DNA-Directed Micropatterning of Cells to Investigate Prostate Cancer Dormancy in the Bone Marrow

虽然大约50%的前列腺癌患者存在播散性肿瘤细胞，但只有1%的前列腺癌患者 播散性肿瘤细胞最终会进展为大转移性病变，其余的要么死亡要么 仍然处于休眠状态。骨髓中多种细胞类型之间的复杂相互作用 微环境既影响存在于造血干细胞生态位中的肿瘤细胞，也受其影响。 骨髓微环境提供了什么线索，使得DTC要么保持休眠，要么 继续增殖对前列腺癌进展和治疗的研究至关重要 抵抗。然而，对肿瘤细胞进入或逃脱休眠状态的研究受到以下因素的限制 目前体内和体外系统的缺点。DNA定向单细胞构图技术在生物医学研究中的应用 在体外构建的骨髓壁龛中绘制六种细胞类型的模式将使我能够研究 前列腺癌常见继发部位肿瘤进展的微环境。这个错综复杂的系统 将使我能够通过以下方式具体询问不同表型对疾病结果的贡献 具体目的如下：1)制作并验证体外骨髓细胞壁龛。2)成立公司及 在体外高度交互作用的骨髓中监测已建立的前列腺癌细胞系的细胞 微环境。3)探讨肿瘤干细胞与上皮间充质的关系 过渡表型与肿瘤细胞休眠。 在优化细胞培养和图案化参数后，我将使用高通量光刻- 以DNA为基础的细胞模式，包括成骨细胞，破骨细胞，骨细胞，血管细胞， 巨噬细胞、肿瘤细胞进入微环境阵列进行关键参数的系统研究。在……里面 具体地说，我将研究不同细胞类型对肿瘤细胞群体和肿瘤细胞的影响 针对特定的干性和/或上皮-间充质转化表型进行预分类。这些研究将 初始化一种研究肿瘤细胞休眠的新工具，并将其应用于 微环境中肿瘤细胞影响因素的系统性和可重复性研究。特别是这一点 解决了前列腺癌的一个问题，但最终可以推广到其他局限于 骨髓或其他肿瘤微环境。 这项提案中的工作将在加州大学伯克利分校进行，由 莉迪亚·孙教授的照片。除了进行上述研究外，我还将追求负责任的 在研究员任期内进行研究和专业发展培训。我也会 通过参加突出研究的会议，交流我的成果并获得专业经验 癌症生物学和生物医学科学领域。我的博士后经历将使我获得技能 和经验，让我为终身教职的长期职业目标做好准备。

While approximately 50% of prostate cancer patients present with disseminated tumor cells, only 1% of those disseminated tumor cells will ultimately progress to macrometastatic lesions, with the rest either dying or remaining dormant. The complex interactions between the multiple cell types in the bone-marrow microenvironment both influence and are influenced by tumor cells present in the hematopoietic stem cell niche. What cues the bone-marrow microenvironment provides such that DTCs either maintain their dormancy or proceed to proliferate are of paramount importance to the study of prostate cancer progression and therapeutic resistance. The study of why tumor cells enter into, or escape from, dormancy, however, are limited by shortcomings in current in vivo and in vitro systems. The application of DNA-directed single cell patterning to pattern six cell types in a fabricated in vitro bone-marrow niche will enable me to study the role of the microenvironment in tumor progression at a common secondary location in prostate cancer. This intricate system will allow me to specifically interrogate the contributions of different phenotypes to disease outcomes through the following specific aims: 1) To fabricate and validate an in vitro bone-marrow niche. 2) To incorporate and monitor cells from established prostate cancer cell lines in a highly interactive in vitro bone-marrow microenvironment. 3) To investigate the relationship between cancer stem cell and epithelial-mesenchymal transition phenotypes and tumor cell dormancy. Following optimization of cell culture and patterning parameters, I will use high-throughput photolithography- based DNA-directed cell patterning to incorporate osteoblasts, osteoclasts, osteocytes, vascular cells, macrophages, and tumor cells into arrays of microenvironments for the systematic study of key parameters. In particular, I will study the influence of different cell types on both the bulk tumor cell population and tumor cells pre-sorted for specific stemness and/or epithelial-mesenchymal transition phenotypes. These studies will initialize a new tool in the study of tumor cell dormancy as well as take the first steps toward its application to the systematic and replicable study of factors affecting tumor cells in the microenvironment. This specifically addresses a problem in prostate cancer, but can ultimately be generalized to other cancers that localize to the bone marrow or other tumor microenvironments. The work in this proposal will be conducted at the University of California, Berkeley, under the sponsorship of Professor Lydia Sohn. In addition to conducting the aforementioned research, I will pursue responsible conduct in research and professional development training over the course of the fellowship tenure. I will also communicate my results and gain professional experience by attending conferences that highlight research in the fields of cancer biology and the biomedical sciences. My postdoctoral experience will allow me to gain skills and experience to prepare me for my long-term career goal of tenure-track academic faculty.