Effect of early psychosine accumulation in Krabbe Disease on CNS progenitor cells

克拉伯病早期精神嘧啶积累对中枢​​神经系统祖细胞的影响

• 批准号: 8313213
• 负责人: Nicole Janet Scott-Hewitt
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8313213
• 关键词: 2 year old; Adolescent; Adult; Affect; Age-Months; Animals; Astrocytes; Attenuated; Behavioral; Brain; Cell Count; Cell Death; Cell Proliferation; Cells; Cellular biology; Cessation of life; Cytometry; Data; Defect; Demyelinating Diseases; Demyelinations; Deterioration; Development; Disease; Ensure; Enzymes; FDA approved; Genes; Genetic; Globoid cell leukodystrophy; Goals; Harvest; Hematopoietic stem cells; Hereditary Disease; In Vitro; Individual; Intervention; Investigation; Length; Libraries; Lipids; Live Birth; Longevity; Microscopy; Molecular; Motor; Mus; Mutation; National Institute of Neurological Disorders and Stroke; Nervous system structure; Neuraxis; Neurodegenerative Disorders; Neurologic; Oligodendroglia; Oxidation-Reduction; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Population; Proteins; Psychosine; Quality of life; Receptor Protein-Tyrosine Kinases; Replacement Therapy; Research; Role; Screening procedure; Severities; Small Interfering RNA; Spinal Cord; Staining method; Stains; Stem cells; Symptoms; System; Testing; Therapeutic; Therapeutic Intervention; Time; Toxic effect; Transplantation; Umbilical Cord Blood; United States; Western Blotting; base; cell motility; commercialization; critical developmental period; cytotoxic; drug candidate; drug discovery; early onset; galactosylceramidase; in vivo; infancy; knock-down; migration; mouse model; mutant; myelination; nervous system development; novel; progenitor

DESCRIPTION (provided by applicant): The molecular effect of early psychosine accumulation in Krabbe disease has not been studied in primary central nervous system (CNS) progenitor cell populations, specifically those of the oligodendroglial lineage responsible for proper myelination. By the time early onset Krabbe disease patients have presented with neurological symptoms, damage to the CNS may be irreparable due to early accumulation of abnormal levels of psychosine and its effect on vulnerable progenitor populations during critical developmental periods. Preliminary data suggests that physiologically relevant concentrations of psychosine are activating the previously described [13] redox/Fyn/c-Cbl pathway in primary oligodendrocyte/type-2 astrocyte progenitor cells (O-2A/OPCs). This pathway plays an important role in normal O-2A/OPC proliferation and migration. Further investigations on this pathway will better define cellular and molecular mechanisms of psychosine toxicity. Moreover, FDA-approved drugs in the NINDS II library will be screened to identify those that either ameliorate or exacerbate the toxic effects of psychosine. Rapid in vitro screening will be conducted using an automated adherent cell cytometry system. Candidate drugs will be further tested in vivo in the early onset Krabbe disease mouse model, the twitcher mouse. Developmental effects will be identified through immunohistochemical staining of brain and spinal cord sections and Western blot analysis. Behavioral and motor function tests, lifespan and symptom severity will be conducted as functional readouts of drug treatment. The goal is to protect early CNS development from effects of endogenous psychosine accumulation in early onset Krabbe patients through drug treatment intervention, ultimately to enable increased efficacy when used in combination with genetic and enzymatic replacement therapies. PUBLIC HEALTH RELEVANCE: Krabbe Disease is a debilitating genetic disorder of the nervous system that affects 1 in 100,000 live births in the United States (however the carrier rate for common mutations is estimated at 1 in 125 individuals) and results in severe neurological degeneration, demyelination, and death. At present there is no cure, and current therapeutic interventions have little beneficial effect in patients that have the more severe early onset (infantile) form of the disease. My research aims to identify abnormalities in early nervous system development due to alterations in normal progenitor cell biology and to screen FDA-approved drugs that ameliorate or exacerbate these effects to contribute to a multi-therapeutic approach that affords increased length and quality of life to patients that currently have no efficacious treatment options.

描述（由申请人提供）：尚未在原代中枢神经系统（CNS）祖细胞群，特别是负责正确髓鞘形成的少突胶质细胞谱系中研究克拉伯病中早期精神嘧啶积累的分子效应。当早期发病的克拉伯病患者出现神经系统症状时，由于精神碱异常水平的早期积累及其对关键发育时期脆弱祖细胞群的影响，对中枢神经系统的损害可能是不可挽回的。初步数据表明，生理相关浓度的精神嘧啶正在激活原代少突胶质细胞/2 型星形胶质细胞祖细胞 (O-2A/OPC) 中先前描述的 [13] 氧化还原/Fyn/c-Cbl 途径。该途径在正常的 O-2A/OPC 增殖和迁移中发挥重要作用。对该途径的进一步研究将更好地定义精神毒理学的细胞和分子机制。此外，还将对 NINDS II 库中 FDA 批准的药物进行筛选，以确定那些可以减轻或加剧精神毒副作用的药物。将使用自动贴壁细胞计数系统进行快速体外筛选。候选药物将在早发克拉伯病小鼠模型（抽搐小鼠）中进一步进行体内测试。将通过脑和脊髓切片的免疫组织化学染色以及蛋白质印迹分析来确定发育影响。行为和运动功能测试、寿命和症状严重程度将作为药物治疗的功能读数进行。目标是通过药物治疗干预，保护早发 Krabbe 患者的早期中枢神经系统发育免受内源性精神碱积累的影响，最终与遗传和酶替代疗法结合使用时提高疗效。 公共健康相关性：克拉伯病是一种使神经系统衰弱的遗传性疾病，在美国影响十万分之一的活产儿（但常见突变的携带率估计为每 125 人中有 1 人），并导致严重的神经退行性变、脱髓鞘和死亡。目前尚无治愈方法，目前的治疗干预措施对于早期病情较严重的患者几乎没有什么有益效果 该疾病的发病（婴儿）形式。我的研究旨在识别由于正常祖细胞生物学改变而导致的早期神经系统发育异常，并筛选 FDA 批准的可改善或加剧这些影响的药物，以促进多种治疗方法，为目前没有有效治疗选择的患者提供更长的寿命和更高的生活质量。