e17548 Background: Clinically actionable variants (CAVs) in BRCA1 and BRCA2 determine eligibility for targeted therapy in high grade serous ovarian cancer (HGSOC). However, RNA based biomarkers indicating BRCA status remain limited. Circular RNAs (circRNAs) are non coding RNAs with regulatory roles in gene expression. Their stability and detectability in human biofluids make circRNAs attractive liquid biopsy candidates. This study examined whether circRNA expression varies by BRCA background and assessed translational potential. Methods: This was a translational biomarker study examining circRNA expression across distinct BRCA backgrounds in high grade serous ovarian cancer. Three HGSOC cell lines representing BRCA1 mutant, BRCA2 mutant, and BRCA wildtype backgrounds were analysed. The primary endpoint was identification of circRNAs differentially expressed according to BRCA status. circRNA profiling was performed using the Arraystar circRNA microarray platform. Differential expression was assessed using normalised signal intensity, a fold change threshold ≥2, and associated statistical outputs, with adjustment for multiple testing. Candidate circRNAs were prioritised based on fold change, reproducibility, and annotation quality. Selected circRNAs were validated using divergent primers and quantitative PCR across the HGSOC panel, a chemotherapy resistant model, and clinical tumour samples stratified by BRCA status. Results: Heatmaps and volcano plots showed circRNA profiles were driven mainly by cell line specific patterns. However, at the individual transcript level, distinct circRNAs demonstrated BRCA associated differences. Five circRNAs were prioritised for validation. Three circRNAs (hsa_circ_400223, hsa_circ_008026, and hsa_circ_003300) were upregulated in both BRCA1 and BRCA2 mutant models compared with wildtype. hsa_circ_100059 was predominantly expressed in the BRCA1 mutant cell line, while hsa_circ_025636 was highly expressed in the BRCA2 mutant cell line. In clinical samples, hsa_circ_025636, a RASSF8 derived circRNA, showed higher expression in a subset of BRCA mutant cases relative to wildtype (p<0.05). Conclusions: This study provides evidence that circRNAs may serve as biomarkers of BRCA status in HGSOC. While global circRNA profiles were not defined by BRCA genotype, specific circRNAs demonstrated BRCA associated expression. The RASSF8 derived circRNA hsa_circ_025636 is a promising candidate and warrants evaluation in larger cohorts alongside functional studies.

e17558 Background: Most patients with high grade serous ovarian cancer (HGSOC) ultimately relapse with chemotherapy resistant disease, and resistance to Poly (ADP ribose) polymerase inhibitors (PARPi) is increasingly recognised. The mechanisms underlying inherent and acquired resistance remain poorly understood, and no validated biomarkers exist to guide treatment beyond BRCA status. circularRNAs (circRNAs) are stable non-coding RNAs with regulatory roles in gene expression and may capture transcriptional reprogramming associated with treatment resistance. This study examined circRNA expression in chemotherapy and PARPi resistant ovarian cancer (OC) models to identify candidate biomarkers of resistance. Methods: Chemotherapy (carboplatin and paclitaxel) and PARPi resistant (olaparib) COV362 (BRCA1 mutant) sublines were generated using progressive dose escalation. Resistance was confirmed using CCK-8 and BrdU proliferation assays. This was a preclinical, exploratory biomarker discovery study. The primary endpoint was the identification of circRNAs differentially expressed between parental (drug-sensitive), chemotherapy-resistant and PARPi-resistant cell lines. circRNA expression profiling was performed using the Arraystar circRNA microarray platform. Differential expression was assessed using normalised signal intensity, fold-change thresholds and associated statistical outputs, with adjustment for multiple testing. Candidate circRNAs were prioritised based on magnitude of differential expression and annotation quality, and were validated using divergent primers and quantitative PCR. Results: Chemotherapy and PARPi resistant COV362 sublines were successfully established over 10 and 14 months, respectively. circRNA profiling revealed distinct segregation of parental (drug sensitive), chemotherapy and PARPi resistant cell lines using hierarchical clustering. Although global circRNA patterns remained broadly conserved, each model showed a distinct set of upregulated transcripts. At the individual transcript level, however, a subset of circRNAs demonstrated resistance-associated differences and were prioritised for further evaluation. hsa_circ_0085803 (p <0.05) was upregulated in the chemotherapy resistant model, whereas hsa_circ_0003258 (p <0.05) was predominately expressed in the PARPi resistant mode compared to matched parental cell line. Conclusions: Distinct circRNA signatures were associated with chemotherapy and PARPi resistance in COV362, with hsa_circ_0085803 and hsa_circ_0003258 identified as candidate biomarkers of resistance. These findings support the potential of circRNAs as indicators of treatment resistance in HGSOC.

