Imaging tumor microenvironment by Optical Fiber-Tethered Simultaneous Lifetime-resolved Autofluorescence-Multiharmonic (OFT-SLAM) microscopy

通过光纤系留同步寿命分辨自体荧光多谐波 (OFT-SLAM) 显微镜对肿瘤微环境进行成像

• 批准号: 10212984
• 负责人: Stephen A Boppart
• 金额: $ 34.16万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212984
• 关键词: Adult; Anabolism; Anatomy; Animal Cancer Model; Animal Model; Animals; Biological; Biological Process; Biology; Blood Vessels; Breast cancer metastasis; Cancer Patient; Cells; Cephalic; Chemicals; Clinical; Consumption; Cooperative Human Tissue Network; Crime; Dependence; Development; Embryo; Engineering; Epithelial Cells; Evolution; Formalin; Funding; Genetic; Goals; Grant; Heterogeneity; Histopathology; Human; Human Resources; Image; Imaging Device; Imaging Techniques; Imaging technology; Immune; Immune response; Immunohistochemistry; Implant; Individual; Inflammatory; Injections; Knowledge; Label; Laboratories; Link; Lung; Malignant Neoplasms; Mammary Gland Parenchyma; Mammary Neoplasms; Mediating; Medicine; Metabolic; Metabolism; Metastatic Neoplasm to the Lung; Metastatic malignant neoplasm to brain; Microscope; Microscopy; Modeling; Modification; Morphogenesis; Mus; Neoplasm Metastasis; Normal Cell; Nude Mice; Operative Surgical Procedures; Optics; Organ; Outcome; Pathologic; Pathologist; Patients; Phenotype; Positioning Attribute; Postoperative Period; Prognosis; Rattus; Recording of previous events; Research; Sampling; Signal Transduction; Slide; Specimen; Stains; System; Technology; Time; Tissue Transplantation; Tissues; Training; Transgenic Mice; Tumor-infiltrating immune cells; United States National Institutes of Health; Validation; Visualization; Visualization software; Warburg Effect; animal tissue; anticancer research; base; cancer diagnosis; cancer imaging; cancer therapy; carcinogenesis; cell type; clinically relevant; comparative; empowered; flexibility; fluorescence lifetime imaging; human tissue; imaging scientist; imaging system; in vivo; individual variation; industry partner; insight; instrumentation; interest; intravital microscopy; malignant breast neoplasm; mammary; molecular imaging; mouse model; movie; multimodality; multiphoton microscopy; neoplastic cell; optical fiber; optical imaging; pre-clinical; therapeutic development; tool; tumor; tumor microenvironment; tumor progression; user-friendly; wound healing

Summary A label-free imaging technology is proposed for general cancer research, termed as Optical Fiber-Tethered Simultaneous Lifetime-resolved Autofluorescence and Multiharmonic (OFT-SLAM) microscopy, to overcome the lack of a versatile tool to simultaneously visualize tumor and non-tumor cells in authentic tumor microenvironment. The non-tumor cells broadly include the fibroblastic cells, angiogenic vascular cells, and infiltrating immune cells that engage normal biological functions such as embryonic/adult development and inflammatory/immune response (e.g. wound healing). However, in the tumor microenvironment where the overall metabolism is known to switch from energy consumption to proliferative biosynthesis (the Warburg effect), these normal (neutral) cells have all been recently recognized as the accessories to the crime (cancer). Thus, the proposed development of this imaging technology will interrogate the interrelations between metastatic tumor cells (principal) and various non-tumor cells (accessories) that conspire to kill a cancer patient (crime). This interrogation will be more comprehensive than imaging-based cancer research that has typically focused on one specific cell type of interest (the principal or one accessory of the crime). Without a label-free imaging technology like OFT-SLAM to avoid cell-specific labeling, simultaneous visualization of various cells would perturb the tumor microenvironment by exogenous staining, cell/tissue transplantation, and genetic modification. We will build the “SLAM” of OFT-SLAM based on multimodal multiphoton microscopy and fluorescence-lifetime imaging, and invoke general intrinsic contrasts of cellular optical heterogeneity and metabolic activity to reveal and differentiate tumor and non-tumor cells. We will then empower the “SLAM” with the “OFT” to flexibly access different anatomical sites in intravital animal/preclinical microscopy and ex vivo human/clinical histopathology. We will subsequently employ the resulting OFT-SLAM to image the formalin-fixed human specimens of breast cancer from Cooperative Human Tissue Network (CHTN), including the primary breast tumors, breast cancer- induced lung and brain metastases, and surrounding peri-tumoral fields at different stages from different patients (n > 200). In parallel, we will apply OFT-SLAM to long-term (imaging window-assisted) intravital microscopy of three prototypical breast cancer rat/mouse models, covering all known steps throughout the invasion-metastasis cascade. With the unique capability of OFT-SLAM to bridge otherwise isolated ex vivo human histopathology (snapshots taken by pathologists in a clinical setting) and intravital animal microscopy (movies acquired by biologists in a laboratory), we will strive to identify various cancer-associated cells and their interrelations in an evolving tumor microenvironment and their dependence on spatial heterogeneity and individual variability. The successful outcome of this project will demonstrate a versatile visualization tool to interrogate tumor microenvironment with built-in translational ability, and thus transform cancer diagnosis and therapy.

