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A novel mutation in the COMP responsible for pseudoachondroplasia

A novel mutation in the COMP responsible for pseudoachondrop...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Li Shan Zhao Xiuli Department of Medical Genetics Institute of Basic Medical Sciences Chinese Academy of Medical Sciences School of Basic Medicine Peking Union Medical College

Objective Pseudoachondroplasia（PSACH） is an autosomal dominant osteochondrodysplasia ch aracterized by short?limb short stature, brachydactyly and early?onset osteoarthropathy. As so fa r, most cases of PSACH were caused exclusively by mutations in the gene for cartilage oligomeri c matrix protein（COMP）. In this study,we report a family with the typical PSACH. The pedigree analysis revealed that the proband and her daughter affected by severe short stature within a spa n of two generations and the disorder exhibited transmission form of the autosomal dominant. Th e aim of this study is to identify the causative mutation in the family. Method Genomic DNA was extracted from peripheral blood by standard phenol/chloroform meth od. Polymerase chain reaction（PCR） was adopted to amplify the all the 19 exons and the exon/intron boundaries of COMP gene. Then the PCR fragments were screened by DNA Sanger sequen cing to find the causative mutation. Result A heterozygous deletion mutation ***1371385 GGACTCAGACCACGA in the exon 13 was identified in the proband, which resulted in a five-amino-acid deletion（Glu457 Aspdel DSDHD） in the sixth calmodulin-like repeat of the COMP protein. Then this mutation was confirmed to cosegregate with PSACH manifestations of the affected in the family, but not detected in the unaffec teds including 3 familial members and 50 population controls. Conclusion The mutation of ***1371385 GGACTCAGACCACGA is a novel mutation respons ible for severe familial PSACH（not reported in HGMD）. Our result expanded the mutation spectru m of the COMP gene, and indicated that molecular analysis will be the most promising method for the clinical diagnosis of PSACH in future.

关键词： A novel mutation in the COMP responsible for pseudoachondroplasia

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Development of a new method for genotype of G6PD deficiency

Development of a new method for genotype of G6PD deficiency

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Li Yinshu Yi Jia Liu Jingjing YanengBIOscience (shenzhen) Co. Ltd

Objective Objective: To establish an effective, feasible, economic, rapid and pollution-free practic al assay for genetic diagnosis of glucose-6-phosphate dehydrogenase(glucose-6-phosphate dehydrogenase, G6 PD) deficiency. Method Methods: A multiple fluorescence PCR system based Taqman MGB probes was perform ed to amplify the target fragments of G6 PD gene, and detect the domestic main 10 mutation sites kinds(1376 G>T, 1388 G>A, 95 A>G, 871 G>A, 392 G>T, 1024 C>T, 1004 C>A, 517 C>T, 592 C>T, 487 G>A). At the same time, the beta actin gene monitors the whole PCR system. Repeatability w as tested in 50 samples with known genotype. A double-blind trial for 346 unknown samples was performed by using multiplex fluorescence PC R assay and DNA sequencing simultaneously. Result Results: The established multiple fluorescence PCR assay based Taqman MGB probes w as able to detect the most common 10 mutations of G6 PD gene simultaneously in 1 hour, and bot h specificity and accuracy reached 100%. Among the 346 unknown samples, 57 mutational cases was detected. All the genotyping results of multiple fluorescence PCR assay based Taqman MG B probes were in complete concordance with direct DNA sequencing, and both specificity and ac curacy reached 100%. Conclusion Conclusion: The established multiple fluorescence PCR assay based Taqman MGB probes, detecting samples in 1 hour without open tubes after PCR, is a rapid, effective and feasibl e method of G6 PD deficiency gene diagnosis.

关键词： Taqman Development of a new method for genotype of G6PD deficiency

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Two novel FBN1 gene splicing mutations of Marfan syndrome

Two novel FBN1 gene splicing mutations of Marfan syndrome

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Cao Yingxi Tan Hu Zhang Rui Wu Lingqian State Key Laboratory of Medical Genetics Central South University

Objective Marfan syndrome(MFS) is an autosomal dominant inherited connective tissue disorde r. The phenotypes are variable from person to person, but mainly affect multiple systems includin g skeletal, cardiovascular, ocular, pulmonary, dura, skin and integument. The Ghent criteria is the internationally accepted criteria for MFS diagnosis. The two patients in this study are both diagno sed with MFS. Thus we performed the genetic testing of FBN1 which is the main disease causing gene of MFS to make a definite diagnosis. Method We collected the peripheral blood samples of patients and their family members, trying to find the causal FBN1 mutation using different methods including high-throughput sequencing, PC R and Sanger sequencing. Bioinformatics websites are essential for our follow up data analysis. Result We identified two novel FBN1 gene splicing mutations in our patients. Both of them are he terozygous splicing mutations. These mutation sites have been reported in the Human Gene Mut ation Database as Marfan syndrome disease causing mutations, but the base changing are differ ent. Based on this results, RNA of one patient was extracted and Sanger sequencing of the RT-P CR product confirm the splicing change. The RT-PCR of the other patient is not finished yet. Conclusion The two novel FBN1 gene splicing mutations of Marfan syndrome are first reported i n our study. However, the pathogenicity of these mutations still need further confirmation. Becaus e of the phenotypic similarity of the two patients, we hypothesise that there is a correlation betwe en the phenotype and mutation type. Our study has confirm the clinical diagnosis, and enlarge th e spectrum of FBN1 mutation. It will be helpful for prenatal diagnosis and genetic counseling.