Supplementary Table 1. All validation results and evaluation of intratumoral heterogeneity.

On the International Agency for Research on Cancer's 60th birthday, we reflect on the continuing importance of the agency's international remit in cancer research, driven by its founding principle that the discovery of carcinogenic agents is relevant to all humans globally. The present narrative elaborates on this continued stimulus through the discussion of three themes: (1) trust and independence, especially amid vested interests and misinformation; (2) equity and efficiency as essential elements of progress in cancer research, achieved by prioritizing research in underserved and understudied areas and locations where exposure doses or contrasts are high; and (3) a commitment to context-relevant research through strong local collaborations and capacity building. We demonstrate that the international research conducted at the International Agency for Research on Cancer is a core means to advance carcinogenic discovery, building reciprocal partnerships where all countries benefit when discoveries are shared across borders. If we do not attempt to understand cancer everywhere, we will never fully understand it anywhere.

Purpose HPV driven oropharyngeal squamous cell carcinoma (OPSCC) proffers a more favorable prognosis in contrast to its HPV negative counterparts. Consequently, studies have analyzed de-intensification measures of current treatment practices to minimize the associated morbidity. However, 25% of patients with HPV positive OPSCC experience a locoregional recurrence (LR) and/or distant metastasis (DM). This study aimed to determine if patient demographics, tumor characteristics and specific HPV genotyping were attributable to HPV positive OPSCC who develop LR and/or DM based on data collated from the largest Head and Neck Cancer center in the Republic of Ireland, with a mean follow up period of 11.16 years. Methods Patient demographics was obtained from HPV positive OPSCC patients from 3 Dublin Hospitals diagnosed over a 10-year period. In addition, HPV genotyping via Ion Torrent Next Generation Sequencing (NGS) and mutations analysis using smMIP panel based NGS was conducted on the cohort archival tumor DNA. Results 67 patients met the criteria for inclusion and had a mean follow-up period of 11.16 years. 28.36% developed LR and/or DM. A statistically significant correlation was identified linking age at diagnosis (p = 0.047), excessive alcohol consumption (p = 0.017) and a higher clinical stage (p = 0.01) with LR and/or DM.HPV 16 was the most prevalent genotype (93.65%), with dual HPV infection illustrated in 4 cases. BRAF, EGFR, ERBB2, KIT, KRAS, NRAS, PDGFRA somatic mutations were not identified in the recurrence cohort. Conclusion With the ongoing focus to reduce treatment for patient with HPV positive OPSCC, upfront recognition of patients at greater risk of LR and/or DM is essential. In the era of personalized medicine, it is hoped that interrogation of archival tumor tissue via similar techniques used in this study may yield pioneering findings with a clinical and prognostic significance.

