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Supplemental material, sj-doc-5-tmj-10.1177_03008916211014954 for PD-L1 SNPs as biomarkers to define benefit in patients with advanced NSCLC treated with immune checkpoint inhibitors by Roberta Minari, Francesco Bonatti, Giulia Mazzaschi, Alessandra Dodi, Francesco Facchinetti, Francesco Gelsomino, Gloria Cinquegrani, Anna Squadrilli, Paola Bordi, Sebastiano Buti, Melissa Bersanelli, Alessandro Leonetti, Agnese Cosenza, Leonarda Ferri, Elena Rapacchi, Federico Quaini, Andrea Ardizzoni and Marcello Tiseo in Tumori Journal

Supplemental material, sj-doc-3-tmj-10.1177_03008916211014954 for PD-L1 SNPs as biomarkers to define benefit in patients with advanced NSCLC treated with immune checkpoint inhibitors by Roberta Minari, Francesco Bonatti, Giulia Mazzaschi, Alessandra Dodi, Francesco Facchinetti, Francesco Gelsomino, Gloria Cinquegrani, Anna Squadrilli, Paola Bordi, Sebastiano Buti, Melissa Bersanelli, Alessandro Leonetti, Agnese Cosenza, Leonarda Ferri, Elena Rapacchi, Federico Quaini, Andrea Ardizzoni and Marcello Tiseo in Tumori Journal

Supplemental material, sj-doc-4-tmj-10.1177_03008916211014954 for PD-L1 SNPs as biomarkers to define benefit in patients with advanced NSCLC treated with immune checkpoint inhibitors by Roberta Minari, Francesco Bonatti, Giulia Mazzaschi, Alessandra Dodi, Francesco Facchinetti, Francesco Gelsomino, Gloria Cinquegrani, Anna Squadrilli, Paola Bordi, Sebastiano Buti, Melissa Bersanelli, Alessandro Leonetti, Agnese Cosenza, Leonarda Ferri, Elena Rapacchi, Federico Quaini, Andrea Ardizzoni and Marcello Tiseo in Tumori Journal

Supplemental material, sj-doc-1-tmj-10.1177_03008916211014954 for PD-L1 SNPs as biomarkers to define benefit in patients with advanced NSCLC treated with immune checkpoint inhibitors by Roberta Minari, Francesco Bonatti, Giulia Mazzaschi, Alessandra Dodi, Francesco Facchinetti, Francesco Gelsomino, Gloria Cinquegrani, Anna Squadrilli, Paola Bordi, Sebastiano Buti, Melissa Bersanelli, Alessandro Leonetti, Agnese Cosenza, Leonarda Ferri, Elena Rapacchi, Federico Quaini, Andrea Ardizzoni and Marcello Tiseo in Tumori Journal

Supplemental material, sj-doc-2-tmj-10.1177_03008916211014954 for PD-L1 SNPs as biomarkers to define benefit in patients with advanced NSCLC treated with immune checkpoint inhibitors by Roberta Minari, Francesco Bonatti, Giulia Mazzaschi, Alessandra Dodi, Francesco Facchinetti, Francesco Gelsomino, Gloria Cinquegrani, Anna Squadrilli, Paola Bordi, Sebastiano Buti, Melissa Bersanelli, Alessandro Leonetti, Agnese Cosenza, Leonarda Ferri, Elena Rapacchi, Federico Quaini, Andrea Ardizzoni and Marcello Tiseo in Tumori Journal

Supplemental material, figure_A3 for CEA and CYFRA 21-1 as prognostic biomarker and as a tool for treatment monitoring in advanced NSCLC treated with immune checkpoint inhibitors by Filippo G. Dall’Olio, Francesca Abbati, Francesco Facchinetti, Maria Massucci, Barbara Melotti, Anna Squadrilli, Sebastiano Buti, Francesca Formica, Marcello Tiseo and Andrea Ardizzoni in Therapeutic Advances in Medical Oncology

Supplemental material, Supplementary_data_PDF for CEA and CYFRA 21-1 as prognostic biomarker and as a tool for treatment monitoring in advanced NSCLC treated with immune checkpoint inhibitors by Filippo G. Dall’Olio, Francesca Abbati, Francesco Facchinetti, Maria Massucci, Barbara Melotti, Anna Squadrilli, Sebastiano Buti, Francesca Formica, Marcello Tiseo and Andrea Ardizzoni in Therapeutic Advances in Medical Oncology

