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

基本信息

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
项目类别：
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

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

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项目摘要

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.

转移造成约90％的与人类癌症相关的死亡，但我们对转移和驱动次级，第三纪等的受调节特征非常缺乏肿瘤。尤其，围绕传播细胞的早期利基对于生存，休眠和/或成功似乎至关重要发展微转移的发展。确实，乳腺癌患者屈服于在治疗后数十年的复发或转移性疾病使该疾病无法检测到。实际上，超过67％的乳腺癌死亡发生在5年生存窗口之外在“无病”十多年后，出现了复发。但是，我们对启动和维护休眠与转移计划的内在和环境因素进展仍然极为有限。在这里，我们寻求阐明基本的物理和分子控制生态部位细胞命运的机制。迄今为止，许多技术障碍阻碍了我们的能够研究休眠和复发的遗传和微环境驱动因素，尤其是体内这些事件很少见，并且不容易控制。确实，允许控制细胞的体外平台基于细胞态（即休眠与进展）的微环境和允许细胞分离才能识别和表征有关这些行为的分子机制，可以验证和针对性体内。为了应对这些重大挑战，该提案利用癌症生物学和癌症的专家通过众多创新技术（例如，微流体生成转移性壁ni，先进的光学成像，使用CRISPR技术等级的尖端细胞工程等）独特地启用我们大大提高了我们对在体内如何调节休眠的理解。在这里，我们假设转移性壁ches中的休眠或菌落增殖取决于癌细胞之间的锁定行为具有特定的遗传和表观遗传信号，以及生态的初始和不断发展的特性微环境。我们的假设将在以下特定目的中进行检验：（1）定义特定的特定目的细胞外基质组合物，可使用高通量微型 - 工程转移环境（MEME）技术；（2）剖析控制分子机制定义的转移性微环境中的生存，休眠或定殖；（3）定义特定骨髓衍生和组织特异性居民巨噬细胞在癌细胞存活中的影响，休眠或定殖。通过这些努力，我们将剖析散布肿瘤的机械驱动器细胞休眠或增殖，这将阐明治疗靶标，以防止从逃避疗法。此外，这些研究将揭示直接杀死休眠细胞的治疗靶标或防止他们摆脱休眠至增殖，以防止复发。