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TCELL分化的发展阶段，在这种阶段中，由于CD4-8+tcralpha /beta sinus阶段，逆转录病毒引起的逆转录病毒插入引起的逆转转化阶段是很明显的。因此，肿瘤在TCRβ重排方面是单克隆的，但表达了多种tcralpha链。肿瘤表现出正常的核型和基因组稳定性（由Sky和CGH验证），但其特征是致癌基因的表达和调节存活率和增殖的基因的表达失调。由于测序尚未揭示遗传 逆转录病毒后2-3周可以早早检测到不稳定性和恶性细胞，我们将专注于逆转录病毒载体作为一种协同事件的插入诱变，因为仅ICN1过度表达并不能导致多克隆，非肿瘤细胞在与现场型型的生存率和繁殖时略有增加。我们还建立了肿瘤表现出miRNA表达的异常模式。因此，我们将解决以下假设：插入诱变和异常表达的miRNA有助于恶性肿瘤，并将尝试干预 通过使用Antagomirs和Mimim的MiRNA以及击倒与恶性生长有关的异常表达基因，通过使用恶性生长。 AIMI：确定肿瘤和非恶性ICN1过表达细胞中逆转录病毒载体的整合位点，以分析插入诱变对肿瘤发育的贡献。 AIM2：T-ALL与表型相似但正常或ICN1过表达非瘤细胞的表观遗传分析。 AIM3：miRNA对恶性转化的贡献。 AIM 4：SH RNA介导的基因的敲低在T-ALL和特异性抑制肿瘤中特异性过表达的基因的敲低。