Introduction

Early detection of oesophageal adenocarcinoma (OAC) will facilitate preventive treatment for a major public health priority. There are major efforts to devise better non-invasive tests for the precursor lesion of OAC which is defined by metaplastic transformation from the squamous oesophageal epithelium to intestinal metaplasia known as Barrett’s oesophagus (BO). The minimally invasive non-endoscopic capsule sponge test collects cells from the lining of the oesophagus. The current laboratory assays rely on processing the cells retrieved from the device for pathology, immunohistochemistry or sequencing.

Aims & Methods

In this proof-of-concept study, we investigated the possibility of using the acellular portion of the sponge sampling to test for secretory protein biomarkers. These proteins are released from the cells collected by the sponge into the preservative fluid and we hypothesised they could be used for a more rapid diagnostic that does not rely on cytological pathology reports.
Biomarker candidates were selected from previously published bulk and single cell-RNA-sequencing (scRNA-seq) data^1,2 based on their differential expression in BO compared to normal oesophagus and gastric cardia, and subsequent validation at protein level with immunohistochemical staining. Enzyme-linked immunosorbent assays (ELISA) were used to quantitatively detect trefoil factor-3 (TFF3) (Bio-Techne, #SPCKB-PS-000530, USA), regenerating family member 4 (REG4) (Biorbyt, orb180655, UK), Meprin A subunit alpha (MEP1a) (Abcam, ab26727), and serine peptidase inhibitor kazal type-4 (SPINK4) (Abbexa, abx528747) using manual sandwich-ELISA from capsule sponge supernatant samples. Six different classification algorithms were tested to distinguish between BO and heartburn controls based on the cell-free protein quantification of protein biomarkers either used singly or in combination to maximise performance.

Results

Data from scRNA-seq analysis highlighted REG4, TFF3, SPINK4, and MEP1a as strong candidate biomarkers related to the characteristic BO goblet cells which are secretory in nature. TFF3 and REG4 were individually detected at a significantly higher concentration in BO sponge supernatants (n=56) compared to controls (n=53) (p = <0.0001, Mann-Whitney test). While the average TFF3 concentration for BO samples was 1.2 times higher (62.5ng/ml) compared to controls (33.1 ng/ml), the average concentration of REG4 in BO samples was 5.9 times higher (2.1 ng/ml) compared to heartburn (0.36 ng/ml). REG4 protein concentration positively correlated with Barrett’s segment length (p < 0.0001, Spearmen’s correlation). A significant difference was also observed with SPINK4 in BO (n=34) compared to controls (n=38) (p = 0.01). Gaussian NB emerged as the best performing prediction model with a specificity of 0.94, sensitivity of 0.70 and area under the curve (AUC) of 0.86 when testing TFF3 and REG4 combined. Although MEP1a concentration could not significantly differentiate between BO and heartburn alone (p = ns), adding MEP1a as an additional marker to the TFF3/REG4 prediction model reached a AUROC of 0.85 (p <0.0001, logistic regression).

Conclusion

A panel of secreted proteins in the Cytosponge supernatant has potential to serve as non-invasive biomarkers for earlier detection and risk stratification of patients with BO. Our initial findings warrant further validation studies to investigate the clinical utility of the capsule sponge supernatant .

References

1 Jammula S, Katz-Summercorn A, Li X, et al. Gastroenterology (2020) 158(6):1682-1697
2 Nowicki-Osuch K, Zhuang L, Cheung TS, et al. Cancer Discovery (2023) 13(6):1346-1363

Disclosure

RCF is named on patents related to the Cytosponge-trefoil factor 3 test. Covidien GI Solutions (now Medtronic) licensed the Cytosponge from the Medical Research Council, and the device has now received the CE mark and is cleared by the US Food and Drug Administration. RCF is a shareholder in Cyted, a company working on early detection technology. MDP reports personal consultant fees from Medtronic, outside of the submitted work. The remaining authors have no conflicts of interest to declare.

Introduction

COVID-19 infection may increase the risk of developing Disorders of Gut-Brain Interaction (DGBI). However, the extent of this effect on a population level is poorly understood. We performed a two-country survey to address this issue.

Aims & Methods

A population-based Internet survey with pre-defined demographic quotas was conducted across the UK and USA in 2017 (pre-pandemic, n=4050) and repeated in 2023 (post-pandemic, n=4002). The surveys included the Rome IV diagnostic questionnaire, and questions about non-gastrointestinal somatic symptoms, anxiety and depression, quality of life, and healthcare utilization. The 2023 survey also included questions regarding COVID-19 infection and illness history.