IntroductionSepsis is responsible for 1 in 5 deaths globally and the majority of those who survive have lasting health issues. A hallmark of sepsis is a deregulated inflammatory response to infection, with leukocytes playing a critical role.MethodsThis study utilised a targeted single-cell multi-omics approach to characterise peripheral blood mononuclear cell (PBMC) populations and their transcriptomic profiles in an Irish cohort of people with (i) sepsis and (ii) bacteraemia without sepsis (defined as clinically significant positive blood culture without sepsis as assessed by the Clinical Microbiology service).ResultsVariable leukocyte distributions were identified, with decreased cytotoxic lymphocytes, including CD8+ T cells, natural killer cells, CD56+ T cells, γδ T cells, mucosal-associated invariant T cells, and increased T helper (Th) cell subsets within sepsis samples. Additionally, PBMCs from sepsis samples displayed an impaired expression profile in effector T cells, resulting in widespread suppression of PBMC cytotoxic activity.DiscussionThese results identify potential mechanisms underlying the functional impairment witnessed in sepsis. Such mechanisms may inform future diagnostic and treatment strategies.

Background Developing artificial intelligence (AI) models for digital pathology requires large datasets from multiple sources. However, without careful implementation, AI models risk learning confounding site-specific features in datasets instead of clinically relevant information, leading to overestimated performance, poor generalizability to real-world data, and potential misdiagnosis. Methods Whole-slide images (WSIs) from The Cancer Genome Atlas (TCGA) colon (COAD), and stomach adenocarcinoma datasets were selected for inclusion in this study. Patch embeddings were obtained using three feature extraction models, followed by ComBat harmonization. Attention-based multiple instance learning models were trained to predict tissue-source site (TSS), as well as clinical and genetic attributes, using raw, Macenko normalized, and Combat-harmonized patch embeddings. Results TSS prediction achieved high accuracy (AUROC > 0.95) with all three feature extraction models. ComBat harmonization significantly reduced the AUROC for TSS prediction, with mean AUROCs dropping to approximately 0.5 for most models, indicating successful mitigation of batch effects (e.g., CCL-ResNet50 in TCGA-COAD: Pre-ComBat AUROC = 0.960, Post-ComBat AUROC = 0.506, p<0.001). Clinical attributes associated with TSS, such as race and treatment response, showed decreased predictability post-harmonization. Notably, the prediction of genetic features like MSI status remained robust after harmonization (e.g., MSI in TCGA-COAD: Pre-ComBat AUROC = 0.667, Post-ComBat AUROC = 0.669, p=0.952), indicating the preservation of true histological signals. Conclusion ComBat harmonization of deep learning-derived histology features effectively reduces the risk of AI models learning confounding features in WSIs, ensuring more reliable performance estimates. This approach is promising for the integration of large-scale digital pathology datasets.

Background: Recent advances in computational pathology have shown potential in predicting biomarkers from haematoxylin and eosin (H&E) whole-slide images (WSI). However, predicting the outcome directly from WSIs remains a substantial challenge. In this study, we aimed to investigate how gene expression, predicted from WSIs, could be used to evaluate overall survival (OS) in patients with lung adenocarcinoma (LUAD). Methods: Differentially expressed genes (DEGs) were identified from The Cancer Genome Atlas (TCGA)-LUAD cohort. Cox regression analysis was performed on DEGs to identify the gene prognostics of OS. Attention-based multiple instance learning (AMIL) models were trained to predict the expression of identified prognostic genes from WSIs using the TCGA-LUAD dataset. Models were externally validated in the Clinical Proteomic Tumour Analysis Consortium (CPTAC)-LUAD dataset. The prognostic value of predicted gene expression values was then compared to the true gene expression measurements. Results: The expression of 239 prognostic genes could be predicted in TCGA-LUAD with cross-validated Pearson’s R > 0.4. Predicted gene expression demonstrated prognostic performance, attaining a cross-validated concordance index of up to 0.615 in TCGA-LUAD through Cox regression. In total, 36 genes had predicted expression in the external validation cohort that was prognostic of OS. Conclusions: Gene expression predicted from WSIs is an effective method of evaluating OS in patients with LUAD. These results may open up new avenues of cost- and time-efficient prognosis assessment in LUAD treatment.

