Generating a Porcine Model for Human Microvillus Inclusion Disease (MVID) by Gene Editing

通过基因编辑生成人类微绒毛包涵体病 (MVID) 猪模型

• 批准号: 9141460
• 负责人: JAMES Richard GOLDENRING
• 金额: $ 39.47万
• 依托单位: RECOMBINETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-05 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9141460
• 关键词: Accounting; Actins; Alleles; Alternative Therapies; Animal Genetics; Animal Model; Animals; Apical; Biotechnology; Breeding; Cells; Cessation of life; Childhood; Chromatin; Chronic; Clinic; Clinical; Cloning; Cytomegalovirus Infections; Cytoskeleton; DNA cassette; Defect; Dehydration; Development; Diagnosis; Diarrhea; Disease; Doctor of Philosophy; Enterocytes; Etiology; Evaluation; Family suidae; Fibroblasts; Founder Generation; Functional disorder; Gastroenterologist; Gastrointestinal Diseases; Generations; Genes; Genetic Models; Genetic Techniques; Genotype; Hereditary Disease; Human; Immunosuppression; Industry; Inherited; Innovative Therapy; Intervention; Intestines; Lead; Leucine; Life; Malabsorption Syndromes; Metabolic; Modeling; Monitor; Monomeric GTP-Binding Proteins; Motor; Mus; Mutation; Myosin ATPase; Navajo; Operative Surgical Procedures; Parenteral Nutrition; Patients; Pattern; Phenotype; Physiology; Proline; Proteins; Research; Site; Small Business Innovation Research Grant; Small Intestines; Testing; Therapeutic; Tight Junctions; Translations; Transplantation; Universities; Work; base; cellular microvillus; cost; curative treatments; disease phenotype; gastrointestinal; human disease; improved; innovation; insight; liver infection; medical schools; mouse model; mutant; neonate; novel; novel therapeutic intervention; novel therapeutics; outcome forecast; professor; protein transport; prototype; public health relevance; trafficking; transcription activator-like effector nucleases

 DESCRIPTION (provided by applicant): Microvillus inclusion disease (MVID) is the most severe cause of Congenital Diarrheal Diseases in neonates. MVID results from mutations that lead to malabsorption and life-threatening, intractable, secretory diarrhea. To date, no curative therapy exists. MVID patients are largely dependent on parenteral nutrition. Prognosis is generally poor, due to metabolic decompensation, dehydration, infections and liver complications associated with parenteral nutrition. The only alternative therapy to parenteral nutrition is intestinal transplantation; MVID accounts for 7% of pediatric bowel transplantation worldwide. However, the overall five year survival after small bowel transplantation is only about 50%. Parenteral nutrition and bowel transplants are non-permanent solutions for treating MVID, further work in deciphering how inactivating mutations in MYO5B lead to aberrant trafficking in enterocytes will provide novel insights into genotype-phenotype relations and pave the way for development of improved diagnosis and viable alternative therapeutic strategies. There is a limited availability of patient material and no suitable animal models for MVID, hampering the thorough understanding of the disease's molecular mechanisms. While we have recently been able to develop a mouse model of germline and intestinally-targeted deletion of MYO5B, these mice die in their first week of life, so any analysis of interventions that might alter the course f disease are not possible due to the small size. To that end, we propose utilizing our state-of-the-art gene-editing platform to develop swine with a specific mutation (P663L) corresponding to the human P660L mutation of the MYO5B gene, present in Navajo MVID patients. We hypothesize that introducing a Proline (P) to Leucine (L) mutation in pigs at a site analogous to the human P660L allele can induce MVID in piglets. Execution of the hypothesis can be accomplished with the following specific aim: develop and evaluate the MYO5B (P663L) mutant pigs as models of human MVID. In addition to state-of-the art gene editing platform with expertise of Drs. Melkamu (Veterinary Physiology) and Carlson (Animal biotechnology) from Recombinetics, we have engaged a world-renown expert in MVID (Dr. James R. Goldenring, MD, PhD; a gastroenterologist and a Professor of Experimental Surgery) from Vanderbilt University School of Medicine. A reliable large animal model of MVID will have tremendous impact on industry and academic research to develop and test new drugs and novel therapeutic approaches to treat this awful disease.

 描述（由申请人提供）：微绒毛包涵体病（MVID）是新生儿先天性腹泻病的最严重原因。MVID由导致吸收不良和危及生命的顽固性分泌性腹泻的突变引起。迄今为止，还没有治愈性疗法。MVID患者在很大程度上依赖于肠外营养。由于与肠外营养相关的代谢失代偿、脱水、感染和肝脏并发症，预后通常较差。肠外营养的唯一替代疗法是肠移植; MVID占全球小儿肠移植的7%。然而，小肠移植后的五年生存率仅为50%左右。 肠外营养和肠移植是治疗MVID的非永久性解决方案，进一步研究MYO 5 B失活突变如何导致肠细胞异常运输将为基因型-表型关系提供新的见解，并为开发改进的诊断和可行的替代治疗策略铺平道路。患者材料有限，没有合适的动物模型用于MVID，阻碍了对疾病分子机制的深入了解。虽然我们最近已经能够开发出一种种系和生殖靶向MYO 5 B缺失的小鼠模型，但这些小鼠在出生后第一周就死亡，因此由于体积小，任何可能改变疾病进程的干预措施的分析都是不可能的。 为此，我们建议利用我们最先进的基因编辑平台开发具有特定突变（P663 L）的猪，该突变对应于Navajo MVID患者中存在的MYO 5 B基因的人类P660 L突变。我们假设在猪中引入脯氨酸（P）至亮氨酸（L）突变的位点类似于人P660 L等位基因，可以诱导仔猪的MVID。该假设的实施可以通过以下具体目标来实现：开发和评估MYO 5 B（P663 L）突变猪作为人MVID模型。 除了拥有Melkamu博士（兽医生理学）和Carlson博士（动物生物技术）专业知识的最先进的基因编辑平台外，我们还聘请了世界知名的MVID专家（James R. Goldenring，MD，PhD;胃肠病学家和实验外科学教授）。可靠的MVID大型动物模型将对工业和学术研究产生巨大影响，以开发和测试治疗这种可怕疾病的新药和新治疗方法。