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

用于捕获和回收 CTC 的热响应聚合物微芯片

基本信息

批准号：
8591278
负责人：
Mitch Garcia
金额：
$ 20.05万
依托单位：
CYTOLUMINA TECHNOLOGIES CORPORATION, INC
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-19 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8591278
关键词：
Affinity Biological Assay Biological Markers Blood Blood Volume Blood donor Blood flow Blood specimen CD44 gene Cancer Biology Cancer Patient Cell Culture Techniques Cell Line Cells Chemistry Clinical Collaborations Color Cytokeratin Cytolysis DNA Data Detection Development Devices Diagnostic Disease E-Cadherin Feasibility Studies Funding Gene Targeting Genes Glutamate Carboxypeptidase II Goals IL8RB gene In Vitro Individual Integrins Joints LNCaP Leukocytes Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Mediator of activation protein Medicine Microfluidics Mixed Function Oxygenases Molecular Motivation N-Cadherin NKX3-1 gene Neoplasm Circulating Cells Neoplasm Metastasis Nuclear PC3 cell line PTPRC gene Patients Performance Phase Phased Innovation Awards Polymers Positioning Attribute Primary Neoplasm Process Prostate Recording of previous events Recovery Research Reverse Transcriptase Polymerase Chain Reaction Running Sampling Signal Transduction Site Slide Small Business Innovation Research Grant Solid Neoplasm Staging Staining method Stains Stream Structure of thyroid parafollicular cell Surface TACSTD2 gene Technology Temperature Therapeutic Tissues Travel Vimentin Work anticancer research base cancer cell castration resistant prostate cancer experience fluorescence microscope immunocytochemistry innovation insight member microchip mutant nanomaterials nanostructured polydimethylsiloxane programs prostate cancer cell prototype public health relevance screening tumor progression

项目摘要

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.

描述（由申请人提供）：该 SBIR 第一阶段提案的目标是 i) 开发一种商业原型 NanoVelcro Assay，该原型使用热敏聚合物捕获和释放循环肿瘤细胞 (CTC)；ii) 验证执行简化的流程从 CTC 捕获、分离释放开始，到回收可行的 CTC 以用于癌症患者血液的分子和功能分析。该提案由 Garcia 博士（PI）领导，他在 TR-NanoVelcro CTC Assay 的早期开发方面拥有丰富的经验，并拥有纳米材料、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，它们通过血流到达不同的组织，预示或永久转移。然而，由于血液细胞中 CTC 丰度极低（~1-100s CTCsmL），CTC 检测和表征在技术上一直具有挑战性。目前，CellSearchTM Assay 是唯一获得 FDA 批准的设备，但其成本高昂且捕获 CTC 效率低下，并且不适合下游分子或功能分析。因此，需要一种超越简单计数的商业 CTC 测定方法，这种测定方法价格低廉，能够回收可行的 CTC 用于分子和功能分析，并且可以轻松集成到临床环境中。在 NCI IMAT 计划的连续资金支持（通过 R21 和 R33）下，CytoLumina 在 UCLA 和 Cedars-Sinai 的学术顾问展示了一种高效、廉价的细胞亲和测定，能够富集、识别、从癌症患者的血液中分离 CTC。该团队率先开发了“NanoVelcro”基材，其中将捕获剂涂覆在纳米结构表面上，以在固定设备环境中吸引 CTC。为了实现更大的血容量吞吐量，从而增加捕获的 CTC 数量，该团队在基板上放置了聚二甲基硅氧烷混沌混合器，以诱导血液在纳米结构表面上垂直流动。首先，CytoLumina 会将 100 个患者来源的前列腺癌细胞掺入 2 mL 健康捐赠者的血液中，生成人工 CTC 样本，用于确定捕获效率和恢复性能。优化后临床可行性将通过收集 10 名前列腺患者的血液样本并通过 TR-NanoVelcro 检测进行评估。最后，我们将生成 14 个基因的 RT-PCR 数据，并获得两种源自患者 CTC 的原代细胞系，以证明 TR-NanoVelcro CTC 检测的诊断和临床价值。该提案的成功论证将实现广泛的应用，例如，生成患者 CTC 衍生细胞系，并使用 CTC 作为个性化医疗潜在疗法的体外筛选方法。