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Numerous variants of unknown significance (VUSs) exist in hereditary breast and ovarian cancers. Although multiple methods have been developed to assess the significance of BRCA1/2variants, functional discrepancies among these approaches remain. Therefore, a comprehensive functional evaluation system for these variants should be established. We performed conventional homologous recombination (HR) assays for 50 BRCA1and 108 BRCA2VUSs and complementarily predicted VUSs using a statistical logistic regression prediction model that integrated six in silico functional prediction tools. BRCA1/2VUSs were classified according to the results of the integrative in vitro and in silico analyses. Using HR assays, we identified 10 BRCA1and 4 BRCA2VUSs as low-functional pathogenic variants. For in silico prediction, the statistical prediction model showed high accuracy for both BRCA1and BRCA2compared with each in silico prediction tool individually and predicted nine BRCA1and seven BRCA2variants to be pathogenic. Integrative functional evaluation in this study and the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG/AMP) guidelines strongly suggested that seven BRCA1variants (p.Glu272Gly, p.Lys1095Glu, p.Val1653Leu, p.Thr1681Pro, p.Phe1761Val, p.Thr1773Ile, and p.Gly1803Ser) and four BRCA2variants (p.Trp31Gly, p.Ser2616Phe, p.Tyr2660Cys, and p.Leu2792Arg) were pathogenic. This study demonstrates that integrative evaluation using conventional HR assays and optimized in silico prediction comprehensively classified the significance of BRCAVUSs for future clinical applications.

We previously reported that L63X and Q934X are BRCA1common founder variants in Japan. So far, there have been no reports of a correlation between such BRCAcommon variants and the risk of BRCA-related cancers. In this analysis, we investigated the correlation between the risk of ovarian cancer (OC) and BRCArecurrent pathogenic variants. We examined the database of the Japanese organization of hereditary breast and ovarian cancer. The database contained 3517 probands who underwent BRCAgenetic testing. Among them, 11.1% (392/3517) had germline BRCA1pathogenic variant, and 8.3% (293/3517) had BRCA2pathogenic variant. We calculated the OC prevalence, breast cancer (BC) prevalence, and the ratio of OC to BC within second-degree relatives. The ratio of OC to BC in Q934X family members was significantly higher than that in the overall BRCA1family members (0.80 vs.0.52: p= 0.038), and the ratio in STOP799 was 0.42, which was relatively lower than the overall BRCA1value. Both Q934X and STOP799 are located in the ovarian cancer cluster region (OCCR), however there seems to be a difference in the risk of OC. R2318X family members had a significant higher ratio of OC to BC at 0.32 than the overall BRCA2value of 0.13 (p= 0.012). R2318X is known to be located in the OCCR. This is the first report to investigate the correlation between BRCArecurrent variants and the risk of OC in Japan. The family members of probands with Q934X or R2318X have a higher risk of OC than that with other BRCAvariants.

This study aimed to clarify the breast cancer prognosis in Japanese patients with BRCA1/2pathogenic variant. We analyzed 2235 women with breast cancer who underwent BRCA1/2genetic testing between 1996 and 2018 using data from the Japanese hereditary breast and ovarian cancer syndrome registry. The cumulative risk for contralateral and ipsilateral breast cancers and time to death since the first breast cancer were stratified based on the BRCA1/2variant status. The median follow-up was 3.0 years (0.1–34.1 years) after the first breast cancer. The annual average risks of contralateral breast cancer in BRCA1and BRCA2and non-BRCA1/2pathogenic variant carriers were 4.0%, 2.9%, and 1.9%, respectively (P= 0.001). The annual average risks of ipsilateral breast cancer in the three groups were 2.7%, 1.4%, and 1.1%, respectively (P= 0.06). BRCA1pathogenic variant carriers had significantly higher risks of contralateral (hazard ratio 1.91, P< 0.001) and ipsilateral (hazard ratio 2.00, P= 0.02) breast cancers than non-BRCA1/2pathogenic variant carriers. The time to death by the BRCA1/2variant status was not significantly difference (P= 0.28). The prognosis of breast cancer patients who received standard treatment was comparable regardless of the BRCA1/2variant status.

