title: An ADAM10 promoter polymorphism is a functional variant in severe sepsis patients and confers susceptibility to the development of sepsis.
abstract: BACKGROUND
  Although genetic variants of the A disintegrin and metalloproteinase 10 (ADAM10) gene have been shown to be associated with susceptibility to several inflammatory-related diseases, to date little is known about the clinical relationship in the development of sepsis.
  
  
  METHODS
  Two genetic variants in the promoter of ADAM10 were selected to analyze the potential association with the risk of sepsis. A total of 440 sepsis patients and 450 matched healthy individuals in two independent Chinese Han population were enrolled. Pyrosequencing and polymerase chain reaction-length polymorphism was used to determine the genotypes of the rs514049 and rs653765. A real-time qPCR method was used to detect the mRNA level of ADAM10. Enzyme-linked immunosorbent assay was used to measure the expression levels of substrates CX3CL1, interleukin (IL)-6R, tumor necrosis factor alpha (TNF-α), and the pro-inflammatory cytokines IL-1β and IL-6. Luciferase assay was used to analyze the activities of the promoter haplotypes of ADAM10.
  
  
  RESULTS
  No statistically significant differences between sepsis cases and controls in the genotype or allele frequencies were observed, suggesting that ADAM10 single nucleotide polymorphisms (SNPs) may not be risk factors for the occurrence of sepsis. A significant difference in the genotype and allele frequencies of the rs653765 SNP between patients with sepsis subtype and severe sepsis (P = 0.0014) or severe sepsis/sepsis shock (P = 0.0037) were observed. Moreover, the rs653765 CC genotype in severe sepsis showed a higher ADAM10 level compared to healthy groups, and the rs653765 CC polymorphism had a strong impact on the production of the ADAM10 substrates CX3CL1, IL-6R and TNF-α. Furthermore, the functional assay showed that ADAM10 C-A haplotype carriers exhibited significantly higher reporter activity compared with the T-A carriers and T-C carriers in human acute monocytic leukemia cell line.
  
  
  CONCLUSIONS
  Our data initially indicated the ADAM10 rs653765 polymorphism was associated with the development of severe sepsis; the risk CC genotype could functionally affect the expression level of ADAM10 mRNA and was accompanied by the up-regulation of its substrates. Thus, ADAM10 might be clinically important and play a critical role in the pathogenesis of the development of sepsis, with potentially important therapeutic implications.
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. lilicui1983@aliyun.com.
authors.forename: Lili
authors.initials: L
authors.lastname: Cui
authors.affiliation: The Intensive Care Unit, the Forth Affiliated Hospital of Harbin Medical University, Harbin, China. gaoyan_1971@126.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.affiliation: The Intensive Care Unit, Affiliated Hospital of Guangdong Medical College, Zhanjiang, PR China. sasa711711@qq.com.
authors.forename: Yuliu
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Clinical Research Center of Guangdong Medical College, Affiliated Hospital of Guangdong Medical College, Zhanjiang, PR China. jojoyan@aliyun.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. 434775643@qq.com.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Cai
authors.affiliation: The Intensive Care Unit, Affiliated Hospital of Guangdong Medical College, Zhanjiang, PR China. 812672728@qq.com.
authors.forename: Xin
authors.initials: X
authors.lastname: Shao
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. mxtgdmc@163.com.
authors.forename: Xiaotang
authors.initials: X
authors.lastname: Ma
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. dreamerly2001@163.com.
authors.forename: You
authors.initials: Y
authors.lastname: Li
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. maguoda@gdmc.edu.cn.
authors.forename: Guoda
authors.initials: G
authors.lastname: Ma
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. 363816619@qq.com.
authors.forename: Gen
authors.initials: G
authors.lastname: Liu
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. 619448969@qq.com.
authors.forename: Wanwen
authors.initials: W
authors.lastname: Cheng
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. 353360830@qq.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. 563876302@qq.com.
authors.forename: Tingting
authors.initials: T
authors.lastname: Liu
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. pqwforeve@163.com.
authors.forename: Qunwen
authors.initials: Q
authors.lastname: Pan
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. taohua1943@126.com.
authors.forename: Hua
authors.initials: H
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authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. liuzhou102@126.com.
authors.forename: Zhou
authors.initials: Z
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authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. zhaobine@vip.tom.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
authors.affiliation: The Intensive Care Unit, Affiliated Hospital of Guangdong Medical College, Zhanjiang, PR China. yimingshao_gmc@126.com.
authors.forename: Yiming
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authors.lastname: Shao
authors.affiliation: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Renmin street south 57, Xiashan district, Zhanjiang City, 524001, Guangdong Province, PR China. keshenli1971@126.com.
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mesh_terms: D051722:ADAM Proteins / Q000235:genetics* / Q000378:metabolism; D000072197:ADAM10 Protein; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics* / Q000378:metabolism; D044466:Asian People / Q000235:genetics; D016022:Case-Control Studies; D002681:China; D016207:Cytokines / Q000378:metabolism; D005260:Female; D005787:Gene Frequency; D006239:Haplotypes; D006801:Humans; D007378:Interleukins / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D018805:Sepsis / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D016207:Cytokines; D007378:Interleukins; D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human