The increasing need to identify new molecular biomarkers in patients affected by non-small cell lung cancer (NSCLC), in order to detect potential therapeutic targets and personalize treatment approaches, has led molecular biology laboratories to increasingly adopt technologies with high sensitivity and specificity. In this context, Next Generation Sequencing (NGS) currently represents the gold standard, as it allows the simultaneous analysis of multiple genes and multiple patients. The aim of this study is to characterize the molecular profile of patients with NSCLC using an extended panel comprising 50 genes. An experimental study was conducted on 154 patients diagnosed with NSCLC, analyzing lung biopsy samples and specimens obtained from lung resections. Molecular analysis was performed using NGS technology with a 50-gene extended panel. Data analysis was carried out using the Myriapod® NGS Data Analysis Software. The results showed that 60.60% of patients presented genetic alterations associated with the availability of targeted therapies currently used in clinical practice, whereas 39.40% exhibited mutations for which no specific treatments are yet available. Among the molecular alterations treatable with targeted therapy, mutations in EGFR (13.64%), KRAS G12C (7.79%), and BRAF V600E (5.19%) were identified. Additionally, 3.90% of patients showed ALK gene fusions, while ERBB2 alterations were detected in 0.65% of cases. Furthermore, 28.78% of patients tested positive for PD-L1 expression, an important predictive biomarker for access to immunotherapy. Among alterations for which no approved targeted therapies are currently available, mutations in STK11 were observed in 2.60% of cases, PIK3CA in 4.55%, and other KRAS variants in 9.09% of patients. The coexistence of TP53 and EGFR alterations was identified in 24.03% of cases. The obtained results confirm the key role of NGS technology in the molecular characterization of NSCLC. Indeed, this approach allows not only the identification of genomic alterations currently amenable to targeted treatment, but also the detection of variants lacking specific therapeutic options. These findings expand current knowledge of the mutational landscape of NSCLC and provide a basis for the future development of new strategies, enabling more accurate clinical and therapeutic patient management.
La crescente esigenza di individuare nuovi biomarcatori molecolari nei pazienti affetti da carcinoma polmonare non a piccole cellule (NSCLC, Non-Small Cell Lung Cancer), al fine di identificare possibili bersagli terapeutici e personalizzare l’approccio terapeutico, ha spinto i laboratori di biologia molecolare a utilizzare sempre più frequentemente tecnologie ad alta sensibilità e specificità. Il tale ambito di sequenziamento di nuova generazione (NGS, Next Generation Sequencing) attualmente rappresenta il gold stardard in quanto consente l’analisi di più geni e di più pazienti contemporaneamente. Il presente lavoro ha lo scopo di caratterizzare il profilo molecolare di pazienti con NSCLC mediante l'utilizzo di un pannello esteso comprendente 50 geni. È stato condotto uno studio sperimentale su 154 pazienti con diagnosi di NSCLC, analizzando campioni bioptici polmonari e campioni derivanti da resezione polmonare. L'analisi molecolare è stata svolta mediante tecnologia NGS utilizzando un pannello esteso di 50 geni. L’analisi dei dati è stata effettuata tramite il software Myriapod® NGS Data Analysis Software. I risultati hanno evidenziato che il 60,60% dei pazienti presentava alterazioni genetiche associate alla disponibilità di terapie target attualmente in uso nella pratica clinica, mentre il 39,40% mostrava mutazioni per le quali non sono ancora disponibili trattamenti specifici. Tra le alterazioni molecolari trattabili con terapia mirata sono state riscontrate mutazioni di EGFR (13,64%), KRAS G12C (7,79%) e BRAF V600E (5,19%). Inoltre, il 3,90% dei pazienti presentava una fusione di ALK, mentre alterazioni di ERBB2 sono state rilevate nello 0,65% dei casi. Il 28,78% dei pazienti è risultato positivo all’espressione di PD-L1, importante biomarcatore predittivo per l'accesso all'immunoterapia. Tra le alterazioni per le quali non sono ancora disponibili terapie target approvate sono state osservate mutazioni di STK11 nel 2,60% dei casi, PIK3CA nel 4,55% dei casi e ulteriori varianti del gene KRAS nel 9,09% dei pazienti e la coesistenza di alterazioni a carico di TP53 ed EGFR nel 24,03% dei casi. I risultati ottenuti confermano il ruolo chiave della tecnologia NGS nella caratterizzazione molecolare del NSCLC. Infatti, tale tecnologia consente non solo l'identificazione di alterazioni genomiche attualmente suscettibili di trattamento mirato, ma anche la rilevazione di varianti prive di specifiche opzioni terapeutiche. Tali evidenze ampliano le conoscenze sul panorama mutazionale del NSCLC e rappresentano un fondamento per lo sviluppo futuro di nuove strategie consentendo il più corretto inquadramento clinico e terapeutico del paziente.
NGS: nuovi marker molecolari nel NSCLC.
D'ANNIBALLE, JESSICA
2025/2026
Abstract
The increasing need to identify new molecular biomarkers in patients affected by non-small cell lung cancer (NSCLC), in order to detect potential therapeutic targets and personalize treatment approaches, has led molecular biology laboratories to increasingly adopt technologies with high sensitivity and specificity. In this context, Next Generation Sequencing (NGS) currently represents the gold standard, as it allows the simultaneous analysis of multiple genes and multiple patients. The aim of this study is to characterize the molecular profile of patients with NSCLC using an extended panel comprising 50 genes. An experimental study was conducted on 154 patients diagnosed with NSCLC, analyzing lung biopsy samples and specimens obtained from lung resections. Molecular analysis was performed using NGS technology with a 50-gene extended panel. Data analysis was carried out using the Myriapod® NGS Data Analysis Software. The results showed that 60.60% of patients presented genetic alterations associated with the availability of targeted therapies currently used in clinical practice, whereas 39.40% exhibited mutations for which no specific treatments are yet available. Among the molecular alterations treatable with targeted therapy, mutations in EGFR (13.64%), KRAS G12C (7.79%), and BRAF V600E (5.19%) were identified. Additionally, 3.90% of patients showed ALK gene fusions, while ERBB2 alterations were detected in 0.65% of cases. Furthermore, 28.78% of patients tested positive for PD-L1 expression, an important predictive biomarker for access to immunotherapy. Among alterations for which no approved targeted therapies are currently available, mutations in STK11 were observed in 2.60% of cases, PIK3CA in 4.55%, and other KRAS variants in 9.09% of patients. The coexistence of TP53 and EGFR alterations was identified in 24.03% of cases. The obtained results confirm the key role of NGS technology in the molecular characterization of NSCLC. Indeed, this approach allows not only the identification of genomic alterations currently amenable to targeted treatment, but also the detection of variants lacking specific therapeutic options. These findings expand current knowledge of the mutational landscape of NSCLC and provide a basis for the future development of new strategies, enabling more accurate clinical and therapeutic patient management.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12075/27845