Mouse models of severe combined immunodeficiencies

严重联合免疫缺陷小鼠模型

• 批准号: 7614101
• 负责人: Frederick W. Alt
• 金额: $ 47.73万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-05 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614101
• 关键词: Abelson murine leukemia virus; Affect; Age; Antibodies; B-Cell Development; B-Lymphocytes; Boston; Cell Count; Cell Line; Cells; Child; Code; Collaborations; Complex; DNA; DNA Damage; DNA ligase IV; Data; Defect; Development; Disease; Embryo; Event; Family Research; Fibroblasts; Floor; Funding; Genes; Genomic Instability; Genomics; Goals; Human; Human Resources; Immune; Immune response; Immune system; Immunity; Immunodeficiency and Cancer; Immunoglobulin Class Switching; Immunoglobulin Switch Recombination; Immunologic Deficiency Syndromes; Incidence; Infection; Instruction; Ionizing radiation; Knock-in Mouse; LIG4 gene; Laboratories; Last Name; Lead; Ligase; Lymphocyte; Malignant Neoplasms; Mature B-Lymphocyte; Modeling; Mus; Mutant Strains Mice; Mutation; Names; Neurologic; Neurons; Oncogenic; Paste substance; Pathway interactions; Patients; Pediatric Hospitals; Phenotype; Point Mutation; Predisposition; Principal Investigator; Process; Proteins; Radiation Tolerance; Radio; Research; Research Personnel; Role; Severe Combined Immunodeficiency; Staging; Stem cells; Syndrome; System; T-Cell Development; T-Cell Leukemia; T-Lymphocyte; Tumor Suppressor Proteins; V(D)J Recombination; artemis; base; cell type; embryonic stem cell; human disease; immunodeficient mouse model; in vivo; interest; lymphoid neoplasm; mouse model; mutant; novel; null mutation; programs; protein function; repaired; response; tumor; tumorigenesis

V(D)J recombination defects underlie diseases ranging from immunodeficiency and auto-immunity to cancer. Human SCIDs result either from defects in T cell development or both T and B cell development. The latter disease, referred to as T'B'SCID, generally results from V(D)J recombination defects. Null mutations in RAG-1 or RAG-2 are the basis for about half of the human TB'SCIDs. Hypomorphic RAG mutations also cause Omenn syndrome (OS), a rare autosomal recessive SCID that often presents as a "complex" immunodeficiency with an auto-immune component. Mechanisms by which RAG mutations lead to OS are unknown. A major aim of this application is to elucidate normal RAG functions and aberrant RAG activities that could lead to complex immunodeficiency and cancer by characterizing RAG mutant mouse lines. We have generated mice that lack both the "non-core" RAG1 and RAG2 regions implicated in suppressing transpositions and a mouse carrying a RAG1 point mutation that has a potential joining defect that may predispose to oncogenic translocations. We will continue analyses of these mice and also generate additional RAG knock-in mutations including several proposed to allow RAG cutting but impaired joining. We also will employ a novel Abelson Murine Leukemia virus (A-MuLV)-transformed pro-B cell line system as a rapid in vivo screen for mutations of interest and as a basis for mechanistic analyses of RAG protein functions. The other half of human TB'SCIDs have normal RAG proteins; but show V(D)J recombination defects and increased ionizing radiation-sensitivity. The defects underlying this subset are mutations in genes that encode the Artemis, XLF and DMALigase 4 non-homologous DMA end-joining factors. Human Artemis mutations completely inactivate the gene; whereas human Lig4 mutations are all hypomorphic. It is not yet clear whether known human XLF mutations completely inactivate the gene. We previously generated a mouse model for Artemis deficiency and recently generated mouse models for both XLF and hypomorphic Ligase 4 deficiency. The other major aims of this proposal are to characterize these new models to elucidate normal functions of these NHEJ factors and, again, to determine how mutations in these genes contribute to various human immunodeficiency syndromes and potentially to cancer. Our immune system recognizes and eliminates a vast array of foreign invaders based on a process in which limitless numbers of antibody genes are formed by cutting and pasting gene cassettes. Defects in human genes that code for proteins that carry out this cutting and pasting process cause immunodeficiency (susceptibility to infection). Our proposed research seeks to make mouse models for such human diseases to better understand the functions of the cutting and pasting proteins and to develop better therapies. Children's Hospital Karp Family Research Laboratories 9th Floor One Blackfan Circle Boston, MA 02115 PHS 398 (Rev. 04/06) Page Form Page 2 Principal Investigator/Program Director (Last, First, Middle): TeitlOrst, ComeliS P.I Alt, Frederick W. KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shownbelow. Start with Principal Investigator(s). List all other key personnel in alphabetical order, last name first. Name eRA CommonsUser Name Organization Role onProject Alt, Frederick W. FREDALT Children'