Supplemental material, Supplementary_table_1 for KRAS and ERBB-family genetic alterations affect response to PD-1 inhibitors in metastatic nonsquamous NSCLC by Marika Cinausero, Noemi Laprovitera, Giovanna De Maglio, Lorenzo Gerratana, Mattia Riefolo, Marianna Macerelli, Michelangelo Fiorentino, Elisa Porcellini, Vanessa Buoro, Francesco Gelsomino, Anna Squadrilli, Gianpiero Fasola, Massimo Negrini, Marcello Tiseo, Manuela Ferracin and Andrea Ardizzoni in Therapeutic Advances in Medical Oncology

Aim: This prospective study aimed to envisage the putative prognostic significance of clinical and hematologic parameters in advanced non-small-cell lung cancer patients treated with nivolumab. Materials & methods: Correlations of several parameters with disease control and survival outcomes were provided. Results: A total of 54 patients were included. An ECOG performance status 0–1, the lack of liver and bone metastases and a timeframe from the last systemic treatment ≥4 months correlated with better disease control. The same was observed for baseline low levels of white blood cells and neutrophils, for high levels of NK cells and a neutrophil/lymphocyte ratio

Background The identification of ALK and ROS1 rearrangements and the availability of an effective target therapy, such as crizotinib, represent a new option in the treatment of advanced non-small cell lung cancer (NSCLC) patients. In light of recent advances in non-invasive diagnostic procedures, we aimed to demonstrate that direct cytological smears are suitable for assessing ALK and ROS1 rearrangements in patients with NSCLC. Methods Fifty-five patients with a cytological diagnosis of lung adenocarcinoma (ADC) were evaluated for ALK rearrangements by fluorescence in situ hybridization (FISH) and 12 patients for ROS1 FISH rearrangements. Seventeen of the 55 cytological samples tested for ALK were obtained from the primary tumor and 38 from metastatic lesions. Ten of 12 samples evaluated for ROS1 were obtained from metastatic sites and two from the primary tumor. Results ALK FISH was successful in 49/55 (89%) cytological ADC samples and ROS1 FISH in all 12 cytological samples. ALK rearrangements were found in 3/13 (23%) primary tumors and 7/36 (19%) metastatic sites. ROS1 rearrangements were found in one of the two primary tumors and in two of the 10 metastases. Two of the three rearranged cases were tested on cytology after knowing that they were rearranged on histology in order to increase representativeness of ROS1 rearranged cases in this study. Conclusion Whenever cytology represents the only available material for diagnosis and biological characterization of NSCLC, minimally invasive procedures may provide an additional important source of cellular material for FISH assessment of ALK and ROS1 rearrangements.

Introduction The exclusion of circulating tumor cells (CTCs) that have lost epithelial antigens during the epithelial-to-mesenchymal transition (EMT) process by using Epithelial Cell Adhesion Molecule (EpCAM) based capture methods is still a matter of debate. In this study, cells obtained after depletion procedure from blood samples of squamous cell lung cancer (SQCLC) patients were identified based on morphology and characterized with the combination of FISH assessment and immunophenotypic profile. Materials and Methods Five mL blood samples, collected from 55 advanced SQCLC patients, were analyzed by a non-EpCAM-based capture method. After depletion of leukocytes and erythroid cells, the negative fraction was characterized by both FISH using a fibroblast growth factor receptor 1 (FGFR1) probe and by immunocytochemistry. Thirty healthy donors were also tested. Results Based on morphology (nuclear dimension ≥10 μm, shape and hypercromatic aspect) suspicious circulating cells clearly distinguishable from contaminant leukocytes were observed in 49/55 (89%) SQCLC patients. Thirty-four of the 44 (77%) samples evaluable for FGFR1 FISH showed ≥ 6 FGFR1 gene copy number on average per cell. Vimentin expression involved 43% (18/42) of pooled circulating SQCLC cells, whereas only 29% (14/48) were EpCAM positive. Confocal microscopy confirmed the localization of FGFR1 probe in suspicious circulating cells. Suspicious circulating elements were also observed in healthy donors and did not show any epithelial associated antigens. A significantly lower number of suspicious circulating cells in healthy donors compared to SQCLC patients was found. Conclusions Among the heterogeneous cell population isolated by depletion procedure, the coexistence of cells with epithelial and/or mesenchymal phenotype suggests that EMT may participate to transendothelial invasion and migration of tumor cells in advanced SQCLC. The finding of cells with neither EpCAM or EMT phenotype, retrieved after non-EpCAM-based systems, underlines the presence of suspicious elements in the blood of both SQCLC patients and healthy donors. Further phenotyping and molecular analyses are necessary to fully characterize these circulating elements.