BACKGROUND: Hepatocellular carcinoma (HCC) is a highly heterogeneous malignancy with a poor prognosis and limited response to current therapies. Cancer stem cells (CSCs) contribute to this heterogeneity, driving tumor progression, immune evasion, and therapeutic resistance. Epigenetic regulators have emerged as pivotal modulators of CSC phenotypes. The histone methyltransferase SETDB1, a key stemness factor in both embryonic and adult stem cells, catalyzes H3K9 trimethylation (H3K9me3) and acts as a major oncogenic driver in a wide range of solid and hematological malignancies. Yet, its specific role in maintaining stemness and contributing to therapeutic resistance in HCC remains poorly defined. METHODS: Integrative analyses of bulk and single-cell transcriptomic datasets from public HCC cohorts were performed. Tumors were stratified by SETDB1 expression, followed by pathway enrichment, immune deconvolution, and stemness scoring. Associations with clinical outcomes, immune phenotypes, drug resistance signatures, and chromatin-binding partners (DNMT3A, TRIM28, HDAC1) were also explored. The role of SETDB1 was then examined in vitro, where a stem-like phenotype was induced in HCC cell lines under hypoxic conditions, and patterns of SETDB1 expression and stemness marker levels were assessed following paclitaxel treatment. RESULTS: SETDB1-low tumors were enriched in well-differentiated, immunologically active subclasses and were associated with increased predicted responsiveness to immune checkpoint blockade (ICB), sorafenib, and transarterial chemoembolization. Conversely, SETDB1-high tumors exhibited transcriptional features of dedifferentiation, elevated stemness scores, and enrichment in aggressive molecular subtypes. These tumors also displayed higher TP53 mutation rates, reduced immune infiltration, immune exclusion signatures, and activation of pathways linked to ferroptosis and ABC transporter dysfunction, consistent with resistance to ICB and conventional therapies. SETDB1 expression was also associated with hypoxia-related pathways, suggesting a role in maintaining CSC niches. In vitro, hypoxia-induced stemness coincided with increased SETDB1 levels, whereas paclitaxel treatment decreased both SETDB1 expression and stemness markers. A chromatin risk score incorporating SETDB1 and its partners predicted poor disease-free survival independently of clinical parameters. CONCLUSIONS: SETDB1 defines a stemness-enriched, immune-resistant HCC subtype associated with poor outcomes and therapeutic failure. Correlations between SETDB1 expression and stem-like features in vitro suggest a potential role in maintaining CSC phenotypes, supporting its relevance as a biomarker and candidate for epigenetic- and CSC-directed therapeutic strategies.

Hepatocellular carcinoma (HCC) is a highly heterogeneous malignancy characterized by poor prognosis and limited therapeutic response. Cancer stem cells (CSCs) contribute to tumor progression, therapeutic resistance, and tumor recurrence. Among transcriptional regulators potentially involved in these processes, Specificity Protein 1 (SP1) has emerged as a candidate integrator of oncogenic and epigenetic signaling networks. However, its contribution to CSC-associated phenotypes and drug resistance in HCC remains incompletely defined. In this study, we combined transcriptomic analyses of TCGA datasets with functional experiments in HCC cell lines (Huh7 and HepG2). SP1-associated transcriptional programs were targeted pharmacologically using mithramycin A (MIT-A) and genetically using siRNA-mediated knockdown. The effects were assessed by RNA sequencing, RT-qPCR, Western blotting, flow cytometry, and functional assays evaluating proliferation, migration, CSC-associated properties, and response to sorafenib. MIT-A treatment markedly reduced the expression of stemness-associated transcription factors (NANOG, OCT4, SOX2) and CSC markers (CD133, CD24), impaired CSC-related functions including ALDH activity and the Side Population phenotype, and inhibited cell proliferation and migration. MIT-A also sensitized both parental and sorafenib-resistant HCC cells to sorafenib, associated with modulation of apoptotic regulators and reduced transporter-mediated efflux activity. SP1 knockdown partially reproduced several of these effects, supporting a contribution of SP1-dependent transcriptional programs to these phenotypes. Overall, these findings identify SP1-associated transcriptional networks as potential regulators of CSC features and therapeutic resistance in HCC and support targeting SP1-associated transcriptional programs as a strategy to enhance sorafenib efficacy.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease, projected to affect 55% globally by 2040. Up to one-third of NAFLD patients develop non-alcoholic steatohepatitis (NASH), with 40% progressing to fibrosis. However, there are currently few reliable tools to predict disease progression. Impaired mitochondrial dynamics, characterized by dysregulated fission, fusion, and mitophagy, have emerged as key events in NAFLD pathophysiology, contributing to hepatocyte death and inflammation. This study explored the transition from steatosis to NASH through transcriptomic analyses, including data from patients with steatosis and those with NASH at different fibrosis stages. By identifying a transcriptomic signature associated with disease progression, the study revealed increased expression of genes involved in mitochondrial dynamics in NASH compared to steatosis and during NASH-related fibrosis. Histological analyses highlighted the central role of Dynamin-related protein 1 (Drp1), a dynamin GTPase essential for mitochondrial fission and mitophagy. In human liver biopsies, Drp1 expression progressively increased from NAFLD to NASH and NASH-related fibrosis and cirrhosis, predominantly in Kupffer cells. These finding suggest Drp1 is a potential driver of the transition to more severe liver damage, making it a promising biomarker for NASH development and progression and a potential therapeutic target in metabolic disorders.

