Somatic alterations of targetable oncogenes are frequently observed in BRCA1/2 mutation negative male breast cancers

Male breast cancer (MBC) is a rare disease. Due to its rarity, MBC research and clinical approach are mostly based upon data derived from its largely known female counterpart. We aimed at investigating whether MBC cases harbor somatic alterations of genes known as prognostic biomarkers and molecular therapeutic targets in female breast cancer.We examined 103 MBC cases, all characterized for germ-line BRCA1/2 mutations, for somatic alterations in PIK3CA, EGFR, ESR1 and CCND1 genes.Pathogenic mutations of PIK3CA were detected in 2% of MBCs. No pathogenic mutations were identified in ESR1 and EGFR. Gene copy number variations (CNVs) analysis showed amplification of PIK3CA in 8.1%, EGFR in 6.8% and CCND1 in 16% of MBCs, whereas deletion of ESR1 was detected in 15% of MBCs. Somatic mutations and gene amplification were found only in BRCA1/2 mutation negative MBCs.Significant associations emerged between EGFR amplification and large tumor size (T4), ER-negative and HER2-positive status, between CCND1 amplification and HER2-positive and MIB1-positive status, and between ESR1 deletion and ER-negative status.Our results show that amplification of targetable oncogenes is frequent in BRCA1/2 mutation negative MBCs and may identify MBC subsets characterized by aggressive phenotype that may benefit from potential targeted therapeutic approaches

L’infezione da Clostridioides difficile tossinogeno: uno stargate per l’invasione da germi nosocomiali? Diagnostica molecolare ed epidemiologia

L’infezione da Clostridioides difficile (CDI) è una delle principali cause di diarrea infettiva ed è responsabile dell’insorgenza della colite pseudomembranosa. Le lesioni anatomiche connesse alla CDI fanno sì che questa possa rappresentare la via di accesso verso la circolazione sistemica per germi multi-antibiotico resistenti (MDR), quali quelli contenuti nel lume intestinale di pazienti ospedalizzati e sottoposti a terapie antibiotiche. L’entità della lesione e la probabilità di traslocazione batterica sarebbero direttamente dipendenti dal pattern tossinogeno espresso dallo specifico ceppo, o ribotipo, di C. difficile coinvolto nell’insorgenza della patologia. Ne deriva che la conoscenza del profilo tossinogeno, dell'incidenza di CDI acquisita in ambito ospedaliero e comunitario e delle infezioni sostenute da germi a prevalente impatto nosocomiale risulta fondamentale per il monitoraggio dell'epidemiologia, per la prevenzione negli ambienti ospedalieri e per una più corretta gestione clinica del paziente affetto. Questo studio si è proposto di analizzare diffusione e incidenza della CDI in un grande ospedale, di valutare il pattern tossinogeno dei ceppi di C. difficile isolati e di valutare l’eventuale correlazione con la colonizzazione da parte di germi responsabili di infezioni nosocomiali quali Candida spp, Enterococchi Vancomicina-resistenti (VRE), Enterobatteriaceae produttrici di carbapenemasi (CPE) e Stafilococchi Meticillino-resistenti (MRS). Tali dati sono stati confrontati con quelli ottenuti dall’analisi di una popolazione controllo, per evidenziare eventuali differenze nella presenza di infezioni sostenute da germi MDR e ottenere una completa visione dell’impatto della CDI a livello ospedaliero. Le analisi sono state effettuate su una serie di 100 campioni positivi per C. difficile tossinogeno (casi) e di 100 campioni negativi allo screening (controlli), pervenuti presso il laboratorio di Microbiologia Clinica dell’Azienda Ospedaliera Universitaria Umberto I di Roma e collezionati consecutivamente. Il 77% dei casi e il 95% dei controlli proveniva da pazienti ospedalizzati. L’analisi caso-controllo ha evidenziato che la colonizzazione intestinale da germi MDR si osservava con una frequenza all’incirca doppia nei casi di CDI rispetto ai controlli (81% vs. 49%), con una differenza statisticamente significativa (p=0.0001). Considerando le singole specie, si è osservata per tutte una maggiore frequenza di colonizzazione nella popolazione dei casi rispetto ai controlli, con differenze nelle frequenze di isolamento che sono risultate essere statisticamente significative (χ2 p compresi tra 0.00001 e 0.0037). Candida ed Enterococco VRE sono emersi quali germi più frequentemente riscontrati sia nei casi che nei controlli. Candida ha inoltre mostrato una diversa distribuzione tra i pazienti a provenienza ospedaliera e comunitaria, con una significativa maggiore presenza nei pazienti ospedalizzati affetti da CDI (p=0.04). Per quanto riguarda la caratterizzazione molecolare del profilo tossinogeno e del ribotipo dei ceppi di C. difficile isolati, il 65% dei ceppi presentava il profilo tossinogeno di “tossina B”, seguiti dai ceppi ipervirulenti NAP1/027 recanti la delezione del gene repressore tcdC e da ceppi con profilo “tossina B e binaria” (19% e 16%, rispettivamente). La distribuzione dei profili tossinogeni tra pazienti ospedalieri e comunitari è risultata essere statisticamente significativa (χ2 p=0.0019), con il profilo «tossina B» più frequente nei pazienti ospedalizzati. Sono stati inoltre identificati 22 diversi ribotipi, con una maggiore frequenza osservata per i ribotipi 018 (28%), 027 (19%) e 078 (8%). Il ribotipo 027 è risultato prevalente nei pazienti con CDI comunitaria, mentre il ribotipo 018 è stato osservato più frequentemente nei pazienti ospedalizzati. Non sono state evidenziate associazioni statisticamente significative tra un particolare germe MDR e un dato ribotipo o profilo tossinogeno. Da quanto osservato nello studio, la colonizzazione intestinale nei pazienti con CDI può essere quindi un fattore da valutare attentamente per identificare tempestivamente pazienti ad «alto rischio» di traslocazione batterica dal tratto intestinale e attuare delle strategie di monitoraggio corretto. Allo stesso tempo, la messa in atto di programmi per il corretto uso di antibiotici e la somministrazione di probiotici permetterebbe un maggiore controllo delle infezioni nosocomiali sostenute da microrganismi antibiotico-resistenti

