A platform to functionally sort and analyze tumor cells within combinatorial metastatic micorenvironments

在组合转移微环境中对肿瘤细胞进行功能分类和分析的平台

• 批准号: 10414891
• 负责人: Paolo Provenzano
• 金额: $ 57.39万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-08 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10414891
• 关键词: 3-Dimensional; Address; Adhesions; Aftercare; Apoptosis; Behavior; Biomedical Engineering; Blocking Antibodies; Bone Marrow; Breast Cancer Patient; Breast Carcinoma; CRISPR screen; CRISPR/Cas technology; Cancer Biology; Cell Line; Cell Separation; Cell Survival; Cells; Cessation of life; Coculture Techniques; Complex; Cues; Cytokine Receptors; Development; Disease; Disease Progression; Disease Resistance; Engineering; Environment; Environmental Risk Factor; Epigenetic Process; Event; Extracellular Matrix; Flow Cytometry; Focal Adhesions; Generations; Genetic; Genetic study; Heterogeneity; Human; In Vitro; Integrins; Intrinsic factor; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Microfluidics; Micrometastasis; Molecular; Mus; Neoplasm Metastasis; Organ; Outcome; Paracrine Communication; Pathway interactions; Patients; Population; Probability; Property; Rapid screening; Recurrence; Signal Pathway; Signal Transduction; Site; Technology; Testing; Therapeutic; Time; Tissues; Work; base; body system; cancer cell; cancer stem cell; cellular engineering; combinatorial; cytokine; experimental study; genome-wide; high throughput screening; human cancer mouse model; human model; improved; in vivo; innovative technologies; macrophage; malignant breast neoplasm; mouse model; neoplastic cell; novel; novel therapeutics; optical imaging; prevent; programs; therapeutic target; therapy resistant; transcriptome; tumor

Metastases are responsible for ~90% of human cancer-related deaths, yet our understanding of the stages of metastasis and the regulating features that drive secondary, tertiary etc. tumors is sorely lacking. In particular, the early niche surrounding disseminated cells appears critical for survival, dormancy, and/or successful development of progressing micrometastases. Indeed, not infrequently, breast cancer patients succumb to recurrent or metastatic disease years to decades after treatment that had rendered the disease undetectable. In fact, greater than 67% of breast cancer deaths occur beyond the 5-year survival window and some patients present with recurrence after more than a decade of being “disease-free”. Yet, our understanding of the intrinsic and environmental factors that initiate and maintain programs of dormancy versus metastatic progression remains extremely limited. Here, we seek to elucidate fundamental physical and molecular mechanisms that govern cell fate in ectopic sites. To date, numerous technical hurdles have impeded our ability to study the genetic and microenvironmental drivers of dormancy and recurrence, particularly in vivo where these events are rare and not easily controlled. Indeed, in vitro platforms that permit control of the cell microenvironment and permit cell isolation based on cell state (i.e. dormant vs. progressing) are required to identify and characterize molecular mechanisms governing these behaviors that can be validated and targeted in vivo. To address these significant challenges, this proposal leverages expertise in cancer biology and cancer bioengineering through numerous innovative technologies (e.g. microfluidic generation of metastatic niches, advanced optical imaging, cutting edge cell engineering with CRISPR technologies etc.) that uniquely enable us to drastically improve our understanding of how dormancy is regulated in vivo. Here, we hypothesize that dormancy or colony proliferation in metastatic niches is dictated by lock-and-key behavior between cancer cells with specific genetic and epigenetic signaling and the initial and evolving properties of the ectopic microenvironment. Our hypotheses will be tested in the following Specific Aims: (1) Define specific extracellular matrix compositions that drive survival, dormancy, or colonization using high-throughput micro- engineering metastatic environments (MEME) technology; (2) Dissect the molecular mechanisms governing survival, dormancy, or colonization in defined metastatic niche microenvironments; (3) Define the specific influence of bone marrow-derived and tissue-specific resident macrophages in carcinoma cell survival, dormancy, or colonization. Through these efforts we will dissect the mechanistic drivers of disseminated tumor cell dormancy or proliferation, which will elucidate therapeutic targets to prevent dormant tumor cells from evading therapy. Additionally, these studies will reveal therapeutic targets to kill dormant cells directly or prevent their escape from dormancy to proliferation in order to prevent recurrence.

约90%的人类癌症相关死亡是由转移引起的，但我们对 转移和驱动继发性、三级等肿瘤的调节功能严重缺乏。特别是， 播散细胞周围的早期生态位似乎对存活、休眠和/或成功至关重要。 进展性微转移的进展。事实上，乳腺癌患者并不罕见地死于 在治疗后数年至数十年内复发或转移性疾病，使疾病无法被检测到。 事实上，超过67%的乳腺癌死亡发生在5年生存窗口之后，一些患者 经过十多年的“无病”治疗后出现复发。然而，我们对 启动和维持休眠与转移程序的内在和环境因素 进展仍然极其有限。在这里，我们试图阐明基本的物理和分子 控制异位部位细胞命运的机制。到目前为止，许多技术障碍阻碍了我们的 能够研究导致休眠和复发的遗传和微环境驱动因素，特别是在体内 在那里这些事件很少发生，也不容易控制。事实上，允许控制细胞的体外平台 需要微环境并允许基于细胞状态(即休眠与进展)的细胞隔离 识别和描述控制这些行为的分子机制，这些机制可以被验证和定向 在活体内。为了应对这些重大挑战，这项建议利用了癌症生物学和癌症方面的专业知识 通过许多创新技术进行生物工程(例如，转移壁龛的微流控生成， 先进的光学成像、采用CRISPR技术的尖端细胞工程等)它独特地实现了 我们需要极大地提高我们对休眠是如何在体内调节的理解。在这里，我们假设 转移的小生境中的休眠或菌落增殖是由癌细胞之间的锁钥行为决定的 具有特定的遗传和表观遗传信号以及异位的初始和进化特性 微环境。我们的假设将在以下具体目标中得到检验：(1)定义具体的 细胞外基质成分，使用高通量的微胶囊驱动存活、休眠或定居 工程转移环境(MEME)技术；(2)剖析 在确定的转移性生态位微环境中存活、休眠或定居；(3)定义特定的 骨髓源性和组织特异性驻留巨噬细胞对癌细胞存活的影响 休眠，或殖民。通过这些努力，我们将剖析播散性肿瘤的机制驱动因素。 细胞休眠或增殖，这将阐明防止休眠肿瘤细胞 逃避治疗。此外，这些研究将揭示直接或间接杀死休眠细胞的治疗目标。 防止它们从休眠状态逃脱到增殖，以防止复发。