Studies of Antibody Diversity and Genome Stability: Regulation of Activation-Indu

抗体多样性和基因组稳定性的研究：激活诱导的调节

• 批准号: 8591624
• 负责人: Yee Ling Wu
• 金额: $ 0.79万
• 依托单位: MEDICAL RES COUNCIL LAB OF MOLEC BIOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-11-24 至 2013-11-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8591624
• 关键词: Address; Antibodies; Antibody Affinity; Antibody Diversity; Antigens; B-Cell Lymphomas; B-Lymphocytes; Biochemical; Cancer Etiology; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromosomal translocation; Complex; Coupled; DNA; DNA Sequence; DNA repair protein; Data; Deamination; Deoxycytidine; Disease; Embryo; Ensure; Enzyme Activation; Enzymes; Equilibrium; Event; Fellowship; Gene Conversion; Gene Expression; Generation of Antibody Diversity; Generations; Genes; Genetic Transcription; Genome; Genome Stability; Genomic Instability; Genomics; Goals; Hereditary Disease; Immune; Immunoglobulin Class Switching; Immunoglobulin Constant Region; Immunoglobulin Somatic Hypermutation; Immunoglobulin Switch Recombination; Immunoglobulins; Immunologics; Immunoprecipitation; Induced Mutation; Infection; Knockout Mice; Knowledge; Lesion; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microbe; Molecular; Molecular Profiling; Mus; Mutate; Mutation; Normal Cell; Organism; Outcome; Physiological; Process; Proteins; Proteomics; Proto-Oncogenes; RNA Splicing; Regulation; Specificity; Tertiary Protein Structure; Tumor Suppressor Genes; Uracil; Vertebrates; activation-induced cytidine deaminase; cis acting element; immune function; insight; leukemia/lymphoma; microbial; mutant; next generation sequencing; novel; novel therapeutics; pathogen; protein activation; research study

DESCRIPTION (provided by applicant): Activation-induced deaminase (AID) is an enzyme that mutates genes for antibodies in vertebrates. These AID-mediated mutations are necessary for an organism to generate specific antibodies to recognize and eradicate great varieties of microbial infections. However, if AID is not properly regulated, it can change DNA sequences of genes important for normal cell functions and cause diseases. Indeed, some lymphomas and leukemias are caused by abnormal activities of AID. How AID can change DNA sequences only in antibody genes but not in other genes is poorly understood. Recently, the catenin 2 like 1 protein (CTNNBL1) has been found to physically interact with AID. Little is known for the physiologic function of CTNNBL1. However, the interaction with CTNNBL1 is required for AID to change DNA sequences specifically in the antibody genes. In this study, a combination of high throughput approaches including next-generation sequencing and microarray analyses and targeted biochemical and immunologic analyses will be used to investigate how AID specifically mutate antibody genes, particularly what genomic regions are accessible to AID, both in the presence and in the absence of CTNNBL1. These experiments will also detect any alterations in transcription and splicing caused by the absence of CTNNBL1, and thereby shed lights on normal cellular functions of CTNNBL1. Mass spectrometry, mutagenic analyses and immunologic experiments will be performed to determine the molecular details of the interaction between AID and CTNNBL1 as well as to elucidate the mechanisms by which CTNNBL1 regulates the targeted activity of AID. It is anticipated that results from this study will be highly relevant for understanding the mechanisms underlying the generation of antibody diversity, and in the creation of new therapeutic strategies to minimize genomic disorders and cancers.

描述（由申请人提供）：激活诱导脱氨酶（AID）是一种使脊椎动物抗体基因突变的酶。这些 AID 介导的突变对于生物体产生特异性抗体来识别和根除多种微生物感染是必需的。然而，如果 AID 没有得到适当的调节，它可能会改变对正常细胞功能很重要的基因的 DNA 序列并引起疾病。事实上，一些淋巴瘤和白血病是由AID的异常活动引起的。 AID 如何仅改变抗体基因中的 DNA 序列而不改变其他基因中的 DNA 序列，人们对此知之甚少。最近，人们发现连环蛋白 2 样 1 蛋白 (CTNNBL1) 与 AID 存在物理相互作用。对于 CTNNBL1 的生理功能知之甚少。然而，AID 需要与 CTNNBL1 相互作用才能特异性改变抗体基因中的 DNA 序列。在本研究中，将结合使用包括下一代测序和微阵列分析以及靶向生化和免疫学分析在内的高通量方法来研究 AID 如何特异性突变抗体基因，特别是在存在和不存在 CTNNBL1 的情况下，AID 可访问哪些基因组区域。这些实验还将检测由于 CTNNBL1 缺失而引起的转录和剪接的任何改变，从而揭示 CTNNBL1 的正常细胞功能。将进行质谱、诱变分析和免疫学实验，以确定 AID 和 CTNNBL1 之间相互作用的分子细节，并阐明 CTNNBL1 调节 AID 靶向活性的机制。预计这项研究的结果将与了解抗体多样性产生的机制以及制定新的治疗策略以最大限度地减少基因组疾病和癌症高度相关。

项目成果

Intrinsic transcriptional heterogeneity in B cells controls early class switching to IgE.

• DOI: 10.1084/jem.20161056
• 发表时间: 2017-01
• 期刊: The Journal of experimental medicine
• 作者: Wu YL;Stubbington MJ;Daly M;Teichmann SA;Rada C
• 通讯作者: Rada C

DNA deaminases induce break-associated mutation showers with implication of APOBEC3B and 3A in breast cancer kataegis.

• DOI: 10.7554/elife.00534
• 发表时间: 2013-04-16
• 期刊: eLife
• 影响因子: 7.7
• 作者: Taylor BJ;Nik-Zainal S;Wu YL;Stebbings LA;Raine K;Campbell PJ;Rada C;Stratton MR;Neuberger MS
• 通讯作者: Neuberger MS