Proffered papers and posters presented at the Sixth International Symposium on Hereditary Breast and Ovarian Cancer— BRCA: Challenges and Opportunities

Poster P100 Whole-Exome Sequencing Revealed a Novel PALB2 Mutation in a Male Breast Cancer Family Objectives Male breast cancer (mbc) is a rare disease, whose causation appears to be largely associated with genetic factors. Whole-exome sequencing (wes) is a powerful tool to explore the heritability of complex diseases, including breast cancer (bca). Our aim was to evaluate whether rare mutations may explain a fraction of mbc not accounted for by BRCA1/2 genes. Here, we applied wes analysis to a high-risk mbc family. Methods A BRCA1/2–negative family with 2 first-degree male and 4 female bca cases was selected for the study. Peripheral-blood dna samples from 1 male and 2 female bca cases were examined. Libraries were prepared and sequenced on the Illumina HiSeq instrument. A bioinformatic pipeline available at https://bioinformatics.cineca.it/wep/ was used. A validation series of 48 high-risk BRCA1/2–negative mbc patients from the Italian Multicenter Study on mbc was analyzed by Sanger sequencing. Results A novel PALB2 truncating mutation, c.419delA (p.K140fsX35), was identified by wes in a female bca case and her paternal uncle, diagnosed with melanoma and bca, but not in her maternal aunt, affected with bca; her father, diagnosed with bca, was an obligate mutation carrier. PALB2 mutational analysis of 48 high-risk mbcs identified another truncating mutation, c.1984A>T (p.K662X), in a man diagnosed with breast, lung, and prostate cancer, and with family history of bca. Overall, 3/50 high-risk mbcs (6%) were carriers of PALB2 pathogenic mutations. Conclusions This study highlights the importance of a particular selection of pedigrees including mbc to better define the fraction of bca attributable to genetic predisposition. Our results add to the accumulating evidence that PALB2 is strongly involved in bca risk in both sexes. Consideration should be given to clinical testing for PALB2 in BRCA1/2– negative families, including not only multiple female bca cases, but also 1 or more members diagnosed with mbc or other cancers

Fast and reliable diagnosis of XDR Acinetobacter baumannii meningitis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Bacterial meningitis is a medical emergency needing quick and timely diagnosis. Even though meningitis caused by Acinetobacter baumannii is relatively rare, it is associated with high mortality rates especially in neurosurgery patients and represents a serious therapeutic problem due to the limited penetration of effective antibiotics into the cerebrospinal fluid. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) has been effectively used as a rapid method for microbial identification. In this case report we identified A. baumanni by MALDI-TOF technique directly from the CSF drawn from the external ventricular drainage of a patient with severe confusional state and signs of meningism. Simultaneously the antibiotic susceptibility test was performed by automated method from the pellet of the broth-enriched sample. The MALDI-TOF technique allowed microbial identification in less than 30 minutes, and the susceptibility test result was available in eight hours, thus allowing a fast diagnosis ready for prompt and targeted antimicrobial therapy

