Lineage tracing and clonal analysis of oral cancer initiating cells

口腔癌起始细胞的谱系追踪和克隆分析

• 批准号: 8889248
• 负责人: Zhe Li
• 金额: $ 21.9万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8889248
• 关键词: Accounting; Adenoviruses; Affect; Behavior; Biological Assay; Breast Epithelial Cells; Cancer Model; Carcinogens; Cell Lineage; Cells; Chimeric Proteins; Collaborations; Daughter; Development; Diagnosis; Disease; Disease Resistance; Distant Metastasis; Ensure; Environment; Epithelial Cells; Erinaceidae; Estrogen Receptors; Exhibits; Fusion Protein Expression; Generations; Genetic Recombination; Goals; Habitats; Head and Neck Squamous Cell Carcinoma; Health; Human; Injection of therapeutic agent; Knowledge; Label; Life; Link; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of prostate; Mediating; Modeling; Molecular; Mus; Natural regeneration; Neoplasm Metastasis; Oral; Outcome; Pathway interactions; Pharmaceutical Preparations; Physiological; Population; Property; Recurrence; Reporter; Reporter Genes; Research; Resistance; Signal Pathway; Squamous Cell Neoplasms; Staging; Stem cells; Survival Rate; Tamoxifen; Testing; Therapeutic Intervention; Tissues; Translating; Transplantation; Tumor Expansion; Ursidae Family; Xenograft procedure; base; cancer cell; cancer stem cell; cancer type; cell type; chemotherapy; daughter cell; experience; genetic approach; in vitro Assay; in vivo; innovation; interest; leukemia; malignant breast neoplasm; malignant mouth neoplasm; medical schools; mouse model; mouth squamous cell carcinoma; new therapeutic target; notch protein; novel; novel strategies; novel therapeutic intervention; promoter; recombinase; reconstitution; research study; self-renewal; smoothened signaling pathway; stem cell fate; therapy resistant; tumor; tumor progression

DESCRIPTION (provided by applicant): Oral Squamous Cell Carcinomas (OSCCs) represent the vast majority cases of squamous cell carcinomas of the head and neck. Currently the 5-year survival rate for OSCCs remains at ~50-60%, largely due to their therapy resistance, recurrence, distant metastasis, and late diagnosis. It has been proposed that in many cancer types, including OSCCs, cancer cells are organized into aberrant cell hierarchies, including cancer stem cells [CSCs, also known as cancer initiating cells (CICs)] and their more differentiated daughter cancer cells. Several recent studies in murine cancer models have used lineage tracing to identify cancer cells that bear markers of tissue stem cells, and which give ris to long-lived clones that drive tumor expansion. This in vivo lineage tracing approach typically involves tamoxifen-induced transient activation of a Cre recombinase-estrogen receptor fusion protein (CreERT), which leads to activation of a conditional Cre reporter (e.g., the Rosa26-Stop-YFP conditional reporter). CreERT expression can be controlled by a cell type-specific promoter and CreERT-expressing cells and their daughter cells can be genetically labeled by the Cre reporter (e.g., YFP) upon Cre-mediated recombination (thus forming a cell lineage). Although these recent lineage tracing studies in murine models have provided compelling evidence to support the existence of CSCs in intact tumors, they give few clues about the molecular mechanisms required to ensure CSC self- renewal, nor about whether different signaling pathways might control distinct CSC fates. It is also unclear whether CSCs form a homogeneous population or whether different subpopulations of CSCs might exhibit different behaviors (e.g., differential survival) upon chemotherapy. In OSCCs, although it is thought that CSCs are responsible for their metastasis, recurrence and therapy resistance, the identity of OSCC CSCs remains elusive. In this R21 project, an innovative approach is proposed to study CSCs during OSCC development and upon therapy in a well-established carcinogen-induced murine OSCC models by a novel pathway-based in vivo lineage tracing strategy. This approach involves genetically marking subsets of cancer cells based on activation of a panel of pathways known to be involved in regulating stem cells (e.g., Axin2-CreERT or Gli1-CreERT for labeling Wnt or Hedgehog signaling-responsive cells, respectively) (Aim 1). By quantitatively comparing clonal outcomes between different pathway-labeled cell populations, one can reveal which pathways are active in CSCs and how cell fate choices are affected by pathway activity. In Aim 2, how oral cancer-initiating cells/CSCs in this murine model (i.e., defined based on their activit for stem cell-related pathway) respond to therapeutic interventions at the clonal level will be further determined by lineage tracing. The long-term goal of this project is to use this pathway-based lineage tracing approach to identify which pathways correlate with generation and regeneration (upon therapy) of CSCs or with therapeutic resistance, and through this to develop and test novel therapy to target OSCC CSCs.

描述（由申请方提供）：口腔鳞状细胞癌（OSCC）代表了头颈部鳞状细胞癌的绝大多数病例。目前，OSCC的5年生存率保持在约50- 60%，主要是由于其治疗抗性、复发、远处转移和晚期诊断。已经提出，在许多癌症类型中，包括OSCC，癌细胞被组织成异常的细胞层次，包括癌症干细胞[CSC，也称为癌症起始细胞（CIC）]及其更分化的子癌细胞。最近在小鼠癌症模型中的几项研究已经使用谱系追踪来识别携带组织干细胞标志物的癌细胞，这些癌细胞会导致长寿命的克隆，从而驱动肿瘤扩张。这种体内谱系追踪方法通常涉及他莫昔芬诱导的Cre重组酶-雌激素受体融合蛋白（CreERT）的瞬时激活，这导致条件性Cre报告基因（例如，Rosa 26-Stop-YFP条件性报道基因）。CreERT表达可由细胞类型特异性启动子控制，并且表达CreERT的细胞及其子代细胞可由Cre报告基因遗传标记（例如，在Cre介导的重组（从而形成细胞谱系）后，尽管这些最近在鼠模型中的谱系追踪研究提供了令人信服的证据来支持CSC在完整肿瘤中的存在，但它们几乎没有提供关于确保CSC自我更新所需的分子机制的线索，也没有提供关于不同信号传导途径是否可以控制不同CSC命运的线索。还不清楚CSC是否形成同质群体或CSC的不同亚群是否可能表现出不同的行为（例如，差异存活）。在OSCC中，尽管认为CSCs负责其转移、复发和治疗抗性，但OSCC CSCs的身份仍然难以捉摸。在这个R21项目中，提出了一种创新的方法来研究CSCs在OSCC的发展和治疗后，在一个完善的致癌物诱导的小鼠OSCC模型通过一种新的途径为基础的体内谱系追踪策略。这种方法涉及基于已知参与调节干细胞的一组途径的激活来遗传标记癌细胞的亚群（例如，Axin 2-CreERT或Gli 1-CreERT分别用于标记Wnt或Hedgehog信号传导应答细胞）（目的1）。通过定量比较不同途径标记的细胞群之间的克隆结果，可以揭示哪些途径在CSC中是活跃的以及细胞命运选择如何受到途径活性的影响。在目的2中，口腔癌起始细胞/CSC如何在该鼠模型（即，基于它们对干细胞相关途径的活性定义）在克隆水平上对治疗干预的响应将通过谱系追踪进一步确定。该项目的长期目标是使用这种基于途径的谱系追踪方法来确定哪些途径与CSC的生成和再生（治疗后）或治疗抗性相关，并通过此开发和测试靶向OSCC CSC的新疗法。