Hepatic small vessel neoplasm (HSVN) and anastomosing hemangioma (AH) are 2 capillary liver neoplasms that are challenging to differentiate in clinical practice. This study aimed to refine their classification through integrated histopathologic and molecular analyses. We conducted a bicentric retrospective study of 25 cases (15 resections and 10 biopsies). Four liver pathologists (A.B., A.L.B., V.P., C.G.) independently reviewed histologic and immunohistochemical features. Targeted DNA next-generation sequencing and RNA sequencing were performed on 21 and 15 cases, respectively. Seven lesions (28%) were diagnosed as AH, presenting as well-demarcated tumors with fibrotic stroma, often associated with cirrhosis (5/7, 71%). The remaining 18 lesions (72%) were identified as HSVN, typically infiltrative and occurring predominantly in noncirrhotic livers (78%). Microscopically, HSVN displayed 2 distinct patterns: (1) a classic pattern (8/18, 44%) composed of small, thin-walled vessels and (2) a hepatocellular-reactive pattern (7/18, 39%) with small vessels interwoven with regenerative hepatocellular trabeculae. A mixed pattern appeared in 3 of 18 cases (17%). No cytologic atypia or mitoses were observed. Tumor cells consistently expressed ERG, CD31, and CD34, with Ki67 indices <10% in all cases. Mutations in GNA genes were found in 20 of 21 cases (95%), distributed between GNA14 (81% overall: 4 AH and 13 HSVN, P = .544) and GNAQ (19% overall: 2 AH and 2 HSVN, P = .544). Transcriptomic analysis revealed no differentially expressed genes between AH and HSVN. After a median follow-up of 18 months, no recurrence was observed in resected tumors. Tumor growth was noted in 3 nonresected or partially resected cases (2 AH and 1 HSVN). In conclusion, despite subtle pathological differences, HSVN and AH exhibit overlapping clinical and molecular features, supporting their reclassification as a single benign entity, namely AH. Notably, some lesions exhibit a hepatocellular-reactive pattern that closely mimics hepatocellular adenoma, posing a significant diagnostic challenge, particularly in biopsy specimens.

