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: 10632016
• 负责人: Paolo Provenzano
• 金额: $ 56.21万
• 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/10632016
• 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; 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; Macrophage; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Microfluidics; Micrometastasis; Molecular; Mus; Neoplasm Metastasis; Organ; Outcome; Paracrine Communication; Pathway interactions; Patients; Peptide Initiation Factors; Population; Probability; Proliferating; Property; Rapid screening; Recurrence; Signal Pathway; Signal Transduction; Site; Sorting; Technology; Testing; Therapeutic; Time; Tissues; Work; 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; malignant breast neoplasm; mouse model; multi-photon; neoplastic cell; novel; novel therapeutics; optical imaging; prevent; programs; receptor; 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造成的，但我们对转移的各个阶段的了解 转移和驱动二级、三级等肿瘤的调节功能非常缺乏。尤其， 播散细胞周围的早期生态位似乎对于生存、休眠和/或成功至关重要 进展性微转移的发展。事实上，乳腺癌患者死于乳腺癌的情况并不少见。 治疗后数年至数十年复发或转移性疾病，导致该疾病无法检测到。 事实上，超过 67% 的乳腺癌死亡发生在 5 年生存期之后，并且一些患者 “无病”十多年后复发。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 控制异位位点细胞命运的机制。迄今为止，许多技术障碍阻碍了我们 研究休眠和复发的遗传和微环境驱动因素的能力，特别是在体内 这些事件很少发生且不易控制。事实上，允许控制细胞的体外平台 需要微环境并允许根据细胞状态（即休眠与进展）进行细胞分离 识别和表征控制这些行为的分子机制，这些机制可以被验证和瞄准 体内。为了应对这些重大挑战，该提案利用了癌症生物学和癌症方面的专业知识 通过众多创新技术进行生物工程（例如转移微流体生成， 先进的光学成像、采用 CRISPR 技术的尖端细胞工程等），使 我们极大地提高了对体内休眠如何调节的理解。在这里，我们假设 转移性微环境中的休眠或集落增殖是由癌细胞之间的锁匙行为决定的 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进行检验：（1）定义具体目标 使用高通量微-驱动生存、休眠或定植的细胞外基质组合物 工程转移环境（MEME）技术； (2) 剖析调控的分子机制 在确定的转移微环境中的存活、休眠或定植； (3) 明确具体 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， 休眠或定殖。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的治疗靶点 逃避治疗。此外，这些研究将揭示直接杀死休眠细胞或 防止它们从休眠状态逃逸到增殖状态，以防止复发。