BRCA1/2pathogenic variant prevalence in Japanese breast cancer is unclear. Here, we analyzed BRCA1/2pathogenic variant prevalence with a particular focus on age factors, using the Japanese HBOC consortium database. All registered subjects were Japanese individuals who underwent BRCA1/2genetic testing from January 1996 to July 2017 according to the Japanese HBOC consortium database. Cases were extracted and analyzed for each evaluation item. Overall BRCA1and BRCA2pathogenic variant prevalence was 11.2% and 9.0% in the cohort of 2366 proband patients, respectively. The age at onset of breast cancer for patients with BRCA1/2pathogenic variants was significantly lower than that for patients without a BRCA1/2pathogenic variant. In both BRCA1/2patients, ages at onset were not statistically significantly different between two subtype groups (ER-positive vs. TNBC). We analyzed the BRCA1/2pathogenic variant prevalence among age groups in patients with no family history of breast or ovarian cancer. In the TNBC group, the rate of genetic variants was more frequent among younger patients. Our results demonstrated that early breast cancer onset is associated with a BRCA1/2pathogenic variant in the Japanese population. Younger TNBC patients were more likely to have a BRCA1/2pathogenic variant irrespective of a family history of breast or ovarian cancer.

Lynch syndrome is a hereditary disease characterized by an increased risk of colorectal and other cancers. Germline variants in the mismatch repair (MMR) genes are responsible for this disease. Previously, we screened the MMR genes in colorectal cancer patients who fulfilled modified Amsterdam II criteria, and multiplex ligation-dependent probe amplification (MPLA) identified 11 structural variants (SVs) of MLH1and MSH2in 17 patients. In this study, we have tested the efficacy of long read-sequencing coupled with target enrichment for the determination of SVs and their breakpoints. DNA was captured by array probes designed to hybridize with target regions including four MMR genes and then sequenced using MinION, a nanopore sequencing platform. Approximately, 1000-fold coverage was obtained in the target regions compared with other regions. Application of this system to four test cases among the 17 patients correctly mapped the breakpoints. In addition, we newly found a deletion across an 84 kb region of MSH2in a case without the pathogenic single nucleotide variants. These data suggest that long read-sequencing combined with hybridization-based enrichment is an efficient method to identify both SVs and their breakpoints. This strategy might replace MLPA for the screening of SVs in hereditary diseases.

Lynch syndrome (LS) is an autosomal dominantly inherited disease predisposed to not only colorectal cancer but also other LS-related tumors. Although the clinical and genetic characteristics of LS in Western countries have been well characterized, the information of Japanese LS is limited. As a collaborative study of Japanese Society for Cancer of the Colon and Rectum (JSCCR), we registered colorectal cancer (CRC) patients who fulfilled the modified Amsterdam II criteria including gastric cancer as an LS-related tumor. Among 4030 CRC patients initially registered in this project, 85 patients (2.1%) fulfilled the modified criteria. An additional 26 patients who met the same criteria were enrolled in the analysis. We analyzed three major responsible genes, MLH1, MSH2, and MSH6by direct sequencing, and further performed multiplex ligation-dependent probe amplification for MLH1and MSH2. Consequently, we identified pathogenic variants in 64 of the 111 patients comprising of 34 patients in MLH1, 28 in MSH2, and 2 in MSH6. It is of note that large structural alterations were found in 17 patients. Among the 64 patients, 11 patients would not have been enrolled in the analysis if gastric cancer were not included in the modified criteria. In addition, 10 of the 64 variant carriers (15.6%) had medical history of gastric cancer. Furthermore, the standardized incidence ratio of gastric cancer in the LS patients to the Japanese population is estimated to be as high as 20.2. These data underscore the importance of gastric cancer in the diagnosis and healthcare of Japanese LS patients.