Candida gut colonization, yeast species distribution, and biofilm production in Clostridioides difficile infected patients. A comparison between three populations in two different time periods

Candida gut colonization and yeast biofilm production capacity were investigated, by means of XTT reduction assay, in Clostridioides difficile infected (CDI) patients, in non-CDI diarrheic patients, and in healthy donors in two different time periods (2013–2015 and 2018–2019 respectively). Candida gut colonization was significantly (p < 0.001) associated to C. difficile infection, and to patients infected with hypervirulent C. difficile strains bearing the tcdC deletion at nucleotide 117 (p = 0.0003). Although there was not a prevalent yeast species in CDI patients, C. albicans was the species significantly (p < 0.001) associated to both the infections sustained by the non-hypervirulent C. difficile strains and those caused by the hypervirulent strain (p = 0.001). The biofilm production by the yeasts isolated from the CDI patients and from non-CDI diarrheic patients did not differ significantly. However, a significantly (p = 0.007) higher biofilm production was observed in the Candida strains, particularly C. albicans, isolated from healthy donors compared to that of the yeasts cultured from CDI patients. Seasonal occurrence was observed in the isolation rate of CDI and non-CDI diarrheic cases (p = 0.0019), peaking in winter for CDI patients and in spring for non-CDI diarrheic patients. Furthermore, seasonality emerged in the gut colonization by Candida of CDI patients in the winter. It seems, therefore, that the reduced capacity of biofilm production by Candida strains isolated from CDI patients might have a role in the development of C. difficile infection, probably facilitating the spread of the bacteria into the gut thus amplifying their pathogenic action

Mutational profiling in melanocytic tumors: multiple somatic mutations and clinical implications.

In this study, we analyzed multiple somatic mutations in 10 genes relevant in melanoma tumorigenesis and targeted therapies. Overall, 45% of the tumors showed mutations and, in particular, 33% had multiple mutations. Based on our results, we conclude that the assessment of mutation status of multiple genes, including CDKN2A, could provide a genetic profile that can be useful as a prognostic and therapeutic marker in melanocytic tumors. © 2014 S. Karger AG, Basel

A-1012G Promoter Polymorphism of Vitamin D Receptor Gene Is Associated with Psoriasis Risk and Lower Allele-Specific Expression

Psoriasis is caused by a combination of genetic, immunologic, and environmental factors. The vitamin D receptor (VDR) is involved in antiproliferative and prodifferentiation pathways in keratinocytes and exerts immunosuppressive effects. We aimed to investigate possible associations between VDR polymorphisms and psoriasis susceptibility and to evaluate functional effects of potential psoriasis-associated polymorphisms. We genotyped 108 patients with psoriasis and 268 healthy controls at 5 VDR polymorphisms (A-1012G, FokI, BsmI, ApaI, and TaqI) by TaqMan allelic-discrimination real-time polymerase chain reaction. We found a significant increased overall risk of psoriasis for the VDR A-1012G promoter polymorphism (odds ratio [OR]=2.43, 95% confidence interval [CI]: 1.15-5.13; p=0.05). A significant higher frequency (p=0.035) of the A allele was found in psoriatic cases compared with controls. In a case-case analysis, a statistically significant association between A-1012G and family history emerged (p=0.033). Furthermore, a significant association of A-1012G risk genotypes with a lower expression of VDR mRNA emerged (p=0.0028). Our data show that VDR promoter A-1012G polymorphism is associated with psoriasis risk and suggest that this polymorphism may modulate psoriasis risk by affecting VDR expression

EMSY copy number variation in male breast cancers characterized for BRCA1 and BRCA2 mutations

