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：目前临床肿瘤成像的局限性阻碍了对药物如何 靶细胞生长对乳腺癌循环肿瘤细胞(CTCs)转移潜能的影响 病人。有超过220万女性退伍军人，目前乳腺癌的发病率预测 至少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 将致命转移扩散的风险降至最低。CTC分析的最新进展表明，聚类物 乳腺癌CTC的转移潜能最高可达50倍。马丁实验室发现了独特的 乳腺肿瘤细胞表面的微触手(McTns)可促进成簇形成，具有指示性作用 促进乳腺癌转移的干细胞特性升高。目前的癌症治疗方法 稳定的微管蛋白(如紫杉烷和伊波硫酮)可以增加McTns，干细胞特性，肿瘤细胞 群集和重新连接。这些结果强调了澄清当前药物如何影响自由漂浮的必要性。 因此，治疗方法可以更好地针对个别患者，并降低长期转移风险。 目标/假设：汇聚一个由肿瘤细胞生物学家、生物工程师、 和乳腺癌临床医生；本项目的目标是使用一种新型的微流控设备来快速成像 并定义了3种可预测转移潜能的表型(McTns， 球体形成、聚集)和关键分子标记。这些表型和分子图谱将是 与临床相关的PDX移植在小鼠体内的转移潜能和药物反应有关。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的假设 细胞可以作为转移潜能的直接指示器，并提供一个平台来快速测试 个体患者肿瘤细胞对抗癌药物的反应。 具体目标： 1)优化微流控细胞系系，测量转移潜能的3种功能表型。 2)建立患者来源的异种移植(PDX)细胞对肿瘤细胞药物反应的分子框架。 3)确定新鲜患者肿瘤细胞、PDX和CTCs的共同分子和功能特征。 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， 和聚集)和一组现有的患者来源的异种移植(PDX)，由 马里兰大学格林鲍姆癌症中心的翻译核心实验室。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记和表型 最终在老鼠体内生长的PDX。PDX重述患者原始肿瘤的转移行为 比任何组织培养模型都更忠实。个体患者的表型和分子标记 肿瘤细胞将与原始患者肿瘤的分子特征(ER/PR/HER2)进行比较，因为 以及在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，以定义 患者肿瘤细胞的表型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，因此这项技术将开启一种新的早期 帮助确保药物治疗避免在靶向肿瘤时无意中增加转移风险的窗口 成长。由于该项目将重点放在FDA批准的乳腺癌药物上，因此发现更容易 转化为通过为个别女性退伍军人量身定做疗法来影响乳腺癌的临床治疗。

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.