Engineered Microenvironments to model effect of size in tumor progression

工程微环境模拟肿瘤进展中大小的影响

• 批准号: 8680848
• 负责人: Shilpa Sant
• 金额: $ 7.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8680848
• 关键词: Affect; Apoptosis; Apoptotic; Area; Basal Cell; Basal Cell Cancer; Basic Science; Biological; Breast Cancer Cell; Cancer Etiology; Cancer Patient; Cell Communication; Cell-Cell Adhesion; Cells; Clinical; Clinical Research; Coupled; Data; E-Cadherin; Engineering; Epithelial; Epithelium; Event; Exhibits; Extracellular Matrix; Generations; Growth; Hormonal; Hydrogels; Hypoxia; Immunohistochemistry; Intercellular Fluid; Kinetics; Link; MCF10A cells; MCF7 cell; Mesenchymal; Metabolic stress; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; N-Cadherin; Necrosis; Neoplasm Metastasis; Nodal; Nutritional; Outcome; Oxygen; Pathway interactions; Patients; Pattern; Phenotype; Preclinical Drug Evaluation; Proliferating; RNA Interference; Radial; Regulation; Role; Signal Transduction; Snails; Spatial Distribution; Stress; Stromal Cells; Technology; Tumor Cell Invasion; Up-Regulation; Vimentin; Work; in vivo; inhibitor/antagonist; insight; interstitial; malignant breast neoplasm; monolayer; mortality; neoplastic cell; novel; novel strategies; pressure; public health relevance; receptor; slug; small molecule; three-dimensional modeling; transcription factor; tumor; tumor growth; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): The objective of this proposal is to model the effect of tumor size and growth in promoting tumor progression. More specifically, the proposal aims to create a micro fabricated platform that will allow generation of uniform, size-controlled micro tumors in a high throughput manner and study size-dependent differences in tumor microenvironment, cell-cell interaction and transcription factors. Recently tumor microenvironment has been considered as an important player in tumorigenesis and progression leading to metastasis, a major cause of cancer-related mortality. Tumor microenvironment consists of tumor & stromal cells, extracellular matrix and non-cellular components that include hypoxia, interstitial pressure and metabolic stresses. Growing tumor exerts forces on the surrounding cells and epithelium creating radial and circumferential stress. Tumor growth also results in metabolic stress, hypoxia, necrosis, increased matrix stiffness and interstitial fluid pressure. Although tumor size has been considered as one of the important predictor of metastasis and patient survival, the mechanistic link between tumor size and the clinical outcome (metastasis, mortality) is not well understood. In breast cancer patients, this scenario is complicated by nodal and hormonal receptor status. This proposal aims 1) to develop three-dimensional (3D) micro fabricated platform to generate size-controlled micro tumors with controlled microenvironments using sub-type specific breast cancer cell lines and 2) to understand the effect of tumor size on microenvironment and delineate the mechanism by which tumor size promotes tumorigenesis and metastasis. Aim 1 will use micro fabrication technologies to pattern cells in non-adhesive hydrogel micro wells and generate size-controlled micro tumors of different sizes. Various sub-type specific cell lines (luminal, basal, triple negative etc.) will be used to investigate the size-dependent differences in their growth kinetics, hormonal status, cell-cell interaction and tumor microenvironment. In Aim 2, size-dependent transcriptional changes in micro tumors will be studied and this information will be used further to understand the mechanism behind tumor size-related tumor progression using RNA interference and small molecule inhibitors. The proposed approach is novel, easy and inexpensive and will allow fabricating 3D micro tumor models in a high throughput manner. Ability to control microenvironments in size-dependent manner will facilitate discovery of key signaling mechanisms involved in the regulation of morphological and transcriptional changes in tumor cells. This approach can potentially be used as a more relevant 3D drug screening platform to replace current 2D cell monolayers.

描述（由申请人提供）：该提案的目的是模拟肿瘤大小和生长对促进肿瘤进展的影响。更具体地说，该提案旨在创建一个微制造平台，允许以高通量方式生成均匀、尺寸受控的微型肿瘤，并研究肿瘤微环境、细胞间相互作用和转录因子的尺寸依赖性差异。最近，肿瘤微环境被认为是肿瘤发生和进展导致转移的重要参与者，转移是癌症相关死亡的主要原因。肿瘤微环境由肿瘤和基质细胞、细胞外基质和非细胞成分组成，包括缺氧、间质压力和代谢应激。生长的肿瘤对周围的细胞和上皮施加力，产生径向和周向应力。肿瘤生长还会导致代谢应激、缺氧、坏死、基质硬度和间质液压力增加。尽管肿瘤大小被认为是转移和患者生存的重要预测因素之一，但肿瘤大小与临床结果（转移、死亡率）之间的机制联系尚不清楚。在乳腺癌患者中，这种情况因淋巴结和激素受体状态而变得复杂。该提案的目的是：1）开发三维（3D）微制造平台，利用亚型特异性乳腺癌细胞系生成具有受控微环境的尺寸控制的微肿瘤；2）了解肿瘤尺寸对微环境的影响，并阐明肿瘤尺寸促进肿瘤发生和转移的机制。目标 1 将使用微制造技术在非粘附性水凝胶微孔中对细胞进行图案化，并生成不同大小的大小受控的微肿瘤。各种亚型特异性细胞系（管腔细胞系、基础细胞系、三阴性细胞系等）将用于研究其生长动力学中大小依赖性的差异， 激素状态、细胞间相互作用和肿瘤微环境。在目标 2 中，将研究微肿瘤中大小依赖性转录变化，并将利用这些信息进一步利用 RNA 干扰和小分子抑制剂了解肿瘤大小相关的肿瘤进展背后的机制。 所提出的方法新颖、简单且廉价，并且允许以高通量方式制造 3D 微肿瘤模型。以大小依赖性方式控制微环境的能力将有助于发现涉及肿瘤细胞形态和转录变化调节的关键信号机制。这种方法有可能用作更相关的 3D 药物筛选平台，以取代当前的 2D 细胞单层。