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: 7260186
• 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/7260186
• 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（由申请人提供）：克拉伯病或球样细胞脑白质营养不良是由编码溶酶体酶半乳糖苷酶（GALC）的基因突变引起的，该酶是消化髓鞘形成少突胶质细胞和许旺细胞中的半乳糖苷所必需的。这种快速进行性脱髓鞘疾病的病理学标志是神经系统中巨噬细胞（球状细胞）的显著积聚。然而，这些细胞的功能尚不清楚。迄今为止，骨髓干细胞和脐带血细胞移植是唯一被证明有益的治疗方法，供体来源的小胶质细胞可能提供酶。然而，大多数婴儿期发病的患者无法治疗，并在两岁时死亡。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，为处于疾病任何阶段的患者设计有效的治疗方法。在GALC基因中有突变的抽搐小鼠（twi）被认为是克拉布病的真正模型。最近，我们创建了巨噬细胞缺陷的twi小鼠（twi+op），通过交叉twi和巨噬细胞缺陷的骨硬化小鼠（op），唯一的克拉伯氏病模型，其中巨噬细胞被淘汰。我们对twi+op的初步研究表明，小胶质细胞/巨噬细胞的缺乏加剧了twi的脱髓鞘病理。我们还表明，twi少突胶质细胞可以维持很长一段时间，当他们有机会在体内酶。指导该提议的具体假设是小胶质细胞/巨噬细胞在twi的脱髓鞘病理学中是有益的，并且这些细胞可以用作治疗载体以向突变髓鞘形成细胞供应GALC。为了验证这些假设，我们提出了三个具体目标。1)确定小胶质细胞/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中的脱髓鞘加重，并探索潜在的分子机制。2)确定局部或全身移植小胶质细胞和巨噬细胞作为酶替代是否可以减轻twi的脱髓鞘。3)确定移植GALC过表达的骨髓干细胞和感染GALC基因编码慢病毒的脐带血细胞是否可以胜过传统的骨髓移植。这些综合数据将为设计治疗方法以治疗疾病各个阶段的患者提供所需的基础。