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（由适用提供）：微绒毛包含疾病（MVID）是新生儿先天性腹泻疾病的最严重原因。 MVID是由于突变导致吸收不良和威胁生命的，棘手的秘书长腹泻而引起的。迄今为止，尚无治愈疗法。 MVID患者在很大程度上取决于父母的营养。由于代谢代谢，脱水，感染和与父母营养相关的肝脏并发症，预后通常很差。父母营养的唯一替代疗法是肠道移植。 MVID占全球小儿肠移植的7％。但是，小肠移植后的整体五年生存率仅约50％。肠胃外营养和肠道移植是用于治疗MVID的非永久解决方案，进一步的努力在解释肌5b中的灭活突变如何导致肠上细胞中的异常贩运将提供新颖的见解，从而为基因型 - 表型关系提供新的见解，并为改进的诊断和可行的替代策略提供开发的方式。患者材料的可用性有限，没有合适的动物模型用于MVID，从而阻碍了对疾病分子机制的彻底理解。虽然我们最近能够开发出肌5b的种系和刻有靶向的删除的小鼠模型，但这些小鼠在生命的第一周就死亡，因此，由于尺寸较小，对可能改变病程F疾病的干预措施的任何分析都是不可能的。为此，我们建议利用我们的最先进的基因编辑平台以特异性突变（P663L）开发猪，该突变（P663L）与纳瓦霍人MVID患者中的MyO5B基因的人类P660L突变相对应。我们假设在类似于人类P660L等位基因类似的部位，将脯氨酸（P）引入猪（L）突变可能会诱导小猪中的MVID。可以通过以下特定目的来实现该假设的执行：开发和评估MyO5b（P663L）突变猪作为人类MVID模型。除了具有DRS专业知识的最先进的基因编辑平台。 Melkamu（兽医生理学）和Carlson（Animal Biotechnology）来自重组研讨会，我们从事MVID的世界知名专家（James R. Goldenring博士，博士学位；胃肠病学家和一名胃肠病学家和实验性外科手术教授）。可靠的大型动物模型将对工业和学术研究产生巨大影响，以开发和测试新药以及新型治疗方法以治疗这种可怕的疾病。