While precision medicine is increasingly available, the first trials produced disappointing results, partly due to limitations of molecular profiling to mutational analyses. Here, we report a monocentric real-life experience of an integrative and interactive approach using a dedicated genomic platform for metastatic cancer patients in a situation of therapeutic failure. From October 2020 to May 2024, 1049 patients with cancers in a situation of treatment failure have been prospectively enrolled in the national program “France Médecine Génomique 2025”, 73 patients from Avicenne Hospital. Data from 20 patients with hepatocellular carcinoma were previously reported. We report here data from 27 among the remaining patients. For each patient, Sequencing Omics Information Analysis platform performed tumor and germline whole-genome, whole-exome sequencing, and RNAseq to receive tailored treatments. Tumor responses and survival data were systematically collected. Twenty-seven patients with locally advanced or metastatic cancers from twenty histological types received 32 treatment lines, 37.5% being ESCAT-I/II (high-proof efficacy level). 40% of lines were guided in part by transcriptomic analyses. Median treatment duration was 15.6 months, and median survival was not linked to ESCAT category. SeqOIA-tailored treatment outperformed previous lines, with a Growth Modulation Index≥ 1.33 for 70% of the patients. 69% of treatment lines were combined therapies, showing greater clinical benefit (86% vs. 54%, P < 0.01) and longer survival than monotherapies (15.8 vs. 13.9 months, HR=0.77, P = 0.01). Our monocentric real-life experience of interactive multidisciplinary care based on integrative genomic analyses improved survival of metastatic cancer patients using off-label combined personalized therapies.

BACKGROUND AND OBJECTIVE:Urothelial bladder cancer (UBC) carries significant mortality and treatment morbidity. Conventional diagnostic and monitoring methods, including cystoscopy and biopsy, are invasive and limited in sensitivity. Circulating tumor DNA (ctDNA), a minimally-invasive "liquid biopsy," has emerged as a promising tool for real-time disease assessment. This study aimed to systematically evaluate the diagnostic, prognostic, and predictive value of plasma and urine ctDNA in localized, locally advanced, and metastatic UBC. METHODS:A systematic review was conducted according to PRISMA guidelines using PubMed, MEDLINE, and Web of Science databases (January 2016-October 2025). Eligible studies evaluated plasma and urine ctDNA. Main outcomes included diagnostic performance, correlation with tissue mutations, disease monitoring, and prognostic or predictive value. Data synthesis focused on ctDNA detection methods, timing relative to treatment, and survival outcomes. KEY FINDINGS AND LIMITATIONS: ctDNA showed strong concordance with tumor tissue mutations and high potential for real-time monitoring of tumor burden. Data are more robust in muscle-invasive bladder cancer (MIBC) than non-muscle-invasive bladder cancer (NMIBC). Postcystectomy ctDNA positivity was an independent predictor of recurrence-free, progression-free, and overall survival. ctDNA clearance correlated with response to neoadjuvant chemotherapy and immune checkpoint inhibitors. In metastatic disease, high ctDNA mutation burden was associated with shorter overall survival and resistance mechanisms, such as PIK3CA-mediated fibroblast growth factor receptor inhibitor resistance. Limitations included heterogeneous methodologies, variable assay sensitivity, and small patient cohorts. CONCLUSIONS AND CLINICAL IMPLICATIONS:ctDNA represents a powerful non-invasive biomarker for diagnosis, surveillance, and treatment guidance in UBC. Ongoing phase III trials (IMvigor011, TOMBOLA, MODERN) may establish its clinical utility as a standard tool for personalized disease management.

