Sarah J. Lord

The clinical management of Barrett’s esophagus (BE) and the risk of esophageal adenocarcinoma (EAC) is dependent on the presence or absence of dysplasia. The histopathological diagnosis of dysplasia is unreliable however, and would benefit from additional diagnostic criteria. Here, we obtained EM-seq whole-genome methylation profiles from 86 samples across the BE/EAC disease spectrum, notably supplementing the scant existing methylation evidence for low-grade and high-grade dysplasia. While the bulk of methylation changes occur in the transition from normal squamous epithelium to intestinal metaplasia (non-dysplastic Barrett’s esophagus), we identified a smaller set of changes that mark the onset of dysplasia and progression to EAC. Methylation changes were broadly independent of copy number variation. They were tightly anticorrelated with RNA-seq expression for some genes. Deconvolution of the bulk methylation signatures into constituent cell types revealed four sample groupings enriched with a particular cell type (squamous, columnar, lymphoid and erythroid cells), somewhat reflecting existing clinical classifications. We outline potential markers that could improve the diagnosis of metaplasia and dysplasia in EAC.

BACKGROUND: Point-of-care high-sensitivity cTn (cardiac troponin; hs-cTn) testing has the potential to expedite decision-making and reduce emergency department (ED) length of stay for patients presenting with possible myocardial infarction by ensuring that results are consistently available when sought by clinicians. We assessed the real-life effectiveness and safety of implementing point-of-care hs-cTn testing in the ED. METHODS: We conducted a pragmatic, stepped-wedge cluster-randomized trial. The control arm was usual care with an accelerated diagnostic pathway using a single-sample rule-out step with a central laboratory hs-cTn assay. The intervention arm used the same pathway with a point-of-care hs-cTnI assay. The primary effectiveness outcome was ED length of stay assessed using a generalized linear mixed model. The safety outcome was myocardial infarction or cardiac death within 30 days. RESULTS: Six sites participated, with 59 980 ED presentations (44 747 individuals; mean 61±19 years of age; 49.5% female) from February 2023 through January 2025; 31 392 presentations were during the intervention arm. After adjustment for covariates associated with length of stay, the intervention reduced length of stay by 13% (95% CI, 9%–16%; P P =0.54). CONCLUSIONS: Implementation of whole-blood hs-cTnI testing at the point of care into an accelerated diagnostic pathway was safe and reduced length of stay in the ED compared with laboratory testing. REGISTRATION: URL: https://www.anzctr.org.au ; Unique identifier: ACTRN12619001189112.

New prognostic biomarkers for oesophageal adenocarcinoma (OAC) may help treatment planning as current staging does not accurately predict survival for many patients. We used a 451 cancer-associated gene panel to assess genomic alterations associated with survival for a cohort of 65 OAC patients who underwent surgical resection without perioperative treatment, reflecting institutional practice at the time. Median age at operation was 72 years, 88% were male, median 32 lymph nodes were removed per patient, and minimum follow-up exceeded 11 years. TNM stages (pTNM, 8 ^th ed.) were 14% stage 1, 14% stage 2, 45% stage 3, and 6% stage 4 (non-locoregional nodal metastases). Copy number changes were common, and somatic mutations were similar to those reported previously. Genetic associations with worse survival included amplification of CCND1 (HR=3.08, p=0.02), EGFR (HR=2.98, p=0.01), loss of ARID2 (HR=2.12, p=0.05), KMT2D (HR=1.99, p=0.07) and missense mutation within RYR1 (HR=2.68, p=0.01). This study identifies candidate genomic prognostic biomarkers in OAC using a cohort treated by surgery alone, enabling assessment of tumour biology independent of the confounding effects of peri-operative systemic therapy and radiotherapy. Several of the identified alterations are potentially targetable, supporting further investigation of genomic profiling to refine prognostication and guide treatment selection.

Esophageal adenocarcinoma (EAC) has a poor five-year survival rate and one of the fastest-rising incidences of any cancer. The presence of dysplasia in Barrett’s esophagus (BE) is the main risk factor for EAC development and guides clinical management. Unfortunately, the current histopathological diagnosis of dysplasia is unreliable, with poor inter-observer agreement, highlighting the need for novel biomarkers that can improve diagnostic accuracy. Here, we performed transcriptome profiling across the full spectrum of BE-related neoplasia in 85 samples to delineate gene expression alterations in progressively worse disease stages and identify biomarkers that could complement histopathology to improve the detection of dysplasia and EAC in endoscopic biopsy specimens. Differential gene expression and pathway analyses revealed that the most extensive transcriptional changes occurred during the transition from normal squamous (NSq) to non-dysplastic BE (NDBE), consistent with metaplastic transformation. Compared to NDBE, dysplasia was characterized by enhanced cellular growth and proliferation; upregulation of immune processes and oncogenic signaling pathways were present in EAC. Using machine learning approaches, we identified a novel five-gene panel suitable for a potential RNAseq-based diagnostic test (SLC11A1, IL36A, LUCAT1, MIR215, RNU6-954P) and performed an initial validation of this signature in an additional 51 samples. We also identified several potential novel immunohistochemical markers that may warrant further evaluation, including TREM1, CXCL5, OSM, and motilin. In summary, by delineating transcriptional changes across the full disease spectrum, this study identifies several candidate biomarkers for improving current diagnostic methods for Barrett’s dysplasia and EAC. Significance statement: Current histopathological assessment of Barrett’s dysplasia has limited diagnostic reproducibility. By defining transcriptional changes across the disease spectrum, this study identifies novel biomarker candidates for the detection of dysplasia and adenocarcinoma. These include a five-gene panel for a potential RNA sequencing- based diagnostic test to complement histopathology.