关键词： MFS Two novel FBN1 gene splicing mutations of Marfan syndrome

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Rare Mutations in SHROOM3 are Associated with Human Neural Tube Defects

Rare Mutations in SHROOM3 are Associated with Human Neural T...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Kuang Lele Wang Hongyan The State Key Laboratory of Genetic Engineering and Ministry of Education Key Laboratory of Contemporary Ant hropology & The Collaborative Innovation Centre of Genetics and Development School of Life ScienceFudan University Jiangwan Campus

Objective to explore the molecular mechanism and function of Shroom3 in human Neural tube d efects Method with the next generation whole exon sequencing,we sequenced the coding region of SH ROOM3 among all cases dan ***,lots of mutations were found including common(MA F>5%) and rare mutations(MAF<1%).As common mutations in human often don't have a good co nsistency with sides,and rare mutations play more and more important roles in NTD ***, we focus the de novo rare *** Sanger sequencing,we identify three case-specific nonsynonymous and one frameshift mutations in human ***,functional studies in vitro such as Co-IP and fluorescent colocalization were taken to explore the effect of mutations. Result In this study,363 miscarried of stillborn Han Chinese NTD fetuses from Jiangsu and Shan xi province, China were collected as *** group contains 215 unrelated normal fetuses born in the same ***,four rare case-specific heterozygous mutations were found in hum an *** vitro functional assays showed that all four mutations increased the interaction with D vl2,and lost the ability to inhibit PCP *** addition,Co-IP and fluorescent colocalization assa ys showed that frameshift mutation also disrupted its binding and colocalization with Rock1 SBD. Conclusion Apical constriction and convergent extension are the most critical processes during n eural tube ***3 mutations may act through PCP and Rock1 pathway together with ot her gene network and environment risk factors to mediate the severity of NTD phenotypes.

关键词： Rare Mutations in SHROOM3 are Associated with Human Neural Tube Defects

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Effects of G6PD activity inhibition on the viability, migration and morphology of cervical cancer cells

Effects of G6PD activity inhibition on the viability, migrat...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Fang ZiShui Jiang ChengRui Feng Yi Chen RiXin Lin XiaoYing Zhang ZhiQiang Han LuHao Chen XiaoDan Li HongYi Guo YiBin Jiang WeiYing Sun Yat-sen University

Objective Glucose-6-phosphate dehydrogenase(G6 PD) deficiency has been revealed to be involved in the efficacy to anti-cancer therapy but the mechanism remains unclear. We aimed to investigate the anti-cancer mechanism of G6 PD deficiency. Method In our study, Dehydroepiandrosterone(DHEA) and sh RNA technology were used for inhibiting the activity of G6 PD of cervical cancer cells. Flow cytometry was used to detect the apoptosis and reactive oxygen species(ROS) generation. Scanning electron microscopy was used to observe cell morphology. And the migration rate of cell was also observed using the imaging system of inverted living cell microscope. Result When G6 PD was inhibited by DHEA or RNA interference, idioblasts were observed in He La cells. Simultaneously, G6 PD deficiency resulted in decreased He La cells migration and proliferation ability but increased ROS generation inducing apoptosis. What‘s more, the inhibition of G6 PD activity caused filopodia and reticulopodium of cells shorten and reduce as well as cells shrinkage. Conclusion Our results indicated the anti-cervix cancer mechanism of G6 PD deficiency may be involved with the decreased cancer cells proliferation caused by the increased ROS and decreased migration ability as a result of abnormal filopodia and reticulopodium of cells. Suppression of G6 PD may be a promising strategy in developing novel therapeutic methods for cervical cancer.

