(PQ7) Quantitative in vivo optical imaging of tumor heterogeneity

(PQ7) 肿瘤异质性的定量体内光学成像

• 批准号: 9323359
• 负责人: Melissa Caroline Skala
• 金额: $ 38.84万
• 依托单位: MORGRIDGE INSTITUTE FOR RESEARCH, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9323359
• 关键词: Address; Alpha Cell; Animals; Antineoplastic Agents; Behavior; Binding; Breast Cancer Model; Breast Cancer Treatment; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Cellular Metabolic Process; Cyclophosphamide; DNA Damage; Data; Disease remission; Enzymes; Equilibrium; FDA approved; Fibroblasts; Flow Cytometry; Fluorescence; Fluorescent Probes; Genomic Instability; Goals; Gold; Growth; Heterogeneity; Image; Immune; Individual; Label; Location; Malignant Neoplasms; Measurement; Measures; Metabolic; Metabolism; Microscopy; Modeling; NADH; Nutrient; Optics; Oxidation-Reduction; Oxidative Stress; Pharmaceutical Preparations; Pharmacotherapy; Polyomavirus; Population; Publishing; Reporting; SDZ RAD; Surface; T-Lymphocyte; Testing; Time; Tumor Subtype; Work; cancer therapy; cell behavior; cell type; cellular imaging; chemotherapy; combat; fluorescence imaging; imaging modality; improved; in vivo; in vivo imaging; in vivo optical imaging; insight; mTOR Inhibitor; macrophage; metabolic imaging; mouse model; neoplastic cell; public health relevance; response; tool; treatment duration; treatment response; tumor; tumor growth; tumor heterogeneity; tumor metabolism

 DESCRIPTION (provided by applicant): The goal of this project is to address PQ7: What in vivo imaging methods can be developed to determine and record the identity, quantity, and location of each of the different cell types that contribute to the heterogeneity of a tumor and it microenvironment? Heterogeneity exists in tumor cell response to treatment, and in the pro-tumor vs. anti-tumor behavior of the cells in the microenvironment (e.g. fibroblasts, immune cells). However, there are a lack of in vivo imaging methods that can quantify this heterogeneity, thus limiting our understanding of cellular-level tumor behavior, and limiting our ability to develop improved cancer treatments. Cellular metabolism provides dynamic insight into individual cell behavior. We and others have shown that tumor cell metabolism reflects anti-cancer drug response, and tumor cells alter their metabolic activities in order to resist drug treatment. The metabolism of tumor-associated fibroblasts also support malignancy, by supplying nutrients to drive tumor growth. Additionally, the metabolism of immune cells is reflective of their pro- and anti-tumor behavior, with distinct changes in metabolic activities between M1-like and M2-like macrophages, between CD4+ and CD8+ T cells, and within the CD4+ T cell subset. Due to the key role of metabolism in maintaining the tumor and its microenvironment, drugs that disrupt the metabolism of tumor cells and cells in the microenvironment have been FDA approved for breast cancer treatment in combination with standard therapies. The goal of this proposal is to develop and validate optical metabolic imaging (OMI) to quantify dynamic metabolic heterogeneity within the tumor cell, fibroblast, macrophage, and T cell populations in tumors in vivo. Multiphoton microscopy will resolve individual cells within the Polyoma middle T (PyMT) mouse model of breast cancer throughout a treatment time-course. Autofluorescence from the metabolic co-enzymes NADH and FAD will quantify cellular metabolism, and report on metabolic heterogeneity within cell populations. Specifically, OMI will measure the optical "redox ratio" of each cell (fluorescence intensity of NADH divided by that of FAD), which reflects redox balance in a cell. OMI also quantifies the fluorescence lifetimes of NADH and FAD, which reflect the enzyme binding activity of these molecules. Our published work and preliminary data demonstrate that OMI is sensitive to heterogeneous drug response in tumors in vivo, and OMI can distinguish sub-types of tumor cells and immune cells. The proposed work will validate in vivo OMI of tumor heterogeneity with ex vivo flow cytometry and in vivo imaging of fluorescent cell surface markers. The proposed aims will test the hypothesis that in vivo OMI can record the identity, quantity, and location of different cell types that contribute to the metabolic heterogeneity of a tumor and its microenvironment. These tools will enable longitudinal quantification of the in vivo metabolic heterogeneity of tumor cells, fibroblasts, macrophages, and T- cells. The insights gained from these measurements can be used to develop improved cancer treatments that combat tumors on a single-cell level in order to achieve remission-free survival.

 描述（由申请方提供）：本项目的目标是解决PQ7：可以开发哪些体内成像方法来确定和记录导致肿瘤及其微环境异质性的每种不同细胞类型的身份、数量和位置？异源性存在于肿瘤细胞对治疗的反应中，以及微环境中细胞（例如成纤维细胞、免疫细胞）的促肿瘤与抗肿瘤行为中。然而，缺乏可以量化这种异质性的体内成像方法，从而限制了我们对细胞水平肿瘤行为的理解，并限制了我们开发改进的癌症治疗方法的能力。细胞代谢提供了对单个细胞行为的动态洞察。我们和其他人已经表明，肿瘤细胞代谢反映了抗癌药物的反应，肿瘤细胞改变其代谢活动，以抵抗药物治疗。肿瘤相关成纤维细胞的代谢也支持恶性肿瘤，通过提供营养物质来驱动肿瘤生长。此外，免疫细胞的代谢反映了它们的促肿瘤和抗肿瘤行为，M1样和M2样巨噬细胞之间、CD4+和CD8+ T细胞之间以及CD4+ T细胞亚群内的代谢活性存在明显变化。由于代谢在维持肿瘤及其微环境中的关键作用，破坏肿瘤细胞和微环境中细胞代谢的药物已被FDA批准与标准疗法联合用于乳腺癌治疗。该提案的目标是开发和验证光学代谢成像（OMI），以量化体内肿瘤中肿瘤细胞、成纤维细胞、巨噬细胞和T细胞群内的动态代谢异质性。多光子显微镜将在整个治疗时间过程中解析乳腺癌的多瘤中T（PyMT）小鼠模型中的单个细胞。来自代谢辅酶NADH和FAD的自发荧光将量化细胞代谢，并报告细胞群内的代谢异质性。具体来说，OMI将测量每个细胞的光学“氧化还原比”（NADH的荧光强度除以FAD的荧光强度），这反映了细胞中的氧化还原平衡。OMI还定量了NADH和FAD的荧光寿命，这反映了这些分子的酶结合活性。我们已发表的工作和初步数据表明，OMI对体内肿瘤的异质性药物反应敏感，并且OMI可以区分肿瘤细胞和免疫细胞的亚型。拟开展的工作将通过离体流式细胞术和荧光细胞表面标记物的体内成像来验证肿瘤异质性的体内OMI。所提出的目标将测试以下假设：体内OMI可以记录不同细胞类型的身份、数量和位置，这些细胞类型有助于肿瘤及其微环境的代谢异质性。这些工具将能够纵向定量肿瘤细胞、成纤维细胞、巨噬细胞和T细胞的体内代谢异质性。从这些测量中获得的见解可用于开发改进的癌症治疗方法，在单细胞水平上对抗肿瘤，以实现无缓解生存。