Elucidating cellular heterogeneity among cancer stem cells by Raman Spectroscopy

通过拉曼光谱阐明癌症干细胞之间的细胞异质性

• 批准号: 8758098
• 负责人: FARIBA BEHBOD
• 金额: $ 17.49万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8758098
• 关键词: 8q24; Award; Biochemical; Biological Assay; Breast Cancer Cell; Breast Cancer Model; CD44 gene; Cancer Relapse; Cell Separation; Cells; Characteristics; Clinical; Clonal Evolution; Cluster Analysis; Communities; Data; Department of Defense; Development; Discriminant Analysis; Discrimination; Drug resistance; Epigenetic Process; Exhibits; Flow Cytometry; Future; Genomics; Goals; Heterogeneity; Human; In Situ Hybridization; Individual; Indolent; Knockout Mice; Lasers; Least-Squares Analysis; Life; Light; Malignant Neoplasms; Mammary Neoplasms; Methodology; Methods; Molecular; Molecular Profiling; Mus; Mutation; Outcome; Patients; Photons; Raman Spectrum Analysis; Research; Research Design; Resistance; Resolution; Role; Sorting - Cell Movement; Source; Specificity; Spectrum Analysis; Surface; Technology; Therapeutic; Time; Transplantation; Tumorigenicity; anti-cancer therapeutic; base; cancer cell; cancer stem cell; cancer therapy; chemotherapy; design; extracellular; in vivo; malignant breast neoplasm; neoplastic cell; new technology; physical property; prevent; public health relevance; therapeutic development; tool; tumor; tumorigenic

DESCRIPTION (provided by applicant): Breast cancer cells often exhibit functional and phenotypic heterogeneity. Cellular heterogeneity may be the source of drug resistance and dormancy promoting cancer relapse. One source of tumor cell heterogeneity may be the existence of cancer stem cells. Based on this hypothesis, the tumorigenic CSCs, herein referred to as tumor initiating cells (TICs), "differentiate" into non-tumorigenic cancer cells creating a hierarchical organization and tumor cell heterogeneity. Breast TICs have been identified by the expression of unique extracellular surface markers such as CD44hi/CD24lo. It is now believed that even TICs may exhibit cellular heterogeneity through clonal evolution. In order to study heterogeneity among breast TICs, we utilized Raman Spectroscopy (RS). RS enables the characterization of intracellular molecular composition of live TICs. By performing RS, we demonstrated that live TICs exhibited both inter- and intra-tumoral heterogeneity based on the expression of a unique intracellular molecular component. Based on these data, we hypothesize that TICs are heterogeneous and Raman Tweezer Spectroscopy (RTS) may be a method by which to sort and functionally evaluate the tumorigenic potential of TICs groups that exhibit unique intracellular molecular profiles (IMPs). Our long-term goal is to identify and eradicate the most resistant, dormant, and aggressive breast TICs in order to prevent cancer relapse. Our objective is to characterize heterogeneity among TICs by IMP using RTS. Our Rationale is that so far, TICs have been isolated and characterized by the expression of unique surface markers. However, a methodology for isolation and characterization of TICs based on combined extracellular and IMP has not been evaluated. RS is a vibrational spectroscopy. Photons of the laser light are absorbed by live cells and then reemitted, providing spectral profiles (wavelengths) that represent the intracellular biochemical signatures of living cells. RTS will be combined with advanced mathematical tools to quantify characteristic IMPs of TICs non-invasively, with high specificity and rapidly. RTS permits sorting cells according to their IMPs. The trapped cells will then be functionally evaluated for their tumorigenic potential by limiting dilution transplantation assays. These studies will provide the basis for future molecular characterization of the most aggressive and indolent breast TICs and design of therapeutic strategies for their eradiation.

描述(申请人提供)：乳腺癌细胞通常表现出功能和表型的异质性。细胞异质性可能是耐药和休眠促进癌症复发的根源。肿瘤细胞异质性的一个来源可能是肿瘤干细胞的存在。基于这一假设，致瘤CSCs，在这里被称为肿瘤起始细胞(TICS)，“分化”成非致癌癌细胞，创造了层次化的组织和肿瘤细胞的异质性。乳腺抽动是通过CD44hi/CD24lo等独特的细胞外表面标志物的表达来鉴定的。现在人们认为，即使是抽动，也可能通过克隆进化表现出细胞的异质性。为了研究乳房痉挛之间的异质性，我们使用了拉曼光谱(RS)。RS能够表征活的痉挛的细胞内分子组成。通过RS，我们证明了基于一种独特的细胞内分子成分的表达，活体抽动表现出了瘤内和瘤内的异质性。基于这些数据，我们假设TICS是异质性的，拉曼光谱分析(RTS)可能是一种对表现出独特细胞内分子特征(IMP)的TICS组进行分类和功能评估的方法。我们的长期目标是识别和根除 最具抵抗力、潜伏性和侵略性的乳房抽搐，以防止癌症复发。我们的目标是通过使用RTS的IMP来表征TICS之间的异质性。我们的理论基础是，到目前为止，抽搐已经被分离出来，并以独特的表面标记的表达为特征。然而，一种基于细胞外和IMP相结合的TICS分离和鉴定方法还没有得到评价。RS是一种振动光谱学。激光的光子被活细胞吸收，然后重新发射，提供代表活细胞内生化特征的光谱轮廓(波长)。RTS将与先进的数学工具相结合，以非侵入性、高特异性和快速的方式量化TICS的特征IMP。RTS允许根据IMP对单元格进行排序。然后，通过限制稀释移植试验，对捕获的细胞进行功能评估，以确定其致瘤潜力。这些研究将为未来最具侵袭性和惰性的乳房抽搐的分子特征和设计治疗策略提供基础。