Article Reply to “Is this quantitative test fit-for-purpose?” was published on March 24, 2025 in the journal Clinical Chemistry and Laboratory Medicine (CCLM) (volume 0, issue 0).

Recent changes in the regulatory assessment of in vitro medical tests reflect a growing recognition of the need for more stringent clinical evidence requirements to protect patient safety and health. Under current regulations in the United States and Europe, when needed for regulatory approval, clinical performance reports must provide clinical evidence tailored to the intended purpose of the test and allow assessment of whether the test will achieve the intended clinical benefit. The quality of evidence must be proportionate to the risk for the patient and/or public health. These requirements now cover both commercial and laboratory developed tests (LDT) and demand a sound understanding of the fundamentals of clinical performance measures and study design to develop and appraise the study plan and interpret the study results. However, there is a lack of harmonized guidance for the laboratory profession, industry, regulatory agencies and notified bodies on how the clinical performance of tests should be measured. The Working Group on Test Evaluation (WG-TE) of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) is a multidisciplinary group of laboratory professionals, clinical epidemiologists, health technology assessment experts, and representatives of the in vitro diagnostic (IVD) industry. This guidance paper aims to promote a shared understanding of the principles of clinical performance measures and study design. Measures of classification performance, also referred to as discrimination, such as sensitivity and specificity are firmly established as the primary measures for evaluating the clinical performance for screening and diagnostic tests. We explain these measures are just as relevant for other purposes of testing. We outline the importance of defining the most clinically meaningful classification of disease so the clinical benefits of testing can be explicitly inferred for those correctly classified, and harm for those incorrectly classified. We introduce the key principles and a checklist for formulating the research objective and study design to estimate clinical performance: (1) the purpose of a test e.g. diagnosis, screening, risk stratification, prognosis, prediction of treatment benefit, and corresponding research objective for assessing clinical performance; (2) the target condition for clinically meaningful classification; (3) clinical performance measures to assess whether the test is fit-for-purpose; and (4) study design types. Laboratory professionals, industry, and researchers can use this checklist to help identify relevant published studies and primary datasets, and to liaise with clinicians and methodologists when developing a study plan for evaluating clinical performance, where needed, to apply for regulatory approval.

Introduction Clinical assessment in emergency departments (EDs) for possible acute myocardial infarction (AMI) requires at least one cardiac troponin (cTn) blood test. The turn-around time from blood draw to posting results in the clinical portal for central laboratory analysers is ~1–2 hours. New generation, high-sensitivity, point-of-care cardiac troponin I (POC-cTnI) assays use whole blood on a bedside (or near bedside) analyser that provides a rapid (8 min) result. This may expedite clinical decision-making and reduce length of stay. Our purpose is to determine if utilisation of a POC-cTnI testing reduces ED length of stay. We also aim to establish an optimised implementation process for the amended clinical pathway. Methods and analysis This quality improvement initiative has a pragmatic multihospital stepped-wedge cross-sectional cluster randomised design. Consecutive patients presenting to the ED with symptoms suggestive of possible AMI and having a cTn test will be included. Clusters (comprising one or two hospitals each) will change from their usual-care pathway to an amended pathway using POC-cTnI—the ‘intervention’. The dates of change will be randomised. Changes occur at 1 month intervals, with a minimum 2 month ‘run-in’ period. The intervention pathway will use a POC-cTnI measurement as an alternate to the laboratory-based cTn measurement. Clinical decision-making steps and logic will otherwise remain unchanged. The POC-cTnI is the Siemens (Erlangen Germany) Atellica VTLi high-sensitivity cTnI assay. The primary outcome is ED length of stay. The safety outcome is cardiac death or AMI within 30 days for patients discharged directly from the ED. Ethics and dissemination Ethics approval has been granted by the New Zealand Southern Health and Disability Ethics Committee, reference 21/STH/9. Results will be published in a peer-reviewed journal. Lay and academic presentations will be made. Māori-specific results will be disseminated to Māori stakeholders. Trial registration number ACTRN12619001189112.