Kouichi Ozaki

ITPKC and CASP3 polymorphisms and risks for IVIG unresponsiveness and coronary artery lesion formation in Kawasaki disease

Functional single-nucleotide polymorphisms (SNPs) in inositol 1,4,5-trisphosphate 3-kinase C (ITPKC) (rs28493229) and caspase-3 (CASP3) (rs113420705; formerly rs72689236) are associated with susceptibility to Kawasakis disease (KD). To evaluate the involvement of these 2 SNPs in the risk for intravenous immunoglobulin (IVIG) unresponsiveness, we investigated 204 Japanese KD patients who received a single IVIG dose of 2u2009gu2009kg−1 (n=70) or 1u2009gu2009kg−1 daily for 2 days (n=134). The susceptibility allele of both SNPs showed a trend of overrepresentation in IVIG non-responders and, in combined analysis of these SNPs, patients with at least 1 susceptible allele at both loci had a higher risk for IVIG unresponsiveness (P=0.0014). In 335 prospectively collected KD patients who were treated with IVIG (2u2009gu2009kg−1), this 2-locus model showed a more significant association with resistance to initial and additional IVIG (P=0.011) compared with individual SNPs. We observed a significant association when all KD patients with coronary artery lesions were analyzed with the 2-locus model (P=0.0031). Our findings strongly suggest the existence of genetic factors affecting patients’ responses to treatment and the risk for cardiac complications, and provide clues toward understanding the pathophysiology of KD inflammation.

Kerr effect in gas and its application to noncontact measurement of electric field

A noncontact measurement of electric field is proposed based on the Kerr effect in gases. With the Kerr sensing technique, the electric field can be measured without causing any field disturbance. Kerr constants of gases are, however, very small compared with those of liquids, and this ideal technique has not been applied to electric field measurement in gas. In this study, a system is developed for a highly sensitive measurement of the small phase retardation caused by the Kerr electro-optic effect in gases. An optical phase modulation technique is effectively adopted to improve the sensitivity of the system. With this optical system, (i) dc stressed uniform electric fields in sulfur hexafluoride, nitrogen, and carbon dioxide gases at the pressure of 0.2–0.5 MPa and (ii) ac stressed uniform electric fields in atmospheric air are measured.

Fine-scale SNP map of an 11-kb genomic region at 22q13.1 containing the galectin-1 gene

AbstractGalectins, a family of animal lectins that bind β-galactoside sugar chains, are thought to have a variety of intra- and extracellular functions. Through a case-control study in the Japanese population and subsequent functional analyses, we previously showed that a functional single nucleotide polymorphism (SNP) in the gene encoding galectin-2 (lectin, galactoside-binding, soluble, 2; LGALS2) is associated with susceptibility to myocardial infarction. As an addition to the genetic information about LGALS2 reported earlier, we provide here a map of polymorphic sites within an 11-kb region containing the gene encoding a closely related molecule, galectin-1 (lectin, galactoside-binding, soluble, 1; LGALS1). The map includes 14 SNPs and two genetic variations of other types detected in a Japanese population sample. Five of the 14 SNPs were not among those deposited in the dbSNP database in the US National Center for Biotechnology Information and appeared to be novel. We also analyzed linkage disequilibrium (LD) using the 12 SNPs in which minor-allele frequencies were >0.20. Investigation of haplotype structure within the LGALS1 locus revealed five common haplotypes covering more than 95% of the test population. One, or a pair, of the SNPs described here might serve as a “tag” for detecting associations between complex diseases and genes in this local segment of chromosome 22q13.1.

