IDENTIFICAZIONE DI UNA FIRMA MOLECOLARE ASSOCIATA ALLA PROGNOSI E ALLA PLATINO SENSIBILITÀ IN PAZIENTI AFFETTE DA CARCINOMA OVARICO AVANZATO

Il carcinoma ovarico è una delle principali cause di morte per cancro nelle donne. L'elevata mortalità è principalmente legata alla diagnosi tardiva, alla mancanza di un test di screening e all’insorgenza di chemioresistenza. I criteri prognostici e predittivi attualmente utilizzati non consentono di predire la recidiva e la responsività ai trattamenti. Risulta necessario identificare nuovi markers utilizzabili nella diagnosi precoce di questa neoplasia e nella selezione di pazienti affette da carcinoma ovarico candidabili a trattamenti personalizzati. Le moderne tecnologie utilizzate in biologia molecolare consentono di studiare il genoma e il trascrittoma. Il mio lavoro si è concentrato sullo studio del trascrittoma nelle pazienti affette da carcinoma ovarico avanzato (stadio III e IV) con l'obiettivo di identificare dei trascritti per distinguere le donne sane da quelle affette da neoplasia ovarica e in questo gruppo quelle con buona e cattiva prognosi. La tesi è articolata in tre parti: la prima parte consiste nell'analisi del miRNoma tissutale per identificare dei miRNA differenzialmente espressi tra le pazienti platino resistenti e sensibili,la seconda parte studia sempre i miRNA ma nel siero di donne sane e donne affette da cancro con l'obiettivo di trovare dei miRNA che possano essere utilizzati per la diagnosi precoce del trattamento,nel siero sono state anche ricercate differenze di espressione con correlazioni con la risposta al trattamento. Nella terza parte della tesi invece oltre ai miRNA abbiamo sequenziato l’intero trascrittoma di un piccolo gruppo di pazienti platino resistenti e di un gruppo analogo di pazienti platino sensibili per identificare trascritti differenzialmente espressi in questi due gruppi di pazienti. Le pazienti arruolate in questo studio sono pazienti affette da carcinoma ovarico avanzato sottoposte a chemioterapia a base di platino, appartengonenti a tre casistiche differenti omogenee per istotipo, stadio e trattamento chemioterapico di prima linea. Di esse è disponibile del tessuto tumorale fresco congelato e per due delle tre coorti anche il siero. Sono stati utilizzati i sieri di alcuni soggetti sani per la seconda parte della tesi. L'analisi del miRNoma è stata effettuata sulla prima coorte mediante microarrray e non evidenziato dei miRNA differenzialmente espressi con correlazioni con platino resistenza e sensibilità. Anche lo studio dei miRNA sierici non ha evidenziato miRNA differenzialmente espressi con valore predittivo o prognostico. Confrontando l'espressione dei miRNA sierici di donne sane e di donne affette da carcinoma ovarico abbiamo identificato in questo secondo gruppo un’aumentata espressione di 3 miRNA. Questi risultati sono stati validati anche in real time PCR confermando l’overespressione dei 3 miR nelle donne affette da carcinoma ovarico. L'ultima parte del lavoro prevedeva lo studio del trascrittoma mediante RNAseq,sono stati identificati in tutto 1371 trascritti differenzialmente espressi in queste pazienti. Gli esperimenti di validazione di questi trascritti mediante real time PCR non hanno avuto esito positivo. In conclusione non sono stati identificati miRNA tissutali e sierici con correlazione con la sensibilità al platino e la sopravvivenza, nel trascrittoma sono stati identificati numerosi trascritti differenzialmente espressi ma tali risultati non sono stati validati. Sono stati identificati 3 miRNA sierici differenzialmente espressi tra le pazienti affette da carcinoma ovarico e donne sane, essi potrebbero essere ulteriormente studiati per il loro utilizzo come marcatori di screening per la neoplasia ovarica, una patologia per cui ad oggi non è riconosciuto un test di screening efficace

Systemic pro-inflammatory response identifies patients with cancer with adverse outcomes from SARS-CoV-2 infection: the OnCovid Inflammatory Score

