Pancreatic Neuroendocrine tumors (pNETs) comprise 1% to 3% of yearly pancreatic tumors diagnosis but, when compared to other types of cancers, they more frequently present themselves as unresectable neoplasms or with distant metastases. Indeed, the only potentially curative treatment for pNETs is surgery, which, due to the inherently malignant potential of these tumors, is not always a suitable option for the patients. The main objective of this study was to identify a novel therapeutic strategy for these tumors. We specifically identified two molecular pathways that are mediating the proliferation and survival of pNET cancer cells (i.e. PI3K pathway and cell cycle regulation), and targeted them. We then assessed the effects of inhibiting these signaling cascades on cell proliferation, viability and downstream pathway suppression. The human pancreatic tumor cell lines BON1, and NT-3 were exposed to two drugs Ribociclib, a CDK4/CDK6 inhibitor, and Buparlisib, a PI3K inhibitor, alone or in combination. BON1 and NT-3 cells were grown as 2D or 3D cultures. After exposure, we measured cell proliferation and apoptosis. RNA and proteins were extracted to then be employed in quantitative PCR and Western Blot analysis, respectively, to verify the inhibition of downstream targets. The results showed a dose-dependent anti-tumor effect of the drugs. Although the single drugs were effective, we observed a stronger anti-tumor effect when the cells were administered the combination of Buparlisib and Ribociclib. This strategy has great potential as it allowed for a drastic reduction of the concentrations of Buparlisib and Ribociclib while still maintaining a potent anti-tumor effect when compared to the use single drugs. Indeed, higher drug concentration would lead to toxicity problems in the patients. Overall, we identified a possible novel treatment strategy for pNETs by combining PI3K and CDK4/CDK6 inhibitors.
Nuovi strategie per il trattamento di Tumori Neuroendocrini del Pancreas. I tumori neuroendocrini pancreatici (pNET) comprendono dall'1% al 3% della diagnosi annuale di tumori pancreatici ma, rispetto ad altri tipi di tumori, si presentano più frequentemente come neoplasie non resecabili o con metastasi a distanza. In effetti, l'unico trattamento potenzialmente curativo per i pNET è la chirurgia, che, a causa del potenziale intrinsecamente maligno di questi tumori, non è sempre un'opzione adatta per i pazienti. L'obiettivo principale di questo studio era identificare una nuova strategia terapeutica per questi tumori. Abbiamo specificamente identificato due percorsi molecolari che mediano la proliferazione e la sopravvivenza delle cellule tumorali pNET (ovvero il percorso PI3K e la regolazione del ciclo cellulare) e li abbiamo presi di mira. Abbiamo quindi valutato gli effetti dell'inibizione di queste cascate di segnalazione sulla proliferazione cellulare, la vitalità e la soppressione del percorso a valle. Le linee cellulari di tumore pancreatico umano BON1 e NT-3 sono state coltivate come colture 2D o 3D esposte a due farmaci Ribociclib, un inibitore CDK4/CDK6, e Buparlisib, un inibitore PI3K, da soli o in combinazione. Dopo l'esposizione, abbiamo misurato la proliferazione cellulare e l'apoptosi. L'RNA e le proteine sono state estratte per poi essere impiegate in PCR quantitativa e analisi Western Blot, rispettivamente, per verificare l'inibizione dei bersagli a valle. I risultati hanno mostrato un effetto antitumorale dose-dipendente dei farmaci. Sebbene i singoli farmaci fossero efficaci, abbiamo osservato un effetto antitumorale più forte quando alle cellule è stata somministrata la combinazione di Buparlisib e Ribociclib. Questa strategia ha un grande potenziale in quanto ha permesso di ridurre drasticamente le concentrazioni di Buparlisib e Ribociclib pur mantenendo un potente effetto antitumorale rispetto all'uso di singoli farmaci. Infatti, una maggiore concentrazione del farmaco porterebbe a problemi di tossicità nei pazienti. Nel complesso, abbiamo identificato una possibile nuova strategia di trattamento per i pNET combinando gli inibitori PI3K e CDK4/CDK6.
Novel treatment strategies for Pancreatic Neuroendocrine Tumors
GENIO, EDOARDO
2021/2022
Abstract
Pancreatic Neuroendocrine tumors (pNETs) comprise 1% to 3% of yearly pancreatic tumors diagnosis but, when compared to other types of cancers, they more frequently present themselves as unresectable neoplasms or with distant metastases. Indeed, the only potentially curative treatment for pNETs is surgery, which, due to the inherently malignant potential of these tumors, is not always a suitable option for the patients. The main objective of this study was to identify a novel therapeutic strategy for these tumors. We specifically identified two molecular pathways that are mediating the proliferation and survival of pNET cancer cells (i.e. PI3K pathway and cell cycle regulation), and targeted them. We then assessed the effects of inhibiting these signaling cascades on cell proliferation, viability and downstream pathway suppression. The human pancreatic tumor cell lines BON1, and NT-3 were exposed to two drugs Ribociclib, a CDK4/CDK6 inhibitor, and Buparlisib, a PI3K inhibitor, alone or in combination. BON1 and NT-3 cells were grown as 2D or 3D cultures. After exposure, we measured cell proliferation and apoptosis. RNA and proteins were extracted to then be employed in quantitative PCR and Western Blot analysis, respectively, to verify the inhibition of downstream targets. The results showed a dose-dependent anti-tumor effect of the drugs. Although the single drugs were effective, we observed a stronger anti-tumor effect when the cells were administered the combination of Buparlisib and Ribociclib. This strategy has great potential as it allowed for a drastic reduction of the concentrations of Buparlisib and Ribociclib while still maintaining a potent anti-tumor effect when compared to the use single drugs. Indeed, higher drug concentration would lead to toxicity problems in the patients. Overall, we identified a possible novel treatment strategy for pNETs by combining PI3K and CDK4/CDK6 inhibitors.È consentito all'utente scaricare e condividere i documenti disponibili a testo pieno in UNITESI UNIPV nel rispetto della licenza Creative Commons del tipo CC BY NC ND.
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https://hdl.handle.net/20.500.14239/15619