PURPOSE:We investigated whether circulating tumor DNA (ctDNA) changes may be useful to assess clinical outcomes in patients with metastatic colorectal cancer (mCRC) randomized in the TIME-PRODIGE-28 trial comparing biweekly maintenance with cetuximab alone with observation after 4-month fluorouracil, folinic acid, and irinotecan (FOLFIRI) plus cetuximab induction chemotherapy. EXPERIMENTAL DESIGN:ctDNA samples were collected at four time points from baseline until disease progression during the first chemotherapy-free interval and analyzed using next-generation sequencing and methylation marker approaches. Progression-free survival (PFS) and overall survival (OS) from randomization were analyzed according to ctDNA kinetics and EGFR-MAPK pathway alterations. RESULTS:Among 139 randomized patients, 104 (74.8%) had paired samples available. Patients with negative baseline ctDNA remaining negative after 4-month induction chemotherapy had significantly longer PFS from randomization (9.6 months) as compared with patients with a ctDNA decrease of ≥80% (3.4 months) or a ctDNA decrease of <80% (2.1 months; P = 0.013). Patients with EGFR-MAPK pathway alterations identified either in tissue or baseline ctDNA had worse PFS and OS from randomization. Acquired alterations found in 17 of 63 (26.9%) patients at disease progression during the first chemotherapy-free interval were associated with worse OS from reintroduction of the full induction chemotherapy (14.9 vs. 19.4 months; P = 0.025). CONCLUSIONS:Our findings show the prognostic impact of both ctDNA kinetics and EGFR-MAPK pathway alteration dynamics following induction chemotherapy with FOLFIRI-cetuximab in patients with mCRC. Prospective studies evaluating ctDNA-guided treatment strategies are needed to validate the clinical utility of ctDNA monitoring to improve patient selection for first-line treatment de-escalation and anti-EGFR-based maintenance regimens, including treatment adaptation over time.

CONTEXT:Bladder cancer ranks among the most common malignancies worldwide, with urothelial carcinoma as the predominant histological type. Its development from normal urothelium to non-muscle-invasive bladder cancer (NMIBC) or muscle-invasive bladder cancer (MIBC) is driven by complex molecular events, including chromosomal aberrations, somatic mutations, and epigenetic dysregulation. Advances in next-generation sequencing (NGS) and transcriptomic profiling have transformed the understanding of urothelial tumor biology and opened new perspectives for precision oncology. OBJECTIVE:To provide an updated synthesis of molecular profiling in urothelial bladder cancer across disease stages and to discuss its diagnostic, prognostic, and therapeutic relevance. EVIDENCE ACQUISITION:We conducted a targeted narrative review based on a structured PubMed search of studies published between January 2000 and February 2026 using the keywords "genomic," "epigenetic," "transcriptomic," and "urothelial bladder cancer." Eligible publications included original research articles, reviews, and meta-analyses in English. Evidence was synthesized across NMIBC, MIBC, and metastatic disease, focusing on molecular alterations and their clinical relevance. EVIDENCE SYNTHESIS:Both NMIBC and MIBC present frequent TERT promoter mutations. NMIBC is further characterized by frequent FGFR3 and PIK3CA mutations, loss of chromosome 9, and widespread chromatin-remodeling gene alterations. MIBC displays high genomic instability, with recurrent mutations in TP53, RB1, and chromatin modifiers, as well as frequent alterations in the RTK/RAS/PI3K pathway. Consensus transcriptomic classifications identify reproducible molecular subtypes (luminal, basal/squamous, stroma rich, and neuroendocrine like), each with distinct biological and clinical behaviors. In metastatic disease, NGS and whole-exome analyses reveal strong concordance with localized tumors, with frequent TP53, ARID1A, KMT2D, RB1, and FGFR3 alterations and APOBEC mutational signatures. Transcriptomic profiling shows subtype heterogeneity and therapy-induced remodeling. Clinically, only FGFR3 alterations currently guide therapy, with erdafitinib approved by the US Food and Drug Administration and the European Medicines Agency for FGFR3-altered metastatic disease. Beyond tumor testing, circulating tumor DNA assays show promise for molecular residual disease detection and adjuvant treatment stratification. CONCLUSIONS:Molecular profiling has deepened the understanding of bladder cancer biology and identified potential biomarkers for diagnosis, prognosis, and treatment. However, clinical implementation remains limited, and prospective biomarker-driven trials are needed to establish the role of genomic and transcriptomic alterations in therapeutic decision-making.