Background The predictive value of C-reactive protein (CRP) for anticipating or ruling out complications after elective abdominal surgery has been extensively studied. However, its role following elective liver resection (LR) remains poorly explored. This study aimed to evaluate the performance of CRP levels after LR. Methods The CRP values on postoperative day (POD) 1, 3 and 5 in a cohort of patients undergoing elective LR at a single tertiary center were retrospectively analyzed. Analyses were performed for laparoscopic and major LR subgroups. Results A total of 448 LR (392 patients), including 134 laparoscopic LR and 169 major LR, were analyzed. Severe morbidity (Clavien-Dindo grade >= III) occurred in 56 cases (12.4%) with an overall 90-day mortality rate of 2% (n = 9). Peak CRP levels were observed on POD 3. On POD 3, CRP levels were lower in the laparoscopic subgroup compared to open LR and lower after major LR compared to limited LR. In the overall cohort, the predictive CRP value on POD 1, 2, 3 remained poor for detecting major complications or any complications, with all calculated area under the curve (AUCs) below 0.65. Among the 69 patients with a POD 3 CRP value < 100 mg/L, only 2 developed major complications. Conclusion Unlike other abdominal procedures, CRP levels on POD 1, 3 and 5 after LR have insufficient predictive value for guiding postoperative management. The role of alternative biomarkers or early imaging strategies should be investigated.

Background: Liver transplantation (LT) remains a potentially haemorrhagic procedure whose perioperative bleeding and transfusion could be better monitored using point-of-care devices. Quantra® is a device based on sonorheometry to assess whole blood clot formation. Our aims were to describe Quantra® parameters during LT and to study their correlations with standard laboratory parameters, and to determine Quantra® cut-off values for thrombocytopenia, hypofibrinogenemia and coagulation factors’ deficit. Methods: In 34 patients undergoing LT, blood samples were collected before surgical incision, 15 min after the beginning of the anhepatic phase, and 15 min after arterial revascularization of the graft. Results: Clotting time (CT) was well correlated with prothrombin (PT) ratio and activated partial thromboplastin time (aPTT) ratio. Platelet contribution to clot stiffness (PCS) was correlated with platelets (ρ = 0.82, p < 0.001) and fibrinogen contribution clot stiffness (FCS) with fibrinogen (Fg) (ρ = 0.74, p < 0.001). CT predicted a PT ratio < 30% with an area under the curve (AUC) of 0.93 (95% CI 0.87–0.98; p < 0.001). PCS predicted a platelet count < 50 G/L with an AUC of 0.87 (95% CI 0.76–0.98, p < 0.001). FCS predicted a Fg < 1.0, 1.2 or 1.5 g/L, with an AUC of 0.86 (95% CI 0.77–094, p < 0.001), 0.82 (95% CI 0.74–0.91, p < 0.001) and 0.88 (95% CI 0.82–0.95, p < 0.001), respectively. Conclusion: Quantra® provides a rapid assessment of haemostasis during LT.

INTRODUCTION:Considering the growing interest in matched cancer treatment, our aim was to evaluate the ability of a comprehensive genomic profiling (CGP) assay to propose at least one targeted therapy given an identified genomic alteration or signature (actionability), and to collect the treatment modifications based on the CGP test results in clinical practise for solid tumors.METHODS:This retrospective, multicentre French study was conducted among 25 centres that participated in a free of charge program between 2017 and 2019 for a tissue CGP test. Data were collected on the patient, disease, tumor genomic profile, treatment suggested in the report (related to the genomic profile results) and subsequent therapeutic decisions according to the physician's declaration.RESULTS:Among the 416 patients, most had lung cancer (35.6%), followed by biliary tract cancer (11.5%) or rare cancers (11.1%); 75% had a metastatic disease. The actionability was 75.0% (95% CI [70.6%-78.9%]) for all patients, 85.1% and 78.4%, respectively in lung cancer and metastatic patients. After exclusion of clinical trial suggestions, the actionability decreased to 62.3% (95% CI [57.5%-66.8%]). Treatment modification based on the test results was observed in 17.3% of the patients and was more frequent in metastatic disease (OR = 2.73, 95% CI [1.31-5.71], p = 0.007). The main reasons for no treatment modification were poor general condition (33.2%) and stable disease or remission (30.2%). The genomic-directed treatment changes were performed mostly during the first six months after the CGP test, and interestingly a substantial part was observed from six to 24 months after the genomic profiling.CONCLUSION:This French study provides information on the real-life actionability of a CGP test based on tissue samples, and trends to confirm its utility in clinical practice across the course of the disease, in particularly for patients with lung cancer and/or advanced disease.