Dopamine (DA) D2 receptors (D2Rs) have 2 isoforms, a long form (D2L) and a short form (D2S). D2L is predominantly postsynaptic in the striatal medium spiny neurons and cholinergic interneurons. D2S is principally presynaptic autoreceptors in the nigrostriatal DA neurons. Recently, we demonstrated that L-3,4-dihydroxyphenylalanine (L-DOPA) augments D2L function through the coupling between D2L and GPR143, a receptor of L-DOPA that was originally identified as the gene product of ocular albinism 1. Here we show that GPR143 modifies the functions of D2L and D2S in an opposite manner. Haloperidol-induced catalepsy was attenuated in DA neuron-specific Gpr143 gene-deficient (Dat-cre;Gpr143flox/y) mice, compared with wild-type (Wt) mice. Haloperidol increased in vivo DA release from the dorsolateral striatum, and this increase was augmented in Gpr143-/ymice compared with Wtmice. A D2R agonist quinpirole-induced increase in the phosphorylation of GSK3β(pGSK3β(S9)) was enhanced in Chinese hamster ovary (CHO) cells coexpressing D2L and GPR143 compared with cells expressing D2L alone, while it was suppressed in cells coexpressing D2S and GPR143 compared with D2S alone, suggesting that GPR143 differentially modifies D2R functions depending on its isoforms of D2L and D2S.

The hereditary breast and ovarian cancer (HBOC) registration system of Japan was established by the Japanese HBOC Consortium. The first trial was registered in 2015 in four institutions to which some registration committee members belonged. We analyzed the information of 830 Japanese pedigrees, who underwent BRCA1/2genetic testing, including mutation carriers with BRCA1(N= 127) and BRCA2(N= 115), and their families. The mutation-positive rate was 19.7%. Variants of uncertain significance were found in 6.5% of all individuals subjected to genetic testing for BRCA1/2. Compared to the United States, Japan had a higher mutation-positive rate in most categories, except for the groups with male breast cancer. Among the intrinsic subtypes of BRCA1-associated breast cancers, 75.8% were triple-negative. The incidence rate of contralateral breast cancer in BRCA1/2mutation carriers was 0.99%/year. Among 240 mutation carriers, 26 and 62 patients underwent risk-reducing mastectomy (RRM) and risk-reducing salpingo-oophorectomy (RRSO), respectively; the respective frequencies of occult cancer were 7.1 and 3.2%. Metachronous breast cancer after RRM or peritoneal cancer after RRSO was not observed during the follow-up period. The nationwide registration system began last year and the system enables follow-up analysis in Japan.

Methylphenidate (MPH) and methamphetamine (METH) are the current treatments of choice for attention deficit/hyperactivity disorder. We previously reported that METH induces the release of dopamine (DA) and of the neurotransmitter candidate L-3,4-dihydroxyphenylalanine (L-DOPA). In contrast, we here found that MPH increased the DA release while it did not affect the L-DOPA release from the dorsolateral striatum. Nevertheless, MPH-induced hyperlocomotion was reduced in Gpr143(L-DOPA receptor) gene-deficient (Gpr143-/y) mice. The rewarding effect and increased c-fos expression induced by MPH were also attenuated in Gpr143-/ymice. Together, these findings suggest that GPR143 is involved in the acute and chronic actions of MPH.