Background:Anaplastic lymphoma kinase (ALK)-positive non-small cell lung cancer (NSCLC) represents a distinct molecular subtype characterized by younger age at diagnosis, prolonged survival with sequential ALK tyrosine kinase inhibitors (TKIs) therapy, and a high incidence of central nervous system (CNS) metastases. Despite major therapeutic advances, metastatic ALK-positive NSCLC remains incurable, yet end-of-life (EOL) outcomes in this population are poorly described. We aimed to characterize causes of death, EOL management, and factors associated with palliative care referral and place of death in patients who died from metastatic ALK-positive NSCLC. Methods:We conducted a retrospective multicentric cohort study including all deceased patients with histologically confirmed metastatic ALK-positive NSCLC treated in seven centers in France between 2009 and 2023. Clinical characteristics, molecular data, treatment sequences, CNS progression, causes of death, and EOL care indicators were collected from medical records. Causes of death were classified as cancer-related or non-cancer-related. Multivariable logistic regression analyses were performed to identify factors associated with palliative care referral and in-hospital death. Results:Among 232 screened patients, 89 deceased patients were included. Median age at diagnosis was 59.9 years, 58.4% were female, and 62.9% were never-smokers. Median overall survival (OS) was 26 months. CNS progression occurred in 59.6% of patients, with a median interval of 11 months between first CNS progression and death. Overall, 79.8% of deaths were cancer-related, mainly due to neurological failure (36.6%), respiratory failure (29.6%), or cachexia (29.6%). In the last 2 weeks of life, 71.9% of patients were still receiving active anti-cancer treatment. Most deaths occurred in hospital settings (73.1%). Only 51.1% of patients were referred to a palliative care specialist. Palliative care referral was associated with lower rates of in-hospital death, earlier treatment discontinuation, more frequent multidisciplinary decisions focused on comfort care, and better documentation of treatment limitation orders. In multivariable analysis, active anti-cancer treatment within the last 2 weeks of life was the strongest independent predictor of in-hospital death. Conclusions:Despite prolonged survival with ALK-targeted therapies, most patients with metastatic ALK-positive NSCLC die from cancer-related causes, frequently after intensive hospital-based care. Palliative care referral remains suboptimal but is associated with improved EOL outcomes. Early and systematic integration of palliative care should be considered an essential component of the management of metastatic ALK-positive NSCLC.

Background: Genome and transcriptome analysis has enhanced the characterisation of pancreatic ductal adenocarcinoma (PDAC), paving the way for targeted therapies. Tumours KRAS wild type (WT) represent a unique subgroup. Objectives: Characterise the population and molecular abnormalities present in KRAS WT PDAC. Design: Clinical and molecular data from a large retrospective cohort of KRAS WT PDAC were analysed. Methods: Next-generation sequencing (NGS) was used to analyse DNAs and RNAs, allowing molecular and transcriptomic characterisation. Results: We identified 93/1059 (9%) KRAS WT PDAC, among which eight had druggable fusions ( n = 8/30 contributive samples), six had BRAF mutations and 19 ( n = 19/47) had mutations in homologous recombination (HR) pathway genes. Potential molecular targets in this series may be underestimated due to many non-contributive results. Clinical characteristics and survival did not differ between patients with KRAS WT and KRAS- mutated tumours. Transcriptomic data were available for 350 samples. Their analysis shows a difference in phenotype between mutated and WT tumours, with a molecular profile that appears to be better prognostic for KRAS WT tumours. Conclusion: KRAS WT tumours are enriched with molecular abnormalities of therapeutic interest. These include oncogene driver alterations (gene fusions and mutations) and mutations in genes of the HR pathway. Targeted therapy strategies for PDAC rely on molecular testing beyond RAS , but further research is needed to identify new therapeutic approaches that improve outcomes in PDAC.

