4D Microfluidic Platforms for Targeting Breast Cancer:Lymphatic Interactions

描述(申请人提供)：淋巴转移是乳腺癌细胞最初扩散的主要途径，它们在区域淋巴结中的存在是临床上最重要的负面预后因素。一旦进入淋巴结，乳腺癌细胞就可以进入循环，扩散到远处。在这种情况下，对我们来说，至关重要的是开发模型，使我们能够定义肿瘤细胞最初侵入淋巴管的机制，并开发针对这些机制的药物筛选平台。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相互作用的3D模型可以用来确定与淋巴管侵袭和淋巴管生成相关的可用药途径。我们的基本原理是，体外确定的通路将至少概括为患者样本中存在的通路的子集，从而允许我们为新的治疗策略定义可用药的结节。我们研究翻译的关键是筛选平台，该平台可用于以3D方式复制生长乳房和淋巴细胞，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)建立总结乳腺癌细胞和淋巴管微血管内皮细胞之间体内相互作用的最佳条件；2)设计、制造和测试可用于成像、识别治疗靶点和药物筛选的新型微流控平台；3)对于原理验证研究，确定候选治疗靶点并在患者样本中进行验证；以及4)进行针对候选治疗靶点的药物的初步筛选。我们将使用创新的4D(3D+TIME)培养来模拟人类乳腺癌细胞和人类淋巴管微血管内皮细胞之间的相互作用。我们将为新的3D培养物的长期生长构建新的微流体平台，通过共聚焦显微镜对培养物成像，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，并针对这些靶点和相关途径筛选药物。有效的筛查要求我们的模型具有高度的重复性，并且我们使用的平台允许我们在多种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微型平台方面拥有丰富的经验，包括用作患者的假肢设备。我们的研究应该加速确定有效的药物，可能是针对个别患者的药物。

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.