Isolation of a Novel Gene Showing Reduced Expression in Metastatic Colorectal Carcinoma Cell Lines and Carcinomas

To investigate genes involved in mctastatic stages of cancer, we analyzed expression of mRN As in three cell lines derived from murine colon adenocarcinoma 26 by means of a differential display method. Each of these lines exhibits distinct metastatic characteristics. Among many bands representing different expression patterns in the display, we confirmed by northern analysis that a gene corresponding to one amplified fragment, termed grm2 (gene related to metastasis 2), was expressed more abundantly in NL4, the derivative with the lowest metastatic potential, than in cell lines NL17, an experimentally metastatic derivative, and in NL22, a spontaneously metastatic derivative. Using thegrm.2 fragment as a probe, we isolated murine cDNA clones and subsequently human cDNA clones corresponding to the GRM2 gene. The human and mouse homologues both encode proteins of 600 amino‐acid residues, which show weak homologies to proteins belonging to the myosin family. When we examined the expression levels of this novel gene in human colon cancers and in corresponding metastatic foci, we found that in more than half of these tissues, expression was significantly reduced in association with malignant potential. Our resultsimply that in humans the GRM2 gene product may regulate the metastatic phenotype of some colorectal cancers.

High-density single-nucleotide polymorphism (SNP) map in the 96-kb region containing the entire human DiGeorge syndrome critical region 2 (DGCR2) gene at 22q11.2

AbstractWe constructed a high-density single-nucleotide polymorphism (SNP) map in the 96-kb region containing the DiGeorge syndrome critical region 2 (DGCR2) gene at chromosome 22q11.2, a human counterpart of mouse seizure-related gene SEZ-12. A total of 102 SNPs were isolated from the region by systematic screening among 48 Japanese individuals: 9 SNPs in the 5′ flanking region, 3 in the 5′ untranslated region, 2 in the coding regions, 77 in introns, 7 in the 3′ untranslated region, and 4 in the 3′ flanking region. By a comparison of our data with SNPs deposited in the dbSNP database in the National Center for Biotechnology Information, 80 SNPs (78.4%) were considered to be novel. The ratio of transition to transversion was 3.08:1. In addition, eight other types of genetic variations (one GA dinucleotide polymorphism and seven insertion/deletion polymorphisms) were discovered. The high-resolution map that we constructed will be a useful resource for analyzing gene scans of complex diseases mapped to this local segment on chromosome 22.

Association of NOD2 Mutations with Aggressive Periodontitis

Aggressive periodontitis (AgP) is characterized by rapid alveolar bone destruction and tooth loss early in life, and its etiology remains unclear. To explore the genetic risk factors of AgP, we performed genome-wide single-nucleotide polymorphism genotyping for identity-by-descent mapping and identified 32 distinct candidate loci, followed by whole exome sequencing with 2 pedigrees of AgP consisting of 3 cases and 1 control in 1 family and 2 sibling cases in the other. After variant filtering procedures and validation by targeted Sanger sequencing, we identified 2 missense mutations at 16q12 in NOD2 (p.Ala110Thr and p.Arg311Trp), which encodes nucleotide-binding oligomerization domain protein 2. We further examined 94 genetically unrelated AgP patients by targeted sequencing of NOD2 and found that 2 patients among them also carried the p.Arg311Trp variant. Furthermore, we found 3 additional missense mutations in this gene (p.His370Tyr, p.Arg459Cys, and p.Ala868Thr). These mutations either had not been previously observed or are extremely rare (frequency <0.001) in Asian populations. NOD2 plays a crucial role in innate immunity as an intracellular receptor initiating nuclear factor κB–dependent and mitogen-activated protein kinase–dependent gene transcription. These results demonstrated NOD2 as a novel gene involved in AgP.

Genetic Background of Myocardial Infarction

Myocardial infarction (MI) is a common disease whose pathogenesis includes genetic factors, and it is among the leading causes of death. In 2000, we started a genome-wide association study (GWAS) for MI using nearly 90,000 gene-based single-nucleotide polymorphisms (SNPs), and identified lymphotoxin-a (LTA) conferring risk of MI in Japanese population. This was the first GWAS that identified a disease susceptibility gene in the world. Moreover, through examining the LTA cascade by combination of biological and genetic analyses, we have identified additional MI-susceptible genes, LGALS2, PSMA6, and BRAP, so far. We present here our recent work focused on identification and functional analyses of genes that confer risk of MI.