Background: Patients with cancer are particularly susceptible to SARS-CoV-2 infection. The systemic inflammatory response is a pathogenic mechanism shared by cancer progression and COVID-19. We investigated systemic inflammation as a driver of severity and mortality from COVID-19, evaluating the prognostic role of commonly used inflammatory indices in SARS-CoV-2-infected patients with cancer accrued to the OnCovid study. Methods: In a multicenter cohort of SARS-CoV-2-infected patients with cancer in Europe, we evaluated dynamic changes in neutrophil:lymphocyte ratio (NLR); platelet:lymphocyte ratio (PLR); Prognostic Nutritional Index (PNI), renamed the OnCovid Inflammatory Score (OIS); modified Glasgow Prognostic Score (mGPS); and Prognostic Index (PI) in relation to oncological and COVID-19 infection features, testing their prognostic potential in independent training (n=529) and validation (n=542) sets. Results: We evaluated 1071 eligible patients, of which 625 (58.3%) were men, and 420 were patients with malignancy in advanced stage (39.2%), most commonly genitourinary (n=216, 20.2%). 844 (78.8%) had ≥1 comorbidity and 754 (70.4%) had ≥1 COVID-19 complication. NLR, OIS, and mGPS worsened at COVID-19 diagnosis compared with pre-COVID-19 measurement (p<0.01), recovering in survivors to pre-COVID-19 levels. Patients in poorer risk categories for each index except the PLR exhibited higher mortality rates (p<0.001) and shorter median overall survival in the training and validation sets (p<0.01). Multivariable analyses revealed the OIS to be most independently predictive of survival (validation set HR 2.48, 95% CI 1.47 to 4.20, p=0.001; adjusted concordance index score 0.611). Conclusions: Systemic inflammation is a validated prognostic domain in SARS-CoV-2-infected patients with cancer and can be used as a bedside predictor of adverse outcome. Lymphocytopenia and hypoalbuminemia as computed by the OIS are independently predictive of severe COVID-19, supporting their use for risk stratification. Reversal of the COVID-19-induced proinflammatory state is a putative therapeutic strategy in patients with cancer

Determinants of enhanced vulnerability to coronavirus disease 2019 in UK patients with cancer: a European study

Despite high contagiousness and rapid spread, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to heterogeneous outcomes across affected nations. Within Europe (EU), the United Kingdom (UK) is the most severely affected country, with a death toll in excess of 100,000 as of January 2021. We aimed to compare the national impact of coronavirus disease 2019 (COVID-19) on the risk of death in UK patients with cancer versus those in continental EU. Methods: We performed a retrospective analysis of the OnCovid study database, a European registry of patients with cancer consecutively diagnosed with COVID-19 in 27 centres from 27th February to 10th September 2020. We analysed case fatality rates and risk of death at 30 days and 6 months stratified by region of origin (UK versus EU). We compared patient characteristics at baseline including oncological and COVID-19-specific therapy across UK and EU cohorts and evaluated the association of these factors with the risk of adverse outcomes in multivariable Cox regression models. Findings: Compared with EU (n = 924), UK patients (n = 468) were characterised by higher case fatality rates (40.38% versus 26.5%, p < 0.0001) and higher risk of death at 30 days (hazard ratio [HR], 1.64 [95% confidence interval {CI}, 1.36-1.99]) and 6 months after COVID-19 diagnosis (47.64% versus 33.33%; p < 0.0001; HR, 1.59 [95% CI, 1.33-1.88]). UK patients were more often men, were of older age and have more comorbidities than EU counterparts (p < 0.01). Receipt of anticancer therapy was lower in UK than in EU patients (p < 0.001). Despite equal proportions of complicated COVID-19, rates of intensive care admission and use of mechanical ventilation, UK patients with cancer were less likely to receive anti-COVID-19 therapies including corticosteroids, antivirals and interleukin-6 antagonists (p < 0.0001). Multivariable analyses adjusted for imbalanced prognostic factors confirmed the UK cohort to be characterised by worse risk of death at 30 days and 6 months, independent of the patient's age, gender, tumour stage and status; number of comorbidities; COVID-19 severity and receipt of anticancer and anti-COVID-19 therapy. Rates of permanent cessation of anticancer therapy after COVID-19 were similar in the UK and EU cohorts. Interpretation: UK patients with cancer have been more severely impacted by the unfolding of the COVID-19 pandemic despite societal risk mitigation factors and rapid deferral of anticancer therapy. The increased frailty of UK patients with cancer highlights high-risk groups that should be prioritised for anti-SARS-CoV-2 vaccination. Continued evaluation of long-term outcomes is warranted

