Engineering the Cancer Microenvironment

项目摘要/摘要 90%的癌症相关死亡是由转移引起的12，17.转移级联是一个复杂的序列 由癌细胞表型变化引发的事件，通过一个过程引起侵袭性 称为上皮间充质转化(MET)7、14、15。一旦转移细胞进入循环 当这些细胞进入全身时，细胞必须向远处的组织渗入，从而形成继发性肿瘤。什么时候 癌细胞到达新的组织，它们的行为在一定程度上是由不可溶的信号驱动的 微环境中的细胞外基质(ECM)蛋白。目前，不存在任何系统可以 重建这一序列的可溶和不可溶的信号事件，减缓有效信号的发展 治疗。因此，在约尔格·拉汉博士的合作下，我提议开发一种工程化癌症 可用于评估特定微环境线索对癌细胞影响的微环境 行为。Lahann实验室最近开发了一种3D聚合物光纤写入系统，它允许 具有图案化纤维结构的超多孔聚合物支架的制备。该系统使精确度 控制一系列特征，包括支架的物理/机械性能，并促进 包被各种ECM蛋白网络(在其3D生物活性构象中)。这些模块 支架可以种植各种类型的细胞，从而能够研究细胞与细胞或细胞与基质的相互作用。 因此，这种可定制的平台技术可以被设计成模仿本地癌症的关键要素 微环境。在此建议书的指导K99阶段，机械性能和ECM 成分将被调整以模拟原发和转移肿瘤的成分。这种变化的效果是 肿瘤微环境对癌细胞增殖、肿瘤干细胞分布及表型均有影响 已评估。此外，这些环境还将用于培养从 胸腔积液。细胞外基质成分对肿瘤干细胞分布和衰老的影响 将对初级患者样本进行评估。癌症微环境的细胞成分和 转移前的生态位也将被调查。在R00阶段过渡到独立，我将评估 工程化癌症微环境促进继发性肿瘤形成的体内疗效。这 K99/R00奖将使一种能够概括转移性癌细胞侵袭的系统的创建成为可能 活体，提供了一种更好地了解癌症的分子和细胞基础的方法。如果 该系统的成功将为组合药物筛选提供平台技术，并可能导致 开发个性化的癌症治疗方法。在密歇根大学，我有机会获得世界级的 设施，以及癌症研究领域无与伦比的领军人物的集合。我已经组装了一个 杰出的顾问委员会，指导我的研究，促进我向独立的过渡。约尔格博士 Lahann，生物界面研究所所长，聚合物纤维和颗粒的流体工艺专家 虚构，将作为我的主要导师。密歇根大学校长马克斯·维查博士 综合癌症中心和肿瘤发生领域的领先者，将担任我的共同导师。其他 我的指导团队的尊敬的成员包括Diane Simeone博士，翻译主任 肿瘤学项目和胰腺癌肿瘤发生专家加里·卢克博士是分子方面的专家 乳腺癌的成像系统和小鼠模型，以及Jan Stegemann博士，开发 3D组织工程环境和ECM信号。本奖项提供的机会，结合 拥有世界一流的设施和密歇根大学的指导团队，将使我能够实现 无论是我成为终身教职助理教授的短期目标，还是我的长期目标 开发技术，帮助患有侵袭性癌症的患者改善他们的 治疗结果和总体生活质量。

PROJECT SUMMARY/ABSTRACT 90% of cancer related deaths occur as a result of metastasis12,17. The metastatic cascade is a complex series of events initiated by changes of the cancer cell phenotype, giving rise to invasive properties through a process known as the epithelial-to-mesenchymal transition (MET)7, 14, 15. Once metastatic cells are in the circulatory system, the cells must extavasate into distant tissues, resulting in the formation of secondary tumors. When cancer cells arrive in the new tissues, their behavior is driven in part by the non-soluble signaling of extracellular matrix (ECM) proteins within the microenvironment10. Currently, no system exists that can recreate this sequence of soluble and non-soluble signaling events, slowing the development of effective therapies. Therefore, in collaboration with Dr. Joerg Lahann, I propose to develop an engineered cancer microenvironment that can be used to evaluate the effects of specific microenvironmental cues on cancer cell behavior. A 3D polymer fiber writing system has recently been developed within the Lahann lab that allows fabrication of hyper-porous polymer scaffolds with patterned fiber architectures. This system enables precise control over a range of characteristics including the scaffold physical/mechanical properties, and facilitates coating with networks of various ECM proteins (in their 3D biologically active conformations). These modular scaffolds can be seeded with a variety of cell types, enabling investigation of cell-cell or cell-matrix interactions. As such, this tailorable platform technology could be designed to mimic key elements of the native cancer microenvironment. During the mentored K99 phase of this proposal, the mechanical properties and ECM composition will be tuned to mimic elements of primary and metastatic tumors. The effect that changes in the cancer microenvironment has on cancer cell proliferation, cancer stem cell distribution, and phenotype will be evaluated. Furthermore, these environments will also be used to culture primary cancer cells obtained from pleural effusions. The effect of the ECM composition on cancer stem cell distribution and senescence of primary patient samples will be evaluated. The cellular components of the cancer microenvironment and the pre-metastatic niche will also be investigated. Transitioning into independence in the R00 phase, I will evaluate the in vivo efficacy of the engineered cancer microenvironments to promote secondary tumor formation. This K99/R00 award would enable the creation of a system that could recapitulate metastatic cancer cell invasion in vivo, providing a means by which the molecular and cellular basis of cancer can be better understood. If successful, this system will provide a platform technology for combinatorial drug screening and may lead to the development of personalized cancer therapies. At the University of Michigan, I have access to world-class facilities as well as an unparalleled collection of leaders in the field of cancer research. I have assembled an distinguished advisory committee to guide my research and facilitate my transition to independence. Dr. Joerg Lahann, Director of the Biointerfaces Institute and expert in fluid-based processes for polymer fiber and particle fabrication, will serve as my primary mentor. Dr. Max Wicha, Director of the University of Michigan Comprehensive Cancer Center and leader in the field of tumorigenesis, will serve as my co-mentor. Other distinguished members of my mentoring team include Dr. Diane Simeone, Director of the Translational Oncology Program and expert in pancreatic cancer tumorigenesis, Dr. Gary Luker, is an expert in molecular imaging systems and mouse models of breast cancer, and Dr. Jan Stegemann, expert in the development of 3D tissue engineered environments and ECM signaling. Opportunities provided by this award, in combination with the world-class facilities and mentoring team at the University of Michigan, would enable me to achieve both my short-term goal of becoming a tenure-track assistant professor, as well as my long-term goal of developing technologies that will help patients suffering from aggressive forms of cancer improve their therapeutic outcomes and overall quality of life.