TGF-betas in breast cancer progression

TGF-β 在乳腺癌进展中的作用

基本信息

批准号：
10014285
负责人：
Lalage Wakefield
金额：
$ 88.17万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10014285
关键词：
Address Allografting Antibodies Biological Biological Models Blood Circulation Breast Cancer Model Cell Compartmentation Cell Differentiation process Cell division Cells Characteristics Clinical Collaborations Complex Development Disease Functional Imaging Genomics Goals Homeostasis Human Image Imagery In Situ In Vitro Individual Lung Malignant Neoplasms Medicine Methods Minority Modeling Molecular Neoplasm Metastasis Normal tissue morphology Oncogenic Pathway interactions Patients Phenotype Play Population Pre-Clinical Model Primary Neoplasm Process Proteins Recurrence Reporter Resistance Role Sampling Signal Transduction Site Specific qualifier value Stem cells Structure System Testing Therapeutic Time Transforming Growth Factor Beta 2 Transforming Growth Factor beta Transgenic Model Tumor Suppressor Proteins Xenograft procedure advanced disease anti-tumor immune response base breast cancer progression cancer stem cell cancer therapy carcinogenicity cell type clinical development college functional genomics genetic regulatory protein imaging approach in vivo intravital imaging lung metastatic macrophage malignant breast neoplasm mature animal metastatic process mouse model neoplastic cell novel preservation prevent promoter response self-renewal stem stem cell biology stem cell population three dimensional cell culture tissue repair transcription factor tumor tumor microenvironment tumor progression tumorigenesis

项目摘要

In FY19, we have continued to address the role of TGF-beta in regulating the cancer stem cell (CSC) compartment using a novel functional imaging approach that we developed to allow visualization of this minority cell population in real time and in situ. Our lentiviral-based CSC reporter uses a synthetic promoter in which expression of a fluorescent protein is driven by the stem cell master transcription factors Oct4 and Sox2. Using this approach, we have developed methods that allow extended cell fate mapping of individual CSCs in 2D and 3D culture systems and we have an ongoing collaboration with the lab of Dr. John Condeelis at the Albert Einstein College of Medicine to perform intravital imaging of the CSC population in the primary tumor and at the lung metastatic site. The intravital imaging approach has allowed us to demonstrate that the CSCs are slow-moving, invasive cells in the primary tumor that are preferentially associated with the microanatomical structure (TMEM) that drives intravasation of tumor cells into the bloodstream. We have shown induction of a stem phenotype in non-stem tumor cells on contact with macrophages, and have characterized changes in CSC representation across the entire metastatic process, demonstrating peak CSC representation on early arrival at the metastatic site. We are now addressing the effect of TGF-beta on CSC biology in breast cancer model systems that show either tumor suppressor or pro-progression responses to TGF-beta. Importantly, we have shown that treatment with neutralizing anti-TGF-beta antibodies in vivo will increase the cancer stem cell population in two xenograft breast cancer models in which TGF-beta functions as a tumor suppressor, while decreasing it in a model in which TGF-beta has pro-progression effects. Furthermore, we have evidence that TGF-beta signal transduction is different in stem vs. non-stem cells. These results have important implications for the ongoing clinical development of TGF-beta pathway antagonists. Studies to understand the detailed mechanisms by which TGF-beta induces either an expansion or a reduction in the cancer stem cell population are ongoing. We are focusing on effects of TGF-beta on phenotypic plasticity and differential effects on self-renewing vs differentiating cell divisions. We do this through imaging of stem and non-stem cells early after arrival at the metastatic site in the lung, as well as in 3D culture in vitro. We are also employing integrated genomic and single cell approaches to address the molecular mechanisms underlying differential effects of TGF-beta on CSC and non-CSC compartments at early and late stages of the disease process. Understanding how CSCs are regulated in vivo will be critical to development of more effective cancer therapies, as these cells are largely resistant to existing therapeutic approaches.

在 2019 财年，我们继续利用一种新颖的功能成像方法来解决 TGF-β 在调节癌症干细胞 (CSC) 区室中的作用，我们开发了这种方法，可以实时、原位可视化该少数细胞群。我们基于慢病毒的 CSC 报告基因使用合成启动子，其中荧光蛋白的表达由干细胞主转录因子 Oct4 和 Sox2 驱动。利用这种方法，我们开发了一些方法，可以在 2D 和 3D 培养系统中对单个 CSC 进行扩展的细胞命运图谱，并且我们与阿尔伯特·爱因斯坦医学院的 John Condeelis 博士实验室持续合作，对 CSC 进行活体成像。原发肿瘤和肺转移部位的 CSC 群体。活体成像方法使我们能够证明，CSC 是原发肿瘤中移动缓慢的侵袭性细胞，它们优先与驱动肿瘤细胞内渗到血流中的微解剖结构 (TMEM) 相关。我们已经展示了在与巨噬细胞接触时非干肿瘤细胞中干表型的诱导，并且表征了整个转移过程中CSC表现的变化，证明了早期到达转移部位时CSC表现的峰值。我们现在正在研究 TGF-β 对乳腺癌模型系统中 CSC 生物学的影响，该系统显示出对 TGF-β 的肿瘤抑制或促进展反应。重要的是，我们已经证明，在两种异种移植乳腺癌模型中，用中和性抗 TGF-β 抗体进行体内治疗会增加癌症干细胞群，其中 TGF-β 充当肿瘤抑制因子，而在 TGF-β 发挥肿瘤抑制因子作用的模型中，则减少癌症干细胞群。 -beta具有促进展作用。此外，我们有证据表明 TGF-β 信号转导在干细胞与非干细胞中是不同的。这些结果对于 TGF-β 途径拮抗剂的持续临床开发具有重要意义。旨在了解 TGF-β 诱导癌症干细胞群扩增或减少的详细机制的研究正在进行中。我们重点关注 TGF-β 对表型可塑性的影响以及对自我更新与分化细胞分裂的差异影响。我们通过在到达肺部转移部位后早期对干细胞和非干细胞进行成像以及体外 3D 培养来实现这一点。我们还采用整合的基因组和单细胞方法来解决 TGF-β 在疾病过程的早期和晚期对 CSC 和非 CSC 区室产生不同影响的分子机制。了解 CSC 在体内的调节方式对于开发更有效的癌症疗法至关重要，因为这些细胞在很大程度上对现有的治疗方法有抵抗力。