Background: Peritoneal carcinomatosis often results in alterations in intestinal peristalsis and recurrent abdominal pain. Pain management in these patients is often unsatisfactory. This study aimed to investigate whether endothelin-1 (EDN1) was involved in pain mediation in peritoneal carcinomatosis, and thus whether the EDN1 pathway could be a new therapeutic target for peritoneal carcinomatosis-associated pain.Methods: EDN1 plasma levels and abdominal pain severity were assessed in patients with abdominal tumors, with or without peritoneal carcinomatosis, and in healthy donors. The effects of EDN1 on the visceromotor response to colorectal distension, and on colonic contractions were then examined in mice, and the mechanism of action of EDN1 was then investigated by measuring the impact of EDN1 exposure on calcium mobilization in cultured neurons. Inhibition studies were also performed to determine if the effects of EDN1 exposure could be reversed by EDN1-specific receptor antagonists.Results: A positive correlation between EDN1 plasma levels and abdominal pain was identified in patients with peritoneal carcinomatosis. EDN1 exposure increased visceral sensitivity and the amplitude of colonic contractions in mice and induced calcium mobilization by direct binding to its receptors on sensory neurons. The effects of EDN1 were inhibited by antagonists of the EDN1 receptors.Conclusions: This preliminary study, using data from patients with peritoneal carcinomatosis combined with data from experiments performed in mice, suggests that EDN1 may play a key role mediating pain in peritoneal carcinomatosis. Our findings suggest that antagonists of the EDN1 receptors might be beneficial in the management of pain in patients with peritoneal carcinomatosis.

The emergence and quick spread of SARS-CoV-2 has pointed at a low capacity response for testing large populations in many countries, in line of material, technical and staff limitations. The traditional RT-qPCR diagnostic test remains the reference method and is by far the most widely used test. These assays are limited to a few probe sets, require large sample PCR reaction volumes, along with an expensive and time-consuming RNA extraction step. Here we describe a quantitative nanofluidic assay that overcomes some of these shortcomings, based on the BiomarkTM instrument from Fluidigm. This system offers the possibility of performing 4608 qPCR end-points in a single run, equivalent to 192 clinical samples combined with 12 pairs of primers/probe sets in duplicate, thus allowing the monitoring of SARS-CoV-2 including the detection of specific SARS-CoV-2 variants, as well as the detection other pathogens and/or host cellular responses (virus receptors, response markers, microRNAs). The 10 nL-range volume of BiomarkTM reactions is compatible with sensitive and reproducible reactions that can be easily and cost-effectively adapted to various RT-qPCR configurations and sets of primers/probe. Finally, we also evaluated the use of inactivating lysis buffers composed of various detergents in the presence or absence of proteinase K to assess the compatibility of these buffers with a direct reverse transcription enzymatic step and we propose several protocols, bypassing the need for RNA purification. We advocate that the combined utilization of an optimized processing buffer and a high-throughput real-time PCR device would contribute to improve the turn-around-time to deliver the test results to patients and increase the SARS-CoV-2 testing capacities.

