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 重组缺陷是免疫缺陷和自身免疫等疾病的基础 到癌症。人类 SCID 是由 T 细胞发育缺陷或 T 细胞和 B 细胞发育缺陷引起的。 后一种疾病称为 T'B'SCID，通常由 V(D)J 重组缺陷引起。无效的 RAG-1 或 RAG-2 突变是大约一半人类 TB'SCID 的基础。亚态RAG 突变还会导致 Omenn 综合征 (OS)，这是一种罕见的常染色体隐性遗传 SCID，通常表现为 具有自身免疫成分的“复杂”免疫缺陷。 RAG突变导致的机制 操作系统未知。该应用的主要目的是阐明正常的 RAG 功能和异常的 RAG 通过表征 RAG 突变小鼠的活动可能导致复杂的免疫缺陷和癌症 线。我们已经培育出缺乏“非核心”RAG1 和 RAG2 区域的小鼠，这些区域与 抑制转座和携带 RAG1 点突变（具有潜在连接缺陷）的小鼠 这可能会导致致癌易位。我们将继续对这些小鼠进行分析 产生额外的 RAG 敲入突变，包括几种提议允许 RAG 切割但受损的突变 加入。我们还将采用新型 Abelson 鼠白血病病毒 (A-MuLV) 转化的亲 B 细胞系 系统作为感兴趣突变的快速体内筛选并作为 RAG 机制分析的基础 蛋白质功能。人类 TB'SCID 的另一半具有正常的 RAG 蛋白；但显示 V(D)J 复合缺陷和电离辐射敏感性增加。该子集的缺陷是 编码 Artemis、XLF 和 DMAligase 4 非同源 DMA 末端连接的基因发生突变 因素。人类阿尔忒弥斯突变使该基因完全失活；而人类 Lig4 突变都是 亚态的。目前尚不清楚已知的人类 XLF 突变是否会完全使该基因失活。我们 之前生成了针对 Artemis 缺陷的小鼠模型，最近又生成了针对这两种缺陷的小鼠模型 XLF 和亚效型连接酶 4 缺陷。该提案的其他主要目标是描述这些 新模型来阐明这些 NHEJ 因子的正常功能，并再次确定突变如何 这些基因会导致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识别并消除大量的外来入侵者 其中通过剪切和粘贴基因盒形成无限数量的抗体基因。缺陷于 编码执行这种剪切和粘贴过程的蛋白质的人类基因会导致免疫缺陷 （对感染的易感性）。我们提出的研究旨在为此类人类疾病建立小鼠模型 更好地了解剪切和粘贴蛋白质的功能并开发更好的疗法。 儿童医院 卡普家族研究实验室 9楼 一黑扇圈 波士顿, 马萨诸塞州 02115 PHS 398（修订版 04/06）页面表格第 2 页 首席研究员/项目总监（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如下所示的格式提供所需信息。 从首席研究员开始。按字母顺序列出所有其他关键人员，姓氏在前。 名称 eRA Commons用户名 项目上的组织角色 Alt, Frederick W. FREDALT 儿童