CTC Purification System Based on Thermoresponsive NanoSubstrates

 描述(由申请人提供)：这项SBIR第二阶段提案的目标是进行第三代温度响应性纳米尼龙搭桥试验的高级开发，以实现从非小细胞肺癌(NSCLC)患者血液样本中快速纯化循环肿瘤细胞(CTCs)，为CTC衍生的分子签名和功能读数铺平道路。该项目由加西亚博士(PI)领导，他在该技术的早期开发方面拥有丰富的经验，并在表面化学、微流体和体外诊断技术方面具有背景。他得到了一个跨学科团队的支持，该团队包括业务开发、FDA的专业知识、QC/QA管理、纳米技术支持、肺癌、临床效用、基因分析、生物统计学、行业合作者和下游潜在客户。非小细胞肺癌占肺癌病例的70%。由于非小细胞肺癌通常对化疗和/或放射不太敏感，表皮生长因子受体(EGFR)抑制剂靶向治疗的出现为携带EGFR基因癌基因突变的NSCLC患者亚群提供了很好的治疗选择。为了指导靶向治疗的实施，采用侵入性活检或手术对非小细胞肺癌组织进行采样，以确定这些EGFR突变的存在。然而，这些侵入性抽样程序给患者带来了巨大的风险。作为另一种选择，CTCs可以被重复捕获并以微创的方式进行分析，从而提供具有高转移能力的恶性克隆的系统图像。建议的温度响应型纳米尼龙搭扣测试由两个单独的组件组成：i)带有嵌入式温度控制模块的数字流体处理器，以及ii)将在CytoLumina设计和制造的定制芯片支架，其中的夹持式设计允许纳米尼龙搭扣衬底与覆盖的PDMS组件的即时组装。为了突出我们下游研究的意义，我们将收集肺癌患者的系列血液样本。我们将收集纯化的CTCs，并对其进行包括egfr在内的8个基因的突变分析。此外，纯化的CTCs将被引入各种体外细胞。 我们的联合团队(加州大学洛杉矶分校和CytoLumina)建立了培养系统(即微流控和3D细胞培养平台)，以创建可行的CTC细胞系。这些个性化的细胞系将被用于研究药物敏感性的模式，这与潜在的基因驱动因素突变有关。

 DESCRIPTION (provided by applicant): The goal of this SBIR Phase II proposal is to conduct the advanced development of the 3rd-gen Thermoresponsive NanoVelcro assay in order to achieve rapid purification of circulating tumor cells (CTCs) from non-small cell lung cancer (NSCLC) patient blood samples, paving the way for CTC-derived molecular signatures and functional readouts. This project is led by Dr. Garcia (PI), who has extensive experience in early-stage development of the technology and has a background in surface chemistry, microfluidics, and in vitro diagnostic technologies. He is supported by an interdisciplinary team comprising business development, FDA expertise, QC/QA management, nanotechnology support, lung cancer, clinical utility, genetic analysis, biostatistics, industrial collaborators, nd downstream potential customers. NSCLC accounts for >70% of lung cancer cases. Since NSCLCs are usually not very sensitive to chemotherapy and/or radiation, the advent of targeted therapy by epidermal growth factor receptor (EGFR) inhibitors offer a great treatment options to a sub-population of NSCLC patients who carry oncogenic driver mutations in their EGFR genes. To guide the implementation of targeted therapy, invasive biopsy or surgery is employed to sample NSCLC tissues for determining the presence of these EGFR mutations. However, these invasive sampling procedures impose significant risk to the patients. As an alternative, CTCs can be captured repetitively and analyzed in a minimally invasive manner thus providing a systemic picture of the malignant clones that possess high metastatic capacity. The proposed Thermoresponsive NanoVelcro assay is composed of two individual components: i) a digital fluidic handler with an embedded temperature control module, and ii) a custom-designed chip holder, in which a clamp-down design allows instant assembly of a NanoVelcro substrate with an overlaid PDMS component, will be designed and fabricated at CytoLumina. To highlight the significance of our downstream studies, we will gather serial blood samples from lung cancer patients. We will collect the purified CTCs and subject them to mutational analysis for eight genes, including EGFR., Further, the purified CTCs will be introduced to a variety of in vitro cell culture systems (i.e., microfluidic and 3D cell-culture platforms) established by our joint team (UCLA and CytoLumina) to create viable CTC cell lines. The personalized cell lines will be used to study patterns in drug susceptibility, which is linked to the underlying genetic driver mutation.