Genetic Control of Phospholipid Biosynthesis and Muscular Dystrophy

磷脂生物合成和肌营养不良症的遗传控制

• 批准号: 8130650
• 负责人: GREGORY A. COX
• 金额: $ 33.26万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-14 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8130650
• 关键词: 22q13.3; Adult; Affect; Alleles; Anabolism; Birth; Brazil; Candidate Disease Gene; Cells; Cessation of life; Choline; Choline Kinase; Choline-Phosphate Cytidylyltransferase; Clinical; Collaborations; DNA; Defect; Disease; Disease Progression; Electron Transport; Enzymes; Eukaryotic Cell; Family; Forelimb; Foundations; Genes; Genetic; Goals; Hindlimb; Histopathology; Human; Japan; Lead; Lecithin; Limb-Girdle Muscular Dystrophies; Link; Lipids; Maintenance; Membrane; Membrane Potentials; Membrane Proteins; Merosin; Mitochondria; Molecular; Molecular Genetics; Mus; Muscle; Muscle Weakness; Muscular Dystrophies; Mutant Strains Mice; Mutation; Myoblasts; Nature; Nuclear Envelope; Pathway interactions; Patients; Phenotype; Phospholipids; Phosphorylation; Phosphorylcholine; Physiology; Population; Process; Proteins; Sampling; Sarcolemma; Severities; Skeletal Muscle; Testing; Tissues; Transgenic Mice; Universities; cohort; congenital muscular dystrophy; disability; disease phenotype; gait examination; genetic linkage; human disease; in vivo; mitochondrial dysfunction; mitochondrial membrane; muscle degeneration; muscle transplantation; mutant; novel; null mutation; premature; protein function; wasting

DESCRIPTION (provided by applicant): Muscular dystrophies include a diverse group of genetically heterogeneous disorders that affect 1:2000 births worldwide. The diseases are characterized by progressive muscle weakness and wasting that lead to severe disability and often premature death. Rostrocaudal muscular dystrophy (rmd) is a new recessive mouse mutation causing a progressive muscular dystrophy with a rostral-to-caudal gradient of severity. We have identified the rmd mutation as a 1.6 kb deletion within the choline kinase beta (Chkb) gene, resulting in loss of CHKB protein and enzymatic activity. CHKB is one of two mammalian choline kinase (CHK) enzymes catalyzing the phosphorylation of choline to phosphocholine in the biosynthesis of the major membrane phospholipid phosphatidylcholine (PC). PC is the most abundant phospholipid in eukaryotic cells and it serves not only as a structural foundation for membrane bilayers, but also as a precursor for several lipid messengers. While mutant rmd mice show dramatically decreased CHK activity in all tissues examined, the disease is only evident in skeletal muscle tissues which display a 30% reduction in total PC content. Several muscular dystrophy mutations have been identified in membrane-associated proteins that function at the sarcolemma or nuclear envelope. However, the rmd mutant mouse is the first to demonstrate that alterations in membrane phospholipid composition can result in a progressive muscular dystrophy phenotype. The aims of our proposal are: 1) To test the hypothesis that rmd muscular dystrophy is due to a cell autonomous loss of CHKB in skeletal muscles. 2) To test the hypothesis that mitochondrial dysfunction underlies the progressive skeletal muscle degeneration in primary mouse myoblast cultures and adult skeletal muscles. 3) To screen a large Brazilian population of congenital muscular dystrophy (CMD) and limb-girdle muscular dystrophy (LGMD) patients for mutations in the orthologous human CHKB gene. In collaboration with Dr. Mayana Zatz, we will test for human CHKB mutations in DNA samples from patients that have previously been screened and eliminated for known molecular causes of muscular dystrophy. The primary goals of this proposal are to use the novel mouse rmd mutation to explore the cell-autonomous nature of this defect in skeletal muscles, to dissect the molecular and genetic mechanisms of muscular degeneration and to determine the range of defects in CHKB that underlie previously unidentified human muscular dystrophies. The rmd mouse offers the first demonstration of a defect in a phospholipid biosynthetic enzyme causing muscular dystrophy and may reveal unique entry points in which to intervene in the disease process.

描述（由申请人提供）：肌营养不良包括一组遗传异质性疾病，影响全球1：2000的出生。这些疾病的特点是进行性肌肉无力和消瘦，导致严重残疾，往往过早死亡。嘴尾型肌营养不良症（rmd）是一种新的隐性基因突变小鼠引起的进行性肌营养不良症的严重程度与嘴尾梯度。我们已经确定rmd突变为胆碱激酶β（Chkb）基因内的1.6 kb缺失，导致CHKB蛋白和酶活性丧失。CHKB是在主要膜磷脂磷脂酰胆碱（PC）的生物合成中催化胆碱磷酸化为磷酸胆碱的两种哺乳动物胆碱激酶（CHK）酶之一。PC是真核细胞中最丰富的磷脂，它不仅是膜双层的结构基础，而且是多种脂质信使的前体。虽然突变rmd小鼠在所有检查的组织中显示出显著降低的CHK活性，但该疾病仅在骨骼肌组织中明显，其显示总PC含量降低30%。已经在肌膜或核膜上起作用的膜相关蛋白中鉴定了几种肌营养不良突变。然而，rmd突变小鼠是第一个证明膜磷脂组成的改变可以导致进行性肌营养不良表型的小鼠。我们的建议的目的是：1）为了测试的假设，rmd肌营养不良症是由于细胞自主丢失的CHKB在骨骼肌。2)在原代小鼠成肌细胞培养物和成人骨骼肌中，验证线粒体功能障碍是进行性骨骼肌变性的基础这一假设。3)筛选巴西大量先天性肌营养不良症（CMD）和肢带型肌营养不良症（LGMD）患者的正向人类CHKB基因突变。与Mayana Zatz博士合作，我们将测试来自先前已筛查并消除已知肌营养不良分子原因的患者的DNA样本中的人类CHKB突变。该提案的主要目标是使用新的小鼠rmd突变来探索骨骼肌中这种缺陷的细胞自主性质，剖析肌肉变性的分子和遗传机制，并确定CHKB中的缺陷范围，这些缺陷是先前未识别的人类肌营养不良症的基础。rmd小鼠首次证明了磷脂生物合成酶的缺陷导致肌营养不良，并可能揭示干预疾病过程的独特切入点。