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，但这些患者中只有1% 扩散的肿瘤细胞最终将进展为大转移病灶，其余的要么死亡，要么死亡。 保持休眠状态骨髓中多种细胞类型之间的复杂相互作用 微环境既影响造血干细胞小生境中存在的肿瘤细胞，又受其影响。 骨髓微环境提供了什么样的线索，使得DTC要么保持休眠状态， 进行增殖对于前列腺癌进展和治疗的研究至关重要。 阻力然而，关于肿瘤细胞为什么进入或摆脱休眠状态的研究受到以下因素的限制： 目前的体内和体外系统的缺点。DNA指导的单细胞模式化在 模式六种细胞类型在体外骨髓龛将使我能够研究的作用， 前列腺癌中常见继发部位的肿瘤进展微环境。这个复杂的系统 将允许我通过以下方式来具体询问不同表型对疾病结果的贡献： 具体目的如下：1）构建并验证体外骨髓龛。2)以掺入和 在高度相互作用的体外骨髓中监测来自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质细胞的关系 转化表型和肿瘤细胞休眠。 在优化细胞培养和图案化参数之后，我将使用高通量光刻- 基于DNA指导的细胞模式化，以整合成骨细胞、破骨细胞、骨细胞、血管细胞， 巨噬细胞和肿瘤细胞进入微环境阵列，用于关键参数的系统研究。在 特别是，我将研究不同类型的细胞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选。这些研究将 启动一个新的工具，在肿瘤细胞休眠的研究，以及采取第一步，其应用于 对影响微环境中肿瘤细胞的因素进行系统和可复制的研究。这具体 解决了前列腺癌的问题，但最终可以推广到其他癌症， 骨髓或其他肿瘤微环境。 本提案中的工作将在加州大学伯克利分校进行，由 Lydia Sohn教授除了进行上述研究外，我还将追究责任。 在研究金任期内进行研究和专业发展培训。我也会 交流我的结果，并通过参加会议，突出研究在专业经验 癌症生物学和生物医学领域。我的博士后经历将使我获得技能 和经验，为我的长期职业目标，终身制的学术教师做好准备。

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.