PURPOSE: Male breast cancer (MBC) is a rare disease that shares some similarities with female breast cancer (FBC). Like FBC, genetic susceptibility to MBC can be referred to mutations in BRCA1 and, particularly, BRCA2 genes. However, only about 10 % of MBCs are caused by BRCA1/2 germ-line mutations, while the largest part are sporadic cancers and may derive from somatic alterations. EMSY, a BRCA2 inactivating gene, emerged as a candidate gene involved in the pathogenesis of sporadic FBC, and its amplification was suggested to be the somatic counterpart of BRCA2 mutations. Considering the relevant role of BRCA2 in MBC, we aimed at investigating the role of EMSY gene copy number variations in male breast tumors. METHODS: EMSY copy number variations were analyzed by quantitative real-time PCR with TaqMan probes in a selected series of 75 MBCs, characterized for BRCA1/2 mutations. RESULTS: We reported EMSY amplification in 34.7 % of MBCs. A significant association emerged between EMSY amplification and BRCA1/2 mutations (p = 0.03). We identified two amplification subgroups characterized by low and high amplification levels, with BRCA2-related tumors mostly showing low EMSY amplification. CONCLUSIONS: Our results show a high frequency of EMSY amplification in MBC, thus pointing to a role of EMSY in the pathogenesis of this disease. EMSY amplification may be a new feature that might uncover underlying molecular pathways of MBCs and may allow for the identification of MBC subgroups with potential clinical implication for targeted therapeutic approaches

<i>Legionella pneumophila</i> Infections during a 7-Year Retrospective Analysis (2016–2022): Epidemiological, Clinical Features and Outcomes in Patients with Legionnaires’ Disease

Legionella pneumophila (LP) is one of the main causative agents of community-acquired pneumonia in Europe and its fifth bacterial cause in Italy (4.9%). We conducted a seven year retrospective analysis of LP infection serogroup 1 in Asti, Piedmont, between 2016 and 2022. Patients were included if they tested positive for the Legionella urinary antigen. Clinical, laboratory, and radiologic data were analyzed to describe the risk factors for mortality. Fifty patients with LD were collected, mainly male, with a median age of 69 years. The main comorbidities were cardiovascular diseases (50%), pulmonary diseases (26%), and neurological diseases (12%). The most common clinical presentations were fever, respiratory, gastrointestinal, and neurologic symptoms. Older age (p = 0.004), underlying cardiovascular diseases (p = 0.009), late diagnosis at admission (p = 0.035), and neurological symptoms at diagnosis (p = 0.046) were more common in the non-survivor group. Moreover, a septic-shock presentation or the need for non-invasive ventilation at admission were associated with a higher mortality. No considerable differences in the biochemical data were found between the two groups except for the median neutrophil count, lymphocyte count, neutrophil-to-lymphocyte ratio, and PCT value. We did not find any differences in mortality related to the choice of antibiotic regimen. Differences in outcome were associated with the median duration of treatment (p =< 0.001) but not to the choice of antibiotic regimen (mainly levofloxacin or azithromycin). In conclusion, early individuation of the wide spectrum of clinical characteristics of LP infection such as respiratory, cardiac, and neurological manifestations of the patient’s comorbidities, and significant biochemical data should help clinicians flag high risk patients and potentially improve their outcome

Whole-exome sequencing and targeted gene sequencing provide insights into the role of PALB2 as a male breast cancer susceptibility gene

BACKGROUND: Male breast cancer (MBC) is a rare disease whose etiology appears to be largely associated with genetic factors.BRCA1 and BRCA2 mutations account for about 10% of all MBC cases. Thus, a fraction of MBC cases are expected to be due to genet-ic factors not yet identified. To further explain the genet ic susceptibility for MBC, whole-exome sequencing (WES) and targeted genesequencing were applied to high-risk, BRCA1/2 mutation–negative MBC cases. METHODS: Germ-line DNA of 1 male and 2 femaleBRCA1/2 mutation–negative breast cancer (BC) cases from a pedigree showing a first-degree family history of MBC was analyzedwith WES. Targeted gene sequencing for the validation of WES results was performed for 48 high-risk, BRCA1/2 mutation–negativeMBC cases from an Italian multicenter study of MBC. A case-control series of 433 BRCA1/2 mutation–negative MBC and female breastcancer (FBC) cases and 849 male and female controls was included in the study. RESULTS: WES in the family identified the partnerand localizer of BRCA2 (PALB2) c.419delA truncating mutation carried by the proband, her father, and her paternal uncle (all affectedwith BC) and the N-acetyltransferase 1 (NAT1) c.97C&gt;T nonsense mutation carried by the proband’s maternal aunt. Targeted PALB2sequencing detected the c.1984A&gt;T nonsense mutation in 1 of the 48 BRCA1/2 mutation–negative MBC cases. NAT1 c.97C&gt;T was notfound in the case-control series. CONCLUSIONS: These results add strength to the evidence showing that PALB2 is involved in BCrisk for both sexes and indicate that consideration should be given to clinical testing of PALB2 for BRCA1/2 mutation–negative fami-lies with multiple MBC and FBC case