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：
8019067
负责人：
CHENGMING ZHU
金额：
$ 28.38万
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 2013-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8019067
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 G1 Phase 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 Phosphorylation Phosphotransferases Play Population Process Proteins RAG1 gene Rag1 Mouse Reaction Receptor Gene Regulation Reporting Role S Phase Signal Transduction Site Source Staging Stem cells Synapses System T-Cell Receptor T-Cell Receptor Genes T-Cell Receptors delta-Chain T-Lymphocyte 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

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 G0/G1 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.

v（d）J重组，一种用于组装抗原的编程DNA重排过程淋巴细胞发育过程中的受体对基因组稳定性构成潜在威胁。长期目的是了解V（d）J重组和抑制的细胞周期控制的机理肿瘤发生在发育中的淋巴细胞中。关键假设是共济失调中突变的基因 Telangictasia（ATM）在V（d）J重组的细胞周期控制中起关键作用，缺乏此控制可能直接导致染色体易位和淋巴瘤。该假设基于1）ATM为物理位于V（d）J重组位点，2）ATM - / - 小鼠屈服于早期的胸腺淋巴瘤，染色体易位和3）消除V（d）J重组根除染色体易位在ATM - / - 小鼠中，并延迟了淋巴瘤的发作。我们的具体目的是： 1）确定ATM在包含V（d）J重组与G0/G1内与DNA断裂的作用细胞周期的阶段。我们将对不同细胞的细胞中的V（D）J特异性DNA断裂使用专门测定周期状态，从野生型和ATM - / - 小鼠以及T和B淋巴细胞发育期间纯化； 2）了解ATM在V（d）J重组期间包含DNA断裂的功能如何，我们将比较在野生型和ATM - / - 小鼠中的RAG核酸酶和组蛋白H2AX磷酸化的调节，这两个因素是生成，包含和/或有效连接DNA断裂的关键调节剂： 3）了解ATM缺乏小鼠的肿瘤发生，我们将完全表征ATM - / - 淋巴瘤建立细胞遗传学特征，易位连接并确定受影响的潜在肿瘤；我们还将设定建立小鼠胚胎干细胞（ESC）培养物，以诱导T和B分化淋巴细胞，表征V（d）J重组，以比较野生型和ATM-ESC的结果，并比较确定DNA断裂如何导致ATM - / - 的易位，并使T和B细胞都转移到免疫缺陷的宿主小鼠，以检查肿瘤性潜力。