A Microchip with Thermally Responsive Polymers for Capture and Recovery of CTCs

描述（由申请人提供）：SBIR I期提案的目标是I)开发商业原型NanoVelcro Assay，使用热响应性聚合物捕获和释放循环肿瘤细胞（CTC）， ii)验证执行从CTC捕获，分离释放到回收可存活的CTC的流线型过程的临床可行性，用于癌症患者血液的分子和功能分析。该提案由Dr. Garcia （PI）领导，他在TR-NanoVelcro CTC检测的早期开发方面拥有丰富的经验，并具有纳米材料，表面化学，微流体和体外诊断技术的背景。ctc是从原发肿瘤或转移部位脱落的癌细胞，在整个恶性转化过程中，它们通过血流进入不同的组织，预示或延续转移。然而，由于血液细胞中CTC丰度极低（~1- 1000s CTCsmL）， CTC检测和表征在技术上具有挑战性。目前，CellSearchTM Assay是唯一获得fda批准的设备，但它在捕获ctc方面成本高且效率低，并且不适合下游分子或功能分析。因此，需要一种超越简单枚举的商业CTC测定方法，这种方法价格低廉，能够回收可行的CTC进行分子和功能分析，并且可以很容易地集成到临床环境中。在NCI IMAT项目的连续资金支持下（通过R21和R33）， CytoLumina在UCLA和Cedars-Sinai的学术顾问展示了一种高效、廉价的细胞亲和力测定方法，能够富集、识别、

DESCRIPTION (provided by applicant): The goal of this SBIR Phase I proposal is i) to develop a commercial prototype NanoVelcro Assay that uses thermoresponsive polymers for circulating tumor cell (CTC) capture and release and ii) to validate the clinical feasibility of performing a streamlined process starting from CTC capture, isolationrelease, and finishing with recovery of viable CTCs for molecular and functional analysis of cancer patient blood. This proposal is led by Dr. Garcia (PI), who has extensive experience in early-stage development of the TR-NanoVelcro CTC Assay and has a background in nanomaterials, surface chemistry, microfluidics, and in vitro diagnostic technologies. CTCs are cancer cells shed from either the primary tumors or metastatic sites, and throughout the malignant transformation process, they travel through the blood stream to different tissues, presaging or perpetuating metastases. However, CTC detection and characterization have been technically challenging due to extremely low CTC abundance (~1-100s CTCsmL) among hematologic cells. Currently, CellSearchTM Assay is the only FDA-cleared device, but it is costly and inefficient in capturing CTCs, and it is not amenable for downstream molecular or functional analysis. Thus, there is a need for a commercial CTC assay that goes beyond simple enumeration - one which is inexpensive, able to recover viable CTCs for molecular and functional analysis, and can be easily integrated into a clinical setting. Under sequential funding support (via R21 and R33) from the NCI IMAT program, CytoLumina's academic consultants at UCLA and Cedars-Sinai demonstrated a highly efficient, inexpensive cell-affinity assay capable of enriching, identifying, and isolating CTCs from cancer patient blood. The team pioneered the development of "NanoVelcro" substrates, in which a capture agent is coated onto a nanostructured surface in order to attract CTCs in a stationary device setting. In order to enable greater blood volume throughput, thereby increasing the number of CTCs captured, the team positioned a polydimethylsiloxane chaotic mixer over the substrate to induce vertical flow of the blood over the nanostructured surface. First, CytoLumina will spike 100 patient derived prostate cancer cells into 2 mL of healthy-donor blood to generate an artificial CTC sample that will be used to determine capture efficiency and recovery performance. After optimization, the clinical feasibility will be assessed by collecting 10 prostate patient blood samples and running them through the TR-NanoVelcro Assay. Finally, we will generate RT-PCR data on 14 genes and obtain two patient-CTC derived primary cell lines to demonstrate the diagnostic and clinical value of the TR-NanoVelcro CTC Assay. The successful demonstration of this proposal will enable a wide range of applications, for example, generating patient CTC-derived cell lines and using CTCs as an in vitro screening approach of potential therapeutics for personalized medicine.