27P * Common gene signature expressed by breast and kidney cancers-derived endothelial colony forming cells

nhibitors of the vascular endothelial growth factor (VEGF) pathway are approved for the treatment of several tumor types but their effectiveness is limited. The heterogeneity of tumor endothelial cells, determined by the dynamics of tumor vascularization not only requires sprouting angiogenesis but impinges on alternative mechanisms such as the recruitment of bone marrow (BM)-derived endothelial progenitor cells (EPCs). We focused on endothelial colony forming cells (ECFCs) the only EPC population truly belonging to the endothelial lineage by comparing normal vs tumor-derived ECFCs phenotype in two distinct, but hypervascularized cancers, such as renal cell carcinoma (RCC) and breast cancer (BC). ECFCs were successfully isolated from healthy donors (N-ECFCs), RCC patients (RCC-ECFCs) and BC patients (BC-ECFCs). ECFC colonies displayed the typical cobblestone-shaped monolayer, both normal and tumor derived ECFCs formed capillary-likes structures, and express classic endothelial markers. There was no difference in growth kinetics or tubulogenic rate between N-, RCC-, and BC-ECFCs. However, RCC-ECFCs were more resistant to rapamycin-induced apoptosis as compared to control cells, while BC-ECFCs were more sensitive. Genomic expression analysis identified a total of 382 differentially expressed genes (DEGs) between N- and BC-ECFCs and of 71 DEGs (33 up-regulated, 38 down-regulated) between N- and RCC-ECFCs. Nevertheless, we identified a common gene signature in BC- and RCC-derived ECFCs due to the presence of 35 DEGs that shared the same direction of regulation, i.e. all genes were either up-regulated or down-regulated in both groups. Ingenuity pathway analysis (IPA) unveiled interactions between these 35 DEGs there were centered on FOS, which plays a key role in the control of cell cycle and apoptosis. These findings provide the first strong molecular evidence that tumor microenviroment may also affect distant tissues and reprogram bone marrow derived-cells, such as ECFCs. However, the common signature identified in ECFCs isolated from patients suffering from two distinct tumor types might be relevant for carcinogenesis and could suggest new targets anti-angiogenic therapies

Regional and temporal heterogeneity of epithelial ovarian cancer tumor biopsies: implications for therapeutic strategies

Stage III/IV epithelial ovarian cancer (EOC) is a systemic disease. The clonal relationship among different tumor lesions at diagnosis (spatial heterogeneity) and how tumor clonal architecture evolves over time (temporal heterogeneity) have not yet been defined. Such knowledge would help to develop new target-based strategies, as biomarkers which can adjudge the success of therapeutic intervention should be independent of spatial and temporal heterogeneity. The work described in this paper addresses spatial and temporal heterogeneity in a cohort of 71 tumor biopsies using targeted NGS technology. These samples were taken from twelve high grade serous (HGS) and seven non HSG-EOC, both at the time of primary surgery when the tumor was na\uefve to chemotherapy and after chemotherapy. Matched tumor lesions growing in the ovary or at other anatomical sites show very different mutational landscapes with branched tumor evolution. Mutations in ATM, ATR, TGFB3, VCAM1 and COL3A1 genes were shared across all lesions. BRCA1 and BRCA2 genes were frequently mutated in synchronous lesions of non HGS-EOC. Relapsed disease seems to originate from resistant clones originally present at the time of primary surgery rather than from resistance acquired de novo during platinum based therapy. Overall the work suggests that EOC continues to evolve. More detailed mapping of genetic lesions is necessary to improve therapeutic strategies

COVID-19 in breast cancer patients: a subanalysis of the OnCovid registry

Background: cancer patients are at higher risk of COVID-19 complications and mortality than the rest of the population. Breast cancer patients seem to have better prognosis when infected by SARS-CoV-2 than other cancer patients. Methods: we report a subanalysis of the OnCovid study providing more detailed information in the breast cancer population. Results: we included 495 breast cancer patients with a SARS-CoV-2 infection. Mean age was 62.6 years; 31.5% presented more than one comorbidity. The most frequent breast cancer subtype was luminal-like (n = 245, 49.5%) and 177 (35.8%) had metastatic disease. A total of 332 (67.1%) patients were receiving active treatment, with radical intent in 232 (47.6%) of them. Hospitalization rate was 58.2% and all-cause mortality rate was 20.3%. One hundred twenty-nine (26.1%) patients developed one COVID-19 complication, being acute respiratory failure the most common (n = 74, 15.0%). In the multivariable analysis, age older than 70 years, presence of COVID-19 complications, and metastatic disease were factors correlated with worse outcomes, while ongoing anticancer therapy at time of COVID-19 diagnosis appeared to be a protective factor. No particular oncological treatment was related to higher risk of complications. In the context of SARS-CoV-2 infection, 73 (18.3%) patients had some kind of modification on their oncologic treatment. At the first oncological reassessment (median time: 46.9 days ± 36.7), 255 (51.6%) patients reported to be fully recovered from the infection. There were 39 patients (7.9%) with long-term SARS-CoV-2-related complications. Conclusion: in the context of COVID-19, our data confirm that breast cancer patients appear to have lower complications and mortality rate than expected in other cancer populations. Most breast cancer patients can be safely treated for their neoplasm during SARS-CoV-2 pandemic. Oncological treatment has no impact on the risk of SARS-CoV-2 complications, and, especially in the curative setting, the treatment should be modified as little as possible

