Adrenocortical cancer (ACC) is a rare malignant neoplasm, with an annual incidence of 0.5-2.0 patients per million in the general population. The prognosis of ACC is variable but more than 60% of patients are diagnosed in stages III and IV with a 5-year survival of <50% and <15%, respectively. For these patients, the proposed algorithm of treatment includes treatment with mitotane alone or in combination with EDP chemotherapy (etoposide, doxorubicin, and cisplatin). In a few cases, locoregional strategies including surgery or radiotherapy are also used. However, the progression of advanced disease occurs almost invariably after less than 18 months and there are no defined follow-up lines of treatment. Therefore, new therapeutic strategies are needed. We focused our attention on the druggable role of the progesterone (Pg)/ Pg receptors (PgR) and estrogens (E)/ E receptors (ER) pathways, WEE1 kinase, and the fibroblast growth factor (FGF)/FGF receptors (FGFR) pathway, trying to demonstrate whether the modulation of their activity could exert antiproliferative effects in the different human ACC cell models now available. We demonstrated that, although ER expression was relatively low, tamoxifen exerted a cytotoxic effect on NCI-H295R but not on metastatic-derived ACC cell models such as MUC-1 and TVBF-7 cells (formerly ACC115m) that expressed very weak levels of ER. On the same line, ER is scarcely expressed in ACC tissues and seemed to decrease as the disease progresses. More interest was raised with results obtained when ACC cells were exposed to Pg, which exerted a cytotoxic effect, although at lower potency in metastatic cell lines compared to the primitive NCI-H295R cell line, probably due to the lower PgR expression in metastatic lines. Autophagy and apoptosis occur hierarchically or independently to contribute to Pg-induced ACC cell death. Notably, drug withdrawal experiments showed that Pg induced a long-lasting effect in metastatic cell models. In the zebrafish model, Pg significantly reduced the xenograft tumor area of each ACC cell line, and the metastasis formation in embryos injected with MUC-1 cells, confirming the in vitro results. This phenomenon is mediated at least in part, by the reduction of MMP2 levels and activity. Regarding WEE1, our results showed that this kinase is overexpressed in ACC and that low WEE1 levels were associated with better overall survival. Interestingly, WEE1 nuclear staining was significantly higher in TP53 mutated samples compared to the wild type. Exposure of different ACC cell models such as NCI-H295R, JIL-2266 and CU-ACC2 cells to adavosertib induced cytotoxicity with IC50 of 1.17, 1.35 and 0.4 µM, respectively. CU-ACC1 and MUC-1 showed less sensitivity to adavosertib alone, but its effect was enhanced by cisplatin and gemcitabine and this additive-synergic effect was observed in all cell lines. Finally, we chose to evaluate the therapeutic potential of the FGF/FGFR pathway in ACC as several studies have demonstrated that FGFR1 and FGFR4 are upregulated in ACC and that their high expression was significantly associated with worse patient prognosis, suggesting that they are potentially interesting therapeutic targets. Evaluating the FGFRs expression in ACC cell lines we found that each cell model tested, namely NCI-H295R, MUC-1, JIL-2266, CU-ACC2 and TVBF-7 cells, showed a particular pattern of FGFRs expression. Exposure of each cell line to increasing concentrations of erdafitinib, rogaratinib and fisogatinib induced different grades of cytotoxicity depending on the cell line. These and other previous results strengthen the role of Pg in ACC, and this hypothesis is now under study in the ongoing randomized phase II clinical trial PESETA. The hope is that the ongoing projects regarding WEE1 inhibitor adavosertib and FGFRs-inhibitors generate robust results that will lead to potential clinical applications for this rare disease with otherwise dismal outcomes.
Il carcinoma della corticale del surrene (AdrenoCortical Carcinoma, ACC) è una rara neoplasia maligna, con una sopravvivenza a 5 anni inferiore al 50% o al 15%, a seconda dello stadio. Il trattamento proposto prevede l'uso di mitotano da solo o in combinazione con la chemioterapia EDP (etoposide, doxorubicina e cisplatino). In alcuni casi, anche la chirurgia e/o radioterapia. Tuttavia, la progressione della malattia si verifica quasi inevitabilmente dopo meno di 18 mesi e non ci sono attualmente linee guida condivise di trattamento. È necessario quindi identificare nuove strategie terapeutiche. Abbiamo focalizzato la nostra attenzione sui recettori del progesterone (Pg/ PgR), sui recettori degli estrogeni (E/ER), sulla chinasi WEE1, e sul pathway mediato dai recettori del fattore di crescita dei fibroblasti (FGF/FGFR), cercando di dimostrare se la modulazione della loro attività possa esercitare effetti antiproliferativi nei diversi modelli cellulari sperimentali di ACC ora disponibili. Abbiamo dimostrato che, sebbene l'espressione di ER sia relativamente bassa, il tamoxifene esercita un effetto citotossico sulla linea cellulare NCI-H295R derivante da un tumore primitivo ma non sulle linee metastatiche che esprimono livelli molto bassi di ER. Allo stesso modo, ER è debolmente espresso nei tessuti di ACC e sembra diminuire con la progressione della malattia. Di maggiore interesse sono invece i risultati riguardanti l’effetto citotossico esercitato dal Pg, sebbene con una potenza inferiore nelle linee metastatiche rispetto alle cellule NCI-H295R, probabilmente a causa di una minore espressione dei PgR. L'autofagia e l'apoptosi si verificano gerarchicamente o