Helicobacter pylori (H. pylori) has been identified as a type I carcinogen and contributes to a high rate of gastric cancer (GC), especially in Eastern Asia. Extracellular vesicles (EVs) have the potential to be used to detect various cancer types and diseases. However, the protein markers in EVs for the prognosis of H. pylori infection and GC are unknown. We aim to identify the proteins within EVs derived from a gastric epithelial cell line (AGS) infected with H. pylori by using LC-MS/MS.
EVs were isolated from AGS cells infected with high- and low-virulence H. pylori (strains TN2wt and Tx30a) by ultracentrifugation. Proteins within these EVs were identified and analyzed for potential marker candidates through bioinformatics. Proteins in H. pylori-derived EVs (HpEVs) from bacterial culture supernatant and HpEVs derived from H. pylori-infected AGS cells were elucidated.
Differentially expressed proteins by proteomic analysis in AGSEVs-Tx30a vs. AGSEVs-noninfected (NI) and AGSEVs-TN2wt vs. AGSEVs-NI were 107 and 55 proteins, respectively. Bioinformatics of these proteomes revealed that essential proteins for H. pylori survival and pathogenicity including outer membrane proteins, metabolism-related, host cell infection-related, and virulence-related proteins were observed in HpEVs. Interestingly, EVs derived from AGS cells infected with H. pylori TN2wt significantly contained multiple proteins related to GC (ATP6V0A1, GAPDH, HINT1, LYZ, and RBX1).
This study provides a comprehensive protein profile of EVs from H. pylori-infected AGS cells and HpEVs, which could serve as liquid-based biomarkers in the future for screening H. pylori infection, especially GC-related.
Helicobacter pylori (H. pylori) is the major cause of gastric mucosal precancerous lesions in adulthood, but its impact on pediatric patients remains unclear. We aimed to investigate H. pylori-induced gastric precancerous lesions in children and adolescents globally and analyze their influencing factors for related disease management and prevention.
We conducted a comprehensive literature search in major databases to identify studies including pediatric patients with gastric precancerous lesions and H. pylori infection status. Prevalence rates were computed using random-effects or fixed-effect models. A stratified analysis was conducted based on location, age, universal health coverage (UHC), and publication time.
Among the 3359 relevant articles screened, 24 studies (7036 participants) met the inclusion criteria. The overall prevalence of precancerous lesions in H. pylori-infected patients was 17.2%, in which atrophic gastritis (AG) and intestinal metaplasia (IM) were 13.5% and 3.6%, respectively. Precancerous lesion rates in infected individuals across different regions were as follows: Africa at 33.8% (AG: 32.6%), Latin America at 22.1% (AG: 17.9%, IM: 4.0%), Asia at 18.1% (AG: 12.4%, IM: 4.4%, Dysplasia: 1.2%), and Europe at 6.3% (AG: 4.3%, IM: 1.7%). Infected adolescents (> 10 years) exhibited a higher prevalence of precancerous lesions than younger children (≤ 10 years) at 14.2% (AG: 9.7%, IM: 2.9%) versus 3.4% (AG: 2.3%, IM: 1.1%), respectively. The prevalence of precancerous lesions in infected patients was higher in areas with low-medium UHC compared with high UHC (24.0% vs. 12.5%).
H. pylori infection causes significant gastric mucosal precancerous lesions in pediatric patients, representing a major concern for this population and a previously neglected area. Future in-depth investigations and proper management for related disease prevention are warranted.
PROSPERO number: CRD42023424683
Stomach cancer is the fourth leading cause of cancer-related deaths worldwide. Helicobacter pylori is the main risk factor for gastric adenocarcinoma (GAC), yet the precise mechanism underpinning this association remains controversial. Gastric intestinal metaplasia (GIM) represents the precancerous stage and follows H. pylori-associated chronic gastritis (CG). Sequencing studies have revealed fewer H. pylori and more non-H. pylori bacteria in GAC. However, the spatial organization of the gastric microbiota in health and disease is unknown.
Here, we have combined RNA in situ hybridization and immunohistochemistry to detect H. pylori, non-H. pylori bacteria, and host cell markers (E-cadherin, Mucins 5AC and 2) on tissue sections from patients with CG (n = 15) and GIM (n = 17).
