Genotypic and phenotypic analysis of familial male breast cancer shows under representation of the HER2 and basal subtypes in BRCA-associated carcinomas

BACKGROUND: Male breast cancer (MBC) is an uncommon and relatively uncharacterised disease accounting for <1% of all breast cancers. A significant proportion occurs in families with a history of breast cancer and in particular those carrying BRCA2 mutations. Here we describe clinicopathological features and genomic BRCA1 and BRCA2 mutation status in a large cohort of familial MBCs. METHODS: Cases (n=60) included 3 BRCA1 and 25 BRCA2 mutation carries, and 32 non-BRCA1/2 (BRCAX) carriers with strong family histories of breast cancer. The cohort was examined with respect to mutation status, clinicopathological parameters including TNM staging, grade, histological subtype and intrinsic phenotype. RESULTS: Compared to the general population, MBC incidence was higher in all subgroups. In contrast to female breast cancer (FBC) there was greater representation of BRCA2 tumours (41.7% vs 8.3%, p=0.0008) and underrepresentation of BRCA1 tumours (5.0% vs 14.4%, p=0.0001). There was no correlation between mutation status and age of onset, disease specific survival (DSS) or other clincopathological factors. Comparison with sporadic MBC studies showed similar clinicopathological features. Prognostic variables affecting DSS included primary tumour size (p=0.003, HR:4.26 95%CI 1.63-11.11), age (p=0.002, HR:4.09 95%CI 1.65-10.12), lymphovascular (p=0.019, HR:3.25 95%CI 1.21-8.74) and perineural invasion (p=0.027, HR:2.82 95%CI 1.13-7.06). Unlike familial FBC, the histological subtypes seen in familial MBC were more similar to those seen in sporadic MBC with 46 (76.7%) pure invasive ductal carcinoma of no special type (IDC-NST), 2 (3.3%) invasive lobular carcinomas and 4 (6.7%) invasive papillary carcinoma. A further 8 (13.3%) IDC-NST had foci of micropapillary differentiation, with a strong trend for co-occurrence in BRCA2 carriers (p=0.058). Most tumours were of the luminal phenotype (89.7%), with infrequent HER2 (8.6%) and basal (1.7%) phenotype tumours seen. CONCLUSION: MBC in BRCA1/2 carriers and BRCAX families is different to females. Unlike FBC, a clear BRCA1 phenotype is not seen but a possible BRCA2 phenotype of micropapillary histological subtype is suggested. Comparison with sporadic MBCs shows this to be a high-risk population making further recruitment and investigation of this cohort of value in further understanding these uncommon tumours

Changes in frailty status and discharge destination post emergency laparotomy

Background: Pre-operative frailty adversely affects morbidity and mortality after emergency laparotomy (EmLap), especially in older adults (65 years and above). Little is known about frailty after EmLap. We explored the change in frailty status from pre- to post-EmLap and any influence on discharge destination. Methods: EmLap patients aged ≥ 65years from an acute surgical site were recruited from May 2022 to April 2023. Prospective data collection included demographics, frailty, mortality and discharge destination. Frailty was assessed using the Rockwood Clinical Frailty Scale at pre-EmLap and day-90 post-EmLap (&lt; 4 as non-frail, 4 as pre-frail and &gt; 4 as frail). EmLap patients with no 90-day follow-up were excluded. A p-value of &lt; 0.05 was considered significant. Results: 63 EmLap patients were included in the study. The median age was 75 years (range 65–91 years) with 36 (57.1%) females. Eleven (17.5%) were living with frailty pre-EmLap, and 10 (15.9%) developed new frailty by day-90 post-EmLap. Pre-EmLap, all patients came from home with 20.6% of the frail and pre-frail group having a package of care service (POC) in place. On 90-day post-EmLap, 1 was still an inpatient but 25.8% had a change in discharge destination: care home (n = 1), home with new POC (n = 2) and home with increased POC (n = 13). Of the 16 patients with change of discharge destination, 9 (56.3%) were frail pre-EmLap. There was a significant association between pre-EmLap frailty and change in home circumstances on discharge (p &lt; 0.00001). Conclusions: Emergency surgery can increase a patient’s frailty status and significantly increases care requirements and social support after hospital discharge. Frailty assessment needs to be performed before and after admission in all EmLap patients to improve post-EmLap care planning and patient expectations

