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：
9343537
负责人：
Lalage Wakefield
金额：
$ 85.82万
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/9343537
关键词：
ATAC-seq Address Allografting Basement membrane Behavior Biological Biological Models Breast Cancer Model Cell Proliferation Cells Characteristics Clinical Collaborations Complex Development Disease Functional Imaging Genomic approach Goals Homeostasis Human Image Imagery In Situ Individual Lung Malignant Neoplasms Maps Methods Minority Mitosis Modeling Molecular Mus Natural Products Normal tissue morphology Pathway interactions Patients Play Population Pre-Clinical Model Preclinical Drug Evaluation Primary Neoplasm Process Proteins Reporter Resistance Role Sampling Signal Transduction Slice Specific qualifier value Staging Stem cells System Testing Therapeutic Time Transforming Growth Factor Beta 2 Transforming Growth Factor beta Transgenic Model Tumor Suppressor Proteins Video Microscopy Work Xenograft Model Xenograft procedure advanced disease base cancer stem cell cancer therapy carcinogenesis cell motility cell type functional genomics genetic regulatory protein in vivo malignant breast neoplasm mature animal member mouse model neutralizing antibody novel prevent promoter repository response self-renewal stem three dimensional cell culture tissue repair transcription factor transcriptome sequencing tumor tumor progression tumorigenesis

ATAC-seq Address Allografting Basement membrane Behavior Biological Biological Models Breast Cancer Model Cell Proliferation Cells Characteristics Clinical Collaborations Complex Development Disease Functional Imaging Genomic approach Goals Homeostasis Human Image Imagery In Situ Individual Lung Malignant Neoplasms Maps Methods Minority Mitosis Modeling Molecular Mus Natural Products Normal tissue morphology Pathway interactions Patients Play Population Pre-Clinical Model Preclinical Drug Evaluation Primary Neoplasm Process Proteins Reporter Resistance Role Sampling Signal Transduction Slice Specific qualifier value Staging Stem cells System Testing Therapeutic Time Transforming Growth Factor Beta 2 Transforming Growth Factor beta Transgenic Model Tumor Suppressor Proteins Video Microscopy Work Xenograft Model Xenograft procedure advanced disease base cancer stem cell cancer therapy carcinogenesis cell motility cell type functional genomics genetic regulatory protein in vivo malignant breast neoplasm mature animal member mouse model neutralizing antibody novel prevent promoter repository response self-renewal stem three dimensional cell culture tissue repair transcription factor transcriptome sequencing tumor tumor progression tumorigenesis

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项目摘要

In FY16, 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 3D culture systems and in lung slice models ex vivo. We are now addressing the effect of TGF-beta superfamily members on CSC proliferation, survival and motility in breast cancer model systems that show either tumor suppressor or pro-progression responses to TGF-beta. Using a xenograft breast cancer model in which TGF-beta functions as a tumor suppressor, we have identified a subpopulation of CSCs with high endogenous TGF-beta signaling that were intrinsically less proliferative than the other CSCs, suggesting that TGF-beta may be important in maintaining a fraction of CSCs in the quiescent state. Exogenous TGF-beta selectively inhibited asymmetric self-renewing mitoses in the CSC compartment, and specifically inhibited the invasion of CSCs through basement membrane. Consistent with these observations, in vivo treatment with TGF-beta neutralizing antibodies increased the CSC content of two xenograft models of well-differentiated breast cancer. Parallel work is ongoing in models of more aggressive advanced disease. We have a major collaboration with the Condeelis Lab at AECOM to use intravital videomicroscopy to observe the localization and behavior of CSCs in real time in the primary tumor and metastatic lungs of mouse xenograft or syngeneic allograft breast cancer models. 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. We are employing integrated genomic approaches including RNA-Seq and ATAC-Seq 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. Finally we are also using our cancer stem cell reporter to develop a medium-to-high throughput drug screen to test the NCI Natural Product Repository for novel agents that selectively inhibit the cancer stem cell.

在2016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，以实时和原位可视化该少数细胞种群。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3D培养系统和肺切片模型中进行单个CSC的扩展细胞命运映射的方法。现在，我们正在解决TGF-BETA超家族成员对乳腺癌模型系统中CSC增殖，生存和运动性的影响，这些系统显示肿瘤抑制剂或对TGF-beta的促进反应。使用异种移植乳腺癌模型，TGF-β充当肿瘤抑制剂，我们已经确定了具有高内源性TGF-β信号传导的CSC的亚人群，这些CSC在本质上比其他CSC的增殖性较小，这表明TGF-beta可能在维持奇异状态的CSC级数可能很重要。外源性TGF-β选择性地抑制了CSC室中的不对称自我更新有丝分裂，并特别抑制了通过基底膜侵袭CSC。与这些观察结果一致，用TGF-β中和抗体进行体内治疗增加了两种分化良好的乳腺癌的异种移植模型的CSC含量。在更具侵略性的晚期疾病模型中，并行工作正在进行中。我们与AECOM的Condeelis Lab进行了主要的合作，使用插入性视频显微镜观察小鼠异种移植物或同种异体同种异体乳腺癌模型的原发性肿瘤和转移性肺中CSC的定位和行为。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。我们正在采用包括RNA-SEQ和ATAC-SEQ在内的综合基因组方法来解决TGF-BETA在疾病过程的早期和晚期对TGF-BETA对CSC和非CSC隔室的差异作用的分子机制。最后，我们还使用癌症干细胞报告基因开发中高吞吐药筛查，以测试NCI天然产物存储库的新型药物，这些新型药物有选择地抑制癌症干细胞。