5576 Background: The PAOLA-1 trial showed that adding ola to bev as maintenance therapy improved overall survival (OS) of HGOC patients with BRCA1/2 mutations (BRCAm) or homologous recombination (HR) deficiency (HRD) defined by the MyChoice HRD Plus assay with a GIS threshold of 42. This post hoc analysis of PAOLA-1 explored if alternative thresholds could better identify patients who benefit most from ola. Methods: New cutoffs were determined through OS analyses using Cox proportional hazards models with an interaction term for GIS. Tumors were categorized into HRP (GIS<42), HRDlow (42–60 for BRCA1/2 wild-type [BRCAwt], 42–67 for BRCAm), and HRDhigh (>60 for BRCAwt, >67 for BRCAm). Genomic analyses included promoter methylation, BRCA loss of heterozygosity (LOH) and HR repair gene mutations. Results: Among 623 patients, 194 (31%) were BRCAm and 429 (69%) BRCAwt. Main clinical prognostic features were well-balanced across BRCAwt/HRDhigh, BRCAwt/HRDlow and BRCAwt/HRP as well as among BRCAm/HRDhigh, BRCAm/HRDlow and BRCAm/HRP. Ola+bev improved progression-free survival (PFS) and OS in BRCAwt/HRDhigh and BRCAm/HRDhigh (Table). Ola+bev improved PFS but not OS in BRCAwt/HRDlow and BRCAm/HRDlow. HRP tumors showed no PFS or OS benefit regardless of BRCA status. BRCA1 / RAD51C promoters were methylated in 75% of BRCAwt/HRDhigh, 47% of HRDlow, and 3% of HRP. HRDlow tumors had fewer HR repair gene mutations than HRDhigh. Among HRP/BRCAm, 37% lacked BRCA LOH, suggesting functional BRCA. Conclusions: Our post-hoc subgroup analyses suggest that refined GIS thresholds identify three distinct populations of HGOC patients with varying survival benefits from ola+bev maintenance. Optimized GIS cutoffs may further improve patient stratification in future PARP inhibitors trials. BRCA WT BRCA Mut HRP HRD low HRD high p HRP HRD low HRD high p N 277 72 80 19 124 51 BRCA mutation 0.36 BRCA1 ‐ ‐ ‐ 11 (57.9%) 82 (66.1%) 38 (74.5%) BRCA2 ‐ ‐ ‐ 8 (42.1%) 42 (33.9%) 13 (25.5%) No BRCA LOH ‐ ‐ ‐ 7 (36.8%) 1 (0.8%) 2 (4.0%) p<0.001 HR gene methylation (NA=134) p<0.001 No 187 (96.9%) 26 (53.1%) 13 (24.5%) ‐ ‐ ‐ BRCA1 1 (0.5%) 15 (30.6%) 34 (64.2%) ‐ ‐ ‐ RAD51C 5 (2.6%) 8 (16.3%) 6 (11.3%) ‐ ‐ ‐ mPFS (95%CI ), months Ola + bev 16.6(15.2-18.2) 28.9(20.3-NR) 38.9(22.1-NR) 21.2(13.9-NR) 51.4(38.9-NR) 75.2(NR-NR) Placebo + bev 16.2(13.9-18.8) 16.4(12.9-27.7) 17.0(12.9-23.4) 20.3(14.7-NR) 19.4(16.6-24.0) 15.5(8.7-NR) HR (95%CI) 1.00(0.76-1.32) 0.51(0.29-0.91) 0.42(0.24-0.72) 0.80(0.28-2.26) 0.39(0.24-0.62) 0.17(0.07-0.41) mOS (95%CI ), months Ola + bev 36.8(30.7-40.9) 54.0(48.3-NR) NR(54.1-NR) 47.0(24.2-NR) NR(NR-NR) 75.2(NR-NR) Placebo + bev 40.4(33.0-53.3) 52.4(45.8-NR) 41.2(34.0-NR) 43.1(29.0-NR) NR(59.8-NR) 55.2(29.8-NR) HR (95%CI) 1.19(0.87-1.62) 1.07(0.55-2.07) 0.49(0.26-0.94) 0.88(0.28-2.79) 0.61(0.32-1.17) 0.15(0.05-0.50)

Colorectal cancer (CRC) is the third leading cause of cancer death and accounts for 10 % of cancer diagnoses worldwide. Despite the advancements achieved over the latest decades, CRC treatments are still based on conventional chemotherapy whose efficacy is limited by acquired resistance and unfavorable toxicity profile, making the search for novel actionable targets a priority. In this context, gene fusions are emerging as promising -albeit very rare - new markers because of their recurrence across different tumor types and their potential actionability. The aim of this review is to investigate the role of gene fusions in CRC by focusing on pathogenesis, screening strategies as well as their clinical implications.

Management of colorectal cancer (CRC) patients relies on the accurate determination of microsatellite instability (MSI) status. MSI status can have an influence on therapy decisions centered on immune checkpoint inhibitors. In this study a novel droplet digital PCR (ddPCR) kit for MSI status determination was validated across 2 separate CRC patient cohorts: 102 tumor samples from the ALGECOLS cohort and 129 plasma samples from the RASANC cohort. Each cohort was assessed for MSI status using the novel ddPCR kit and compared to historical and/or newly obtained results, (either immunohistochemistry analysis or PCR amplification). Concordance between ddPCR and conventional MSI determination methods for the analysis of tissue samples was 97.1% for ALGECOLS. When looking at positive ctDNA samples, a strong concordance was observed for the RASANC cohort. This study illustrates that ddPCR MSI testing represents a rapid, sensitive and accurate method with a strong correlation to established methods. Moreover, the ability of the described approach to monitor MSI status in both tumor and plasma enhances the potential for the use of MSI status in longitudinal monitoring of CRC patients.### Competing Interest StatementThe authors declare a potential conflict of interest and state it below MH, AC, JL, JY are employees of Bio-Rad Laboratories ### Funding StatementThis work was supported by the APHP (grant no. CRC06043), the Poitiers University Hospital, the INSERM-DGOS, the Ministere Enseignement Superieur et de la Recherche, the University Paris City, the CNRS, the INSERM, the Centre de Recherche des Cordeliers and the ligue nationale contre le cancer (no. EL2016.LNCC). JL, JY, AC and MH are Bio-Rad employees. ### Author DeclarationsI confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.YesThe details of the IRB/oversight body that provided approval or exemption for the research described are given below:Cohort 1: Ninety-six (96) FF and 6 FFPE CRC samples with known MSI status were retrospectively selected from the ALGECOLS cohort (according to availability of samples) approved by the Ile-de-France ethics committee number 2 ([NCT01198743][1]). Cohort 2: From the RASANC protocol, approved by the Ile-de-France IV ethics committee, 129 cell-free DNA (cfDNA) samples were analyzed (according to availability)I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.YesI understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).YesI have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.YesAll data produced in the present study are available upon reasonable request to the authors [1]: /lookup/external-ref?link_type=CLINTRIALGOV&access_num=NCT01198743&atom=%2Fmedrxiv%2Fearly%2F2024%2F04%2F15%2F2024.04.12.24305349.atom

