ELUCIDATING THE ROLE OF COATOMER COMPLEX COPI IN SKELETAL DYSPLASIA

项目总结 这份建议书描述了一项为期五年的指导培训经验，旨在为申请者的职业生涯做好准备。 在基础科学和翻译科学研究方面。申请人拥有医学博士和博士学位，并由 美国医学遗传学和基因组学委员会。申请者的长期目标是成为一名医生- 研究骨骼发育不良分子遗传学基础的科学家。职业发展计划包括 指导、正式课程、实验室会议、研讨会、国家会议以及与咨询人员的会议 委员会审议阶段。该计划旨在扩大申请者的研究技能，包括课程和 在模型生物、糖蛋白组学、显微成像和生物统计学方面的实验学习。此外, 拟议的计划将提供领导力、指导、实验室管理、科学写作和 编辑、拨款撰写和研究的伦理影响。分子和人类遗传学系 在贝勒医学院，在培养非常成功的内科科学家方面有着长期的记录。这个 Mentor，Brendan Lee博士，是骨骼发育不良领域的领先专家，一直是 为超过9名K和VA职业发展奖获得者提供指导，所有这些人都已进入独立实验室- 以研究为基础的职业。选择咨询委员会是为了补充导师的专业知识，并 为申请人提供重要的研究和职业指导。拟议的研究将调查 骨中COPB2单倍体功能不全的功能、细胞和生化后果。功能丧失 在患有发育障碍的儿童中发现了COPB2的变异，COPB2是COPI辅原子复合体的一个亚单位 延迟和骨骼脆性。COPI在ER和高尔基人之间以及高尔基人内部的贩运中发挥作用 水池。囊泡运输缺陷，包括COPI功能障碍，已被认为与骨骼发育不良有关。 初步数据显示，Copb2+/-小鼠表现出低骨量表型，而Copb2缺失的斑马鱼 胚胎表现出异常的前胶原分泌。申请人提出COPB2缺乏导致骨骼 通过导致胶原运输延迟，高尔基体功能障碍和改变的自噬，导致脆性，导致 破坏成骨细胞的分化。这项拟议的研究涉及分析中国人的骨骼表型。 COPB2缺陷小鼠模型，包括在特定阶段选择性删除骨骼中的Copb2 成骨细胞分化(“条件基因敲除”)。通过采用这种方法，申请人将在 在骨骼发育的哪个时间点，COPB2的缺乏变得至关重要。建议进行的研究 还旨在检查COPB2缺乏是否会改变蛋白质糖基化，并导致内质网高尔基体功能障碍，以及 这些变化是否可以接受治疗。这项研究将涉及COPb2缺陷的分析 小鼠模型和细胞。这个应用程序提供了广泛的研究经验，并利用先进的 技术，拟议的职业发展计划将为申请人的职业生涯做好准备 独立研究科学家。

PROJECT SUMMARY This proposal describes a five-year mentored training experience designed to prepare the applicant for a career in basic and translational science research. The applicant holds M.D. and Ph.D. degrees, and is certified by the American Board of Medical Genetics and Genomics. The applicant’s long-term goal is to become a physician- scientist studying the molecular genetic basis of skeletal dysplasias. The career development plan includes mentorship, formal coursework, lab meetings, seminars, national conferences, and meetings with the advisory committee. The plan is designed to broaden the applicant’s research skills, including coursework and experimental learning in model organisms, glycoproteomics, microscopy imaging, and biostatistics. In addition, the proposed plan will provide training in leadership, mentorship, laboratory management, scientific writing and editing, grant writing, and the ethical implications of research. The Department of Molecular and Human Genetics at Baylor College of Medicine has a long track record of training highly successful physician scientists. The mentor, Dr. Brendan Lee, is a leading expert in the field of skeletal dysplasia and has been the primary research mentor for over 9 K and VA career development awardees, all of whom have gone onto independent laboratory- research based careers. The advisory committee was selected to complement the mentor’s expertise and to provide important research and career guidance for the applicant. The proposed study will investigate the functional, cellular, and biochemical consequences of COPB2 haploinsufficiency in bone. Loss-of-function variants in COPB2, a subunit of the COPI coatomer complex, were identified in children with developmental delay and bone fragility. The COPI functions in trafficking between the ER and Golgi, and within the Golgi cisternae. Vesicular trafficking defects, including COPI dysfunction, have been implicated in skeletal dysplasia. Preliminary data demonstrated that Copb2+/- mice exhibit a low bone mass phenotype, and copb2-null zebrafish embryos show abnormal secretion of procollagen. The applicant proposes that COPB2 deficiency leads to bone fragility by causing delayed collagen trafficking, Golgi-ER dysfunction, and altered autophagy, resulting in disruption of osteoblast differentiation. The proposed study involves analysis of the skeletal phenotype in COPB2-deficient mouse models, including selective deletion of Copb2 in bone during specific stages of osteoblast cell differentiation (“conditional knock-out”). By taking this approach, the applicant will determine at which time point during skeletal development the deficiency of COPB2 becomes critical. The proposed study also aims to check if COPB2 deficiency alters protein glycosylation, and causes an ER-Golgi dysfunction, and whether these changes may be amenable to therapy. The study will involve the analysis of COPB2-deficient mouse models and cells. This application, which provides a broad research experience and utilizes advanced technologies, and the proposed career development plan will prepare this applicant for a career as an independent research scientist.