Determinants of enhanced vulnerability to coronavirus disease 2019 in UK patients with cancer: a European study

Background: Despite high contagiousness and rapid spread, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to heterogeneous outcomes across affected nations. Within Europe (EU), the United Kingdom (UK) is the most severely affected country, with a death toll in excess of 100,000 as of January 2021. We aimed to compare the national impact of coronavirus disease 2019 (COVID-19) on the risk of death in UK patients with cancer versus those in continental EU. Methods: We performed a retrospective analysis of the OnCovid study database, a European registry of patients with cancer consecutively diagnosed with COVID-19 in 27 centres from 27th February to 10th September 2020. We analysed case fatality rates and risk of death at 30 days and 6 months stratified by region of origin (UK versus EU). We compared patient characteristics at baseline including oncological and COVID-19especific therapy across UK D.J. Pinato et al. / European Journal of Cancer 150 (2021) 190e202 191 and EU cohorts and evaluated the association of these factors with the risk of adverse outcomes in multivariable Cox regression models. Findings: Compared with EU (n Z 924), UK patients (n Z 468) were characterised by higher case fatality rates (40.38% versus 26.5%, p < 0.0001) and higher risk of death at 30 days (hazard ratio [HR], 1.64 [95% confidence interval {CI}, 1.36e1.99]) and 6 months after COVID-19 diagnosis (47.64% versus 33.33%; p < 0.0001; HR, 1.59 [95% CI, 1.33e1.88]). UK patients were more often men, were of older age and have more comorbidities than EU counterparts (p < 0.01). Receipt of anticancer therapy was lower in UK than in EU patients (p < 0.001). Despite equal proportions of complicated COVID-19, rates of intensive care admission and use of mechanical ventilation, UK patients with cancer were less likely to receive anti eCOVID-19 therapies including corticosteroids, antivirals and interleukin-6 antagonists (p < 0.0001). Multivariable analyses adjusted for imbalanced prognostic factors confirmed the UK cohort to be characterised by worse risk of death at 30 days and 6 months, independent of the patient\u2019s age, gender, tumour stage and status; number of comorbidities; COVID- 19 severity and receipt of anticancer and antieCOVID-19 therapy. Rates of permanent cessation of anticancer therapy after COVID-19 were similar in the UK and EU cohorts. Interpretation: UK patients with cancer have been more severely impacted by the unfolding of the COVID-19 pandemic despite societal risk mitigation factors and rapid deferral of anticancer therapy. The increased frailty of UK patients with cancer highlights high-risk groups that should be prioritised for antieSARS-CoV-2 vaccination. Continued evaluation of long-term outcomes is warranted

Time-Dependent COVID-19 Mortality in Patients with Cancer:An Updated Analysis of the OnCovid Registry