OBJECTIVE:Recent evidence suggests that elevated levels of PD-L1 expression may be linked to early resistance to TKI and reduced survival in NSCLC with EGFR mutations. This study aimed to characterize the clinical and molecular features of EGFR-mutated lung adenocarcinomas and determine the prognostic significance associated with high PD-L1 expression. MATERIALS AND METHODS:We conducted a retrospective chart review of 103 consecutive patients with advanced EGFR-mutated NSCLC, who received treatment between 01/01/2016 and 30/12/2020, at our institution. RESULTS:Among the tumors, 17% (n = 18) exhibited high PD-L1 expression (≥50% tumor proportion score), which was associated with a lower prevalence of common EGFR mutations (56% vs. 82%, p = 0.03) and a higher frequency of complex EGFR mutations (28% vs. 7%, p = 0.02). Univariate analysis did not reveal any significant differences in first-line response, progression-free survival, or overall survival between the PD-L1 ≥50% and <50% groups. However, multivariate analysis demonstrated that PD-L1 ≥50% was independently associated with shorter survival (HR = 2.57; 95%CI[1.20-5.55]; p = 0.02), along with male gender (HR = 2.77; 95%CI[1.54-4.19]; p<0.005), presence of liver metastases (HR = 5.80; 95%CI[2.86-11.75]; p<0.005) or brain metastases (HR = 1.99; 95%CI[1.13-3.52]; p = 0.02), and poor general condition at diagnosis (ECOG 3 and 4) (HR = 10.69; 95% CI[4.42-25.85]; p<0.005). Additionally, a trend towards a higher frequency of de novo resistance was observed in the PD-L1 >50% group (7% vs. 17%, p = 0.19). CONCLUSION:High PD-L1 expression was more commonly found in lung adenocarcinomas with uncommon and complex EGFR mutations. Furthermore, high PD-L1 expression independently predicted poor survival. These findings warrant validation through prospective studies.

Aim: Immune checkpoint inhibitors improved the survival of advanced non-small cell lung cancer. However, only 20% of patients respond to these treatments and the search for predictive biomarkers of response is still topical. The objective of this work is to analyze the anti-PD-1 monotherapy benefit based on genetic alterations diagnosed by next generation sequencing (NGS), in advanced non-small cell lung cancer. Methods: Patients with advanced non-small cell lung cancer treated with immunotherapy were retrospectively included in this monocentric study. Clinical data, immunohistochemical expression of PD-L1 and molecular data, with a 22-genes NGS panel, were collected. Results: 107 patients were included. The median age was 65 years [59; 73], 70 were men (65%), 96 had adenocarcinoma (90%), 33 were receiving a first line (31%). 54 patients had KRAS mutation (50%) and 56 had TP53 mutation (52%). The remaining mutations were present in fewer than 10 patients. There was no statistically significant differences in median of progression-free or overall survival based on KRAS-only, TP53-only or KRAS-TP53 mutations co-mutated compared to double wild-type patients (P = 0.46 and P = 0.72 respectively). Conclusions: The search for a predictive composite biomarker remains a major issue in the coming years.

