Multi-scale Molecular Imaging of the Degradome in Breast Tumors

乳腺肿瘤降解组的多尺度分子成像

• 批准号: 8455704
• 负责人: Kristine Glunde
• 金额: $ 31.99万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455704
• 关键词: Affect; Breast Cancer Cell; Breast Cancer Model; Cancer cell line; Cathepsins; Cell Line; Characteristics; Collagen Type I; Complex; Cultured Tumor Cells; Data; Development; Diagnostic Imaging; Dyes; Enzymes; Extracellular Matrix; Fiber; Fluorescent Probes; Functional Imaging; Generations; Genetic Engineering; Growth; Housing; Human; Image; Imaging Techniques; Invaded; Lead; Lesion; Lysosomes; Malignant Neoplasms; Mammary Neoplasms; Medical; Metalloproteases; Metastatic Lesion; Microscopic; Microscopy; Modeling; Monitor; Neoplasm Metastasis; Nodule; Nonmetastatic; Optics; Organelles; Peptide Hydrolases; Play; Primary Neoplasm; Proteins; Research; Role; Shapes; Signal Transduction; Solid Neoplasm; Structure; Surrogate Markers; System; Therapeutic; Tumor Cell Invasion; Tumor Tissue; Urokinase; Xenograft Model; Xenograft procedure; base; cancer cell; cancer therapy; in vivo; in vivo Model; innovation; insight; malignant breast neoplasm; molecular imaging; novel; optical fiber; optical imaging; second harmonic; tumor; tumor microenvironment; tumor progression; tumor xenograft

DESCRIPTION (provided by applicant): Lysosomes are one of the most important cellular organelles in breast cancer invasion, metastasis, and extracellular matrix (ECM) remodeling, because they contain several degradative enzymes that can influence the integrity and structure of the ECM. The roles of lysosomes in the breast cancer degradome in vivo are virtually unexplored, due, in part, to a lack of noninvasive imaging capabilities available to visualize lysosomes. We recently developed and validated novel near-infrared fluorescent (NIRF) probes for noninvasive imaging of lysosomes in cell culture and tumors in vivo. Preliminary data obtained using these probes support the importance of lysosomes in breast cancer invasion and metastasis. We therefore intend to use these probes to understand and define the influence of lysosomes in cancer cells on invasion, metastasis, and ECM remodeling. Collagen type I (Col1) fibers are an important component of the ECM that can facilitate breast tumor formation, invasion, and metastasis. In this application, we will perform multi-scale molecular and functional imaging studies to characterize the Col1 fiber matrix in primary tumors and metastatic nodules to understand how Col1 fibers are shaped by the cancer degradome, and how this promotes invasion and metastasis. We will microscopically image Col1 fibers using innovative and clinically translatable optical second harmonic generation (SHG) microscopy, which detects an intrinsic optical signal from Col1 fibers. We will utilize the lysosome-specific NIRF probe IR-2 to image lysosomes in breast tumor models in vivo. We will optically image protease activities in vivo using MMPSense to detect matrix metalloprotease activities and ProSense to detect cathepsin activities. We will focus on understanding the role of lysosomes in breast tumor invasion in vivo in Aim 1, and in breast cancer metastasis in Aim 2 using multi-scale optical imaging approaches. In Aim 3, we will determine the influence of lysosomes and lysosomal enzymes on Col1 fiber structure and integrity in solid tumors and metastatic nodules in vivo. These three Aims will provide novel insights into the role of lysosomes and lysosomal enzymes within the overall degradome in breast cancer models. The proposed research will further our understanding of the role of lysosomes in cancer invasion, metastasis, and Col1 fiber remodeling, which may eventually lead to novel lysosome-based therapeutic strategies to treat cancer. Understanding the role of lysosomes and lysosomal enzymes in invasion, metastasis, and Col1 fiber remodeling may lead to the development of innovative diagnostic imaging techniques that noninvasively detect lysosomal probes as surrogate markers to assess invasiveness, metastatic potential, and Col1 fiber structure in these tumors.

描述（由申请人提供）：溶酶体是乳腺癌侵袭、转移和细胞外基质（ECM）重塑中最重要的细胞器之一，因为它们含有几种可影响ECM完整性和结构的降解酶。溶酶体在体内乳腺癌降解组中的作用几乎未被探索，部分原因是缺乏可用于可视化溶酶体的非侵入性成像能力。我们最近开发并验证了新型近红外荧光（NIRF）探针，用于细胞培养和体内肿瘤中溶酶体的非侵入性成像。使用这些探针获得的初步数据支持溶酶体在乳腺癌侵袭和转移中的重要性。因此，我们打算使用这些探针来理解和定义癌细胞中溶酶体对侵袭、转移和ECM重塑的影响。I型胶原（Col 1）纤维是ECM的重要组成部分，可以促进乳腺肿瘤的形成，侵袭和转移。在本申请中，我们将进行多尺度分子和功能成像研究，以表征原发性肿瘤和转移性结节中的Col 1纤维基质，以了解Col 1纤维如何通过癌症降解蛋白形成，以及如何促进侵袭和转移。我们将使用创新的和临床上可平移的光学二次谐波发生（SHG）显微镜对Col 1纤维进行显微成像，该显微镜检测Col 1纤维的固有光信号。我们将利用溶酶体特异性NIRF探针IR-2在体内乳腺肿瘤模型中对溶酶体进行成像。我们将使用MMPSense检测基质金属蛋白酶活性，使用ProSense检测组织蛋白酶活性，对体内蛋白酶活性进行光学成像。我们将重点了解溶酶体的作用，在乳腺肿瘤的侵袭在体内的目标1，并在乳腺癌转移的目标2使用多尺度光学成像方法。在目标3中，我们将确定溶酶体和溶酶体酶对体内实体瘤和转移性结节中Col 1纤维结构和完整性的影响。这三个目标将为乳腺癌模型中溶酶体和溶酶体酶在整体降解组中的作用提供新的见解。拟议的研究将进一步了解溶酶体在癌症侵袭，转移和Col 1纤维重塑中的作用，这可能最终导致基于溶酶体的新治疗策略来治疗癌症。了解溶酶体和溶酶体酶在侵袭，转移和Col 1纤维重塑中的作用可能会导致创新的诊断成像技术的发展，这些技术可以非侵入性地检测溶酶体探针作为替代标记物，以评估这些肿瘤的侵袭性，转移潜力和Col 1纤维结构。