Molecular Pathways in T Cell Development and T-ALL

T 细胞发育和 T-ALL 的分子途径

• 批准号: 7780947
• 负责人: HARALD VON BOEHMER
• 金额: $ 21.23万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7780947
• 关键词: Acute; Address; Affect; Appearance; Blood; Cell Line; Cells; Characteristics; Collaborations; DNA Sequence Rearrangement; Data; Development; Disease; Disease Management; Epigenetic Process; Etiology; Event; Exhibits; Face; Funding; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genome Stability; Goals; Growth; Growth and Development function; Human; Insertional Mutagenesis; Karyotype; Lead; Ligation; Lymphoblastic Leukemia; Malignant - descriptor; Malignant Neoplasms; Mediating; MicroRNAs; Modality; Modeling; Molecular; Molecular Analysis; Molecular Target; Mus; Mutation; Non-Malignant; Normal Cell; Nude Mice; Oncogenes; Pathway interactions; Pattern; Pharmacotherapy; Play; RNA; Retroviral Vector; Retroviridae; Role; Signal Transduction; Site; Sorting - Cell Movement; Staging; T-Cell Development; T-Lymphocyte; Testing; Transcript; Transplantation; Tumor Stem Cells; cancer cell; cellular transduction; comparative; cyclin D3; design; genetic analysis; in vivo; insight; knock-down; mouse genome; neoplastic cell; notch protein; overexpression; retroviral transduction; tumor; tumor growth; tumor progression; tumorigenesis; tumorigenic; vector

In the past five years we have identified the developmental stage of Tcell differentiation at which malignant transformation caused by retroviral insertion of a vector containing ICN 1 becomes first apparent as the CD4-8+TCRalpha /beta minus stage that follows pre-TCR signaling. Consequently, the tumors are monoclonal with regard to TCR beta rearrangement but express diverse TCRalpha chains. The tumors exhibit a normal karyotype and genomic stability (verified by SKY and CGH),but are characterized by dysregulated expression of oncogenes and genes regulating survival and proliferation. Since sequencing has not revealed genetic instability and malignant cells can be detected early, 2-3 weeks after retroviral transduction , we will focus on insertional mutagenesis by the retroviral vector as a synergizing event since ICN1 overexpression alone does not result in tumors but in polyclonal, non-tumorigenic cells with slightly increased survival and proliferation when compared to phenotypically identical normal cells. We also established tht tumors exhibit an abnormal pattern of miRNA expression. We therefore will address the hypothesis that insertional mutagenesis and abnormally expressed miRNA contribute to the malignancy and will attempt to interfere with malignant growth by using antagomirs and mimics of miRNA and by knocking down abnormally overexpressed genes that are implicated in malignant growth. AIMI: Determine integration sites of retroviral vector in tumor and non-malignant ICN1 overexpressing cells to analyze contribution of insertional mutagenesis to tumor development. AIM2: Epigenetic analysis of T-ALL versus phenotypically similar but normal or ICN1 overexpressing nontumorigenic cells. AIM3: Contribution of miRNA to malignant transformation. AIM 4: Sh RNA mediated knockdown of genes specifically overexpressed in T-ALL and overexpression of genes specifically repressed in tumors.

在过去的五年中，我们已经确定了T细胞分化的发育阶段，在该阶段，由含有ICN 1的载体的逆转录病毒插入引起的恶性转化首先变得明显，作为前TCR信号传导之后的CD 4 -8+ TCR α/β负阶段。因此，肿瘤在TCR β重排方面是单克隆的，但表达不同的TCR α链。肿瘤表现出正常的核型和基因组稳定性（通过SKY和CGH验证），但其特征在于癌基因和调节存活和增殖的基因的表达失调。由于测序还没有揭示基因 不稳定性和恶性细胞可以在逆转录病毒转导后2-3周早期检测到，我们将集中于逆转录病毒载体的插入诱变作为协同事件，因为ICN 1单独过表达不导致肿瘤，但与表型相同的正常细胞相比，多克隆的非致瘤细胞具有略微增加的存活和增殖。我们还证实了肿瘤表现出异常的miRNA表达模式。因此，我们将阐述插入突变和异常表达的miRNA导致恶性肿瘤的假设，并试图干扰 通过使用miRNA的抑制剂和模拟物以及通过敲除与恶性生长有关的异常过度表达的基因来抑制恶性生长。 AIMI：确定逆转录病毒载体在肿瘤和非恶性ICN 1过表达细胞中的整合位点，以分析插入突变对肿瘤发展的贡献。 AIM 2：T-ALL与表型相似但正常或ICN 1过表达的非致瘤细胞的表观遗传学分析。 AIM 3：miRNA对恶性转化的贡献。 目的4：Sh RNA介导的T-ALL中特异性过表达基因的敲除和肿瘤中特异性抑制基因的过表达。