概括 提出了一种无标签的成像技术，用于一般癌症研究，称为光纤束缚 同时通过寿命排列的自荧光和多哈罗式（通常（Slam）显微镜）来克服 缺乏多功能工具可以同时可视化正宗肿瘤微环境中的肿瘤和非肿瘤细胞。 非肿瘤细胞广泛包括成纤维细胞，血管生成血管细胞和浸润的免疫细胞 参与正常的生物学功能，例如胚胎/成人发育和炎症/免疫 反应（例如伤口愈合）。但是，在肿瘤微环境中，已知总代谢 要从能耗转换为增殖的生物合成（Warburg效应），这些正常（中性）细胞 最近都被认为是犯罪的配件（癌症）。那，提议的发展 这种成像技术将询问转移性肿瘤细胞（主）和各种之间的相互关系 共同杀死癌症患者（犯罪）的非肿瘤细胞（配件）。这种审讯将更多 比基于成像的癌症研究全面，该研究通常集中在一种特定细胞类型上 兴趣（犯罪的本金或一项附件）。没有像oft-slam这样的无标签成像技术 避免使用细胞特异性标记，对各种细胞的简单可视化将扰动肿瘤微环境 通过外源染色，细胞/组织移植和遗传修饰。 我们将基于多模式多光子显微镜和荧光寿命，建立经常使用的“大满贯” 成像，并引起细胞光学异质性和代谢活性的一般固有对比 并区分肿瘤和非肿瘤细胞。然后，我们将用“ oft”授权“大满贯”以灵活访问 插入动物/临床前显微镜和体内人/临床组织病理学中的不同解剖部位。 随后，我们将采用由此产生的slam来对福尔马林固定的乳房标本进行想象 来自合作人组织网络（CHTN）的癌症，包括原发性乳腺肿瘤，乳腺癌 - 诱导肺和脑转移，以及不同患者不同阶段的肿瘤周围场 （n> 200）。同时，我们将在长期（成像窗口辅助）的浸润室显微镜上应用大量 - 链接 三种典型的乳腺癌大鼠/小鼠模型，涵盖了整个入侵 - 纳斯塔斯群岛的所有已知步骤 级联。 Fast-slam具有独特的能力，可以桥接其他孤立的离体组织病理学 （病理学家在临床环境中拍摄的快照）和插入动物显微镜（电影获得的电影 实验室的生物学家），我们将努力鉴定各种与癌症相关的细胞及其相互关系 不断发展肿瘤微环境及其对空间异质性和个体变异性的依赖。 该项目的成功结果将展示一种多功能可视化工具来询问肿瘤 具有内置翻译能力的微环境，从而改变癌症的诊断和治疗。