NanoFlares for the Detection of Circulating Cancer Stem Cells

人类癌症是由异质性的恶性细胞集合代表的，必须根除这些细胞才能使治疗成功。对于许多实体器官肿瘤，癌症干细胞（CSCs）被认为是治疗失败和不良临床结果的原因。因此，CSCs的检测在预测、治疗选择和评估患者对意向的反应方面具有重要的优先地位。用于检测和分离CSCs的传统工具几乎完全是蛋白质有限的。细胞内mRNA靶点已广泛用于癌细胞亚群表型；然而，mRNA靶点的检测需要破坏宝贵的候选细胞进行RT-PCR和牺牲吞吐量。无论是基础科学研究还是临床应用，在活癌细胞群中同时实时检测蛋白质和mRNA标记物的能力都将具有重要意义。该项目旨在引入一种新的和可行的技术，“NanoFlares”，用于检测和分离表型不同的癌症干细胞（CSC）。利用表面被DNA功能化的金纳米颗粒（DNA Au-NPs）的独特特性，NanoFlare技术提供了在蛋白质和mRNA水平上同时对细胞亚群进行表型分析的独特能力。使用共聚焦光显微镜可以进行单细胞分析，而使用流式细胞术（FC）和荧光活化细胞分选（FACS）可以进行高通量的数千个单个细胞测量。首先关注乳腺癌，项目的成功将从CSCs背景下的基础科学发现的角度产生重大影响，并最终对所有形式的局部和播散性癌症患者产生重大影响，其中CSC的检测和根除可能对患者的预后产生重大影响。

Human cancers are represented by heterogeneous collections of malignant cells that must be eradicated for treatment to be successful. For many solid organ tumors, cancer stem cells (CSCs) are thought to be responsible for treatment failure and poor clinical outcomes. Accordingly, the detection of CSCs represents a significant priority with regard to prognostication, choice of treatment, and for assessing patient responses to inten/ention. Convenfional tools for the detecfion and isolation of CSCs are near exclusively protein limited. Intracellular mRNA targets have been used extensively for cancer cell sub-population phenotyping; however, detecfion of mRNA targets requires destruction of precious candidate cells for performing RT-PCR and sacrifices throughput. Either for basic science research or clinical use, the capability of simultaneous detecfion of both protein and mRNA markers in live cancer cell populafions, and in real fime, would be significantly enabling. This project aims to bring a new and enabling technology, 'NanoFlares', to bear on the detection and isolation of phenotypically distinct cancer stem cells (CSC). Taking advantage of the unique properties of gold nanoparticles surface functionalized with DNA (DNA Au-NPs), the NanoFlare technology provides the unique capability of phenotyping cell sub-populations simultaneously at the protein and mRNA level. Single-cell analysis is possible using confocal light microscopy while thousands of individual cell measurements can take place in high throughput using flow cytometry (FC) and fiuorescence activated cell sorting (FACS). Focusing initially on breast cancer, project success will have significant impact from the standpoint of basic science discovery in the context of CSCs and, ultimately, for pafients with all forms of localized and disseminated cancer where CSC detection and eradication could have a dramatic impact on patient outcomes.