En France, 10-11 % des patients avec un cancer broncho-pulmonaire non à petites cellules (CBNPC) sont diagnostiqués avec une mutation EGFR (EGFRm + ). Les traitements standards de première ligne des CBNPC EGFRm+ sont des inhibiteurs de tyrosine kinase (TKI). L’objectif de cette étude était de décrire la prise en charge en vie réelle des patients CBNPC EGFRm+. Le programme Épidémio-Stratégie Médico-Économique (ESME) de Unicancer est une cohorte rétrospective nationale qui recueille des données sur le diagnostic, la prise en charge thérapeutique et le suivi de patients (pts) traités pour un CBP parmi 33 établissements de santé. Les principaux critères d’inclusion de cette analyse sont: ≥ 18 ans, confirmation histologique du CBNPC, diagnostic de maladie avancée ou métastatique entre le 1er janvier 2015 et 31 décembre 2016, EGFRm+, sans traitement systémique préalable. Cut-off: 15 mars 2019. La recherche de mutation EGFR était effectuée chez 54 % des 13537 pts CBNPC inclus dans la base et 56 % des cas non-épidermoïdes localement avancé ou métastatique. Parmi les testés, 1171 pts (16 %) était EGFRm+. 560 pts répondaient aux critères d’inclusion. Cette population était composée à 65 % de femmes, âge médian 66 ans, 99 % d’histologie non-épidermoïde, 85 % Stade IV de novo. Parmi eux 65 % étaient identifiés EGFRm+ avant initiation d’une première ligne de traitement (42 % Exon19Del, 33 % L858R, 2 % T790 M, 11 % autre mutation, [catégories non-exclusives]). Un traitement de première ligne a été administré à 534 pts (95.4 %). La majorité a reçu en première ligne un traitement par un EGFR-TKI en mono-thérapie: 46 % gefitinib ou erlotinib, 15 % afatinib; 18 % un traitement par chimiothérapie (CT), principalement à base de sel de platine. 355 pts (77.0 %) ont reçu une deuxième ligne de traitement, dont 29 % CT, 29 % osimertinib et 23 % un autre EGFR-TKI. Parmi les CBNPC EGFRm+ avec T790 M (138 pts), 87 (63 %) ont reçu osimertinib en deuxième ligne. Après un suivi médian de 31.8 mois (95 %CI: 29.0; 33.6), 266 patients sont décédés. La médiane de survie globale est de 30.1 mois (95 %CI: 28.3; 33.7). Pour la période de sélection, seule la moitié des patients CBNPC non-épidermoïdes ont fait l’objet d’une recherche de mutation EGFR. L’efficacité des EGFR TKIs apparaît similaire en vie réelle à celle rapportée dans les essais cliniques. Les Résultats suggèrent que plus de patients pourraient bénéficier de cette recherche et d’un TKI en première ligne conformément aux recommandations.

Le programme Épidémio-Stratégie Médico-Économique (ESME) de Unicancer est une cohorte rétrospective nationale qui recueille des données sur le diagnostic, la prise en charge thérapeutique et le suivi de patients (pts) traités pour un cancer bronchopulmonaire (CBP) dans 33 établissements de santé. L’objectif de cette analyse est de décrire les modalités de prise en charge des cancers bronchiques à petites cellules (CBPC) métastatiques en vie réelle. Les critères d’inclusions étaient : âge > 18 ans, confirmation histologique du CBPC, diagnostic de maladie métastatique (de novo ou rechute), recevant une première cure par etoposide + chimiothérapie à base de sel de platine (PT) à partir du 1er janvier 2015. Parmi 2374 pts CBPC dans la base de données ESME CBP, 1315 pts (55%) répondaient aux critères d’inclusions : hommes : 64%, âge médian : 64 ans, ECOG 0-1 : 57% (quand disponible) et fumeurs ou ex-fumeurs : 92%, stade IV de novo : 90%. 24% des pts avaient ≥ 3 sites métastatiques au diagnostic. Les sites métastatiques les plus fréquents étaient hépatiques (43%), osseux (36%), cérébraux (32%), et surrénaliens (24%). Parmi les pts avec métastases cérébrales, 35% étaient symptomatiques au diagnostic. 49% des patients ont reçu une seule ligne de traitement, tandis que 30% ont reçu deux lignes et 21% ≥ 3 lignes. En première ligne, la majorité des patients ont reçu étoposide + PT seuls (67% étoposide + carboplatine ; 20% étoposide + cisplatine) avec une durée médiane de traitement similaire : 3.6 mois (IC95% : 2,5–4,0) avec étoposide + carboplatine, 3,6 mois (IC95% : 2,3–4,0) étoposide + cisplatine. Après un suivi médian de 20,1 mois (IC95% : 19,0–22,0) dans la population d’intérêt, 1052 pts (80,0%) ont progressé ou sont décédés. La médiane de survie sans progression (SSP) selon l’investigateur était de 5,8 mois (IC95% : 5,4–6,0) et 6,1 mois (IC95% : 5,6–6,7) respectivement. La médiane de survie globale était de 11,0 mois (IC95% : 10,4–11,8) avec étoposide + carboplatine et 14,2 mois (IC95% : 13,1–16,5) avec étoposide + cisplatine (HR = 1,302, IC95% : 1,196–1,621, p < 0,001). Les résultats montrent que la plupart des patients avec CBPC de stade IV reçoit un traitement par étoposide + sel de platine avec une proportion beaucoup plus importante de patients traités par carboplatine que cisplatine. Les résultats obtenus avec ces traitements en termes de durée de traitement, de survie sans progression, et survie globale sont similaires à ceux reportés dans les essais cliniques.