Changes in frailty status and discharge destination post emergency laparotomy

Background: Pre-operative frailty adversely affects morbidity and mortality after emergency laparotomy (EmLap), especially in older adults (65 years and above). Little is known about frailty after EmLap. We explored the change in frailty status from pre- to post-EmLap and any influence on discharge destination. Methods: EmLap patients aged ≥ 65years from an acute surgical site were recruited from May 2022 to April 2023. Prospective data collection included demographics, frailty, mortality and discharge destination. Frailty was assessed using the Rockwood Clinical Frailty Scale at pre-EmLap and day-90 post-EmLap ( 4 as frail). EmLap patients with no 90-day follow-up were excluded. A p-value of < 0.05 was considered significant. Results: 63 EmLap patients were included in the study. The median age was 75 years (range 65–91 years) with 36 (57.1%) females. Eleven (17.5%) were living with frailty pre-EmLap, and 10 (15.9%) developed new frailty by day-90 post-EmLap. Pre-EmLap, all patients came from home with 20.6% of the frail and pre-frail group having a package of care service (POC) in place. On 90-day post-EmLap, 1 was still an inpatient but 25.8% had a change in discharge destination: care home (n = 1), home with new POC (n = 2) and home with increased POC (n = 13). Of the 16 patients with change of discharge destination, 9 (56.3%) were frail pre-EmLap. There was a significant association between pre-EmLap frailty and change in home circumstances on discharge (p < 0.00001). Conclusions: Emergency surgery can increase a patient’s frailty status and significantly increases care requirements and social support after hospital discharge. Frailty assessment needs to be performed before and after admission in all EmLap patients to improve post-EmLap care planning and patient expectations

An optimised monophasic faecal extraction method for LC-MS analysis and its application in gastrointestinal disease

Liquid chromatography coupled with mass spectrometry (LC-MS) metabolomic approaches are widely used to investigate underlying pathogenesis of gastrointestinal disease and mechanism of action of treatments. However, there is an unmet requirement to assess faecal metabolite extraction methods for large-scale metabolomics studies. Current methods often rely on biphasic extractions using harmful halogenated solvents, making automation and large-scale studies challenging. The present study reports an optimised monophasic faecal extraction protocol that is suitable for untargeted and targeted LC-MS analyses. The impact of several experimental parameters, including sample weight, extraction solvent, cellular disruption method, and sample-to-solvent ratio, were investigated. It is suggested that a 50 mg freeze-dried faecal sample should be used in a methanol extraction (1:20) using bead beating as the means of cell disruption. This is revealed by a significant increase in number of metabolites detected, improved signal intensity, and wide metabolic coverage given by each of the above extraction parameters. Finally, we addressed the applicability of the method on faecal samples from patients with Crohn’s disease (CD) and coeliac disease (CoD), two distinct chronic gastrointestinal diseases involving metabolic perturbations. Untargeted and targeted metabolomic analysis demonstrated the ability of the developed method to detect and stratify metabolites extracted from patient groups and healthy controls (HC), highlighting characteristic changes in the faecal metabolome according to disease. The method developed is, therefore, suitable for the analysis of patients with gastrointestinal disease and can be used to detect and distinguish differences in the metabolomes of CD, CoD, and HC

Investigation of frailty markers including a novel biomarker panel in emergency laparotomy: protocol of a prospective cohort study

