4D Microfluidic Platforms for Targeting Breast Cancer:Lymphatic Interactions

描述（由申请人提供）：摘要淋巴转移是乳腺癌细胞最初扩散的主要途径，并且它们在区域淋巴结中的存在是临床上最重要的阴性预后因素。一旦进入淋巴结，乳腺癌细胞就可以进入循环并扩散到远处。在这种情况下，开发模型对我们来说至关重要，这些模型将使我们能够定义肿瘤细胞最初侵入淋巴管的机制，并开发筛选针对这些机制的药物的平台。我们的工作假设是，概括乳腺癌细胞和淋巴管微血管内皮细胞之间的相互作用的3D模型可用于识别与淋巴管和淋巴管生成的侵袭相关的药物途径。我们的基本原理是，体外鉴定的途径将至少概括患者样本中存在的途径的一个子集，从而使我们能够为新的治疗策略定义可药物化的节点。翻译我们的研究的关键将是筛选平台，这些平台可用于在3D中可重复地生长乳腺和淋巴细胞，并实时测试和量化活细胞对治疗干预的动态反应。 我们的具体目标是：1）建立用于重现乳腺癌细胞和淋巴微血管内皮细胞之间的体内相互作用的最佳条件; 2）设计、制造和测试可用于成像、治疗靶点鉴定和药物筛选的新型微流体平台; 3）对于原理验证研究，鉴定候选治疗靶点并在患者样品中验证;和4）对靶向候选治疗靶标的药剂进行初始筛选。我们将使用创新的4D（3D +时间）培养来模拟人乳腺癌细胞和人淋巴管微血管内皮细胞之间的相互作用。我们将构建新型微流体平台，用于新型3D培养物的长期生长，通过共聚焦显微镜对培养物进行成像，分析这些培养物中动态相互作用改变的生化和免疫化学途径，识别潜在的治疗靶点并筛选针对这些靶点的药物。目标和相关途径。有效的筛选要求我们的模型具有高度可重复性，并且我们使用的平台允许我们在多个浓度下测试许多不同的药物。我们在构建可用于细胞培养的小型化平台方面拥有丰富的经验，包括作为患者的假体设备。我们的研究应该加速有效药物的鉴定，这些药物可能是针对个体患者的。

DESCRIPTION (provided by applicant): Abstract Lymphatic metastasis is the predominant route of the initial spread of breast cancer cells and their presence in regional lymph nodes is the single most clinically important negative prognostic factor. Once in lymph nodes, breast cancer cells can find their way into the circulation and spread to distant sites. This being the case it is critically important for us to develop models that will allow us to define mechanisms fo the initial invasion of tumor cells into lymph vessels and to develop platforms for screening drugs that target those mechanisms. Our working hypothesis is that 3D models that recapitulate the interactions between breast carcinoma cells and lymphatic microvascular endothelial cells can be used to identify druggable pathways associated with invasion into lymphatic vessels and lymphangiogenesis. Our rationale is that the pathways identified in vitro will recapitulate at leas a subset of those present in patient samples and thereby allow us to define druggable nodes for novel therapeutic strategies. Critical to translation of our studies will be screening platforms tht can be used to reproducibly grow breast and lymphatic cells in 3D and test and quantify dynamic responses of live cells to therapeutic interventions in real-time. Our Specific Aims are to: 1) Establish optimal conditions for recapitulating in vivo interactions between breast carcinoma cells and lymphatic microvascular endothelial cells; 2) Design, fabricate and test novel microfluidic platforms that can be used for imaging, identification of therapeutic targets and drug screening; 3) For proof-of-principle studies, Identify a candidate therapeutic target and validate in patient samples; and 4) Perform initial screening of agents that target the candidate therapeutic target. We will use innovative 4D (3D + time) cultures to model the interactions between human breast carcinoma cells and human lymphatic microvascular endothelial cells. We will construct novel microfluidic platforms for long-term growth of the novel 3D cultures, image the cultures by confocal microscopy, analyze biochemical and immunochemical pathways that are altered by the dynamic interactions in these cultures, identify potential therapeutic targets and screen drugs directed against those targets and associated pathways. Effective screening requires that our models be highly reproducible and that we use platforms that allow us to test many different drugs at multiple concentrations. We have extensive experience in constructing miniaturized platforms that can be used for cell culture, including as prosthetic devices in patients. Our studies should accelerate the identification of effective drugs potentially ones targeted to individual patients.