Novel single cell assay for quantitative analysis of cell heterogeneity by noninvasive probing of molecular composition of specific organelles in individual cells

项目摘要 ACIS，LLC提出了一种基于共聚焦拉曼的概念上新的细胞异质性分析 光谱和生物分子成分分析(BCA)。 第二阶段的目标是在商用共焦微拉曼仪器中实现BCA工具箱 用于细胞异质性的定量评估和潜在的定量分类 基于特定细胞器中的大分子组成的细胞状态，单细胞分析 在第一阶段开发，第二阶段专注于将这种新设备推向市场。 这个项目的动机是，目前还没有任何商业工具， 提供活细胞中局部生物分子浓度的直接探测。与此同时，这一点 工具对于解决许多问题非常有价值，这些问题需要确定 不同细胞状态的定量标记(I)病态细胞群体 (癌症-非癌症、不同癌症阶段等)，(Ii)在细胞-药物相互作用期间，以及(Iii)在 细胞内过程(细胞凋亡、增殖、分化等)。 该检测的第一阶段研究和开发已经达到了规定的技术里程碑。在……里面 特别是，发现BCA势可以表征细胞的异质性 大大超过了常规使用的主成分分析标准数学工具 对于拉曼光谱的分析，。 该方案的具体目的是：(1)改装Thermo-Fisher微拉曼系统DXR2 适应生物化学分析应用需求的模型；(2)生物分子成分分析的发展 C++和/或JAVA语言的软件包及其与Thermo-Fisher微拉曼DXR2模型的结合 (3)与Thermo-Fisher微拉曼系统DXR2集成的BCA工具箱验证 正在进行的生物医学研究的模型(A)IDH和IDH之间的异质性比较分析 突变型和WT-IDH胶质瘤细胞及IDH突变细胞与药物相互作用的研究 与NCI合作，(马里兰州贝塞斯达)(B)细胞细胞器的分子图谱 用RPCI法对前列腺癌进行分类(纽约州布法罗)；(4)上市前评估 Thermo-Fisher DXR2微拉曼系统，配备BCA工具箱。 该项目的成果将是一个商用的共焦微拉曼系统，并实现了 BCA工具箱，以及用于定量评估细胞异质性的定制软件， 它可用于许多领域的广泛生物医学应用，如临床 实验室，生物医学细胞研究实验室，制药行业，国家癌症研究所， 等。

Project Summary ACIS, LLC proposes a conceptually novel cellular heterogeneity assay, based on confocal Raman spectrometry and Biomolecular Component Analysis (BCA). The goal of Phase II is to implement a BCA toolbox in a commercial confocal micro- Raman instrument for quantitative assessment of cellular heterogeneity and potential quantitative classification of cellular states based on macromolecular compositions in specific organelles, a single cell assay developed during the Phase I, Phase II focuses on bringing this new device to the market. This project is motivated by the fact, that there is no currently any commercial tool, which provides direct probing of local biomolecular concentration in live cells. At the same time, this tool is extremely valuable for solving a number of problems, which require determination of quantitative markers for different cellular states (i) in diseased cell population (cancer-non-cancer, different cancer stages etc), (ii) during cell-drug interaction, and (iii) in the intracellular processes (apoptosis, proliferation, differentiation, etc). The Phase I research and development of this assay have met stated technical milestones. In particular, it is found that the BCA potential for characterization of cellular heterogeneity significantly exceeds that of the standard math tool of Principal Component Analysis routinely used for analysis of Raman spectra,. The specific aims of this proposal are: (1) Adaptation of a Thermo-Fisher micro-Raman system DXR2 model to meet requirements for BCA applications; (2) Development of Biomolecular Component Analysis Software package in C++ and/or Java and incorporation to DXR2 model of Thermo-Fisher micro-Raman system; (3) Validation of the BCA toolbox integrated with Thermo-Fisher micro-Raman system DXR2 model for ongoing biomedical research (a) Comparative analysis of heterogeneity between IDH- mutated and WT-IDH glioma cells and assessment of cell-drug interaction in IDH-mutant cells in collaboration with NCI, (Bethesda, MD) (b) Molecular profiling of cellular organelles for categorization of Prostate Cancer with RPCI (Buffalo, NY); (4) Pre-market evaluation of the Thermo-Fisher DXR2 micro-Raman system, equipped by the BCA toolbox. The outcome of this project will be a commercial confocal micro-Raman system with an implemented BCA toolbox, along with customized software for quantitative assessment of cellular heterogeneity, which can be used for a broad range of biomedical applications in many sectors such as clinical labs, Biomedical cellular Research labs, Pharmaceutical industries, National Cancer Institutes , etc.