國立成功大學「發展國際一流大學及頂尖研究中心計畫」
延攬優秀人才工作報告表
NCKU Project of Promoting Academic Excellence & Developing World Class Research Centers
Work Report Form for Distinguished Scholars
□續聘
continuation of employment
離職
resignation99 年 4 月 20 日更新 受聘者姓名
Name of the Employee
許淑美 男
女 Male Female 聘 期 Period of Employment From 99 年(y) 4 月(m) 1 日(d) to 100 年(y) 3 月(m) 31 日(d) 研究或教學或科技研發與 管理計畫名稱 The project title of research,teaching, technology development and management
幽門桿菌致病因子與宿主基因易 感 性於 調 控Wnt-1/COX-2 路徑 對胃癌前病變腸上皮化生之影響 計畫主持人 （申請單位主管） Project Investigator (Head of Department/Center)
Bor-Shyang Sheu
補助延聘編號Grant Number
HUA
-一、
研究、教學、科技研發與管理工作全程經過概述。（由受聘人填寫）Please summarize the entire research, teaching, or science and technology R&D and management work process ( To be
completed by the employee)
H. pylori infects more than half human population in the world and is the risk factor of peptic
ulcers and gastric malignancy, implicating that the importance of understanding the bacterial virulence and pathogenesis. Last year, the following objects were investigated: (1) To analyze the genetic profiles of babAB in the strains from different clinical diseases and the antrum and corpus isolates from the patients with single infection; (2) To analyze the transcription regulation of sialic acid-binding adhesin (SabA); (3) To determine the role of putative outer membrane protein during H.
pylori infection. (4) To determine the phosphorylation level of CagA of strains from mixed infection
patients.
(1) To analyze the genetic profiles of babAB in the strains from different clinical diseases and the antrum and corpus isolates from the patients with single infection.
Intragenomic recombination between babA and babB mediates antigenic variation, which
may help H. pylori persistent colonization. It is still unclear how variable genotypes of babA and
babB are generated in different niches within the human stomach. We analyzed isolates from
the antrum and corpus of 19 patients. The forward primers designed from the upstream region of the babA or babB genes, combined with the reverse primer designed from the variable region of both genes, were applied to determine whether babA or babB was at the babA and babB loci. Four genotypes (AB, AB B, A AB, AB AB) were detected, and 3 of them were mixed genotypes, indicating a subpopulation at the babA or babB loci within the strain. Fifteen of nineteen patients (79.0%) were infected with strains having one major babAB genotype across the antrum and corpus, but more than one genotype in either one or both gastric niches was detected in 12 patients. Among them, there was a major dominant genotype found in the isolates from the antrum, which was not found in corpus isolates. The sequence results showed that the main difference in babA and babB between individual patients’ strains was in the CT repeats of babB in the babB locus, largely in the corpus isolates. These results suggest that genetic polymorphism of babAB may allow H. pylori to successfully colonize the individual stomach, and
further variation in babAB genotypes and CT repeats controlling babB more frequently occur in the isolates from corpus than from the antrum.
In western countries, the presence of babA was reported to be significantly associated with peptic ulcer diseases (PUD). However, the association was not found in East Asia and some western countries and whether the genetic profiles from recombination between babA and babB is associated with clinical diseases is still not investigated. In our study, 92 strains were from the patients including gastritis (n = 27), duodenal ulcer (n = 18), gastric ulcer (n = 27) and gastric cancer (n = 20). The PCR reactions described as previously were also used to analyze the strains from different clinical diseases. Four genotypes of babAB (A B, AB B, A AB, and AB AB) were found and the distribution of each genotype was respectively as 46 (50%), 21 (22.8%), 10 (10.9%) and 15 (16.3 %) of all 92 strains. In statistic analysis, AB AB genotype was significantly associated with gastric cancer and the intensity of intestinal metaplasia (IM, p < 0.05). However, each of the genotypes did not associated with the inflammatory scores. The AB AB genotype was first identified and it may have the adhesion ability to adapt the environment of gastric cancer. Because we have published that the patients with mixed infection were significantly associated with IM in the antrum, it will be interesting to investigate the babAB distribution in the mixed infection isolates.
(2) To analyze the transcription regulation of sialic acid-binding adhesin (SabA)
Helicobacter pylori exploit sialyl acid binding adhesin (SabA) to interact with sialyl Lewis x to
establish persistent colonization on inflamed gastric mucosal surface. The CT repeat number can change due to slipped strand mispairing (SSM) during DNA replication and influence the translation of SabA. The analysis of transcriptional patterns showed that sabA mRNA had more expression in log phase. The results of 5’ RACE revealed that the transcriptional start site of
sabA was 66 bp upstream of ATG in reference strain J99 and 64 bp upstream in clinical strain
Hp258. The sequence results of 115 clinical strains revealed that there was a variably homopolymeric thymidine tract (T tract) present closed to the promoter region of sabA. Further analysis showed that the sabA gene also displayed different number of T tract to generate variation in the strain’s population. The cat reporter was used to detect the transcription activity of polyT tract (n=17 and 18) by RT-RCR. The results showed that the polyT 17 and 18 had similar transcription activity. In the future, more polyT number could be selected to analyze the polyT tract effect in sabA transcription.