BACKGROUND:STK11/LKB1 mutations have been associated with primary resistance to PD-1 axis inhibitors and poor prognosis in advanced KRAS-mutant lung adenocarcinoma. This study aimed to assess the prognostic significance of STK11/LKB1 alterations in localized non-squamous non-small cell lung carcinoma (non-sq NSCLC). PATIENTS AND METHODS:Surgical samples from patients undergoing complete resection for stage IIa, IIb, or IIIa (N2 excluded) non-sq NSCLC in the randomized adjuvant phase II trial (NCT00775385 IFCT-1801 TASTE trial) were examined. Patients received either standard chemotherapy (Pemetrexed Cisplatin) or personalized treatment based on EGFR mutation (Erlotinib) and ERCC1 expression. Tumor molecular profiles were analyzed using targeted NGS and correlated with overall survival (OS) and disease-free survival (DFS), adjusting for relevant clinical variables. Additionally, interactions between treatment groups and molecular alterations on OS, PD-L1 expression, and tumor-circulating DNA in post-operative plasma samples were evaluated. RESULTS:Among 134 patients (predominantly male smokers with adenocarcinoma), KRAS mutations were associated with shorter DFS (HR: 1.95, 95 % CI: 1.1-3.4, p = 0.02) and OS (HR: 2.32, 95 % CI: 1.2-4.6, p = 0.014). Isolated STK11/LKB1 mutations (n = 18) did not significantly impact DFS or OS. However, within KRAS-mutated samples (n = 53), patients with concurrent STK11/LKB1 mutations (n = 10) exhibited significantly shorter DFS (HR: 3.85, CI: 1.5-10.2, p = 0.006) and a trend towards shorter OS (HR: 1.80, CI: 0.6-5.3, p = 0.28). No associations were found between PD-L1 expression, other gene mutations, progression-free survival (PFS), or OS. CONCLUSION:This analysis reinforces KRAS mutations as predictive factors for relapse and poor survival in localized non-sq NSCLC. Furthermore, the presence of concomitant STK11/LKB1 mutations exacerbated the prognosis within the KRAS-mutated subset. These findings emphasize the clinical relevance of these molecular markers and their potential impact on treatment strategies in non-sq NSCLC.

Background: Few epidemiological data are available on surgically treated Caucasian patients with non-small-cell lung cancers (NSCLCs) harboring epidermal growth factor receptor ( EGFR ) mutations. The main objective of this study was to describe, in the real-world setting, these patients’ incidence, clinical, and tumoral characteristics. Methods: The participating centers included all consecutive localized non-squamous NSCLC patients undergoing surgery between January 2018 and December 2019 in France. EGFR status was determined retrospectively when not available before surgery. Results: The study includes 1391 no squamous NSCLC patients from 16 centers; EGFR status was determined before surgery in 692 (49.7%) of the cases and conducted as part of the study for 699 (50.3%); 171 (12.3%) were EGFR mutated; median age: 70 (range: 36–88) years; female: 59.6%; never smokers: 75.7%; non-squamous histology 97.7%, programmed death ligand-1 expression 0%/1–49%/⩾50 in 60.5%/25.7%/13.8%, respectively. Surgery was predominantly lobectomy (81%) or segmentectomy (14.9%), with systematic lymph node dissection in 95.9%. Resection completeness was R0 for 97%. Post-surgery staging was as follows: IA: 52%, IB: 16%, IIA: 4%, IIB: 10%, IIIA: 16%, and IIIB: 0.05%; EGFR mutation exon was Del19/exon 21 ( L858R )/20/18 in 37.4%/36.8%/14%, and 6.4% of cases, respectively; 31 (18%) patients received adjuvant treatment (chemotherapy: 93%, EGFR tyrosine kinase inhibitor: 0%, radiotherapy: 20%). After a median follow-up of 31 (95% confidence interval: 29.6–33.1) months, 45 (26%) patients relapsed: 11/45 (24%) locally and 34 (76%) with metastatic progression. Median disease-free survival (DFS) and overall survival were not reached and 3-year DFS was 60%. Conclusion: This real-world analysis provides the incidence and outcomes of resected EGFR -mutated NSCLCs in a European patient cohort. Keywords mutation , lung cancer management , surgery

The prognostic significance of positive peritoneal cytology still varied between cancer types and geographical origin. However, because of the lack of sensitivity of this biomarker, conventional cytology is not routinely performed in every country. Here, we wanted to test a new biomarker, peritoneal tumour DNA, using NGS technique, in order to compare it with the historical one, in patients having peritoneal metastases of gastrointestinal or ovarian cancer.