Abstract Background Emergency laparotomy (EmLAP) is one of the commonest emergency operations performed in the United Kingdom (approximately 30, 000 laparotomies annually). These potentially high-risk procedures can be life changing with frail patients and/ or older adults (≥ 65 years) having the poorest outcomes, including mortality. There is no gold standard of frailty assessment and no clinical chemical biomarkers existing in practice. Early detection of subclinical changes or deficits at the molecular level are essential in improving our understanding of the biology of frailty and ultimately improving patient outcomes. This study aims primarily to compare preoperative frailty markers, including a blood-based biomarker panel, in their ability to predict 30 and 90-day mortality post-EmLAP. The secondary aim is to analyse the influence of perioperative frailty on morbidity and quality of life post-EmLAP. Methods A prospective single centred observational study will be conducted on 150 patients ≥ 40 years of age that undergo EmLAP. Patients will be included according to the established NELA (National Emergency Laparotomy Audit) criteria. The variables collected include demographics, co-morbidities, polypharmacy, place of residence, indication and type of surgery (as per NELA criteria) and prognostic NELA score. Frailty will be assessed using: a blood sample for ultra-high performance liquid chromatography mass spectrometry analysis; preoperative CT abdomen pelvis (sarcopenia) and Rockwood Clinical Frailty Scale (CFS). Patients will be followed up for 90 days. Variables collected include blood samples (at post operative day 1, 7, 30 and 90), place of residence on discharge, morbidity, mortality and quality of life (EQ-5D-5 L). The frailty markers will be compared between groups of frail (CFS ≥ 4) and non-frail using statistical methods such as regression model and adjusted for appropriate confounding factors. Discussion This study hypothesises that frailty level changes following EmLAP in frail and non- frail patients, irrespective of age. We propose that non- frail patients will have better survival rates and report better quality of life compared to the frail. By studying the changes in metabolites/ biomarkers in these patients and correlate them to frailty status pre-surgery, this highly novel approach will develop new knowledge of frailty and define a new area of clinical biomolecular research. Trial registration ClinicalTrials.gov: NCT05416047. Registered on 13/06/2022 (retrospectively registered)

Additional file 1 of Investigation of frailty markers including a novel biomarker panel in emergency laparotomy: protocol of a prospective cohort study

Additional File 1: NELA inclusion and exclusion criteri

PIK3CA mutations are frequently observed in BRCAX but not BRCA2-associated male breast cancer

INTRODUCTION: Although a substantial proportion of male breast cancers (MBCs) are hereditary, the molecular pathways that are activated are unknown. We therefore examined the frequency and clinicopathological associations of the PIK3CA/mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK) pathways and their regulatory genes in familial MBC. METHODS: High resolution melting analysis and confirmatory sequencing was used to determine the presence of somatic mutations in PIK3CA (exon 9 and 20), AKT1 (exon 4), KRAS (exon 2) and BRAF (exon 15) genes in 57 familial MBCs. Further analysis of the PIK3CA/mTOR pathway was performed using immunohistochemistry for the pAKT1, pS6 and p4EBP1 biomarkers. RESULTS: PIK3CA somatic mutations were identified in 10.5% (6 of 57) of cases; there were no AKT1, KRAS or BRAF somatic mutations. PIK3CA mutations were significantly more frequent in cancers from BRCAX patients (17.2%, 5/29) than BRCA2 (0%, 0/25) carriers (P = 0.030). Two BRCAX patients had an E547K mutation which has only been reported in one female breast cancer previously. PIK3CA mutation was significantly correlated with positive pS6 (83.3% vs. 32.0%, P = 0.024) and negative p4EBP1 (100% vs. 38.0%, P = 0.006) expression, but not pAKT expression. Expression of nuclear p4EBP1 correlated with BRCA2 mutation carrier status (68.0% vs. 38.7%, P = 0.035). CONCLUSIONS: Somatic PIK3CA mutation is present in familial male breast cancer but absent in BRCA2 carriers. The presence of two of the extremely rare E547K PIK3CA mutations in our cohort may have specific relevance in MBCs. Further study of PIK3CA in MBCs, and in particular BRCAX patients, may contribute to further establishing the relevance of specific PIK3CA mutations in MBC aetiology and in the identification of particular patient groups most likely to benefit from therapeutic targeting with the novel PIK3CA inhibitors that are currently in development