e15574 Background: This pilot study aimed at the evaluation of the efficacy of anti-EGFR therapy in patients with initially RAS-mutated metastatic colorectal cancer, whose liquid biopsy found no RAS mutation after prior chemotherapy failure. Methods: Sixteen patients with RAS-mutated metastatic colorectal cancer in the solid tumor were included, after they had received 1 to 3 prior chemotherapy lines without anti-EGFR. Before inclusion, RAS genotyping was performed in circulating tumor DNA (ctDNA) from liquid biopsy, after a median duration of ̃24 months after the initial RAS status determination. No RAS mutation was detected in the circulating tumor DNA from 9 patients (56%), who then received cetuximab-FOLFIRI. The 7 patients who had persistent RAS mutation in the liquid biopsy (44%) were treated according to standard recommendations without anti-EGFR. Results: The median progression free survival was 8.2 months [95%CL, 4.5 – 11.8] in patients without detectable ctDNA RAS mutation, as compared to 3.5 months [2.1 – 4.9] in the ctDNA mutated patients (p < 0.001). The median overall survival was 22.3 months [17.3 - 27.3] in the patients with undetectable ctDNA RAS mutation, whereas it was 4.7 months [2.6 - 6.7] in those with ctDNA RAS mutation (p = 0.013). These results suggested the efficacy of cetuximab-based chemotherapy in patients with initially RAS mutated metastatic colorectal cancer, who later displayed no detectable ctDNA RAS mutation. Conclusions: The introduction of an anti-EGFR could provide an additional treatment option for patients with metastatic colorectal cancer with apparent conversion of initial RAS mutation, based on ctDNA assessment after prior failure on anti-EGFR-free chemotherapy.

In this article, we studied geographic variation in the use of personalized genetic testing for advanced non-small cell lung cancer (NSCLC) and we evaluated the relationship between genetic testing rates and local socioeconomic and ecological variables. We used data on all advanced NSCLC patients who had a genetic test between April 2012 and April 2013 in France in the frame of the IFCT Biomarqueurs-France study (n = 15814). We computed four established measures of geographic variation of the sex-adjusted rates of genetic testing utilization at the "départment" (the French territory is divided into 94 administrative units called 'départements') level. We also performed a spatial regression model to determine the relationship between département-level sex-adjusted rates of genetic testing utilization and economic and ecological variables. Our results are the following: (i) Overall, 46.87% lung cancer admission patients obtained genetic testing for NSCLC; département-level utilization rates varied over 3.2-fold. Measures of geographic variation indicated a relatively high degree of geographic variation. (ii) there was a statistically significant relationship between genetic testing rates and per capita supply of general practitioners, radiotherapists and surgeons (negative correlation for the latter); lower genetic testing rates were also associated with higher local poverty rates. French policymakers should pursue effort toward deprived areas to obtain equal access to personalized medicine for advanced NSCLC patients.