(3) To determine the role of putative outer membrane protein during H. pylori infection
Helicobacter pylori is an important human gastric pathogen. Bacterial adhesion is a
prerequisite for colonizing the gastric mucosal surfaces. Many adhesin molecules, such as heat shock proteins, BabA, AlpAB, Hop proteins family, OipA, SabA and Lewis x (Lex) suggested to be involved in the initial colonization of H. pylori. Most of them are outer membrane protein (OMP) and several investigators have extensively studied the relation between BabA and Leb antigen of host cell. Based on the sequence of amino acids of BabA from different clinical isolates, it can not conclude the association with the binding affinity of Leb, but babA exist in
multiple loci of H. pylori chromosome. In addition, the babA and babB genes have the high sequence homology, those two genes could occur gene recombination to increase the variety of
babA, which involves in the adhesion ability of H. pylori. We found a smaller molecular weight
protein detected by anti-BabA polyclonal Ab, which could play a role in the H. pylori adhesion by using the gene disruption. The gene that express SMP (smp) has the structural specificity, that is quite different from the gene recombination between the omp family to change the antigenic
property of adhesin in previous literatures. However, it encodes by the structural gene of hook associated protein (flgK) and omp, which is worth studying in the future. (1) understand the gene structure of smp by using 5’ RACE and reporter assay, confirm its role in the H. pylori adhesion and investigate the prevalence of smp in clinical strains and its association with disease pathology; (2) express and purify SMP to produce the polyclonal Ab and proceed the immunoglod analysis to analyze whether SMP is the OMP, and its ability to stimulate cytokine secretions; (3) confirm whether smp also involves in the adhesion ability and the cytokine stimulation of other clinical H. pylori strains and search for the possible receptor of SMP.
(4) To determine the phosphorylation level of CagA of strains from mixed infection patients
Our previous study have shown that the patients with mixed infections had marginally higher CIS and HPD than those with single infection (p = 0.062 and p = 0.095) in the corpus and had a significantly higher rate of the appearance of intestinal metaplasia (IM) in the antrum (p = 0.005). In order to understand whether the strains from mixed infection are more virulence, we first analyze the phosphorylation level of CagA injected by their type IV secretion system to AGS cells. We have found that the antrum isolates seem to induce lower phosphorylated CagA in AGS cells. Moreover, non-phosphorylated CagA have been demonstrated to deregulate the -catenin signal that promotes intestinal transdifferentiation in gastric epithelial cells. Therefore, it still needs to include more mixed infection patients’ strains to demonstrate the possibility.
二、研究或教學或科技研發與管理成效評估（由計畫主持人或單位主管填寫）
Please evaluate the performance of research, teaching or science and technology R&D and management Work: ( To be
completed by Project Investigator or Head of Department/Center)
(1)是否達到延攬預期目標？
Has the expected goal of recruitment been achieved?
Dr. Sheu SM has conducted the H. pylori virulence factor cagA and its type IV secretion system. Moreover, she has devoted to conduct the mixed infection of H. pylori and its role to have different risk of H. pylori precancerous changes. These study outcomes are now in hand to submit to several SCI journals.
(2)研究或教學或科技研發與管理的方法、專業知識及進度如何？
What are the methods, professional knowledge, and progress of the research, teaching, or R&D and management work?
The study of this post doctor is highly original to answer why H. pylori has mixed isolates infection in clinical setting, it will help the current know-how to improve the resolution of the reason why some infections could be more toxic and related with more adverse outcome after H. pylori infection.
(3)受延攬人之研究或教學或科技研發與管理成果對該計畫(或貴單位)助益如何？
How have the research, teaching, or R&D and management results of the employed person given benefit to the project (or your unit)?
The post doctor is highly valuable to assist the laboratory technique teaching to the young
investigator and the assistants, including both master and PhD program students. (4)受延攬人於補助期間對貴單位或國內相關學術科技領域助益如何？
How has the employed person, during his or her term of employment, benefited your unit or the relevant domestic academic field? She has published several papers and has improved the molecular microbiology teaching for our lab and our session.
(5)具體工作績效或研究或教學或科技研發與管理成果：
Please describe the specific work performance, or the results of research, teaching, or R&D and management work: The publications during her post doctor status have included as the following:
Sheu BS, Odenbreit S, Hung KH, Liu CP, Sheu SM, Yang HB, Wu JJ. 2006 Jan. Interaction between host gastric Sialyl-Lewis X and H. pylori SabA enhances H. pylori density in patients lacking gastric Lewis B antigen. Am J Gastroenterol. 101(1):36-44. Sheu SM, Sheu BS, Yang HB, Lei HY, Wu JJ. 2007 Jun. Anti-Lewis X antibody
promotes Helicobacter pylori adhesion to gastric epithelial cells. Infect Immun. 75(6):2661-7.
Sheu SM, Hung KH, Sheu BS, Yang HB, Wu JJ. 2009 Jan. Association of
nonsynonymous substitutions in the intermediate region of the vacA gene of
Helicobacter pylori with gastric diseases in Taiwan. J Clin Microbiol. 47(1):249-51.
Sheu SM, Sheu BS, Lu CC, Yang HB, Wu JJ. 2009 Mar. Mixed infections of Helicobacter pylori: tissue tropism and histological significance. Clin Microbiol Infect.
15(3):253-9.
Sheu SM, Sheu BS, Wu HM, Wu JJ. Topographic distribution of Helicobacter pylori
strains differing in babA and babB genotypes between the antrum and corpus of the human stomach. J Clin Microbiol. (revised)
(6)是否續聘受聘人? Will you continue hiring the employed person?
■續聘
Yes □不續聘No※此報告表篇幅以三～四頁為原則。This report form should be limited to 3-4 pages in principle.
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This report form can be downloaded in http://www.ncku.edu.tw/~top/top_web/C/lawcom.H