In contrast to the origins of colorectal carcinomas, the mechanisms of carcinogenesis in the small intestine remain unclear. We therefore analyzed the mutational status of the Ki-ras, p53, and adenomatous polyposis coli (APC) genes in primary carcinomas of the small intestine and compared the mutation patterns with those established for colorectal cancers. DNA was extracted from 15 formalin-fixed, paraffin-embedded lesions. Codons 12, 13 and 61 of the Ki-ras gene, exons 5–8 of the p53 gene, and codons 1268–1569, which contain the mutation cluster region (MCR) of the APC gene, were amplified by means of PCR, subcloned and sequenced. Mutations of the Ki-ras and p53 genes were observed in 8 (53.3%) and 4 lesions (26.7%), respectively. The mutational frequency of the Ki-ras gene in the present series of small intestinal carcinomas was similar, while that of the p53 gene was slightly lower than the reported frequencies for colorectal carcinomas. Only one case showed a mutation of the APC gene, involving an insertional mutation of an adenine at codons 1554–1556 with formation of a stop codon immediately downstream. Since the occurrence of an APC mutation is considered an early event in colorectal carcinogenesis, our findings indicating an extremely low frequency of such changes in and around the MCR suggest that carcinomas of the small intestine arise via a genetic pathway distinct from that involved in the development of carcinomas of the colorectum. Int. J. Cancer, 70:390–395, 1997. © 1997 Wiley-Liss, Inc.

Both 2,6-dideoxy-6-fluoro-2-[(3R)- and (3S)-3-hydroxytetradecanamido]-3-O-[(3R)-3-(tetradecanoyloxy)etradecanoyl]-d-glucopyranose 4-phosphate, 11and 11′ (6-fluoro GLA-60 and its 3′-epimer), and 2-deoxy-2-[(3R)- and (3S)-(3-hydroxytetradecanamido]-3-O-[(3R)-3-(tetradecanoyloxy)tetradecaonyl]-α-d-glucopyranosyl fluoride 4-(dihydrogen phosphate), 18and 18′ (1-fluoro GLA-60 and its 3′-epimer), were synthesized from allyl 2-deoxy-4,6-O-isopropylidene-2-trifluoroacetamido-α-d-glucopyranoside.

Lipid A 3-ether analogues were synthesized from allyl 2-deoxy-4,6-O-isopropylidene-2-trifluoroacetamido-α-d-glucopyranoside and 3,4,6-tri-O-acetyl-2-trifluoroacetamido-α-d-glucopyranosyl bromide. The compound lost completely the endotoxic activity.

Both a 3-O-alkyl lipid X analogue and its 4-O-phosphono isomer were synthesized from allyl 2-deoxy-4,6-O-isopropylidene-2-trifluoroacetamido-α-d-glucopyranoside.

Six GLA-60 positional isomers (8, 14, 14′, 20, 26, and 26′) were synthesized to investigate their biological activities. Compound 8exhibited potent agonistic activity, while compounds 26and 26′ exhibited slight agonistic activity on TNFαproduction toward human monoblastic U937 cells. TNFαproduction (% control; 10 ng ml−1of LPS = 100) of compound 8in the concentration of 10 μM was 611, and that of lipid A in the same concentration was 651. In contrast, the difluorinated compounds 14, 14′, and 20showed little agonistic activity on TNFαproduction. And neither compound 8nor compounds 26and 26′ showed antagonistic activity. On the other hand, the difluorinated compounds 14, 14′, and 20showed potent antagonistic activity, and inhibited the LPS-induced TNFαproduction dose-dependently. Compound 14′ (10 μM) inhibited in excess of 80% of the LPS-induced TNFαproduction.

2,3-Dideoxy-2-[(3R)-3-hydroxytetradecanoylamino]-3-C-[(3R)-(3-hydroxytetradecanoyloxy)methyl]-4-O-phosphono-d-mannopyranose was synthesized from (1R,2S,4S,5S)-6-(2,4-dimethoxybenzyl)-4-hydroxy-2-[(1R)-1,2-isopropyridenedioxyethyl]-3-oxa-6-azabicyclo[3.2.0]heptan-7-one.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Correction to: Journal of Human Genetics advance online publication, 08 November 2017; 10.1038/s10038-017-0355-1

Compound 10was synthesized from β-d-glucose pentaacetate in a stereocontrolled manner. It unexpectedly showed LPS-agonistic activity much stronger than that of GLA-60 toward human monoblastic U937 cells in the TNFα production.

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ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.