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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：口腔鳞状细胞癌（OSCC）代表头颈部鳞状细胞癌的绝大多数病例。目前，口腔鳞癌的 5 年生存率仍保持在 50-60% 左右，这主要是由于其治疗耐药、复发、远处转移和诊断较晚。有人提出，在包括 OSCC 在内的许多癌症类型中，癌细胞被组织成异常细胞层次结构，包括癌症干细胞 [CSC，也称为癌症起始细胞 (CIC)] 及其分化程度更高的子癌细胞。最近对小鼠癌症模型的几项研究使用谱系追踪来识别带有组织干细胞标记的癌细胞，这些细胞会产生驱动肿瘤扩张的长寿命克隆。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，从而导致条件性 Cre 报告基因（例如 Rosa26-Stop-YFP 条件报告基因）的激活。 CreERT 表达可以由细胞类型特异性启动子控制，表达 CreERT 的细胞及其子细胞可以在 Cre 介导的重组后由 Cre 报告基因（例如 YFP）进行基因标记（从而形成细胞谱系）。尽管这些最近在小鼠模型中进行的谱系追踪研究提供了令人信服的证据来支持完整肿瘤中 CSC 的存在，但它们几乎没有提供有关确保 CSC 自我更新所需的分子机制的线索，也没有提供有关不同信号通路是否可能控制不同 CSC 命运的线索。。目前还不清楚 CSC 是否形成同质群体，或者不同的 CSC 亚群是否可能在化疗后表现出不同的行为（例如，生存差异）。在 OSCC 中，尽管人们认为 CSC 是造成其转移、复发和治疗耐药的原因，但 OSCC CSC 的身份仍然难以捉摸。在这个 R21 项目中，提出了一种创新方法，通过基于新途径的体内谱系追踪策略，在 OSCC 发育过程中以及在完善的致癌物诱导的小鼠 OSCC 模型中进行治疗后研究 CSC。该方法涉及基于已知参与调节干细胞的一组通路的激活对癌细胞子集进行遗传标记（例如，Axin2-CreERT 或 Gli1-CreERT 分别用于标记 Wnt 或 Hedgehog 信号响应细胞）（目标 1））。通过定量比较不同通路标记细胞群之间的克隆结果，我们可以揭示哪些通路在 CSC 中处于活跃状态，以及细胞命运选择如何受到通路活性的影响。在目标 2 中，该小鼠模型中的口腔癌起始细胞/CSC（即，根据其干细胞相关途径的活性定义）如何响应克隆水平的治疗干预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通路的谱系追踪方法来确定哪些通路与 CSC 的生成和再生（治疗后）或治疗耐药性相关，并通过此开发和测试针对 OSCC 的新疗法CSC。