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（由申请人提供）：乳腺癌细胞通常表现出功能和表型异质性。细胞异质性可能是耐药性和休眠性促进癌症复发的根源。肿瘤细胞异质性的一种来源可能是癌症干细胞的存在。基于这一假设，在此称为肿瘤的CSC，称为肿瘤引发细胞（TICS），“区分”到非肿瘤性癌细胞中，形成了分层组织和肿瘤细胞的异质性。乳房抽搐已经通过独特的细胞外表面标记（例如CD44HI/CD24LO）的表达来鉴定。现在，人们认为，即使抽搐也可能通过克隆进化表现出细胞异质性。为了研究乳房抽搐之间的异质性，我们利用了拉曼光谱法（RS）。 RS实现活抽动的细胞内分子组成的表征。通过执行RS，我们证明了活的抽动基于独特的细胞内分子成分的表达表现出肿瘤间和肿瘤内异质性。基于这些数据，我们假设抽动是异质的，拉曼镊子光谱（RTS）可能是一种方法，可以通过该方法进行分类和功能评估表现出独特细胞内分子特征（IMP）（IMPS）的TICS基团的致瘤潜力。我们的长期目标是确定和消除 最具耐药性，休眠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来表征TICS之间的异质性。我们的理由是，到目前为止，TIC已被隔离和以独特的表面标记的表达为特征。但是，尚未评估基于组合细胞外和IMP的抽动隔离和表征的方法。 RS是振动光谱。激光光的光子被活细胞吸收，然后再重新组成，提供了代表活细胞内细胞内生化特征的光谱谱（波长）。 RT将与先进的数学工具结合使用，以非侵入性，具有高特异性和快速的性质来量化抽动的特征。 RTS允许根据其Imps对细胞进行分类。然后，将通过限制稀释移植测定法对捕获的细胞进行功能评估其致瘤电位。这些研究将为最具侵略性和懒惰的乳房抽搐以及治疗策略设计的未来分子表征提供基础。