Genetic disorders of immunodeficiency and inflammatory disease

免疫缺陷和炎症性疾病的遗传性疾病

• 批准号: 8939435
• 负责人: Richard M Siegel
• 金额: $ 50.59万
• 依托单位: NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8939435
• 关键词: Affect; Arthritis; Biochemical; Biological Assay; Blood Cells; Candidate Disease Gene; Cell Line; Cell Nucleus; Cells; Characteristics; Clinical; Code; Complex; Defect; Development; Disease; Engineering; Evaluation; Event; Family; Family member; Genes; Genetic Transcription; Genome; Hereditary Disease; Human; IKK alpha; Immune response; Immune system; Immunologic Deficiency Syndromes; Individual; Inflammatory; Interferons; Laboratories; Laboratory Finding; Lead; Lymphoid; Mesenchymal Stem Cells; Molecular; Monitor; Mutation; NF-kappa B; NFKB Signaling Pathway; Organism; Pathway interactions; Patients; Phenocopy; Phenotype; Phosphorylation; Population; Protein Isoforms; Proteins; Protocols documentation; RNA Splicing; Rare Diseases; Recruitment Activity; Regulation; Role; Secondary to; Serine; Serum; Signal Transduction; Signaling Protein; Structure; Syndrome; System; T-Lymphocyte; TBK1 gene; Techniques; Therapeutic; Ubiquitination; United States National Institutes of Health; Vasculitis; Work; base; clinical phenotype; congenital immunodeficiency; conserved helix-loop-helix ubiquitous kinase; disease characteristic; exome sequencing; genetic regulatory protein; improved; induced pluripotent stem cell; insight; monocyte; mutant; receptor; reconstitution; response

NEMO, a non-catalytically active component of the IkappaB alpha kinase (IKK) complex, is required for IKK enzymatic activity and localization. If NEMO is defective, NF-kB transcription factor family members do not translocate to the nucleus to regulate gene transcription, and normal development of the organism, including the immune system, does not occur. In addition to these developmental defects, the immune system does not function correctly if NEMO is defective. Individuals with specific mutations that result in NEMO immunodeficiency and inflammatory disease are investigated using molecular, cellular and biochemical assays/techniques. Because NEMO syndrome is associated with defects in development of ectodermal structures in addition to secondary lymphoid structures in association with certain mutations, we seek to understand the function of NEMO in both of these roles in addition to its role in hematopoietically derived cells. In addition to clinical and laboratory evaluation of human patients, these studies are conducted using patient-derived peripheral blood cells, induced pluripotent stem cells and their derivatives, such as mesenchymal stem cells. A reconstitution system utilizing NEMO-deficient Jurkat T cells and THP-1 monocytes is also employed in order to study protein interactions biochemically. NEMO is required for canonical IKK activation, and in addition to IKK, recruits other signaling proteins such as TBK1 to activated receptors in order to induce the type I IFN response. The regulation of NF-kB activation by NEMO is complex: NEMO splice isoforms exist, and NEMO is post-translationally modified by serine phosphorylation, non-degradative ubiquitination, and sumoylation. We evaluate patients at the NIH Clinical Center to be able to comprehensively characterize the spectrum of phenotypes, and are working to develop protocols to offer advanced treatment based on our clinical and laboratory findings. In a similar fashion, we are studying individuals with NEMO-Like Syndrome who have suspected monogenic diseases which phenocopy NEMO syndrome in some aspects. We hypothesize that these individuals will have defects in signaling proteins or other regulatory proteins that interact in the NF-kB signaling pathway. Our general approach has been to discover candidate gene genes that may underly disease by sequencing exomes or genomes of affected individuals and their families. Candidate genes identified are evaluated using assays that interrogate different checkpoints along the NF-kB activation pathway. Insights into the role of proteins gained from study of patients with rare diseases will be applied to evaluation of the regulation of the immune system and immune response as it functions in the population, and in common inflammatory diseases such as arthritis and vasculitis.

Nemo是IKAPABα激酶（IKK）复合物的非催化活性成分，是IKK酶活性和定位所必需的。如果NEMO有缺陷，则NF-KB转录因子家族成员不会转移到细胞核以调节基因转录，并​​且包括免疫系统在内的生物体的正常发育不会发生。除了这些发育缺陷外，如果NEMO有缺陷，免疫系统也无法正常运行。使用分子，细胞和生化测定/技术研究了具有导致NEMO免疫缺陷和炎症性疾病的特定突变的个体。由于Nemo综合征与与某些突变相关的继发性淋巴样结构外，与外胚层结构的发育缺陷有关，因此我们试图了解Nemo在这两个角色中的功能，除了其在造血细胞中的作用外。除了对人类患者的临床和实验室评估外，这些研究还使用患者来源的外围血细胞，诱导多能干细胞及其衍生物（例如间充质干细胞）进行。 还采用了一种利用难题的Jurkat T细胞和THP-1单核细胞的重构系统，以便在生化上研究蛋白质相互作用。 NEMO是典型IKK激活所必需的，除了IKK外，还需要将其他信号蛋白（例如TBK1）募集到活化受体中，以诱导I型IFN响应。 NEMO对NF-KB激活的调节很复杂：存在NEMO剪接同工型，而NeMo是通过丝氨酸磷酸化，非降解性泛素化和Sumoylation在后翻译后修饰的。我们评估NIH临床中心的患者，以便能够全面地表征表型的光谱，并正在努力根据我们的临床和实验室发现，以提供方案以提供高级治疗。以类似的方式，我们正在研究患有线性综合征的个体，他们怀疑某些方面的表征疾病是单基因疾病。我们假设这些个体在信号蛋白或其他在NF-KB信号通路中相互作用的调节蛋白方面存在缺陷。我们的一般方法是发现候选基因可能通过对受影响个体及其家人的外体或基因组进行测序的基因。使用沿NF-KB激活途径询问不同检查点的测定法对鉴定的候选基因进行了评估。从罕见疾病患者研究中获得的蛋白质作用的见解将应用于评估免疫系统的调节和免疫反应，因为它在人群中起作用，以及在常见的炎性疾病中，例如关节炎和血管炎。