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新颖的细胞异质性测定 生物分子成分分析（BCA）。 第二阶段的目标是在商业共焦显微拉曼仪器中实现BCA工具箱 用于定量评估细胞异质性和潜在的定量分类， 基于特定细胞器中大分子组成的细胞状态，单细胞测定 在第一阶段开发的基础上，第二阶段的重点是将这种新设备推向市场。 这个项目的动机是，目前没有任何商业工具， 提供活细胞中局部生物分子浓度的直接探测。同时这 工具是非常有价值的解决一些问题，这需要确定 （i）患病细胞群体中不同细胞状态的定量标志物 （ii）在细胞-药物相互作用期间，和（iii）在细胞-药物相互作用期间， 细胞内过程（凋亡、增殖、分化等）。 该检测试剂盒的I期研究和开发已达到规定的技术里程碑。在 特别是，发现BCA表征细胞异质性的潜力 大大超过了常规使用的标准数学工具主成分分析 为了分析拉曼光谱， 本建议的具体目标是：（1）Thermo-Fisher显微拉曼系统DXR 2的适应性 生物分子成分分析的发展 C++和/或Java软件包，并与Thermo-Fisher显微拉曼的DXR 2模型结合 系统;（3）验证与Thermo-Fisher显微拉曼系统DXR 2集成的BCA工具箱 正在进行的生物医学研究的模型（a）IDH- 突变的和WT-IDH胶质瘤细胞中的细胞-药物相互作用以及IDH突变细胞中的细胞-药物相互作用的评估。 与NCI合作，（Bethesda，MD）（B） 使用RPCI对前列腺癌进行分类（纽约州布法罗）;（4）上市前评估 Thermo-Fisher DXR 2显微拉曼系统，配备BCA工具箱。 该项目的成果将是一个商业共焦显微拉曼系统， BCA工具箱，沿着用于定量评估细胞异质性的定制软件， 其可用于许多领域中的广泛生物医学应用， 实验室，生物医学细胞研究实验室，制药工业，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.