关键词： Effects of G6PD activity inhibition on the viability migration and morphology of cervical cancer cells

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Detection of fetal subchromosomal aberration with cell-free DNA screening led to diagnosis of parental translocation: review of 4992 consecutive cases in a university hospital

Detection of fetal subchromosomal aberration with cell-free ...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Qian Ye-Qing Chen Min Luo Yu-Qin Yan Kai Yang Yan-Mei Liu Bei Wang Li-Ya Huang Ying-Zhi Li Hong-Ge Jin Fan Dong Min-Yue Women's Hospital School of Medicine Zhejiang University Key Laboratory of Reproductive Genetics (Zhejiang University) Ministry of Education Key Laboratory of Women's Reproductive Health of Zhejiang Province

Objective The objective of this study is to report the feasibility of cell-free DNA screening(cf DNA screening) as an indicator of parental balanced chromosome translocation. Method From February 2015 to September 2015, cf DNA screening was offered to 4992 pregnant women. Positive cf DNA screening results for aneuploidies were confirmed by karyotyping, while 4796/4932 negative results were interviewed after delivery. To validate the subchromosomal copy number variations(CNVs), we collected fetal amniotic fluids and tested the samples by chromosome microarray analysis(CMA) and karyotyping. Result In the 4992 cases, cf DNA screening was positive for common trisomies in 33 cases(25 trisomy 21, seven trisomy 18 and one trisomy 13). In addition, 22 cases were positive for sexchromosomal abnormalities, five cases were positive for other deletion/duplication in which two were identified as terminal duplication and deletion on different chromosomes. The CMA results or karyotyping confirmed the cf DNA screening findings and the origin of CNVs were validated afterward by karyotyping or fluorescence in situ hybridization(FISH) using parental blood samples. Conclusion Cf DNA screening may help identify deletions and duplications in fetus, which in some cases may indicate risk of a parent being a balanced rearrangement carrier, and that the diagnostic follow-up testing is necessary.

关键词： Detection of fetal subchromosomal aberration with cell-free DNA screening led to diagnosis of parental translocation review of 4992 consecutive cases in a university hospital DNA

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TFPI affects the migration of Smooth muscle cells through P38/HSP27 pathway

TFPI affects the migration of Smooth muscle cells through P3...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Xiao Jiajun Jin Kaiyue Yang Jichun Zhang Jin Jiang Nan Yang Xiao Ma Duan Key Laboratory of Metabolism and Molecular Medicine Ministry of Education Collaborative Innovation Center of Genetics and Development Department of Biochemistry and Molecular Biology Institute of Biomedical Sciences School of Basic Medical Sciences Fudan University Institute of Geriatrics PLA Postgraduate School of Medicine PLA General Hospital Cardiovascular Center Children‘s Hospital Affiliated to Fudan University

Objective Tissue factor pathway inhibitor(TFPI) is a major physiological inhibitor of TF-initiated c oagulation, but its precise role and signal pathway in vascular smooth muscle cells(SMCs) during the development of atherosclerosis are not clear. Method We constructed short hairpin RNA targeting TFPI gene(TFPI-sh RNA) in SMCs and the TFPI conditional knockout in SMCs of mouse. TFPI knockdown promoted migration of SMCs through a wound healing assay and a modified Boyden`s chamber method. A mouse model of special TFPI knockdown in SMCs was generated, and the mice were fed a high fat diet for 18 weeks and they were then euthanized. Aortas were stained wi th Sudan IV and more atherosclerotic lesions were found in the TFPI conditional knockout mice th an the control mice, but there were not different among the TFPIfl/fl Sma-Cre, TFPIfl/fl and TFPI+/+ mice. Result The result emphasized that the TFPI deficiency promoted the development and not the oc currence of atherosclerosis. In vitro and in vivo experiments identified that TFPI knockout in SMC s enhanced the phosphorylation of P38/HSP27, and it was more important that genes regulated by the p38 MAPK signaling pathway, such as IL-1β, IL-6, IL-8 and CXCL-10, further confirmed th e tendency. Conclusion TFPI deficiency in SMCs promoted the migration of SMCs and accelerated atheroscl erotic development through the phosphorylation of P38/HSP27.

关键词： TFPI affects the migration of Smooth muscle cells through P38/HSP27 pathway HSP

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Novel mutations in cardiac transcriptional factors contribute to Tetralogy of Fallot

Novel mutations in cardiac transcriptional factors contribut...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Xiao Deyong Qian Yanyan Guo Xiao Ma Xiaojing Huang Guoying Wang Huijun Ma Duan Key Laboratory of Metabolism and Molecular Medicine Ministry of Education Department of Biochemistry and Molecular Biology School of Basic Medical Sciences Fudan University Children Hospital of Fudan University Institute of Biomedical Sciences Collaborative Innovation Center of Genetics and Development School of Basic Medical Sciences Fudan University

