High Throughput 3D Cell Assay for Metastatic Prostate Cancer

转移性前列腺癌的高通量 3D 细胞检测

基本信息

批准号：
8652646
负责人：
SHUICHI TAKAYAMA
金额：
$ 3.46万
依托单位：
3D BIOMATRIX, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8652646
关键词：
3-Dimensional Address Aleurites Antineoplastic Agents Biological Biological Assay Bioluminescence Bone Marrow Bone Tissue Cell Culture Techniques Cells Coculture Techniques Complex Development Disseminated Malignant Neoplasm Drops Endothelial Cells Engineering Environment Fluorescence Grant Harvest Hematopoietic stem cells Human In Vitro Light Malignant Neoplasms Malignant neoplasm of prostate Marrow Metastatic Prostate Cancer Methods Michigan Microfluidic Microchips Molecular Nature Neoplasm Metastasis Optics Osteoblasts Parasites Performance Pharmaceutical Preparations Phase Physiological Preclinical Drug Evaluation Preparation Procedures Publishing Resistance Resources Small Business Technology Transfer Research Sterility Stromal Cells System Technology United States Universities Validation bone cancer cell design effective therapy evaporation high throughput screening men mortality novel prostate cancer cell stem cell niche therapy resistant tumor microenvironment

项目摘要

DESCRIPTION (provided by applicant): Prostate cancer is the most common non-skin cancer in United States men. Despite recent advances, prostate cancer mortality still remains high due to the emergence of therapy-resistant cancer cells that metastasize. This lack of effective therapies against metastatic cancer exists, at least in part, because of lack of drug screening platforms that address the unique nature of metastastatic prostate cancer cells and their microenvironment. Recently, our collaborators have found that prostate cancer (PCa) cells act as molecular parasites as they metastasize to bone and harvest resources from the hematopoietic stem cell (HSC) niche environment where they stay dormant and resistant to conventional anti-cancer drugs. If the dormant metastasized PCa cells that parasitize the HSC niche could be recreated in vitro, they would serve as ideal platforms to screen specifically for anti-metastatic PCa drugs. We have recently shown that 3D co-culture spheroids can mimic the PCa cell parasitized HSC niche thereby maintaining PCa cells in a physiological, more quiescent state. What is required to take advantage of the metastatic PCa microtissue engineering capability we have acquired for drug screening applications is to develop a high throughput format of these types of co-culture spheroids. Towards this end, this Phase I STTR proposal will: Aim 1: Validate a 384 array hanging drop plate system for preparation of the co-culture spheroids. This is a major focus of 3-D Biomatrix, LLC. We have already modified our published platform (Tung et al. 2011) to: enhance droplet stability, facilitate pipette tip insertin, mitigate evaporation, and preserve sterility during optical analysis (Figure 1A). We will perform high throughput performance validations for fluorescence, transmitted light, and bioluminescence assays (Z'- factor), and further refine the design as necessary. Aim 2: Create arrays of microengineered 3D tissues of bone metastasized PCa in the quiescent state. This will be the focus of the Takayama lab at the University of Michigan. To develop procedures to utilize the 384 array hanging drop plate to form co-culture spheroids of PCa cells, marrow stromal cells (MSCs), and human bone marrow endothelial cells (HBMECs) similar to those prepared previously in low throughput microfluidic devices. We will also confirm quiescence of the PCa cells in these co-culture spheroids. The biological hypothesis that motivates this study is: We can treat metastatic PCa better by developing drugs that specifically target dormant metastatic PCa cells that are parasitizing the HSC niche. PHS 398 (Rev. 11/07) Page 1

描述（由申请人提供）：前列腺癌是美国男性中最常见的非皮肤癌。尽管最近取得了进展，但由于出现转移的耐药癌细胞，前列腺癌死亡率仍然很高。缺乏针对转移性癌症的有效疗法，至少部分是因为缺乏能够解决转移性前列腺癌细胞及其微环境的独特性质的药物筛选平台。最近，我们的合作者发现，前列腺癌 (PCa) 细胞在转移到骨骼时充当分子寄生虫，并从造血干细胞 (HSC) 利基环境中获取资源，在造血干细胞 (HSC) 利基环境中，它们保持休眠状态并对传统抗癌药物产生耐药性。如果寄生在 HSC 微环境中的休眠转移 PCa 细胞可以在体外重建，那么它们将成为专门筛选抗转移 PCa 药物的理想平台。我们最近表明，3D 共培养球体可以模拟 PCa 细胞寄生的 HSC 生态位，从而将 PCa 细胞维持在生理、更静止的状态。要利用我们在药物筛选应用中获得的转移性 PCa 微组织工程能力，需要开发此类共培养球体的高通量形式。为此，第一阶段 STTR 提案将：目标 1：验证用于制备共培养球体的 384 阵列悬滴板系统。这是 3-D Biomatrix, LLC 的主要关注点。我们已经修改了我们发布的平台（Tung 等人，2011），以：增强液滴稳定性，促进移液器吸头插入，减轻蒸发，并在光学分析过程中保持无菌（图 1A）。我们将对荧光、透射光和生物发光测定（Z'因子）进行高通量性能验证，并根据需要进一步完善设计。目标 2：创建静态骨转移 PCa 的微工程 3D 组织阵列。这将是密歇根大学高山实验室的重点。开发利用 384 阵列悬滴板形成 PCa 细胞、骨髓基质细胞 (MSC) 和人骨髓内皮细胞 (HBMEC) 共培养球体的程序，类似于之前在低通量微流体装置中制备的球体。我们还将确认这些共培养球体中 PCa 细胞的静止状态。推动这项研究的生物学假设是：我们可以通过开发专门针对寄生于 HSC 生态位的休眠转移性 PCa 细胞的药物来更好地治疗转移性 PCa。 PHS 398（2007 年 11 月修订）第 1 页