OBJECTIVE:To evaluate the prognostic impact of circulating tumor DNA (ctDNA) detection at diagnosis (T0) and its early decrease after one cycle (T1) of neoadjuvant chemotherapy (NACT) in patients with advanced epithelial ovarian cancer (EOC) included in the CHIVA trial (NCT01583322). METHODS:Blood samples were collected at T0 and before each administration of NACT. Circulating tumor DNA detection was performed by next-generation sequencing. Multivariate analysis was performed. A p-value of 0.05 was considered significant. Progression-free survival (PFS) and overall survival (OS) were compared between groups defined by ctDNA kinetic profile. Cox survival model was used to search variables associated with PFS and OS. Kaplan-Mayer curve was used to graphically express the differences in PFS and OS. A log-rank test compared the two curves. RESULTS:188 patients were included. Blood samples were available for 168 patients at T0 and for 160 patients at T0 and T1 to assess ctDNA ratio kinetics. At T0, 107 patients (63.7 %) had detectable ctDNA. At T1, 137 (85.6 %) patients had negative ctDNA or a decrease of more than 80 %. There was a significant benefit in either PFS (p = 0.0017) or OS (p = 0.0036) in favor of early decrease of ctDNA ratio. A favorable decrease was associated with a greater likelihood of being able to perform CRS (OR: 3.94 (CI95 % 1.45-10.70), p = 0.0074). CONCLUSIONS:Early decrease of ctDNA ratio can provide prognostic information early in the management of patients, allowing a more accurate information to patients and an early preparation for CRS (prehabilitation).

Patients with EGFR-mutated non-small cell lung cancer (NSCLC) benefit from treatment with tyrosine kinase inhibitors (TKI) targeting EGFR. Despite improvements in patient care, especially with the 3rd generation TKI osimertinib, disease relapse is observed in all patients. Among the various processes involved in TKI resistance, epithelial-to-mesenchymal transition (EMT) is far from being fully characterized. We hypothesized that the cellular prion protein PrPC could be involved in EMT and EGFR-TKI resistance in NSCLC. Using 5 independent lung adenocarcinoma datasets, including our own cohort, we document that the expression of the PRNP gene encoding PrPC is associated with EMT. By manipulating the levels of PrPC in different EGFR-mutated NSCLC cell lines, we firmly establish that the expression of PrPC is mandatory for cells to maintain or acquire a mesenchymal phenotype. Mechanistically, we show that PrPC operates through an ILK-RBPJ cascade, which also controls the expression of EGFR. Our data further demonstrate that PrPC levels are elevated in EGFR-mutated versus wild-type tumours or upon EGFR activation in vitro. In addition, we provide evidence that PRNP levels increase with TKI resistance and that reducing PRNP expression sensitizes cells to osimertinib. Finally, we found that plasma PrPC levels are increased in EGFR-mutated NSCLC patients from 2 independent cohorts and that their longitudinal evolution mirrors that of disease. Altogether, these findings define PrPC as a candidate driver of EMT-dependent resistance to EGFR-TKI in NSCLC. They further suggest that monitoring plasma PrPC levels may represent a valuable non-invasive strategy for patient follow-up and warrant considering PrPC-targeted therapies for EGFR-mutated NSCLC patients with TKI failure.

BackgroundThere is increased evidence that the effects of stem cells can mostly be duplicated by administration of their secretome which might streamline the translation towards the clinics.MethodsThe 12-patient SECRET-HF phase 1 trial has thus been designed to determine the feasibility and safety of repeated intravenous injections of the extracellular vesicle (EV)-enriched secretome of cardiovascular progenitor cells differentiated from pluripotent stem cells in severely symptomatic patients with drug-refractory left ventricular (LV) dysfunction secondary to non-ischemic dilated cardiomyopathy. Here we report the case of the first treated patient (baseline NYHA class III; LV Ejection Fraction:25%) in whom a dose of 20 × 109 particles/kg was intravenously infused three times three weeks apart.FindingsIn addition to demonstrating the feasibility of producing a cardiac cell secretome compliant with Good Manufacturing Practice standards, this case documents the excellent tolerance of its repeated delivery, without any adverse events during or after infusions. Six months after the procedure, the patient is in NYHA Class II with improved echo parameters, a reduced daily need for diuretics (from 240 mg to 160 mg), no firing from the previously implanted automatic internal defibrillator and no alloimmunization against the drug product, thereby supporting its lack of immunogenicity.InterpretationThe rationale underlying the intravenous route is that the infused EV-enriched secretome may act by rewiring endogenous immune cells, both circulating and in peripheral organs, to take on a reparative phenotype. These EV-modified immune cells could then traffic to the heart to effect tissue repair, including mitigation of inflammation which is a hallmark of cardiac failure.FundingThis trial is funded by the French Ministry of Health (Programme Hospitalier de Recherche Clinique AOM19330) and the “France 2030” National Strategy Program (ANR-20-F2II-0003). It is sponsored by Assistance Publique-Hôpitaux de Paris.

