Elucidating cellular heterogeneity among cancer stem cells by Raman Spectroscopy

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的来源，促进癌症复发。肿瘤细胞异质性的一个来源可能是癌症干细胞的存在。基于该假设，致瘤性CSC（本文中称为肿瘤起始细胞（TIC））“分化”成非致瘤性癌细胞，从而产生分级组织和肿瘤细胞异质性。乳腺TIC已通过表达独特的细胞外表面标志物如CD 44 hi/CD 24 lo来鉴定。现在认为，即使是TIC也可能通过克隆进化表现出细胞异质性。为了研究乳腺TIC之间的异质性，我们利用拉曼光谱（RS）。RS能够表征活TIC的细胞内分子组成。通过进行RS，我们证明了基于独特的细胞内分子组分的表达，活TIC表现出肿瘤间和肿瘤内异质性。基于这些数据，我们假设TIC是异质性的，并且拉曼镊子光谱（RTS）可能是一种方法，通过该方法对表现出独特的细胞内分子谱（IMP）的TIC组进行分类和功能性评价。我们的长期目标是确定和消除 大多数耐药、休眠和侵袭性乳腺TIC，以防止癌症复发。我们的目标是通过使用RTS的IMP来表征TIC之间的异质性。我们的基本原理是，到目前为止，TIC已被分离，并通过表达独特的表面标志物来表征。然而，还没有评估基于组合的细胞外和IMP的TIC的分离和表征的方法。RS是一种振动光谱。激光的光子被活细胞吸收，然后重新发射，提供代表活细胞的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，以无创、高特异性和快速的方式量化TIC的特征性IMP。RTS允许根据IMP对细胞进行分选。然后通过有限稀释移植试验对捕获的细胞的致瘤潜力进行功能性评价。这些研究将为未来最具侵袭性和惰性的乳腺TIC的分子特征和设计其放射治疗策略提供基础。

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.