The role of microglia/macrophages and their therapeutic use in Krabbe's disease

小胶质细胞/巨噬细胞的作用及其在克拉伯病中的治疗用途

• 批准号: 7359648
• 负责人: IAN DAVID DUNCAN
• 金额: $ 25.73万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7359648
• 关键词: Ablation; Age-Years; Biochemical; Blood Cells; Blood Circulation; Bone Marrow Stem Cell; Bone Marrow Stem Cell Transplantation; Bone Marrow Transplantation; Cell physiology; Cells; Cessation of life; Clinical; Code; Data; Demyelinating Diseases; Demyelinations; Development; Disease; Down-Regulation; Enzymes; Failure; Foundations; Functional disorder; Future; Galactosylceramides; Gene Transfer; Genes; Genetic; Globoid cell leukodystrophy; Goals; Human; Lipids; Longevity; Mass Spectrum Analysis; Microglia; Modeling; Molecular; Motor; Mus; Mutation; Myelin; Natural regeneration; Nervous system structure; Neuraxis; Neurologic; Oligodendroglia; Onset of illness; Pathology; Patients; Peripheral Nervous System; Protein Overexpression; Psychosine; Research; Research Personnel; Role; Scheme; Schwann Cells; Staging; Subfamily lentivirinae; Symptoms; Testing; Therapeutic; Therapeutic Effect; Therapeutic Uses; Time; Transplantation; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; Western Blotting; base; cellular transduction; design; galactocerebroside; galactosylceramidase; improved; in vivo; infancy; interdisciplinary approach; macrophage; mutant; myelination; progenitor; programs; ranpirnase; response; success; tool

DESCRIPTION (provided by applicant): Krabbe's disease, or globoid cell leukodystrophy, is caused by mutations in the gene coding for the lysosomal enzyme, galactocerebrosidase (GALC), which is required to digest galactocerebroside in myelin forming oligodendrocytes and Schwann cells. A pathological hallmark of this rapidly progressive demyelinating disease is the marked accumulation of macrophages (globoid cells) in the nervous system. However, the function of these cells is unclear. To date, transplantation of bone marrow stem cells and umbilical cord blood cells are the only treatments that have proven beneficial, with donor-derived microglia potentially supplying enzyme. However, the majorities of patients with infantile onset are untreatable and succumb by two years of age. Our long term goal is to devise an efficient therapy for patients at any stage of the disease by enhancing enzyme replacement and by extending the time window for successful therapy. The twitcher mouse (twi), which has a mutation in the GALC gene, is considered a bonafide model of Krabbe's disease. Recently we created macrophage-deficient twi mice (twi+op) by crossing twi and macrophage-deficient osteopetrotic mice (op), the only model of Krabbe's disease from which macrophages are eliminated. Our preliminary study of twi+op indicates that the absence of microglia/macrophages exacerbates the demyelinating pathology of twi. We have also shown that twi oligodendrocytes can be maintained for long periods of time when they have access to enzyme in vivo. The specific hypotheses guiding this proposal are that microglia/macrophages are beneficial in the demyelinating pathology of twi and these cells can be used as a therapeutic vehicle to supply GALC to mutant myelinating cells. To test these hypotheses, we propose three specific aims. 1) Determine the roles of microglia/macrophages in the demyelinating pathology of twi. Using a multidisciplinary approach, the exacerbated demyelination in twi+op will be analyzed and the underlying molecular mechanisms will be explored. 2) Determine whether microglia and macrophages transplanted locally or systemically as a means of enzyme replacement can alleviate the demyelination of twi. 3) Determine whether transplantation of GALC-overexpressing bone marrow stem cells and umbilical cord blood cells infected with a GALC-gene-encoding lentivirus can outperform the conventional bone marrow transplantation in twi. These combined data will provide the foundation required to devise therapeutic approaches to treating patients at all stages of the disease.

描述(申请人提供)：克拉贝氏病，或球状细胞白质营养不良，是由溶酶体酶半乳脑苷酶(GALC)的基因编码突变引起的，半乳脑苷酶是消化髓鞘中半乳糖脑苷脂所必需的，形成少突胶质细胞和雪旺细胞。这种快速发展的脱髓鞘疾病的一个病理特征是神经系统内巨噬细胞(球状细胞)的显著聚集。然而，这些细胞的功能尚不清楚。到目前为止，骨髓干细胞和脐血细胞移植是唯一被证明有效的治疗方法，捐赠者来源的小胶质细胞可能提供酶。然而，大多数婴儿发病的患者是无法治疗的，并在两岁前死亡。我们的长期目标是通过加强酶替代和延长成功治疗的时间窗口，为疾病任何阶段的患者设计一种有效的治疗方法。具有GALC基因突变的抽动鼠被认为是Krabbe病的真正模型。最近，我们通过将TWI和巨噬细胞缺陷的骨质疏松症小鼠(OP)杂交，创造了巨噬细胞缺陷的TWI小鼠(TWI+OP)，这是唯一一种去除巨噬细胞的Krabbe病模型。我们对TWI+OP的初步研究表明，小胶质细胞/巨噬细胞的缺失加剧了TWI的脱髓鞘病理。我们还表明，当两个少突胶质细胞在体内获得酶时，它们可以长时间保持。指导这一提议的具体假设是，小胶质细胞/巨噬细胞在TWI的脱髓鞘病理中是有益的，这些细胞可以作为一种治疗载体，向突变的髓鞘细胞供应GALC。为了检验这些假设，我们提出了三个具体目标。1)探讨小胶质细胞/巨噬细胞在TWI脱髓鞘病理中的作用。使用多学科方法，将分析TWI+OP中加剧的脱髓鞘，并探索其潜在的分子机制。2)确定局部或系统移植小胶质细胞和巨噬细胞作为酶替代手段是否能减轻TWI的脱髓鞘。3)确定GALC高表达的骨髓干细胞和感染GALC基因编码慢病毒的脐血细胞移植能否在两个月内优于传统的骨髓移植。这些合并的数据将为设计治疗方法以治疗疾病所有阶段的患者提供所需的基础。