Engineered Hydrogel Elucidates the Contribution of ECM Stiffness to Barrett's Esophagus Pathogenesis

项目摘要 Barrett食管（BE）是一种食管鳞状上皮被食管上皮替代的疾病， 柱状肠上皮（称为化生），并且据信在美国影响3-4百万人。被 患者发生食管腺癌（EAC）的风险高30-125倍， 等级和高度发育不良状态，与一般人群相比。BE的发病机制 涉及表观遗传/基因组畸变（例如TP 53突变），以及与微环境线索的相互作用。 最近的研究表明，ECM硬化与化生（或BE）-发育不良- 腺癌序列，我的新数据表明，增加ECM刚度，一个关键的生物力学， 性，是BE发病的特征。这突出表明迫切需要了解机制 在ECM僵硬的情况下促进BE化生和发育不良。因此，该提案将整合 我在生物材料工程方面的独特博士前培训，以及我新提出的职业发展培训， 上皮细胞和类器官生物学，以利用工程化的HA基水凝胶来阐明 ECM硬度在BE发病机制，并确定基质激活的治疗靶点。这种生物工程 将结合TP 53突变（在BE的上皮中）来了解BE的发病机制。的 我的建议的首要假设是ECM刚度调节可以重现发育不良的BE状态， 揭示了疾病的新的潜在机制。我将通过以下内容来探讨这一假设 相互关联的具体目标：目标1侧重于建立一个水凝胶平台，以研究促异型增生 基质性质在患者源性BE类器官中的贡献。目标2侧重于体外鉴定和 在暴露于硬基质的患者源性BE类器官中的治疗靶点的体内评价。目标3侧重于 利用BE的hiPSC衍生平台来阐明内在和外在因素之间的合作 有助于BE发病机制。这项工作是创新的，因为它使用了先进的生物工程方法 阐明ECM硬度在人类BE发病机制中的功能作用。成功完成本 通过定义生物力学对BE的贡献，该提案将具有科学和临床意义 并为破坏BE基质硬度的新疗法奠定基础。研究计划， 培训/职业发展/导师计划，咨询委员会，RCR培训，保护研究时间， 和机构支持将为我过渡到最终独立的NIH提供基础 终身教职研究员，谁融合了基础生物学，生物工程和翻译的原则 药

Project Summary Barrett’s esophagus (BE) is a disease in which the squamous epithelium of the esophagus is replaced by a columnar intestinal epithelium (termed metaplasia), and is believed to affect 3-4 million people in the US. BE patients have a 30-125-fold greater risk of developing esophageal adenocarcinoma (EAC) via intermediate low- grade and high-grade dysplastic states, when compared to the general population. The pathogenesis of BE involves epigenetic/genomic aberrations (e.g. TP53 mutations), and an interplay with microenvironmental cues. Recent studies have demonstrated that ECM stiffening is associated with metaplasia (or BE)-dysplasia- adenocarcinoma sequence, and my novel data reveal that increased ECM stiffness, a critical biomechanical property, is characteristic of BE pathogenesis. This underscores a compelling need to understand mechanisms that foster BE metaplasia and dysplasia in the context of ECM stiffness. Therefore, this proposal will integrate my unique predoctoral training in biomaterial engineering with my new proposed career development training in epithelial cell and organoid biology to utilize an engineered HA-based hydrogel to elucidate the contribution of ECM stiffness in BE pathogenesis, and to identify matrix-activated therapeutic targets. This bioengineering approach will be combined with TP53 mutation (in the epithelium of BE) to understand BE pathogenesis. The overarching hypothesis of my proposal is that ECM stiffness modulation can recapitulate a dysplastic BE state, revealing novel underlying mechanisms of the disease. I will pursue this hypothesis through the following interrelated Specific Aims: Aim 1 focuses on establish a hydrogel platform to study the pro-dysplastic contribution of matrix properties in patient-derived BE organoids. Aim 2 focuses on in vitro identification and in vivo evaluation of therapeutic targets in stiff matrix-exposed patient-derived BE organoids. Aim 3 focuses on utilizing an hiPSC-derived platform of BE to elucidate the cooperation between intrinsic and extrinsic factors contributing to BE pathogenesis. This work is innovative because it uses an advanced bioengineering approach to elucidate the functional roles of ECM stiffness in human BE pathogenesis. Successful completion of this proposal will be both scientifically and clinically significant by defining the contribution of biomechanics to BE and laying a foundation for novel therapies to disrupt matrix stiffness in BE. The research proposal, training/career development/mentorship plans, advisory committee, RCR training, protected time for research, and institutional support will provide the foundation for me to transition to an eventual independent NIH funded tenure-track faculty investigator, who blends principles of basic biology, bioengineering, and translational medicine.