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BackgroundSurvival after curative resection of early-stage lung adenocarcinoma (LUAD) varies and prognostic biomarkers are urgently needed.MethodsLarge-format tissue samples from a prospective cohort of 200 patients with resected LUAD were immunophenotyped for cancer hallmarks TP53, NF1, CD45, PD-1, PCNA, TUNEL and FVIII, and were followed for a median of 2.34 (95% CI 1.71–3.49) years.ResultsUnsupervised hierarchical clustering revealed two patient subgroups with similar clinicopathological features and genotype, but with markedly different survival: “proliferative” patients (60%) with elevated TP53, NF1, CD45 and PCNA expression had 50% 5-year overall survival, while “apoptotic” patients (40%) with high TUNEL had 70% 5-year survival (hazard ratio 2.23, 95% CI 1.33–3.80; p=0.0069). Cox regression and machine learning algorithms including random forests built clinically useful models: a score to predict overall survival and a formula and nomogram to predict tumour phenotype. The distinct LUAD phenotypes were validated in The Cancer Genome Atlas and KMplotter data, and showed prognostic power supplementary to International Association for the Study of Lung Cancer tumour–node–metastasis stage and World Health Organization histologic classification.ConclusionsTwo molecular subtypes of LUAD exist and their identification provides important prognostic information.

Introduction: Lung adenocarcinoma (LUAD) is the most frequent lung cancer in non-smokers. Although carcinogens other than smoking (i.e., radiation) likely cause LUAD, their molecular imprints are obscure. Objectives: To define the molecular imprints of smoking, its carcinogens, and radiation in LUAD RNA. Materials and Methods: FVB mice were exposed to tobacco smoke (500 mg/m3; 2 hrs/day; 5 months on/5 months off), its chemicals ethyl carbamate (1 g/Kg), diethylnitrosamine (20 mg/Kg), and methylnitrosourea (50 mg/Kg), or X-irradiation (15 Gy X-rays delivered to a 100 mm3 tissue voxel of the right upper lobe). Total lung tumor RNA was extracted using Trizol/RNAeasy, was analyzed on an Illumina HiSeq4000, and was interrogated employing STAR, DeSeqR, Pathvisio, GSEA, Samtools, SnpEff, and R*. Results: FVB mice developed LUAD in response to the carcinogens used. RNAseq of tumor tissues provided robust transcript abundance and sequence information, including single nucleotide variation at the mononucleotide, trinucleotide, gene, and chromosome levels, as well as insertions and deletions. The five different carcinogens produced markedly distinct mutational imprints on the exposed tissues. Interestingly, the signatures of smoking and its carcinogens were highly similar, contained KRAS and other point mutations, and were enriched in human KRAS-mutant LUAD, while the signatures of irradiated tissues displayed EGFR point mutations, a preponderance of transversions, and a distinct gene expression set. Conclusions: Bulk RNAseq of environmentally-induced mouse tumors can reveal carcinogenic exposures and causes. We are currently working to identify cellular origin-restricted signatures and to translate murine imprints to human LUAD.

Background: The tobacco carcinogen urethane causes lung adenocarcinomas (LUAD) in mice. They show high similarity to human LUAD of smokers, including driver KrasQ61R mutations. However, the time line of KRASQ61R mutation acquisition as well as the affected cell lineages are still obscure. Objectives: With this study we aim to identify which cells of the lung gain KrasQ61R mutations and when they occur in response to urethane treatment. Methods: Airway and alveolar GFP-lineage-marked mice received single urethane hits (1 g/Kg). Lungs were harvested at 0, 1, 2, 4, and 8 weeks and LUAD tumors were collected at 16, 24, and 32 weeks post-urethane. The DNA was subjected to digital droplet PCR interrogating the coexistence of GFP and KrasQ61R in the same DNA copy. RNA of tumors was submitted to sequencing for gene expression analysis. Results: Starting from even 1 week post-urethane, both airway and alveolar lineages suffered KrasQ61R mutations (2.57% of copies examined). In airway-labelled cells KrasQ61R mutations increased by 4.06% and 2.55% at 4 and 32 weeks post-urethane, respectively. In contrast, in alveolar-labelled cells KrasQ61R mutations declined by 5.92% and 6.12%. Strikingly, gene expression analysis on tumors revealed an over expression of alveolar markers whereas expression of airway marker genes decreased. Conclusion: After pulmonary tumor initiation by urethane, airway cells accumulate KrasQ61R mutations over time, whereas alveolar cells tend to lose them. However, tumor cells are over expressing alveolar cell markers and lose airway markers. Altogether, these results indicate that smoking-induced LUAD develops from club cells which acquire alveolar cell characteristics. These data provide further insights into the mechanisms underlying LUAD evolution in patients.