Quantitative analysis of whole slide scans revealed significant correlations of H. pylori and other bacteria in CG and GIM. In contrast to sequencing studies, significantly fewer non-H. pylori bacteria were detected in H. pylori-negative patients. Importantly, whilst H. pylori exclusively colonized the gastric glands, non-H. pylori bacteria invaded the lamina propria in 6/9 CG and 8/10 GIM H. pylori-positive patients. A rapid and cost-effective modified Gram stain was used to confirm these findings and enabled detection of non-H. pylori bacteria in GIM samples.
The invasion of the gastric lamina propria by non-H. pylori bacteria during H. pylori-associated CG and GIM represents an overlooked phenomenon in cancer progression. Further work must determine the mechanisms underlying the synergistic roles of H. pylori and other bacteria in carcinogenesis. This observation should redirect attempts to prevent, diagnose, and treat GAC.
We aimed to assess the effects of Helicobacter pylori (H. pylori) eradication on the rebound of reflux-related symptoms among gastroesophageal reflux disease (GERD) patients.
This prospective randomized study recruited patients with typical reflux symptoms and reflux esophagitis on esophagogastroduodenoscopy (NCT02934152). Patients positive for H. pylori via a urea breath test (UBT) were randomly assigned to receive bacterial eradication with triple therapy for 2 weeks either before or after proton-pump inhibitor (PPI) treatment for 4 weeks. Follow-up was implemented with serial GerdQ evaluation and a subsequent UBT. The primary outcome was the incidence rates of symptom rebound between patients with and without H. pylori infection. The secondary outcomes included the severity of symptom rebound, incidence rates of symptom rebound, and successful eradication rates between the early and late eradication groups.
A total of 248 patients were enrolled, of whom 107 (43.1%) tested positive for H. pylori infection. All patients with and without concurrent H. pylori infection had significant symptom improvement over the entire treatment. Patients with H. pylori infection had significantly lower rates of symptom rebound (19.8% vs. 34.2%, p = 0.034) and rebound severity (1.8 ± 0.7 vs. 2.8 ± 1.6, p = 0.031) 4 weeks after eradication and PPI treatment than those without. The incidence rates of symptom rebound and successful eradication rates were not significantly different between the early and late eradication groups.
GERD patients with concurrent H. pylori infection were less susceptible to symptom rebound after H. pylori eradication compared to those without.
ClinicalTrial.gov (NCT02934152)
Vonoprazan is a novel potassium-competitive acid blocker (P-CAB) that offers several advantages, such as fast onset time and strong acid inhibition, in the treatment of Helicobacter pylori infection. This study aims to evaluate the efficacy, adverse reactions, and compliance of the 14-day vonoprazan-amoxicillin dual therapy versus drug sensitivity-based individualized therapy in the retreatment of H. pylori infection.
This multicenter, open-label, randomized, controlled non-inferiority study enrolled 240 adult patients who previously failed anti-H. pylori treatment. These patients were randomly assigned to receive the 14-day vonoprazan-amoxicillin dual therapy or drug sensitivity-based individualized therapy. The primary outcome was the eradication rate, and the secondary outcomes mainly included adverse events, patient compliance, antibiotic resistance rates, and risk factors that affected the eradication rate.
The intention-to-treat (ITT) and per-protocol (PP) analyses revealed that the eradication rates for the vonoprazan-amoxicillin dual therapy and drug sensitivity-based individualized therapy were comparably high, with rates of 87.50% and 83.33%, respectively. Furthermore, the vonoprazan-amoxicillin dual therapy fulfilled the criteria for the non-inferiority test, when compared to individualized therapy. The incidence of adverse reactions was significantly lower in the vonoprazan-amoxicillin dual therapy group. Both groups showed similarly good compliance and comparable rates of antibiotic resistance. The previous treatment with a clarithromycin-containing regimen was identified as an independent risk factor for clarithromycin resistance.
The 14-day vonoprazan-amoxicillin dual therapy exhibits high eradication rates and low incidence of adverse reactions in retreated patients, indicating its effectiveness and safety as a rescue regimen for patients with H. pylori infection.
This study evaluates the accuracy of melting curve–based multiplex real-time PCR (multiplex rt-PCR) on stool samples for diagnosing antibiotic resistance in Helicobacter pylori (H. pylori) compared to E-test and sequencing.
Gastric biopsies and stool samples were collected from 385 H. pylori-infected patients. A total of 325 strains were isolated, and genomic DNA was extracted from all 385 stool samples. E-tests were conducted to detect phenotypic resistance for clarithromycin and levofloxacin. Sanger sequencing and multiplex rt-PCR were employed to identify H. pylori 23S rRNA and GyrA mutations.