Objective Congenital heart disease is the most common birth defects in the world. Cardiac transc ription factors play critical roles in the heart development. Abnormalities in those transcriptional fa ctors are the major contributors to the pathogenesis of CHD. Especially, genetic defects in those genes commonly result in CHD but largely remain unknown. Method In this study, twenty six candidate genes for CHD was screened for mutation analysis in 103 patients with TOF using next-generation sequencing *** Results showed novel pathogenic mutations concentrated in GATA family members, GAT A5(c.943 T>A p.S315 T and c.274 G>T p.A92 S) and GATA6(c.331 G>A p.D111 N and c.972 C>G p.H324 Q), their partner FOG2(c.3442 G>A p.E1148 K and c.3014 A>G p.E1005 G) and TBX5(c.G259 T, p.V87 L). Overexpression of the mutant of FOG2(***1148 Lys) in zebrafish leaded to ca rdiac abnormalities. Moreover, the mutant impaired the interaction with GATA4 in vitro. It proved t he mutation in FOG2 is pathogenic. Conclusion Therefore, our results suggested defects in the network involved in GATA family me mbers and their partners are the high risk for CHD.

关键词： FOG Novel mutations in cardiac transcriptional factors contribute to Tetralogy of Fallot CHD

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Two de novo overlapping interstitial duplications at 10q22 associated with intellectual disability, developmental delay, speech impairments and behavior anomalies

Two de novo overlapping interstitial duplications at 10q22 a...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Yuan Haiming Liang Liyang Guangzhou kingmed center for clinical laboratory Co. Ltd. King Med School of Laboratory Medicine Guangzhou Medical University Sun Yat-Sen Memorial Hospital Sun Yat-Sen University

Objective Copy number variants(CNVs) involving chromosome 10 q22 region are rarely reported. At present, only seven patients with deletions at this locus have been reported. The reciprocal du plications are far more rare and have never been described. Method High-resolution chromosomal microarray analysis(CMA) was employed for our patients‘ tests. Result Here, we report two unrelated patients with de novo overlapping duplications at 10 q22.1 q22.3 interval detected by CMA. The CMA showed the two duplications : arr[hg19] 10 q22.1 q22.3(72,331,092-78,710,233)x3 dn and arr[hg19] 10 q22.1 q22.3(70,742,930-80,565,963)x3 dn respectively. Intellectual disability, developmental delay, speech impairments, distinctive facial features and behavior problems were observed in both patients. Conclusion The shared genomic region and the similar clinical features suggest a novel contiguo us gene duplication syndrome at 10 q22. Based on all pathogenic CNVs delineated and candidate genes identified in this interval, the critical region for the novel genomic duplication syndrome is l ocated on 10 q22.2 locus.

关键词： CMA Two de novo overlapping interstitial duplications at 10q22 associated with intellectual disability developmental delay speech impairments and behavior anomalies

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Improved assay performance of SNP array over conventional karyotyping in analyzing products of conception

Improved assay performance of SNP array over conventional ka...

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中华医学会第十五次全国医学遗传学学术会议暨中国医师协会医学遗传医师分会第一届全国学术会议暨2016年浙江省医学遗传学年会

作者： Lin Shaobin Zhou Yi Luo Yanmin Fang Qun Chen Baojiang Fetal Medicine Center Department of Obstetrics and Gynecology First Affiliated Hospital of Sun Yat-Sen University

Objective Conventional karyotyping has been a routine method to identify chromosome abnormalities in products of conception(POC). However, this manner is being transformed by single nucleotide polymorphism(SNP) array, which has advantages over karyotyping including higher resolution, dispensing with cell culture and so on. This study was to evaluate the advantage of high-resolution SNP array in identifying genetic aberrations in POC. Method 155 POC specimens including 139 from first-trimester miscarriage and 16 from secondtrimester miscarriage were collected consecutively. SNP array was performed on these samples in parallel with G-banded *** SNP array obtained test success rate of 98.1%(152/155) that was higher than that in karyotyping(133/155, 85.8%). It yielded a 63.8%(97/152) abnormality rate, and the frequency of various chromosome abnormalities was agreement with other previous studies. Results between array and karyotyping demonstrated 94.0%(125/133) concordance. SNP array obtained additional aberrations in 3.8%(5/133) cases unidentified by karyotyping, which included 3 cases with whole-genome uniparental disomy(UPD), 1 case with pathogenic copy number variation(CNV) and 1 case with del(4)(q35.1 q35.2) and dup(12)(q24.31 q24.33). However, chromosome translocations presented in 2 cases and tetraploidy presented in 1 case were detected by karyotyping instead of array. Additionally, 2 out of 3 cases with mosaic trisomy were revealed by array but recognized as pure trisomy by karyotyping. Conclusion This study demonstrated that SNP array had certain advantages over G-banded karyotyping, including higher success rate, additional detection of CNVs and UPD, and improved sensitivity to mosaicism, so it would be an alternative method to karyotyping in clinical genetic practice.

关键词： SNP Improved assay performance of SNP array over conventional karyotyping in analyzing products of conception

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