IMPORTANCE: Whether the severity and mortality of COVID-19 in patients with cancer have improved in terms of disease management and capacity is yet to be defined. OBJECTIVE: To test whether severity and mortality from COVID-19 among patients with cancer have improved during the course of the pandemic. DESIGN, SETTING, AND PARTICIPANTS: OnCovid is a European registry that collects data on consecutive patients with solid or hematologic cancer and COVID-19. This multicenter case series study included real-world data from 35 institutions across 6 countries (UK, Italy, Spain, France, Belgium, and Germany). This update included patients diagnosed between February 27, 2020, and February, 14, 2021. Inclusion criteria were confirmed diagnosis of SARS-CoV-2 infection and a history of solid or hematologic cancer. EXPOSURES: SARS-CoV-2 infection. MAIN OUTCOMES AND MEASURES: Deaths were differentiated at 14 days and 3 months as the 2 landmark end points. Patient characteristics and outcomes were compared by stratifying patients across 5 phases (February to March 2020, April to June 2020, July to September 2020, October to December 2020, and January to February 2021) and across 2 major outbreaks (February to June 2020 and July 2020 to February 2021). RESULTS: At data cutoff, 2795 consecutive patients were included, with 2634 patients eligible for analysis (median [IQR] age, 68 [18-77] years ; 52.8% men). Eligible patients demonstrated significant time-dependent improvement in 14-day case-fatality rate (CFR) with estimates of 29.8% (95% CI, 0.26-0.33) for February to March 2020; 20.3% (95% CI, 0.17-0.23) for April to June 2020; 12.5% (95% CI, 0.06-22.90) for July to September 2020; 17.2% (95% CI, 0.15-0.21) for October to December 2020; and 14.5% (95% CI, 0.09-0.21) for January to February 2021 (all P < .001) across the predefined phases. Compared with the second major outbreak, patients diagnosed in the first outbreak were more likely to be 65 years or older (974 of 1626 [60.3%] vs 564 of 1008 [56.1%]; P = .03), have at least 2 comorbidities (793 of 1626 [48.8%] vs 427 of 1008 [42.4%]; P = .001), and have advanced tumors (708 of 1626 [46.4%] vs 536 of 1008 [56.1%]; P < .001). Complications of COVID-19 were more likely to be seen (738 of 1626 [45.4%] vs 342 of 1008 [33.9%]; P < .001) and require hospitalization (969 of 1626 [59.8%] vs 418 of 1008 [42.1%]; P < .001) and anti–COVID-19 therapy (1004 of 1626 [61.7%] vs 501 of 1008 [49.7%]; P < .001) during the first major outbreak. The 14-day CFRs for the first and second major outbreaks were 25.6% (95% CI, 0.23-0.28) vs 16.2% (95% CI, 0.13-0.19; P < .001), respectively. After adjusting for country, sex, age, comorbidities, tumor stage and status, anti–COVID-19 and anticancer therapy, and COVID-19 complications, patients diagnosed in the first outbreak had an increased risk of death at 14 days (hazard ratio [HR], 1.85; 95% CI, 1.47-2.32) and 3 months (HR, 1.28; 95% CI, 1.08-1.51) compared with those diagnosed in the second outbreak. CONCLUSIONS AND RELEVANCE: The findings of this registry-based study suggest that mortality in patients with cancer diagnosed with COVID-19 has improved in Europe; this improvement may be associated with earlier diagnosis, improved management, and dynamic changes in community transmission over time

Prevalence and impact of COVID-19 sequelae on treatment pathways and survival of patients with cancer who recovered from SARS-Cov-2 infection: results from the OnCovid registry.

retrospective, registry study David J Pinato, Josep Tabernero, Mark Bower, Lorenza Scotti, Meera Patel, Emeline Colomba, Saoirse Dolly, Angela Loizidou, John Chester, Uma Mukherjee, Alberto Zambelli, Alessia Dalla Pria, Juan Aguilar-Company, Diego Ottaviani, Amani Chowdhury, Eve Merry, Ramon Salazar, Alexia Bertuzzi, Joan Brunet, Matteo Lambertini, Marco Tagliamento, Anna Pous, Ailsa Sita-Lumsden, Krishnie Srikandarajah, Johann Colomba, Fanny Pommeret, Elia Segui, Daniele Generali, Salvatore Grisanti, Paolo Pedrazzoli, Gianpiero Rizzo, Michela Libertini, Charlotte Moss, Joanne S Evans, Beth Russell, Nadia Harbeck, Bruno Vincenzi, Federica Biello, Rossella Bertulli, Raquel Lilian, Sabrina Rossi, Maria Carmen Carmona-Garcia, Carlo Tondini, Laura Fox, Alice Baggi, Vittoria Fotia, Alessandro Parisi, Giampero Porzio, Maristella Saponara, Claudia Andrea Cruz, David Garcia-Illescas, Eudald Felip, Ariadna Roque Lloveras, Rachel Sharkey, Elisa Roldan, Roxana Reyes, Irina Earnshaw, Daniela Ferrante, Javier Marco-Hernandez, Isabel Ruiz-Camps, Gianluca Gaidano, Andrea Patriarca, Riccardo Bruna, Anna Sureda, Clara Martinez-Vila, Ana Sanchez de Torre, Luca Cantini, Marco Filetti, Lorenza Rimassa, Lorenzo Chiudinelli, Michela Franchi, Marco Krengli, Armando Santoro, Aleix Prat, Mieke Van Hemelrijck, Nikolaos Diamantis, Thomas Newsom-Davis, Alessandra Gennari, Alessio Cortellini, on behalf of the OnCovid study group