E-test results indicated that 203 (62.5%) were susceptible to both clarithromycin and levofloxacin, 33 (10.2%) exhibited mono-resistance to clarithromycin, 48 (14.8%) showed mono-resistance to levofloxacin, and 41 (12.6%) had dual resistance to both antibiotics. Compared to E-test results, the sensitivity and specificity of the multiplex rt-PCR method for detecting clarithromycin resistance mutation were 93.2 (95% CI 84.3–97.5) and 87.1% (95% CI 82.2–90.9), respectively. For levofloxacin resistance mutation, the multiplex rt-PCR method showed a sensitivity of 80.7 (95% CI 70.3–88.3) and a specificity of 93.0% (95% CI 88.7–95.8). Compared to Sanger sequencing, the sensitivity and specificity of the multiplex rt-PCR method for detecting clarithromycin resistance mutation were 95.8 (95% CI 90.0–98.4) and 96.0% (95% CI 92.6–98.0), respectively. For levofloxacin resistance mutation, the multiplex rt-PCR method showed a sensitivity of 91.3% (95% CI, 83.1–95.9) and a specificity of 96.1% (95% CI, 92.7–98.0).
Genotypic methods, including Sanger sequencing and multiplex rt-PCR, were rapid and reliable for diagnosing clarithromycin and levofloxacin resistance in the stool samples.
Rising antimicrobial resistance has significantly challenged the eradication rates of Helicobacter pylori. Due to its invasive nature, susceptibility testing based on endoscopic biopsy is controversial, while few studies have focused on the efficacy of tailored bismuth quadruple therapies (BQT) based on fecal susceptibility testing as a first-line treatment.
In this multicenter study, 598 H. pylori-positive patients without previous eradication treatment were recruited and randomly assigned to three groups: empirical BQT, BQT-tailored based on the history of clarithromycin use and tailored based on fecal molecular susceptibility testing where furazolidone was substituted for clarithromycin when there was clinical or laboratory evidence of clarithromycin resistance. This study defines BQT as regimens comprising rabeprazole, colloidal bismuth, amoxicillin, and one additional antibiotic (furazolidone or clarithromycin). The study assessed eradication rates using intention-to-treat (ITT), modified intention-to-treat (mITT), and per-protocol (PP) analyses.
The eradication rates of three groups were 82.00%, 80.90%, and 87.44% in the ITT analysis; 82.41%, 83.42%, and 89.23% in the mITT analysis; and 85.86%, 87.50%, and 94.57% in the PP analysis, respectively. Tailored BQT based on fecal susceptibility testing was not inferior to empirical BQT (all p values for noninferiority < 0.001) and demonstrated greater efficacy in the PP analysis (difference [95% CI]: 7.07% [0.90%, 13.25%]). The incidence of adverse events and treatment compliance did not differ significantly among the groups.
Tailored BQT based on fecal susceptibility testing is an effective regimen for H. pylori eradication, with no increase in adverse events or treatment noncompliance compared with empirical BQT. Therefore, we recommend tailored BQT based on fecal susceptibility testing as a first-line treatment.
EudraCT number: NCT05718609; ClinicalTrials.gov
Helicobacter pylori (H. pylori), a proven carcinogenic microbe, necessitates antimicrobial treatment once infected. However, H. pylori worldwide currently faces serious antibiotic resistance (AMR), requiring infected patients to undergo antibiotic susceptibility testing (AST) to guide therapy. Currently, the recommended ASTs for H. pylori are culture-based methods, which are time-consuming, complicated, and expensive, impeding their widespread application. With in-depth researches on the AMR mechanisms of H. pylori, specific gene mutations and novel proteins have been confirmed as the cause of AMR and can serve as targets of ASTs. Accordingly, molecular biology detection has been developed and tremendously shortened the time and reduced difficulty of AST. However, these assays still struggle to meet the enormous testing demand and need for even faster, simpler, and more accurate methods. In recent years, researchers have developed various new platforms based on biosensors, transcriptomics, proteomics, and single-cell analysis. This review introduces the AMR mechanisms of H. pylori and summarizes the current ASTs from the working principles to application characteristics. Additionally, we draw attention to the potentially applicable techniques for AST of H. pylori from DNA, RNA, protein, and cell perspectives. By systematically recapitulating the past, present, and future of AST for H. pylori, this review provides valuable insights for developing novel assays.
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