrane Proteins Metabolic Metabolic Diseases Metabolic Pathway Metabolism Mission Mitochondria Modeling Mutation National Institute of Neurological Disorders and Stroke Neonatal Screening Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurologic Neurons Patients Phenotype Plasma Procedures Protein Import Public Health Regulation Reporting Research Respiration Risk Role Siblings Source Specific qualifier value Sphingolipids Spinal Cord Diseases Stimulus Supplementation Testing Therapeutic Therapeutic Intervention Time Tissues Translating Up-Regulation Very Long Chain Fatty Acid axonopathy base clinical development clinical predictors comparative course development cytokine disease phenotype drug development effective therapy expectation improved in vivo induced pluripotent stem cell innovation male metabolomics mitochondrial dysfunction motor disorder nervous system disorder new therapeutic target novel peroxisome prognostic indicator response screening panel screening program stem cell gene therapy therapeutically effective transcriptome white matter

项目摘要

PROJECT SUMMARY/ABSTRACT The mechanism of disease progression from benign to fatal phenotypes in X-linked adrenoleukodystro- phy remains unknown and there is no satisfactory cure for the disease. 60% of male X-ALD patients develop fatal cerebral disease (cALD) while the remaining 40% develop milder adrenomyeloneuropathy (AMN) charac- terized by axonopathy. The primary genetic defect in X-ALD (mutation/deletion in ABCD1 gene) and the bio- chemical defect (accumulation of very long chain fatty acid; C>22:0 in plasma and tissues) cannot predict the onset of AMN or cALD. The long-term goal is to contribute to the development of novel clinically useful, mecha- nism-based prognostic indicators and therapeutic options for X-ALD. The overall objective for this application is to determine differential metabolic energy metabolism underlying phenotype variability (AMN vs cALD) in the human astrocytes of male X-ALD phenotypes. The central hypothesis is that altered metabolic reprogramming underlies the differential phenotype development in AMN and cALD astrocytes. These astrocytes were differen- tiated from induced pluripotent stem cells (iPSCs), which, in turn, were generated by reprogramming of human control, AMN and cALD patient-derived fibroblasts. This hypothesis is supported by untargeted metabolomics pilot data identifying metabolites altered between healthy-control and cALD phenotype postmortem brain and between AMN and cALD astrocytes. Within the cALD brain white matter, unique metabolite changes were rec- orded between distant normal looking areas and areas adjacent to the plaque suggesting an association with disease progression. OXPHOS and glycolysis were found to be decreased (low metabolic state) in human cALD astrocytes. This low metabolic state suggests a role for novel alternative source(s) of fuel driving the progression to cALD phenotype in astrocytes. The rationale for the proposed research is that a mechanistic modelling of aberrant energy metabolism in AMN and cALD astrocytes will provide a basis for predicting disease progression and new opportunities for identification of targets for novel therapeutic drug design. The central hypothesis will be tested by pursuing two specific aims: 1) Determine the role of metabolic reprogramming in newly “forged” AMN and cALD astrocytes; and 2) Determine the contribution of mitochondrial dysfunction in metabolic repro- gramming in AMN and cALD astrocytes. The approach will take advantage of control, AMN and cALD astrocytes recently generated from iPSCs in the laboratory. This proposal is innovative because it departs from the status quo by identifying for the first time, metabolic pathways differentially altered in human AMN and cALD astrocytes. The proposed research is significant because the cellular mechanism(s) that lead to less severe AMN or fatal cALD phenotype in response to same ABCD1 mutation remain unknown even four decades after the identifica- tion of gene defect in X-ALD. Successful completion of the proposed research is expected to provide a necessary conceptual framework for the subsequent development of clinically effective strategies for predicting disease progression and improving current limited treatment options.

项目概要/摘要 X连锁肾上腺脑白质营养不良疾病从良性表型进展到致命表型的机制 phy 仍然未知，并且没有令人满意的治愈方法，60% 的男性 X-ALD 患者会出现这种疾病。致命性脑病 (cALD)，而其余 40% 则出现较轻的肾上腺脊髓神经病 (AMN) 特征由轴突病引起的主要遗传缺陷（ABCD1 基因突变/缺失）和生物- 化学缺陷（极长链脂肪酸的积累；血浆和组织中 C>22:0）无法预测 AMN 或 cALD 的发生长期目标是促进新型临床有用的机械的开发。基于 Nism 的 X-ALD 预后指标和治疗选择该应用的总体目标是。确定表型变异性（AMN 与 cALD）背后的差异代谢能量代谢男性 X-ALD 表型的人类星形胶质细胞的中心假设是改变代谢重编程。 AMN 和 cALD 星形胶质细胞的不同表型发育是这些星形胶质细胞不同的基础。源自诱导多能干细胞 (iPSC)，而 iPSC 又是通过对人类细胞进行重新编程而产生的对照、AMN 和 cALD 患者来源的成纤维细胞这一假设得到非靶向代谢组学的支持。试验数据确定了健康对照和 CALD 表型死后大脑之间代谢物的变化在 AMN 和 cALD 星形胶质细胞之间，发现了 cALD 脑白质中独特的代谢变化。在遥远的正常区域和邻近斑块的区域之间排序，表明与发现人类 cALD 中 OXPHOS 和糖酵解降低（低代谢状态）。这种低代谢状态表明新的替代燃料来源在推动进展中发挥着作用。该研究的基本原理是建立星形胶质细胞中的 cALD 表型。 AMN 和 cALD 星形胶质细胞异常的能量代谢将为预测疾病进展提供基础以及确定新型治疗药物设计靶标的新机会。通过追求两个具体目标进行测试：1）确定代谢重编程在新“锻造”中的作用 AMN 和 cALD 星形胶质细胞；2) 确定线粒体功能障碍在代谢生殖中的作用 AMN 和 cALD 星形胶质细胞的编程该方法将利用对照、AMN 和 cALD 星形胶质细胞。该提案是最近从实验室的 iPSC 中产生的，因为它偏离了现状。通过首次确定人类 AMN 和 cALD 星形胶质细胞中代谢途径的差异改变。拟议的研究意义重大，因为导致不太严重的 AMN 或致命的细胞机制即使在鉴定后四十年，对相同 ABCD1 突变的 cALD 表型仍然未知。 X-ALD 中基因缺陷的分析预计成功完成拟议的研究将提供必要的支持。后续开发临床有效预测疾病策略的概念框架进展和改善目前有限的治疗选择。