BackgroundA high density of resident memory T cells (TRM) in tumors correlates with improved clinical outcomes in immunotherapy-treated patients. In most clinical studies, TRMare defined by the CD103 marker. However, it is clearly established that not all TRMexpress CD103, but can be defined by other markers (CD49a, CD69, etc). The frequency of these subpopulations of TRMexpressing or not CD103 varies according to the location of the cancer. Little is known about their functionality and their predictive impact on response to immunotherapy. In preclinical models, only some subpopulations of TRMare associated with cancer vaccine efficacy.MethodsMultiparametric cytometry analyses were used to demonstrate the presence of TRMsubpopulations in the lung in mice after vaccination and in fresh ex vivo human non-small cell lung cancer (NSCLC). An analysis of the T-cell repertoire of these TRMwas conducted to search for their relationships. Multiplex immunofluorescence techniques were used to quantify intratumor infiltration of TRMsubpopulations in two cohorts of patients with NSCLC. The impact on the clinical outcome of the TRMtumor infiltration was also investigated.ResultsWe identified two main TRMsubpopulations in tumor-infiltrating lymphocytes derived from patients with NSCLC: one co-expressing CD103 and CD49a (double positive (DP)), and the other expressing only CD49a (simple positive (SP)); both exhibiting additional TRMsurface markers like CD69. Despite higher expression of inhibitory receptors, DP TRMexhibited greater functionality compared with SP TRM. Analysis of T-cell receptor (TCR) repertoire and expression of the stemness marker TCF1 revealed shared TCRs between populations, with the SP subset appearing more progenitor-like phenotype. In the training cohort, PD-L1 (Programmed Death-Ligand 1) and TCF1+CD8+T cells predict response to anti-PD-1. In patient with NSCLC validation cohorts, only DP TRMpredicted PD-1 blockade response. Multivariate analysis, including various biomarkers associated with responses to anti-PD-(L)1, such as total CD8, TCF1+CD8+T cells, and PD-L1, showed that only intratumoral infiltration by DP TRMremained significant.ConclusionsThis study highlights the non-equivalence of TRMsubpopulations. The population of TRMco-expressing CD103 and CD49a appears to be the most functional and has the most significant capacity for predicting response to immunotherapy in multivariate analysis in patients with NSCLC.

OBJECTIVE:BRCA1 promoter methylation (BRCA1pm) is suspected to alter prognosis of patients with epithelial ovarian cancer (EOC). We aimed to evaluate the prognostic impact of this epigenetic modification. METHODS:We conducted a retrospective, monocentric study from 11/2006 to 08/2018. Patients with EOC and available status concerning somatic BRCA1/2 mutation and BRCA1pm were included. Three groups were defined: patients without BRCA1/2 mutation or BRCA1pm, patients with BRCA1/2 mutation and patients with BRCA1pm. BRCA1/2 mutations were analyzed in current care settings by next-generation sequencing (NGS). BRCA1pm analysis was assessed and quantified from bisulfite converted DNAs using fluorescent methylation specific polymerase chain reaction (PCR) and fragment analysis. All patients signed a consent form and the study was authorized by a Personal Protection Committee. Descriptive statistics were used to describe groups. Multivariate analysis was performed using the logistic regression model and including the variables that could be known at the time of diagnosis and that were significant at univariate analysis. Survival was compared between the groups. Kaplan-Mayer curves were used to express the differences in survival that were compared using log rank tests. RESULTS:145 patients were included: 95 (65.5 %) patients without BRCA1/2 mutation or BRCA1pm, 32 (22.1 %) patients with BRCA1/2 mutation, 18 (12.4 %) patients with BRCA1pm. Median survival was decreased in patients with BRCA1pm. Comparison of survival revealed a significant difference in overall survival (p = 0.0078) with a worse prognosis for patients with a BRCA1pm. CONCLUSION:BRCA1pm in patients with EOC is an independent factor associated with a decreased overall survival. SYNOPSIS:BRCA1 promotor methylation in patients with epithelial ovarian cancer is an independent factor associated with a decreased overall survival.