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

• 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
• 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

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（由申请方提供）：本SBIR I期提案的目标是：i）开发一种商业原型NanoVelcro检测试剂盒，该试剂盒使用温敏聚合物捕获和释放循环肿瘤细胞（CTC）; ii）验证从CTC捕获、分离和释放开始，最后回收活性CTC，用于癌症患者血液分子和功能分析的简化工艺的临床可行性。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是从原发性肿瘤或转移部位脱落的癌细胞，并且在整个恶性转化过程中，它们通过血流行进到不同的组织，预示或永久转移。然而，由于血液细胞中的CTC丰度极低（约1- 100 s CTCsmL），CTC检测和表征在技术上具有挑战性。目前，CellSearchTM Assay是唯一一种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，需要一种商业化的CTC测定法，其超越简单的计数-一种便宜的、能够回收有活力的CTC用于分子和功能分析的测定法，并且可以容易地整合到临床环境中。 在NCI IMAT计划的连续资金支持下（通过R21和R33），CytoLumina在UCLA和Cedars-Sinai的学术顾问展示了一种高效，廉价的细胞亲和测定法，能够富集，鉴定， 以及从癌症患者血液中分离CTC。该团队率先开发了“NanoVelcro”基材，其中捕获剂被涂覆在纳米结构表面上，以在固定设备设置中吸引CTC。为了实现更大的血液容量吞吐量，从而增加捕获的CTC数量，该团队在基底上放置了一个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的垂直流动。 首先，CytoLumina将100个患者来源的前列腺癌细胞掺入2 mL健康供体血液中，以生成人工CTC样品，该样品将用于确定捕获效率和回收性能。优化后，临床可行性 将通过收集10个前列腺患者血液样本并通过TR-NanoVelcro测定进行评估。最后，我们将生成14个基因的RT-PCR数据，并获得两个患者CTC来源的原代细胞系，以证明TR-NanoVelcro CTC Assay的诊断和临床价值。这一提议的成功证明将使广泛的应用成为可能，例如，产生患者CTC衍生的细胞系，并使用CTC作为个性化医疗的潜在治疗剂的体外筛选方法。