International audience; A comprehensive characterization of lung adenocarcinoma (LADC) clinical features is currently missing. We prospectively evaluated Caucasian patients with early-stage LADC. Patients with LADC diagnosed between 2011 and 2015 were prospectively assessed for lung resection with curative intent. Fifty clinical, pathologic, radiologic, and molecular variables were recorded. Patients were followed till death/study conclusion. The main findings were compared to a separate cohort from France. Of 1943 patients evaluated, 366 were enrolled (18.8%; 181 female; 75 never-smokers; 28% of registered Bavarian cases over the study period). Smoking and obstruction were significantly more prevalent in GLAD compared with adult Bavarians (P < 0.0001). Ever-smoker tumors were preferentially localized to the upper lobes. We observed 120 relapses and 74 deaths over 704 cumulative follow-up years. Median overall and disease-free survival were >7.5 and 3.6 years, respectively. Patients aged 65 years, resected >60 days postdiagnosis, with abnormal FVC/DLCO VA , N2/N3 stage, or solid histology had significantly decreased survival estimates. These were fit into a weighted locoregional LADC death risk score that outperformed pTNM7 in predicting survival in the GLAD and in our second cohort. We define the clinical gestalt of locoregional LADC and provide a new clinical tool to predict survival, findings that may aid future management and research design

Oncolytic viruses represent an exciting new aspect of the evolving field of cancer immunotherapy. We have engineered a novel hybrid vector comprising vesicular stomatitis virus (VSV) and Newcastle disease virus (NDV), named recombinant VSV-NDV (rVSV-NDV), wherein the VSV backbone is conserved but its glycoprotein has been replaced by the hemagglutinin-neuraminidase (HN) and the modified, hyperfusogenic fusion (F) envelope proteins of recombinant NDV. In comparison to wild-type VSV, which kills cells through a classical cytopathic effect, the recombinant virus is able to induce tumor-specific syncytium formation, allowing efficient cell-to-cell spread of the virus and a rapid onset of immunogenic cell death. Furthermore, the glycoprotein exchange substantially abrogates the off-target effects in brain and liver tissue associated with wild-type VSV, resulting in a markedly enhanced safety profile, even in immune-deficient NOD.CB17-prkdc(scid)/NCrCrl (NOD-SCID) mice, which are highly susceptible to wild-type VSV. Although NDV causes severe pathogenicity in its natural avian hosts, the incorporation of the envelope proteins in the chimeric rVSV-NDV vector is avirulent in embryonated chicken eggs. Finally, systemic administration of rVSV-NDV in orthotopic hepatocellular carcinoma (HCC)-bearing immune-competent mice resulted in significant survival prolongation. This strategy, therefore, combines the beneficial properties of the rapidly replicating VSV platform with the highly efficient spread and immunogenic cell death of a fusogenic virus without risking the safety and environmental threats associated with either parental vector. Taking the data together, rVSV-NDV represents an attractive vector platform for clinical translation as a safe and effective oncolytic virus. IMPORTANCE The therapeutic efficacy of oncolytic viral therapy often comes as a tradeoff with safety, such that potent vectors are often associated with toxicity, while safer viruses tend to have attenuated therapeutic effects. Despite promising preclinical data, the development of VSV as a clinical agent has been substantially hampered by the fact that severe neurotoxicity and hepatotoxicity have been observed in rodents and nonhuman primates in response to treatment with wild-type VSV. Although NDV has been shown to have an attractive safety profile in humans and to have promising oncolytic effects, its further development has been severely restricted due to the environmental risks that it poses. The hybrid rVSV-NDV vector, therefore, represents an extremely promising vector platform in that it has been rationally designed to be safe, with respect to both the recipient and the environment, while being simultaneously effective, both through its direct oncolytic actions and through induction of immunogenic cell death.