Cellular models of microvillus inclusion disease

微绒毛包涵体病的细胞模型

• 批准号: 8368111
• 负责人: H Lee Sweeney
• 金额: $ 20万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8368111
• 关键词: Actins; Appearance; Binding; Biochemical; Biological Assay; Caco-2 Cells; Cell Culture Techniques; Cell Line; Cell model; Cell physiology; Cells; Cellular Assay; Cessation of life; Code; Cytomegalovirus Infections; Cytoskeleton; Development; Disease; Electron Microscopy; Epithelial Cells; Future; Genes; Goals; Heterozygote; Homozygote; Human; Inclusion Bodies; Intestinal Absorption; Intestines; Kidney Diseases; Kinetics; Lead; Liver diseases; Maintenance; Missense Mutation; Modeling; Molecular Motors; Motor; Movement; Mutation; Myosin ATPase; Outcome; Parenteral Nutrition; Pathogenesis; Pathologic Processes; Proteins; Recycling; Regulation; Role; Staging; Therapeutic Intervention; Time; Vascular Diseases; apical membrane; base; cellular microvillus; design; disease-causing mutation; early onset; functional loss; insight; knock-down; mutant; outcome forecast; research study

DESCRIPTION (provided by applicant): We have developed a cellular model of microvillus inclusion disease. Microvillus inclusion disease (MVID or MID~ MIM#251850) is characterized by the loss of microvilli and the appearance of microvillus inclusion bodies in intestinal epithelial cells. This results in a lss of intestinal absorption, rapidly leading to death without parenteral nutrition (TPN). Te only documented gene involved in this disease in the gene encoding the molecular motor, myosin Vb. The majority of the mutations result in loss of functional myosin Vb protein, while a subset of the mutations are missense mutations that likely alter motor function. We have generated a line of intestinal epithelial cells (Caco-2 cells) in which myosin Vb levels are <5% of wild type and can no longer maintain microvilli when they are polarized. This will serve as a cell culture model for a subset of microvillus inclusion disease, and can be used as a cellular assay for myosin Vb function. We will characterize the impact of MVID-causing missense mutations on the kinetics of the myosin myosin Vb motor. Based on the outcome of the biochemical characterizations, we will create a number of new lines of Caco-2 cells that express representative mutant myosin Vb proteins in the near null (knock down) background, thus creating new cellular models of MVID. Upon differentiation of these cells lines, we will examine the impact the myosin Vb missense mutations on microvilli development/maintenance. The creation of these lines will set the stage for future studies to understand the role of myosin Vb in apical membrane recycling as well as provide a better understanding of the pathogenesis of MVID. PUBLIC HEALTH RELEVANCE: A number of mutations in the gene that codes for the unconventional myosin, myosin Vb, have been found to cause microvillus inclusion disease (MVID). This project seeks to create cell models of the MVID that will allow studies on the effects of the mutations in myosin Vb in sufficient detail to delineate the pathological processes that takes place. This is the first step toward developing strategies for future therapeutic interventions.

描述（由申请人提供）：