Rapid analysis of patient tumor cell drug responses to reduce metastatic risk

快速分析患者肿瘤细胞药物反应以降低转移风险 背景：目前临床癌症成像的局限性阻碍了对药物如何作用的清晰理解。 目的是细胞生长影响乳腺癌中循环肿瘤细胞（CTC）的转移潜力 患者有超过220万女性退伍军人，目前的乳腺癌发病率预测， 至少有275，000名女性退伍军人将面临乳腺癌治疗，并需要有效的治疗， 最大限度地降低致命转移扩散的风险。CTC分析的最新进展表明， 乳腺癌CTC具有高达50倍的高转移潜力。马丁实验室发现了 乳腺肿瘤细胞表面的微触角（McTNs）增加了簇的形成， 促进乳腺癌转移的干细胞特性的升高。目前的癌症治疗， 稳定微管蛋白（如紫杉烷和埃博霉素）可以增加McTN、干细胞特性、肿瘤细胞 集群和再附着。这些结果强调需要澄清目前的药物如何影响自由浮动 肿瘤细胞，使治疗可以更好地适应个体患者，并降低长期转移风险。 目的/假设：汇集肿瘤细胞生物学家，生物工程师， 和乳腺癌临床医生;该项目的目的是使用一种新的微流体设备， 自由漂浮的乳腺肿瘤细胞并定义了3种预测转移潜能的表型（McTNs， 球体形成、聚类）和关键分子标记。这些表型和分子特征将被 在小鼠中使用临床相关的PDX移植与转移潜力和药物反应相关。本研究 将检验新分离的乳腺肿瘤的关键功能表型和分子标志物 细胞可以作为转移潜能的直接指示物，并提供一个平台来快速检测细胞的转移潜能。 个体患者肿瘤细胞对癌症药物的反应。 具体目标： 1)优化微流控细胞系留以测量转移潜能的3种功能表型。 2)在患者来源的异种移植（PDX）细胞中建立肿瘤细胞药物应答的分子框架。 3)定义新鲜患者肿瘤细胞、PDX和CTC的共有分子和功能特征。 方法：本项目将使用共聚焦显微镜检查3种表型（McTNs，球体形成， 和成簇）和一组现有的患者来源的异种移植物（PDX）， 马里兰州格林鲍姆癌症中心的转化核心实验室。同时，我们 将收集新鲜的患者肿瘤样本，以比较新鲜细胞中的分子标志物和表型， 最终在小鼠体内生长的PDX。PDX概括了患者原发肿瘤的转移行为 比任何组织培养模型都要忠实得多。个体患者的表型和分子标志物 将肿瘤细胞与原始患者肿瘤的分子特征（ER/PR/HER2）进行比较， 以及PDX模型中的生长和转移。 影响：该项目的完成将建立一个框架， 患者肿瘤细胞的表型可预测转移潜能和对乳腺癌治疗的反应。 目前的治疗策略主要集中在抑制肿瘤生长上，因此这项技术将开辟一个新的早期 窗口，以帮助确保药物治疗避免无意中增加转移风险，同时靶向肿瘤 增长由于该项目将重点关注FDA批准的乳腺癌药物， 翻译为影响乳腺癌的临床治疗量身定制治疗个别女性退伍军人。

Rapid analysis of patient tumor cell drug responses to reduce metastatic risk Background: The current limitations of clinical cancer imaging prevent a clear understanding of how drugs aimed at cell growth affect the metastatic potential of circulating tumor cells (CTCs) in breast cancer patients. With more than 2.2 million female Veterans, the current incidence of breast cancer predicts that at least 275,000 female Veterans will confront breast cancer treatment and require effective treatments that minimize the risk of lethal metastatic spread. Recent advances in CTC analysis have shown that clusters of breast cancer CTCs have up to 50x higher metastatic potential. The Martin lab discovered unique microtentacles (McTNs) on the surface of breast tumor cells that increase cluster formation, and are indicative of the elevated stem cell characteristics that promote breast cancer metastasis. Current cancer therapies that stabilize tubulin (like taxanes and epothilones) can increase McTNs, stem cell characteristics, tumor cell clustering, and reattachment. These results emphasize the need to clarify how current drugs affect free-floating tumor cells so that therapies can be better tailored to individual patients and reduce long-term metastatic risk. Objective/Hypothesis: Bringing together a multidisciplinary team of tumor cell biologists, bioengineers, and breast cancer clinicians; the objective of this project is to use a novel microfluidic device to rapidly image free-floating breast tumor cells and define 3 phenotypes that are predictive of metastatic potential (McTNs, sphere formation, clustering) and key molecular markers. These phenotypes and molecular profiles will be related to metastatic potential and drug response using clinically-relevant PDX transplants in mice. This study will test the hypothesis that key functional phenotypes and molecular markers of freshly-isolated breast tumor cells can serve as immediate indicators of metastatic potential and provide a platform to rapidly test the responses of individual patient tumor cells to cancer drugs. Specific Aims: 1) Optimize microfluidic cell tethering to measure 3 functional phenotypes of metastatic potential. 2) Establish molecular framework for tumor cell drug responses in patient-derived xenograft (PDX) cells. 3) Define shared molecular and functional characteristics of fresh patient tumor cells, PDX and CTCs. Methods: This project will use confocal microscopy to examine 3 phenotypes (McTNs, sphere formation, and clustering) in breast tumor cell lines and a panel of existing patient-derived xenografts (PDX) supplied by the Translational Core Laboratory at the University of Maryland Greenebaum Cancer Center. In parallel, we will collect fresh patient tumor samples to compare molecular markers and phenotypes in the fresh cells with the PDX that eventually grow in mice. PDX recapitulate the metastatic behavior of the patient’s original tumor far more faithfully than any tissue culture model. Phenotypes and molecular markers of individual patient’s tumor cells will be compared to the molecular characteristics (ER/PR/HER2) of the original patient’s tumor, as well as growth and metastasis in the PDX model. Impact: The completion of this project will establish a framework for defining how the functional phenotypes of patient tumor cells predict metastatic potential and responses to breast cancer therapies. Current treatment strategies focus largely on inhibiting tumor growth, so this technology will open a new early window to help ensure drug treatments avoid inadvertently increasing metastatic risk while targeting tumor growth. Since this project will focus on FDA-approved breast cancer drugs, the findings can be more easily translated to impact the clinical treatment of breast cancer by tailoring therapies for individual female Veterans.