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：
10262017
负责人：
Lalage Wakefield
金额：
$ 93.15万
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/10262017
关键词：
Address Allografting Biological Blood Circulation Cell Compartmentation Cell Count Cell Differentiation process Cell division Cells Characteristics Chromatin Clinical Collaborations Complex Development Enzymes Functional Imaging Genomics Goals Human In Situ In Vitro Individual Mammary Neoplasms Medicine Methods Minority Molecular Pathway interactions Patients Phenotype Play Population Pre-Clinical Model Primary Neoplasm Process Proteins Reporter Resistance Role Sampling Signal Transduction Site Structure Testing Therapeutic Time Transforming Growth Factor Beta 2 Transforming Growth Factor beta Transgenic Model Tumor Suppressor Proteins Visualization Xenograft procedure base breast cancer progression cancer stem cell cancer therapy carcinogenesis carcinogenicity cell type clinical development college epigenetic regulation functional genomics genetic regulatory protein imaging approach in vivo intravital imaging lung metastatic macrophage malignant breast neoplasm metastatic process mouse model neoplastic cell notch protein novel preservation prevent promoter self-renewal stem stem cell population stem cells stemness transcription factor tumor tumor progression tumorigenesis

项目摘要

In FY20, 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 vitro and in vivo, including 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 primary breast 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. Importantly, we have shown induction of a stem phenotype in non-stem tumor cells on contact with macrophages, via a Notch-dependent mechanism in vitro and in vivo. We have characterized changes in CSC representation across the entire metastatic process, demonstrating peak CSC numbers on early arrival at the metastatic site. W have shown that TGF-beta effects and TGF-beta signal transduction are different in stem vs. non-stem cells, and also that TGF-beta regulates the CSC compartment differently depending on whether it is functioning as tumor suppressor or pro-progression factor. These results have important implications for the ongoing clinical development of TGF-beta pathway antagonists. Studies to understand the detailed mechanisms regulating the size of the cancer stem cell population are ongoing. We have identified the chromatin-modifying enzyme PADI4 as having novel tumor suppressive activity on the CSC population through epigenetic regulation of the expression of master transcription factors of stemness. We have developed fate-mapping approaches to address the effects of TGF-beta on phenotypic plasticity and on self-renewing vs differentiating cell divisions. We are also integrating these analyses with genomic and single cell approaches to address underlying molecular mechanisms. 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.

在第20财年，我们继续使用一种新型的功能成像方法来解决TGF-beta在调节癌症干细胞（CSC）室中的作用，我们开发了该方法，以实时和原位对这种少数细胞种群的可视化。我们的基于慢病毒的CSC报告基因使用合成启动子，其中荧光蛋白的表达是由干细胞主转录因子Oct4和Sox2驱动的。使用这种方法，我们开发了允许在体外和体内对单个CSC进行扩展的细胞命运映射，包括与阿尔伯特·爱因斯坦医学院的John Condeelis博士实验室进行的持续合作，以对原发性乳腺肿瘤和肺转移性部位的CSC进行室内成像。插入成像方法使我们能够证明CSC是缓慢移动的原发性肿瘤中的侵入性细胞，这些细胞优先与微解剖结构（TMEM）相关，该结构（TMEM）将肿瘤细胞插入到血液中。重要的是，我们通过体外和体内的凹入依赖性机制显示了与巨噬细胞接触的非茎肿瘤细胞中茎表型的诱导。我们已经表征了整个转移过程中CSC表示的变化，表明CSC峰值在到达转移部位时数量。 w表明，TGF-β效应和TGF-β信号转导在茎和非茎细胞中是不同的，并且TGF-beta也根据肿瘤抑制剂还是促产生因子的作用，对CSC室的调节不同。这些结果对TGF-β途径拮抗剂的持续临床发展具有重要意义。了解调节癌症干细胞种群大小的详细机制的研究正在进行中。我们已经通过表观遗传学调节了干剂的表达，使染色质改良酶PADI4在CSC种群中具有新颖的肿瘤抑制活性。我们已经开发了命运方法来解决TGF-β对表型可塑性以及自我更新与区分细胞分裂的影响。我们还将这些分析与基因组和单细胞方法整合在一起，以解决潜在的分子机制。了解如何在体内调节CSC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具有抵抗力。