N-cadherin in Extraskeletal Osteolineage Cells Modulates Tumor Growth

骨骼外骨细胞中的 N-钙粘蛋白调节肿瘤生长

• 批准号: 10204978
• 负责人: Roberto Civitelli
• 金额: $ 39.89万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10204978
• 关键词: Ablation; Affect; Attention; Bone Marrow; Breast Cancer Cell; Cadherins; Calcium; Cause of Death; Cell Adhesion Molecules; Cell-Cell Adhesion; Cells; Cessation of life; Clinical; Clinical Pathology; Data; Development; Diphtheria Toxin; Engraftment; Environment; Excision; Fibroblasts; Foundations; Genes; Growth; Human; Injections; Internal Breast Prosthesis; Interruption; MAP Kinase Gene; Malignant Neoplasms; Metastatic Neoplasm to the Lung; Methods; Modification; Molecular; Mus; N-Cadherin; Neoplasm Metastasis; Osteolytic; Parabiosis; Pathway interactions; Phenotype; Population; Prevalence; Prognosis; Relapse; Reporting; Research; Resistance; Role; Signal Pathway; Signal Transduction; Site; Soil; Structure of parenchyma of lung; Testing; Translating; Tumor Tissue; Woman; base; bone; bone cell; cancer cell; cancer seeding; cell transformation; combat; epithelial to mesenchymal transition; expectation; human disease; in vivo; indexing; malignant breast neoplasm; mortality; neoplastic cell; novel marker; novel therapeutic intervention; osteogenic; p38 Mitogen Activated Protein Kinase; recruit; single-cell RNA sequencing; subcutaneous; transcriptomics; tumor; tumor growth; tumor microenvironment; tumor progression; tumorigenesis; tumorigenic

Abstract The host microenvironment is necessary for tumor growth and metastasis, and a major determinant of resistance to treatment and relapse. Expression of N-cadherin (Ncad), a calcium-dependent cell-cell adhesion molecule, in cancer associated fibroblasts (CAF) has been reported to favor tumor growth. Ncad is the main cadherin expressed in bone cells, where it functions in cell-cell adhesion, but also regulates signaling and differentiation. In preliminary studies we found that, contrary to expectations, ablation of the Ncad gene (Cdh2) in osteolineage cells – expressing the osteogenic marker, Osterix (Osx+) – does not affect bone engraftment of breast cancer cells; however, subcutaneous tumors grow faster and lung metastases develop earlier than in wild type littermates. We also find, unexpectedly, that Ncad is present in previously unrecognized Osx+ cells in extra-skeletal tumors. These cells have a transcriptomic profile more similar to osteogenic cells than to CAF, and favor tumor growth. Furthermore, Ncad in Osx+ cells down-regulates p38 responsive genes, a pro- tumorigenic pathway. In human breast cancer, Osx+ are an index of poor prognosis. These preliminary results demonstrate that Ncad in Osx+ cells is a negative regulator of cancer progression, an effect opposite to Ncad reported action in CAF. We contend that Ncad exerts multiple and even opposite actions on tumorigenesis depending on the cell context where it is expressed, via modulation of specific signaling pathways. Based on these preliminary data, our central hypothesis is that Ncad in pro-tumorigenic Osx+ cells restrains tumor growth by regulating signals that reprogram the tumor microenvironment. To test this hypothesis, we propose the following Specific Aims: Specific Aim 1 – Modulation of extra-skeletal tumor growth by Ncad in Osx+ cells; testing the hypothesis that Ncad in Osx+ cells restrains tumor growth; loss of Ncad in TAOC increases tumor growth and metastases in mice. Osx+ Ncad+ cells correlate with tumor grading in human breast cancer. Specific Aim 2 – Mechanisms of Ncad modulation of pro-tumorigenic signals in tumor-associate Osx+ cells; testing the hypothesis that Ncad in Osx+ cells is an upstream regulator of p38 and Pten signaling; loss of Ncad in Osx+ cells results in accentuated expression of p38-dependent pro- tumorigenic factors and decreased Pten dependent signals, leading to tumor microenvironment modification and enhanced tumorigenesis. We will use in vivo approaches, including diphtheria toxin-induced selective cell ablation, parabiosis, lineage tracking, as well as non-biased transcriptomic approaches (single cell RNAseq) to unlock the cellular and molecular mechanisms by which Ncad in extraskeletal Osx+ cells affects tumor growth and metastasis. We will also determine the clinical pathology correlates of Ncad expression in Osx+ cells in human tumors. Results of the proposed studies will lay the foundations for the development of new markers of tumor progression and/or new therapeutic strategies aimed at interrupting environmental support of cancer growth and metastasis by targeting specific cells in the tumor stroma.

摘要 宿主微环境是肿瘤生长和转移所必需的，并且是肿瘤生长和转移的主要决定因素。 对治疗的抵抗和复发。钙依赖性细胞间粘附分子N-cadherin（Ncad）的表达 据报道，在癌症相关成纤维细胞（CAF）中，这种分子有利于肿瘤生长。Ncad是主要的 钙粘蛋白在骨细胞中表达，在细胞间粘附中起作用，但也调节信号传导， 分化在初步研究中，我们发现，与预期相反，Ncad基因（Cdh 2）的切除， 在表达成骨标志物Osterix（Osx+）的骨系细胞中， 然而，皮下肿瘤生长得更快，肺转移发展得更早， 野生型同窝仔。我们还意外地发现，Ncad存在于以前未被识别的Osx+细胞中 在骨外肿瘤中。这些细胞具有与成骨细胞比与CAF更相似的转录组学特征， 有利于肿瘤生长此外，Osx+细胞中的Ncad下调了p38应答基因，这是一种促凋亡基因。 致瘤途径在人类乳腺癌中，Osx+是预后不良的指标。这些初步结果 表明Osx+细胞中的Ncad是癌症进展的负调节剂，与Ncad相反 在CAF中报告的行动。我们认为Ncad在肿瘤发生中发挥着多重甚至相反的作用 这取决于其表达的细胞环境，通过调节特定的信号传导途径。 基于这些初步数据，我们的中心假设是促肿瘤发生Osx+细胞中的Ncad 通过调节重新编程肿瘤微环境的信号来抑制肿瘤生长。为了验证这一 假设，我们提出了以下具体目标：具体目标1 -调节骨外肿瘤 Osx+细胞中Ncad的生长;检验Osx+细胞中Ncad抑制肿瘤生长的假设; TAOC中的Ncad增加小鼠中的肿瘤生长和转移。Osx+ Ncad+细胞与肿瘤相关 人类乳腺癌的分级特定目的2 -Ncad调节促肿瘤发生信号的机制 在肿瘤相关Osx+细胞中;检验Osx+细胞中的Ncad是p38的上游调节因子的假设 和Pten信号传导; Osx+细胞中Ncad的缺失导致p38依赖性蛋白原的表达增强。 致瘤因子和减少Pten依赖性信号，导致肿瘤微环境改变 和增强的肿瘤发生。我们将使用体内方法，包括白喉毒素诱导的选择性细胞 消融、联体共生、谱系追踪以及非偏倚转录组学方法（单细胞RNAseq）， 解开外骨骼肌Osx+细胞中Ncad影响肿瘤生长的细胞和分子机制 和转移。我们还将确定Ncad在Osx+细胞中表达的临床病理学相关性， 人类肿瘤这些研究结果将为开发新的遗传标记物奠定基础。 肿瘤进展和/或旨在中断癌症的环境支持的新治疗策略 通过靶向肿瘤间质中的特定细胞来抑制肿瘤生长和转移。