PURPOSE:Combining cetuximab with chemotherapy provides clinical benefit to 60% of the patients with RAS wild-type (RAS-wt) metastatic colorectal cancer (mCRC). This pilot study investigated the efficacy of cetuximab-based chemotherapy in a sample of patients (40%) with RAS mutation (RAS-mt) in their primary tumor whose circulating tumor DNA (ctDNA) was RAS-wt.MATERIALS AND METHODS:The occurrence of Kirsten rat sarcoma viral oncogene homolog (KRAS), neuroblastoma rat sarcoma viral oncogene homolog (NRAS), V-raf murine sarcoma viral oncogene homolog B1 (BRAF), and PI3KCA mutations was determined in ctDNA by using a new ultrasensitive analysis based on mass spectrometry detection. All consenting patients with confirmed RAS-mt mCRC had disease progression on previous chemotherapy that contained no anti-epidermal growth factor receptor (EGFR). The patients with RAS-wt ctDNA received cetuximab + fluorouracil, leucovorin, and irinotecan (FOLFIRI), whereas those with RAS-mt ctDNA were treated with the oncologist's choice of therapy.RESULTS:Of 16 registered patients, 11 were male and five female. They were age 48 to 81 years, and they had unresectable metastatic adenocarcinoma from the colon (n = 11) or rectum (n = 5), with a median of two metastatic sites. They had received a median number of three previous chemotherapy protocols. Plasma genotyping identified RAS-mt in seven patients (44%) and RAS-wt in nine patients (56%). In the patients with wt ctDNA, objective tumor response rate was 50.0%, including one complete response and four partial responses after a median number of 6 courses of cetuximab + FOLFIRI (range, 1 to 16 courses). Two of the nine patients had stable disease, and two had progressive disease. No grade 3 to 4 toxicities were encountered. One-year survival rates were 60.0% for the patients with RAS-wt ctDNA and 17.9% for those with RAS-mt ctDNA. Median overall survival times were not reached and 4.7 months, respectively.CONCLUSION:Patients with RAS-mt mCRC whose plasma biopsies contained RAS-wt could benefit from cetuximab-based therapy, a hypothesis to be tested in a prospective randomized trial.

The Forensic Science Institute of the French “Gendarmerie Nationale” (IRCGN™) developed in 2015 an ISO 17025 certified mobile DNA laboratory for genetic analyses. This Mobil’DNA laboratory is a fully autonomous and adaptable mobile laboratory to perform genetic analyses in the context of crime scenes, terrorism attacks or disasters. To support the hospital task force in Paris during the peak of the COVID-19 epidemic, we adapted this mobile genetic laboratory to perform high-throughput molecular screening for coronavirus SARS-CoV-2 by real-time PCR. We describe the adaptation of this Mobil’DNA lab to assist in Coronavirus SARS-CoV-2 diagnosis.

IntroductionKRAS mutations are detected in 20% to 30% of NSCLC. However, KRAS mutation subtypes may differently influence the outcome of patients with advanced NSCLC.MethodsIn the Biomarkers France study, 4894 KRAS mutations (26.2%) were detected in 4634 patients from the 17,664 enrolled patients with NSCLC. Survival and treatment data on noncurative stage III to IV NSCLC were available for 901 patients. First- and second-line treatment effects on progression-free survival and overall survival were analyzed according to the KRAS mutations subtype.ResultsOver 95% of patients with KRAS mutation were smokers or former smokers who were white (99.5%), presenting with adenocarcinoma (82.5%). The most common KRAS mutation subtype was G12C (374 patients; 41.5%), followed by G12V (168; 18.6%), G12D (131; 14.5%), G12A (62; 6.9%), G13C (45; 5.0%), G13D (31; 3.4%), and others (10; 1%). Approximately 21% of patients had transition mutation and 68.2% had a transversion mutation. G12D and transition mutations were predominant in never-smokers. The median overall survival for patients with KRAS-mutated NSCLC was 8.1 months (95% confidence interval [CI]: 7.5–9.5), without any differences according to the different KRAS subtypes mutations. The median progression-free survival was 4.6 months (95% CI: 4.2–5.1) for first-line treatment and 4.8 months (95% CI: 4.3–6.8) for second-line treatment, without any differences according to the different KRAS subtypes mutations.ConclusionsKRAS mutation subtypes influenced neither treatment responses nor outcomes. The KRAS G12C mutation was detected in 41.5% of patients, who are now eligible for potent and specific G12C inhibitors.