Objectives To examine the feasibility of implementing a 10-week exercise-based cancer rehabilitation programme in a national cancer centre.Design A single-arm prospective feasibility study.Setting An outpatient physiotherapy department.Participants Forty de-conditioned cancer survivors (< 1 year post completion of treatment).Interventions A 10-week programme of twice weekly group-based supervised exercise sessions.Main outcome measures A mixed methods approach was used. The primary outcome of the study was feasibility, evaluated in terms of recruitment, adherence, attrition and stakeholder acceptance of the programme. Secondary outcomes examined the effect of the exercise intervention on physical function and quality of life measures.Results Forty patients (age 60 (SD 10.6) years; n = 12 breast cancer, n = 11 lung cancer, n = 7 prostate cancer, n = 5 colorectal cancer, n = 5 other) participated. In total 82% (n = 33) participants completed the post-programme assessment. Deterioration of health and concerns over COVID-19 were the most common reasons for dropout (both n = 2). Adherence to both the supervised exercise classes and home exercise programme was high (78% and 94% respectively). No adverse events were recorded during the intervention or assessments. Qualitative feedback from stakeholders highlighted the acceptability of the programme as well as many perceived benefits of the exercise programme. Improvements in three quality of life sub-scales (physical function, role function and emotional function), physical activity levels and aerobic fitness levels were found post-intervention.Conclusion It appears feasible to offer a 10-week exercise programme to patients attending a national cancer centre, with adequate recruitment, retention and adherence rates and high acceptability among stakeholders.

5031 Background: Mismatch repair (MMR) deficiency and microsatellite instability (MSI) are predictive biomarkers for immunotherapy response. The best approach to identify patients with such tumors is unclear in prostate cancer. Methods: This study included men diagnosed with primary prostate cancer during prospective follow-up of the Health Professionals Follow-up Study (HPFS) and Physicians’ Health Study (PHS). The highest-grade/index lesions of tumor tissue from radical prostatectomy (95%) or transurethral resections of the prostate were mounted in triplicate on tissue microarrays. Immunohistochemistry for the MMR proteins MLH1, MSH2, MSH6, and PMS2 was performed, with scoring as MMR-deficient requiring a visible staining in a non-tumor internal positive control of the same case. For validation, a polymerase chain reaction-based MSI assay (Idylla MSI Test, Biocartis) was performed on tumor DNA of MMR-deficient cases and a selection of MMR-intact cases. Results: The study included 1016 men with prostate cancer. Tumor stage was predominantly pathologically localized (71% stage pT1/2, 18% T3a/b) with a full distribution of Gleason scores, including 20% Gleason score 6 (grade group 1), 36% 3+4 (grade group 2), 23% 4+3 (grade group 3), 8% 8 (grade group 4) and 13% 9-10 (grade group 5). MMR tumor scoring could be performed for MLH1 in 747 cases (75% of those with tumor tissue), for MSH2 in 903 cases (90%), for MSH6 in 708 cases (74%), and for PMS2 in 703 cases (72%). The remaining tumors were unevaluable due to lack of non-tumor tissue necessary as an internal positive control. Of the 903 tumors evaluable for MSH2 protein loss, 4 tumors had loss of MSH2 (prevalence 0.4%, 95% confidence interval [CI] 0.2–1.1%), and 3 of 708 evaluable tumors had concomitant loss of MSH6 (prevalence 0.4%, 95% CI 0.1–1.2%). No tumor had loss of MLH1 or PMS2. The MMR-deficient tumors had Gleason scores of 3+4, 8 (two cases), and 9–10. Tumor DNA of 1 of the 4 MMR-deficient tumors met the manufacturer’s cut-off for MSI-high; two additional MMR-deficient cases had non-zero repeats, with good reproducibility between tumor cores and technical replicates. One MSH2-deficient tumor and 6 DNA samples from 3 MMR-intact cases had repeat scores of zero. Conclusions: In this nationwide prospective study, MMR deficiency was rare in primary, surgically treated prostate cancer. This low prevalence contrasts with hospital-based studies of MMR deficiency and MSI that may have represented tumors with certain clinical features. The low prevalence and the need for an internal positive control for reliable scoring calls into question to what extent immunohistochemistry-based screening for MMR deficiency on limited tissue specimens, such as prostate biopsies, is routinely feasible.

