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SCID由T细胞发育缺陷或T和B细胞发育缺陷引起。 后一种疾病称为T 'B'SCID，通常由V（D）J重组缺陷引起。Null RAG-1或RAG-2中的突变是约一半人TB'SCID的基础。亚形RAG 突变也会导致Omenn综合征（OS），这是一种罕见的常染色体隐性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 OS是未知的。本申请的主要目的是阐明正常RAG功能和异常RAG 通过表征RAG突变小鼠可能导致复杂免疫缺陷和癌症的活性 线我们已经产生了缺乏“非核心”RAG 1和RAG 2区域的小鼠， 抑制转座和携带具有潜在连接缺陷的RAG 1点突变的小鼠 可能导致致癌基因易位我们将继续分析这些小鼠， 产生额外的RAG敲入突变，包括几个建议允许RAG切割，但受损 加入我们还将采用一种新的Abelson鼠白血病病毒（A-MuLV）转化的pro-B细胞系 系统作为快速体内筛选感兴趣突变和作为RAG机制分析基础 蛋白质功能另一半人TB的SCID具有正常的RAG蛋白;但显示V（D）J 复合缺陷和增加的电离辐射敏感性。这个子集的缺陷是 编码Artemis、XLF和DMALigase 4非同源DNA末端连接的基因突变 因素人类Artemis突变完全破坏了该基因;而人类Lig 4突变则完全破坏了该基因。 亚形的目前尚不清楚已知的人类XLF突变是否完全消除了该基因。我们 先前生成的Artemis缺陷小鼠模型和最近生成的Artemis缺陷小鼠模型 XLF和亚型连接酶4缺乏症。该提案的其他主要目的是描述这些 新的模型来阐明这些NHEJ因子的正常功能，并再次确定突变是如何发生的。 这些基因导致各种人类免疫缺陷综合征并可能导致癌症。 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 人类基因编码的蛋白质进行这一剪切和粘贴过程造成免疫缺陷 （易感染）。我们提出的研究旨在为这些人类疾病建立小鼠模型 以更好地了解剪切和粘贴蛋白的功能，并开发更好的治疗方法。 儿童医院 卡普家族研究实验室 9楼 一个黑扇圈 Boston，MA PHS 398（Rev. 04/06）Page Form第2页 首席研究员/项目主任（最后，第一，中间）：TeitlOrst，ComeliS P.I Alt，Frederick W. 关键人员。参见说明。根据需要使用续页，以下列格式提供所需信息。 从主要研究者开始。按字母顺序列出所有其他关键人员，姓在前。 名称eRA公用用户名组织项目角色 弗雷德里克·阿尔特FREDALT儿童