Mutation Analysis Of Selected Lymphoid Immune Disorders

该项目是为更好地描述和了解免疫缺陷而开展的一系列长期合作研究的延伸。目前正在利用荧光探针进行直接基因测序，对涉及共同γ链（X-SCID）和Fas （ALPS）基因的突变进行评估。这些研究继续在这两种疾病中确定了许多新的突变，这些数据已被输入NIH NHGRI网站，以支持这两种疾病。这始于对FAS编码基因（TNFRSF6）的广泛评估，确认了9个snp，并在评估的对照DNA样本中发现了两个新的snp。随后纳入了针对编码CD40L和NEMO基因的高IgM综合征患者的突变分析，后者侧重于高IgM综合征之外的许多临床表型，这为基因型-表型变异性提供了见解。为了处理NIAID目前的方案，增加了免疫缺陷相关突变的额外测序，重点是宿主防御缺陷与复发性感染，涉及机会性细胞内生物，包括编码干扰素γ受体1和2，IL-12P40和IL-12受体β 1基因的基因。在目前正在测序的所有基因中，已经发现了许多新的突变，在过去的财政年度中，还增加了一些其他基因，包括编码AIRE、ARTEMIS、BTK、FOXP3、ICOS、IL-7Ralpha、JAK3、mu重链、SAP、WASp的基因。最近增加的突变分析菜单代表了与NIAID的LHD和LCID研究人员的合作努力，包括在NIAID科学主任的资助下获得额外的毛细管测序仪。该计划已将评估的与原发性免疫缺陷相关的基因数量扩大到43个：AID、CD40L、FAS、FASLG、KRAS、NRAS、IFNGR1、IFNGR2、IL12RB1、IL12B、IL2RG、NEMO、AIRE、BTK、CASP8、CASP9、CASP10、DKC1、ELA2、FOXP3、ICOS、IKBA、IRAK4、ITK、IL2RA、JAK3、LIG4、MYD88、NBN、NHEJ1、PRF1、RAG1、RAG2、STAT1、STAT3、STIM1、STX11、UNC13D、WAS、XIAP、SAP/SH2D1A、TLR3、ORAI1。此外，我们现在正在研究NextGen方法，以筛选具有常见临床表型的疾病的特定免疫缺陷——这种方法可以证明提高在新患者中识别突变的速度，并降低评估的总体成本。与原发性免疫缺陷疾病相关的其他遗传缺陷已经得到证实，现在被纳入具有未知免疫疾病的CC患者的评估中。此外，Next Gen重测序平台Ion Torrent已被收购，目前正在验证用于筛选原发性免疫缺陷的150基因芯片。

This project represents an extension of a long-standing series of collaborative studies performed to better characterize and understand immune deficiency. Mutations involving the genes for the common gamma chain (X-SCID) and Fas (ALPS) are being evaluated using direct gene sequencing with fluorescent probes. These studies have continued to identify a number of new mutations in both diseases and these data have been entered into the NIH NHGRI web site supporting each of these two disorders. This begane with an extensive evaluation of the gene encoding FAS (TNFRSF6) that confirmed 9 SNPs and identified two new SNPs among the control DNA samples evaluated. This was followed by the inclusion of mutation analysis of patients with hyper IgM syndrome directed at the genes encoding CD40L and NEMO with the latter focusing on a host of clinical phenotypes beyond the hyper IgM syndrome that has provided insight into genotype-phenotype variability. Additional sequencing for immune deficiency associated mutations has been added to deal with current protocols within NIAID focused on host defense defects with recurrent infections involving opportunisitc intracellular organisms including genes encoding the interferon gamma receptor 1 and 2, the IL-12P40 and IL-12 receptor beta 1 genes. A host of new mutations have been identified among all genes that are now being sequenced and over the past fiscal year a number of additional genes have been added to the repertoire including genes encoding: AIRE, ARTEMIS, BTK, FOXP3, ICOS, IL-7Ralpha, JAK3, mu heavy chain,SAP, WASp. The most recent additions to the mutation analysis menu represent a collaborative effort with investigators from the LHD and LCID in NIAID including the acquisition of an additional capillary sequencer supported by funds from the Scientific Director of NIAID. This program has expanded the number of genes evaluated associated with primary immunodeficiencies to 43: AID, CD40L, FAS, FASLG, KRAS, NRAS, IFNGR1, IFNGR2, IL12RB1, IL12B, IL2RG, NEMO, AIRE, BTK, CASP8, CASP9, CASP10, DKC1, ELA2, FOXP3, ICOS, IKBA, IRAK4, ITK, IL2RA, JAK3, LIG4, MYD88, NBN, NHEJ1, PRF1, RAG1, RAG2, STAT1, STAT3, STIM1, STX11, UNC13D, WAS, XIAP, SAP/SH2D1A, TLR3, ORAI1. In addition, we are now working on a NextGen approach to screening for specific immune defects focusing on disorders with common clinical phenotypes - this approach can prove to increase the speed with which a mutation is identified in a new patient as well as decrease the overall cost for this evaluation. Additional genetic defects associated with primary immunodeficiency disorders have been validated and are now included in the evaluation of CC patients with unknown immune disorders. In addition, the Next Gen resequencing platform, Ion Torrent, has been acquired and is in the process of validating a 150 gene chip to screen for primary immunodeficiencies.