Results

The overall DGBI prevalence, i.e., meeting diagnostic criteria for at least one DGBI, has significantly increased from the pre- to post- pandemic era (38.3% vs. 42.6%, OR 1.20, 95% C.I 1.09-1.31), with similar findings independently noted in the UK and USA. The rise in DGBI was observed within the oesophageal (8.8% vs. 10.1%, OR 1.16), gastroduodenal (11.9% vs. 16.4%, OR 1.45), and bowel domains (30.1% vs. 32.5%, OR 1.12). The two most widely investigated DGBI showed large post-pandemic prevalence increases, with functional dyspepsia rising from 8.3% to 11.9% (OR 1.48) and irritable bowel syndrome from 4.7% to 6.0% (OR 1.31).
In multivariable analysis, factors significantly associated with having DGBI in the post-pandemic era included younger age, female sex, anxiety, depression, medium-to-high somatic symptom severity, increasing number of COVID-19 infections, experiencing abdominal pain or diarrhoea during COVID-19 infection, and suffering with long-COVID.
Individuals with DGBI in the post-pandemic era, in particular those with long-COVID, reported reduced quality of life and higher mood disturbances, somatic symptom reporting and healthcare use than individuals with DGBI in the pre-pandemic era.

Conclusion

The population prevalence and burden of DGBI have increased following the COVID-19 pandemic. Healthcare services and research funding bodies need to adapt to this post-COVID surge in DGBI and address how to best manage this patient group.

Introduction

Colorectal cancer (CRC) remains the second most common cause of cancer-related mortality worldwide (1). The introduction of the Danish national CRC screening program in 2014 aimed at reducing mortality rates (2) has marrkedly increased colonoscopies and polypectomies (3,4). A a comprehensive description of the lesions removed has not yet been investigated.

Aims & Methods

Nationwide, cross‑sectional study of all colorectal lesions resected endoscopically in patients ≥18 years between January 2014 and December 2021. Data were obtained from the Danish National Patient Register, Pathology Register and Screening Database. Prevalence of histological sub‑types and predictors of high‑grade dysplasia (HGD) and adenocarcinoma were analyzed with multivariable logistic regression.

Results

Among 663,624 individuals undergoing 1,074,236 colonoscopies or sigmoidoscopies, 194,941 patients (median age 67 years, 58.6% male) had a total of 504,386 lesions removed. Tubular adenomas (TA) accounted for 66.8% of lesions.
The proportion of advanced colorectal polyps—defined as lesions ≥10 mm, adenomas with high-grade dysplasia, or sessile serrated lesions with any dysplasia—declined from 22.7% in 2014 to 10.2% in 2021.
High-grade dysplasia (HGD) was strongly associated with size ≥10 mm (OR 27.1 for 10–19 mm; OR 71.1 for 30–39 mm), and in diminutive lesions (1–5 mm), with tubulovillous or villous histology (OR 6.09).
Adenocarcinoma was identified in 0.65% of lesions and was predominantly left-sided (83.9%), with highest odds observed in the sigmoid colon (OR 6.33) and rectum (OR 6.00), compared to the caecum.
Sex was significantly associated with the risk of adenocarcinoma at the lesion level, but this effect was age-dependent. At age 40, women had significantly higher odds than men (adjusted OR 1.48), but the difference diminished with increasing age, and from age 75 onward, no statistically significant difference between sexes was observed.

Conclusion

Lesion size was the strongest predictor of high-grade dysplasia, outweighing histologic subtype once lesions exceeded 10 mm. At lesion level, the risk of malignancy was higher in younger women than in men of the same age, with the difference progressively diminishing with age. The introduction of the screening programme coincided with a marked reduction in advanced lesions over time. The high predominance of left-sided cancerous lesions highlights the need to optimize the detection of right-sided neoplasia.

References

1. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin [Internet]. 2021 May 1 ;71(3):209–49. Available from: https://onlinelibrary.wiley.com/doi/full/10.3322/caac.21660
2. Kanth P, Inadomi JM. Screening and prevention of colorectal cancer. BMJ [Internet]. 2021 Sep 15;374. Available from: https://www-bmj-com.ez.statsbiblioteket.dk/content/374/bmj.n1855
3. Adamsen S, Mosbach KKT, Milliam PB, Hansen OH. Injuries by colonoscopy and sigmoidoscopy requiring laparotomy. Claims to the Patient Insurance Corporation. Ugeskr Laeger. 2007 May 7;169(19):1788–93.
4. Baatrup G. Lokal resektion af tidlige kolorektale cancere - og hvad gør vi så?. Ugeskr Laeger. 2021 Jun 28;183(26).