Higher Education Institutions (HEIs) have the potential to impact positively on the health and wellbeing of their staff and students. Using and expanding on the 'health promoting university' (HPU) platform within HEIs, this article provides a description of 'Healthy Trinity', which is an initiative underway in Trinity College Dublin, the University of Dublin. First, Healthy Trinity is contextualized in background literature including international and national policy and practice. Second, an overview of Healthy Trinity is provided including its vision and goals. Third, the article describes the steps taken relating to the identification of stakeholders and use of a network and a co-lead model. Within this approach, the article describes a partnership approach whereby responsibilities regarding health and wellbeing are shared by individuals and the institution. Fourth, the design and implementation of Healthy Trinity is discussed by taking a 'settings approach', in which the emphasis for change is placed on individual behaviours, environment, policy and organizational culture. Consideration is given to the interplay between intervention, implementation strategy and context for successful systemic implementation. The fifth element presented is the early-stage challenges encountered during implementation, such as the need to secure recurrent funding and the importance of having a direct input to the governance of the University to enable systemic change. The sixth and final component of the article is an outline of Healthy Trinity's intention to utilize a process evaluation of the early implementation phases of this complex intervention within a settings approach. Potential deliverables and impacts of this HPU initiative are presented and discussed.

Chronic obstructive pulmonary disease (COPD) and lung cancer 17 are two of the most prevalent and debilitating respiratory diseases worldwide, both associated with high morbidity and mortality rates. As major global health concerns, they impose a substantial burden on patients, healthcare systems, and society at large. Despite their distinct aetiologies, lung cancer and COPD share common risk factors, clinical features, and pathological pathways, which have spurred increasing research interest in their co-occurrence. One area of particular interest is the role of the lung microbiome in the development and progression of these diseases, including the transition from COPD to lung cancer. Exploring novel therapeutic strategies, such as metal-based drugs, offers a potential avenue for targeting the microbiome in these diseases to improve patient outcomes. This review aims to provide an overview of the current understanding of the lung microbiome, with a particular emphasis on COPD and lung cancer, and to discuss the potential of metal-based drugs as a therapeutic strategy for these conditions, specifically concerning targeting the microbiome.

Supplementary Data from FOXA1 Is a Potential Oncogene in Anaplastic Thyroid Carcinoma

Purpose All patients living with cancer, including those with metastatic cancer, are encouraged to be physically active. This paper examines the secondary endpoints of an aerobic exercise intervention for men with metastatic prostate cancer. Methods ExPeCT (Exercise, Prostate Cancer and Circulating Tumour Cells), was a multi-centre randomised control trial with a 6-month aerobic exercise intervention arm or a standard care control arm. Exercise adherence data was collected via heart rate monitors. Quality of life (FACT-P) and physical activity (self-administered questionnaire) assessments were completed at baseline, at 3 months and at 6 months. Results A total of 61 patients were included (69.4 ± 7.3 yr, body mass index 29.2 ± 5.8 kg/m 2 ). The median time since diagnosis was 34 months (IQR 7–54). A total of 35 (55%) of participants had > 1 region affected by metastatic disease. No adverse events were reported by participants. There was no effect of exercise on quality of life (Cohen’s d = − 0.082). Overall adherence to the supervised sessions was 83% (329 out of 396 possible sessions attended by participants). Overall adherence to the non-supervised home exercise sessions was 72% (months 1–3) and 67% (months 3–6). Modelling results for overall physical activity scores showed no significant main effect for the group ( p -value = 0.25) or for time ( p -value = 0.24). Conclusion In a group of patients with a high burden of metastatic prostate cancer, a 6-month aerobic exercise intervention did not lead to change in quality of life. Further exercise studies examining the role of exercise for people living with metastatic prostate cancer are needed. Trial Registration The trial was registered at clinicaltrials.gov (NCT02453139) on May 25th 2015.