v（d）J重组缺陷是从免疫缺陷和自身免疫性等疾病的基础 癌症。人类SCID是由T细胞发育缺陷或T和B细胞发育的缺陷所致。 后一种疾病，称为T'b'scid，通常是由V（d）J重组缺陷引起的。无效的 RAG-1或RAG-2中的突变是大约一半人类TB'SCID的基础。肌片 突变还引起Omenn综合征（OS），这是一种罕见的常染色体隐性SCID，通常以 具有自动免疫成分的“复杂”免疫缺陷。抹布突变引导的机制 到OS是未知的。该应用的主要目的是阐明正常的抹布功能和异常抹布 通过表征抹布突变小鼠来导致复杂的免疫缺陷和癌症的活动 线。我们已经产生了缺乏涉及涉及的“非核” rag1和rag2区域的小鼠 抑制具有潜在连接缺陷的RAG1点突变的换位和携带RAG1点突变的小鼠 这可能易于致癌易位。我们将继续分析这些小鼠以及 产生额外的抹布敲入突变，包括提出的几种允许切割但受损的抹布 加入。我们还将采用一种新型的Abelson Murine白血病病毒（A-Mulv）转换的Pro-B细胞系 系统作为感兴趣的突变的快速体内筛选，也是抹布的机械分析的基础 蛋白质功能。人类TB'SCID的另一半具有正常的抹布蛋白。但是显示V（d）j 重组缺陷并增加电离辐射敏感性。该子集的基础缺陷是 编码Artemis，XLF和Dmaligase 4非同源DMA端连接的基因突变 因素。人artemis突变完全使基因失活；而人lig4突变都是 型肌。尚不清楚已知的人XLF突变是否完全使基因失活。我们 先前生成了用于Artemis缺乏症的鼠标模型，并为两者都生成了鼠标模型 XLF和肌电连接酶4缺乏症。该提议的另一个主要目的是将这些特征描述 阐明这些NHEJ因子的正常功能的新模型，并再次确定突变如何 这些基因有助于各种人类免疫缺陷综合症，并可能导致癌症。 我们的免疫系统根据一个过程承认并消除了大量外国入侵者 其中通过切割和粘贴基因盒形成了无限的抗体基因。缺陷 人类基因代码为执行这种切割和粘贴过程的蛋白质代码引起免疫缺陷 （感染的敏感性）。我们拟议的研究旨在为这种人类疾病制作鼠标模型 更好地了解切割和粘贴蛋白质的功能并开发出更好的疗法。 儿童医院 KARP家庭研究实验室 9楼 一个黑凡圈 波士顿，马萨诸塞州02115 PHS 398（修订版04/06）页面2页 首席调查员/计划主管（最后，第一，中间）：Teitlorst，Comelis P.I Alt，Frederick W. 关键人员。请参阅说明。根据需要使用延续页面，以显示的格式显示所需的信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，首先姓氏。 名称ERA COMMONSUSER名称组织角色OnProject alt，弗雷德里克·W·弗雷达特（Frederick W. Fredalt）儿童