Targeting cancer stem cell self-renewal and proliferation mechanisms using in vitro microscale models

项目概述全世界每年诊断出近50万新的结直肠癌(CRC)病例。不幸的是，绝大多数患者无法通过目前的化疗治愈。最近的临床数据导致了分层肿瘤模型，该模型认为少数肿瘤细胞(癌症起始细胞)负责大多数转移潜力。结直肠癌起始细胞(CCIC)对当前的结直肠癌化疗高度耐药，但它们与正常的结肠干细胞有许多相似之处。寻找对正常结肠干细胞没有毒性的具有抗CCIC活性的药物是一个重要的、高影响的目标，可以改善结直肠癌化疗和患者生存。主要有两种方法用于筛选针对CCIC的药物，但两种方法都有其局限性。体外测试速度很快，但在人体内测试时，大多数“命中”都失败了，主要是由于对正常肠道和其他器官的剂量限制毒性。体内动物试验对于筛选大量化合物来说是昂贵和耗时的。它们在发现副作用方面做得更好，但仍然没有将人类细胞置于适当的微环境中，以研究对化疗耐药至关重要的细胞间相互作用。智慧价值在这项提议中，该团队将结合癌症生物学和物理科学来开发一种新的药物筛选平台，以识别专门针对CCIC的药物。在目标1中，该团队将使用体外胃肠道(GI)微型细胞培养模拟物(？CCá)结合代表体循环的多室硅PK-PD模型来表征药物的有效性和毒性。与目前的体外技术相比，该设备为药物测试提供了一个更具生理学意义的环境。在目标2中，研究小组将使用模仿人类肠道隐窝几何形状的微型3D水凝胶GI结构来检查药物对肠道稳态的影响。在目标3中，有希望的候选药物将在最先进的活体CCIC小鼠模型中进行测试。我们将依次使用这三种检测方法从化合物库中鉴定安全的CCIC药物。更广泛的影响拟议的研究试图开发几项基本技术，这些技术可能会彻底改变结直肠癌药物的发现过程。如果证明成功，该团队将把这种方法传播到结肠癌研究社区，并对数百万人的生活产生影响。这份职业提案的教育部分致力于为未来的研究人员提供接触生物医学工程的机会。康奈尔大学的本科生iGEM团队将把基因工程与微流控设备结合起来，参加他们的年度项目竞赛。将为生物教师研究所组织年度研讨会，向初中和高中教师介绍生物医学设备。该团队还将为居里学院开发一个为期一周的研究研讨会，这是一个夏季沉浸项目，旨在吸引对工程感兴趣的高中女孩。

Project SummaryAlmost half a million new cases of colorectal cancer (CRC) worldwide are diagnosed each year. Unfortunately, the great majority of patients are not cured by current chemotherapy. Recent clinical data has led to the hierarchical tumor model, which proposes that a minority of tumor cells (cancer initiating cells) are responsible for the majority of the metastatic potential. Colorectal cancer initiating cells (CCIC) are highly resistant to current CRC chemotherapy, but they share many features with normal colon stem cells. Identifying drugs with anti-CCIC activity that are not toxic to normal colon stem cells is an important, high-impact goal that can improve CRC chemotherapy and patient survival.There are primarily two kinds of assays for screening drugs that specifically target CCIC, but both have their limitations. In vitro assays are fast but most "hits" fail when tested in human, mainly due to dose-limiting toxicities against normal intestine and other organs. In vivo animal assays are expensive and time consuming for screening a large number of compounds. They are better for uncovering side effects, but still do not place human cells in the appropriate microenvironment to study intercellular interactions important for chemoresistance.Intellectual MeritIn this proposal, the team will combine cancer biology and physical science to develop a novel drug-screening platform to identify drugs that specifically target CCIC. In Aim 1, the team will characterize drug effectiveness and toxicity using an in vitro gastrointestinal tract (GI) microscale cell culture analog (ìCCÁ), coupled with a multi-chamber silicon PK-PD model representing the systemic circulation. This device provides a much more physiological environment for drug testing than current in vitro technologies. In Aim 2, the team will examine the drug effects on intestinal homeostasis using a microscale 3D hydrogel GI structure that mimics the geometry of human intestinal crypts. In Aim 3, the promising drug candidates will be tested in a state-of-the-art in vivo CCIC mouse model. We will use these three assays sequentially to identify safe CCIC drugs from a library of compound. Broader ImpactThe proposed study attempts to develop several fundamental technologies that can potentially revolutionize the process of CRC drug discovery. If proven successful, the team will disseminate this method to the colon cancer research community and make an impact on millions of lives.The educational part of this career proposal is devoted to providing future researchers with exposure to Biomedical Engineering. The undergraduate Cornell iGEM team will combine genetic engineering with microfluidic devices for their annual project competition. An annual workshops will be organized for the Institute for Biology Teachers to introduce Biomedical Devices to the middle school and high school teachers. The team will also develop a week-long research workshop for the CURIE Academy, a summer immersion program for engaging high school girls interested in engineering.