OBJECTIVES:Tumor mutation screening is standard of care for patients with stage IV NSCLC. Since a couple of years, widespread NGS approaches used in routine diagnostics to detect driver mutations such as EGFR, KRAS, BRAF or MET allows the identification of other alterations that could modulated the intensity or duration of response to targeted therapies. The prevalence of co-occurring alterations that could affect response or prognosis as not been largely analyzed in clinical settings and large cohorts of patients. Thanks to the IFCT program "Biomarkers France", a collection of samples and data at a nation-wide level was available to test the impact of co-mutations on first line EGFR TKI in patients with EGFR mutated cancers.MATERIALS AND METHODS:Targeted NGS was assessed on available (n = 208) samples using the Ion AmpliSeq™ Cancer Hotspot Panel v2 to screen for mutations in 50 different cancer genes.RESULTS:This study showed that PTEN inactivating mutations, ATM alterations, IDH1 mutations and complex EGFR mutations were predictors of short PFS in patients with a stage 4 lung adenocarcinoma receiving first line EGFR TKI and that PTEN, ATM, IDH1 and KRAS mutations as well as alterations in the MAPK pathway were related to shorter OS.CONCLUSION:These findings may lead to new treatment options in patients with unfavorable genotypes to optimize first line responses.

Aberrant cell death signaling and oxidative stress are implicated in myriad of human pathological states such as neurodegenerative, cardiovascular, metabolic and liver diseases, as well as drug-induced toxicities. While regulated cell death and mild oxidative stress are essential during normal tissue homeostasis, deregulated signaling can trigger massive depletion in a particular cell type and/or damage tissues and impair organ function with deleterious consequences that manifest as disease states. If regeneration cannot restore tissue homeostasis, the severity of the disease correlates with the extent of cell loss. Cell death can be executed via multiple modalities such as apoptosis, necrosis, pyroptosis, necroptosis and ferroptosis, depending on cell autonomous mechanisms (e.g., reactive oxygen species production, calcium overload and altered proteostasis) and/or non-cell autonomous processes (e.g., environmental stress, irradiation, chemotherapeutic agents, inflammation and pathogens). Accordingly, the inhibition of aberrant cell death and oxidative stress together with activation of autophagy, a regulated self-degradation process, are progressively emerging as relevant cytoprotective strategies to sustain homeostasis. In this review, we summarize the current literature on the crosstalk between cellular redox state and cell fate signaling, specifically from the standpoint of autophagy and its role in the maintenance of tissue/organ homeostasis via regulating oxidative stress and the potential implications for the design of novel therapeutic strategies.

The aberrant hypermethylation of BRCA1 promoter CpG islands induces the decreased expression of BRCA1 (Breast Cancer 1) protein. It can be detected in sporadic breast cancer without BRCA1 pathogenic variants, particularly in triple-negative breast cancers (TNBC). We investigated BRCA1 hypermethylation status (by methylation-specific polymerase chain reaction (MS-PCR) and MassARRAY® assays), and BRCA1 protein expression using immunohistochemistry (IHC), and their clinicopathological significance in 248 chemotherapy-naïve TNBC samples. Fifty-five tumors (22%) exhibited BRCA1 promoter hypermethylation, with a high concordance rate between MS-PCR and MassARRAY® results. Promoter hypermethylation was associated with reduced IHC BRCA1 protein expression (p = 0.005), and expression of Programmed death-ligand 1 protein (PD-L1) by tumor and immune cells (p = 0.03 and 0.011, respectively). A trend was found between promoter hypermethylation and basal marker staining (p = 0.058), and between BRCA1 expression and a basal-like phenotype. In multivariate analysis, relapse-free survival was significantly associated with N stage, adjuvant chemotherapy, and histological subtype. Overall survival was significantly associated with T and N stage, histology, and adjuvant chemotherapy. In addition, patients with tumors harboring BRCA1 promoter hypermethylation derived the most benefit from adjuvant chemotherapy. In conclusion, BRCA1 promoter hypermethylation is associated with TNBC sensitivity to adjuvant chemotherapy, basal-like features and PD-L1 expression. BRCA1 IHC expression is not a good surrogate marker for promoter hypermethylation and is not independently associated with prognosis. Association between promoter hypermethylation and sensitivity to Poly(ADP-ribose) polymerase PARP inhibitors needs to be evaluated in a specific series of patients.