Neoglycosylation Epitopes in Metaplasia and Cancer

项目摘要/摘要 本申请提出了一个为期五年的研究职业发展计划，重点是确定如何 在化生和癌症过程中表达的糖基化表位有助于这些具有临床意义的细胞 变形。申请人Jeffrey W.Brown，M.D.，Ph.D.，是 华盛顿大学医学院的胃肠病学博士。自从完成胃肠病研究后， 布朗博士曾在杰森·米尔斯的实验室工作，在那里他发现了一种新的细胞过程，他 称为宣泄细胞作用[希腊语：细胞清洗]，由细胞用来有效地在 化生和癌症的过程。他随后确定宣泄细胞增多症是由 Glycan 3‘-Sulfo-Lea/C和小鼠优先结合该表位的Galectins为空都不能执行 疏泄细胞或不能将磺基粘蛋白包装成成熟颗粒。由于糖生物学对布朗博士来说是一个新领域， 斯图尔特·科恩菲尔德将担任他的主要导师，并得到杰森·米尔斯(共同导师)的持续支持。一起， 应聘者将处于独特的地位，能够获得开发 调查特定糖基化表位如何调控组织转化的独立研究计划 以及在化生和癌症中的分化状态。 硫粘蛋白的表达和分泌在Barrett‘s食道和食道中均匀存在 癌症，是III型[高危]胃肠化生的定义特征，目前是最好的 检测胰腺囊性病变中高度不典型增生和癌的生物标志物。使用全面的 涉及细胞系、器官培养和小鼠模型的方法，这里提出的实验将 确定携带3‘-Sulfo-Lea/C的蛋白质组，细胞信号和转录调控其 合成和分泌，以及特定Galectins调节细胞的分子机制 差异化。最终，在斯图尔特·科恩菲尔德、杰森·米尔斯和这项研究的指导下， 咨询委员会，从拟议的实验中获得的知识和技术技能，以及完成 在概述的职业发展计划中，布朗博士将做好充分准备，建立独立的研究 计划，预计将在R01资金方面具有很强的竞争力。

PROJECT SUMMARY / ABSTRACT This application proposes a five-year research career development program focused on determining how glycosylation epitopes expressed during metaplasia and cancer contribute to these clinically significant cellular transformations. The applicant, Jeffrey W. Brown, M.D., Ph.D., is an Instructor of Medicine in the Division of Gastroenterology at Washington University School of Medicine. Since completing gastroenterology fellowship, Dr. Brown had worked in the laboratory of Jason Mills, where he has discovered a novel cellular process that he calls cathartocytosis [Greek: cellular cleansing] which is used by cells to efficiently dedifferentiate in the processes of metaplasia and cancer. He has subsequently determined that cathartocytosis is annotated by the glycan 3’-Sulfo-LeA/C and mice null for galectins that preferentially bind this epitope either fail to perform cathartocytosis or fail to package sulfomucins into mature granules. As glycobiology is a new field for Dr. Brown, Stuart Kornfeld will serve as his primary mentor with continued support from Jason Mills (Co-Mentor). Together, the candidate will be uniquely positioned to acquire the knowledge and skill set necessary to develop an independent research program investigating how specific glycosylation epitopes modulate tissue transformation and differentiation states in metaplasia and cancer. The expression and secretion of sulfomucins is uniformly present in Barrett’s esophagus and esophageal cancer, is the defining feature of type III [high-risk] gastric intestinal metaplasia, and is currently the best biomarker for detecting high-grade dysplasia and cancer in pancreatic cystic lesions. Using a comprehensive approach involving cell lines, organoid culture, and murine models, the experiments proposed herein will determine the proteome carrying 3’-Sulfo-LeA/C, the cellular signaling and transcriptional profile regulating its synthesis and secretion, as well as the molecular mechanism by which specific galectins modulate cellular differentiation. Ultimately, with the mentorship provided by Stuart Kornfeld, Jason Mills, and the research advisory committee, the knowledge and technical skills derived from the proposed experiments, and completion of the outlined career development plan, Dr. Brown will be well-prepared to establish an independent research program and is expected to be highly-competitive for R01 funding.