Preventing Tumorigenesis in Developing Lymphocytes

预防淋巴细胞发育中的肿瘤发生

基本信息

批准号：
7595835
负责人：
CHENGMING ZHU
金额：
$ 29.26万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至 2012-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7595835
关键词：
ATM Signaling Pathway ATM deficient ATM function Address Adoptive Transfer Affect Antigen Receptors Apoptosis Ataxia Telangiectasia B-Cell Development B-Cell Lymphomas B-Lymphocytes Biological Assay Bone Marrow Bone Marrow Cells Breeding Cell Culture System Cell Culture Techniques Cell Cycle Cell Cycle Checkpoint Cell Cycle Regulation Cell Differentiation process Cell surface Cells Chromatin Chromosomal Breaks Chromosomal translocation Chromosomes, Human, Pair 14 Cleaved cell Coculture Techniques Code Complex Containment Cultured Cells Cytogenetics DNA DNA Damage DNA Double Strand Break DNA Repair DNA-dependent protein kinase Development Diagnostic Double Strand Break Repair Event Exhibits Failure Family Fetal Liver Gene Rearrangement Genes Genetic Recombination Genome Genome Stability Genomic Instability Genotype Goals Harvest IgK Immune system Immunoglobulin Class Switching Immunoglobulin Switch Recombination Immunotherapy Injection of therapeutic agent Joints Lead Length Lymphocyte Lymphoid Lymphoma Lymphomagenesis Maintenance Modeling Monitor Mus Mutate Nonhomologous DNA End Joining Oncogenes Oncogenic Pathway interactions Pattern Peptide Signal Sequences Phase Phosphorylation Phosphotransferases Play Population Process Proteins Reaction Receptor Gene Regulation Reporting Research Role Signal Transduction Site Source Staging Stem cells Synapses System T-Cell Receptor T-Cell Receptor Genes T-Cell Receptors delta-Chain T-Lymphocyte TP53 gene Testing Therapeutic Thymic Lymphoma Thymus Gland Time Tumor Antigens Ubiquitination V(D)J Recombination Wild Type Mouse Work base blastocyst chromatin immunoprecipitation design differentiated B cell ds-DNA embryonic stem cell human H2AX protein in vivo insight interest member mutant nuclease precursor cell prevent programs protein complex reconstitution repaired research study response thymocyte tumor tumorigenesis tumorigenic

项目摘要

DESCRIPTION (provided by applicant): Project Summary: V(D)J recombination, a programmed DNA rearrangement process for assembling antigen receptors during the development of lymphocytes, poses a potential threat to genomic stability. The long-term objective is to understand the mechanism of cell cycle control of V(D)J recombination and suppression of tumorigenesis in developing lymphocytes. The key hypothesis is that the gene mutated in ataxia telangiectasia (ATM) plays a key role in cell cycle control of V(D)J recombination, lack of this control may directly lead to chromosomal translocations and lymphoma. The hypothesis is based on 1) ATM is physically located at the V(D)J recombination sites, 2) ATM-/- mice succumb to early thymic lymphomas with chromosomal translocations and 3) eliminating V(D)J recombination eradicates chromosomal translocation in ATM-/- mice and delays the onset of lymphoma. Our specific aims are: 1) to establish the role of ATM in containing V(D)J recombination related DNA breaks within the G07G1 phase of the cell cycle. We will use specialized assay for V(D)J specific DNA breaks in cells at different cell cycle status, purified from wild-type and ATM-/- mice, and during T and B lymphocyte development; 2) to understand how ATM functions in containing DNA breaks during V(D)J recombination, we will compare regulation of Rag nuclease, and histone H2AX phosphorylation in wild-type and ATM-/- mice, both factors are key regulators in generating, containing and/or efficient joining of DNA breaks: 3) to understand the tumorigenesis in ATM deficient mice, we will fully characterize ATM-/- lymphomas, to establish cytogenetic profiles, translocation junctions and to determine the potential oncogenes affected; We will also set to establish a mouse embryonic stem cell ESC) culture, to induce differentiation into T and B lymphocytes, to characterize V(D)j recombination, to compare results from wild-type and ATM-ESC, and to determine how DNA breaks lead to translocation in ATM-/-, and to adoptive transfer both T and B cells into immunodeficient host mice, to examine the tumorigenic potential. Relevance: V(D)J recombination, if not properly controlled, may lead to develop of lymphoma. The study is designed to understand how genome instability occurs, and how this instability leads to tumor, and to understand how ATM functions in maintaining genome stability and suppressing tumorigenesis.

描述（由申请人提供）：项目摘要：V(D)J 重组是一种在淋巴细胞发育过程中组装抗原受体的程序化 DNA 重排过程，对基因组稳定性构成潜在威胁。长期目标是了解 V(D)J 重组的细胞周期控制和抑制发育中淋巴细胞肿瘤发生的机制。关键假设是共济失调毛细血管扩张症（ATM）中的突变基因在V（D）J重组的细胞周期控制中起着关键作用，缺乏这种控制可能直接导致染色体易位和淋巴瘤。该假设基于：1) ATM 物理上位于 V(D)J 重组位点，2) ATM-/- 小鼠因染色体易位而死于早期胸腺淋巴瘤，3) 消除 V(D)J 重组可根除染色体易位ATM-/- 小鼠并延迟淋巴瘤的发作。我们的具体目标是：1) 确定 ATM 在细胞周期 G07G1 期内包含 V(D)J 重组相关 DNA 断裂中的作用。我们将使用专门的检测方法来检测处于不同细胞周期状态的细胞中的 V(D)J 特异性 DNA 断裂，这些 DNA 断裂是从野生型和 ATM-/- 小鼠中纯化的，以及在 T 和 B 淋巴细胞发育过程中进行的； 2) 为了了解 ATM 如何在 V(D)J 重组过程中抑制 DNA 断裂，我们将比较野生型和 ATM-/- 小鼠中 Rag 核酸酶和组蛋白 H2AX 磷酸化的调节，这两个因素都是产生、包含和/或有效连接 DNA 断裂：3) 为了了解 ATM 缺陷小鼠的肿瘤发生，我们将充分表征 ATM-/- 淋巴瘤，建立细胞遗传学谱、易位连接和确定受影响的潜在致癌基因；我们还将建立小鼠胚胎干细胞（ESC）培养物，诱导分化为T和B淋巴细胞，表征V（D）j重组，比较野生型和ATM-ESC的结果，并确定DNA如何断裂导致 ATM-/- 易位，并将 T 细胞和 B 细胞过继转移到免疫缺陷宿主小鼠，以检查致瘤潜力。相关性：V(D)J 重组如果控制不当，可能会导致淋巴瘤的发生。该研究旨在了解基因组不稳定性是如何发生的，以及这种不稳定性如何导致肿瘤，并了解ATM如何在维持基因组稳定性和抑制肿瘤发生方面发挥作用。