indipendentemente per contribuire alla morte cellulare indotta dal Pg. Inoltre, gli esperimenti di sospensione del trattamento hanno mostrato che l’effetto del Pg è duraturo nei modelli metastatici. Nel modello zebrafish, Pg ha ridotto significativamente l'area della massa tumorale iniettata con ciascuna linea cellulare di ACC, e la formazione di metastasi negli embrioni iniettati con cellule MUC-1, confermando i risultati in vitro. Questo fenomeno è mediato almeno in parte, dalla riduzione dei livelli e dell'attività di MMP2. Riguardo WEE1, nostri risultati hanno mostrato che questa chinasi è maggiormente espressa nei tessuti di ACC e che bassi livelli di WEE1 sono associati a una migliore sopravvivenza globale. È interessante notare che la colorazione nucleare di WEE1 è significativamente più alta nei campioni con mutazioni note in TP53 rispetto a quelli wild tipe. L’esposizione delle cellule NCI-H295R, JIL-2266 e CU-ACC2 all’inibitore di WEE1, adavosertib, ha indotto la citotossicità con valori di IC50 rispettivamente di 1.17, 1.35 e 0.4 µM. Le cellule CU-ACC1 e MUC-1 hanno mostrato una minore sensibilità per adavosertib da solo, ma il suo effetto è stato migliorato se combinato con il cisplatino o la gemcitabina. Questo effetto additivo/sinergico è stato osservato in tutte le linee cellulari. Infine, abbiamo scelto di valutare il potenziale terapeutico della via FGF/FGFR in ACC poiché diversi studi hanno dimostrato che FGFR1 e FGFR4 sono overespressi negli ACC e che la loro alta espressione è significativamente associata a una prognosi peggiore. L'esposizione delle cellule di ACC a concentrazioni crescenti di tre differenti inibitori degli FGFR erdafitinib, rogaratinib e fisogatinib, ha indotto un effetto citotossico a seconda della linea cellulare, e correlato con l’espressione dei recettori. In conclusione, questi risultati rafforzano il possibile ruolo del Pg nell’ACC; questa ipotesi è al momento l’oggetto dello studio clinico PESETA. Inoltre, la speranza è che i progetti in corso, riguardanti l'uso dell'inibitore di WEE1 adavosertib e degli inibitori degli FGFR, generino risultati solidi che possano fornire la base per futuri studi clinici.
Carcinoma of the human adrenal cortex: identification and in vitro and in vivo study of molecular alterations as potential therapeutic targets / Tamburello, Mariangela. - (2023 Feb 02).
Carcinoma of the human adrenal cortex: identification and in vitro and in vivo study of molecular alterations as potential therapeutic targets
Tamburello, Mariangela
2023-02-02
Abstract
Adrenocortical cancer (ACC) is a rare malignant neoplasm, with an annual incidence of 0.5-2.0 patients per million in the general population. The prognosis of ACC is variable but more than 60% of patients are diagnosed in stages III and IV with a 5-year survival of <50% and <15%, respectively. For these patients, the proposed algorithm of treatment includes treatment with mitotane alone or in combination with EDP chemotherapy (etoposide, doxorubicin, and cisplatin). In a few cases, locoregional strategies including surgery or radiotherapy are also used. However, the progression of advanced disease occurs almost invariably after less than 18 months and there are no defined follow-up lines of treatment. Therefore, new therapeutic strategies are needed. We focused our attention on the druggable role of the progesterone (Pg)/ Pg receptors (PgR) and estrogens (E)/ E receptors (ER) pathways, WEE1 kinase, and the fibroblast growth factor (FGF)/FGF receptors (FGFR) pathway, trying to demonstrate whether the modulation of their activity could exert antiproliferative effects in the different human ACC cell models now available. We demonstrated that, although ER expression was relatively low, tamoxifen exerted a cytotoxic effect on NCI-H295R but not on metastatic-derived ACC cell models such as MUC-1 and TVBF-7 cells (formerly ACC115m) that expressed very weak levels of ER. On the same line, ER is scarcely expressed in ACC tissues and seemed to decrease as the disease progresses. More interest was raised with results obtained when ACC cells were exposed to Pg, which exerted a cytotoxic effect, although at lower potency in metastatic cell lines compared to the primitive NCI-H295R cell line, probably due to the lower PgR expression in metastatic lines. Autophagy and apoptosis occur hierarchically or independently to contribute to Pg-induced ACC cell death. Notably, drug withdrawal experiments showed that Pg induced a long-lasting effect in metastatic cell models. In the zebrafish model, Pg significantly reduced the xenograft tumor area of each ACC cell line, and the metastasis formation in embryos injected with MUC-1 cells, confirming the in vitro results. This phenomenon is mediated at least in part, by the reduction of MMP2 levels and activity. Regarding WEE1, our results showed that this kinase is overexpressed in ACC and that low WEE1 levels were associated with better overall survival. Interestingly, WEE1 nuclear staining was significantly higher in TP53 mutated samples compared to the wild type. Exposure of different ACC cell models such as NCI-H295R, JIL-2266 and CU-ACC2 cells to adavosertib induced cytotoxicity with IC50 of 1.17, 1.35 and 0.4 µM, respectively. CU-ACC1 and MUC-1 showed less sensitivity to adavosertib alone, but its effect was enhanced by cisplatin and gemcitabine and this additive-synergic effect was observed in all cell lines. Finally, we chose to evaluate the therapeutic potential of the FGF/FGFR pathway in ACC as several studies have demonstrated that FGFR1 and FGFR4 are upregulated in ACC and that their high expression was significantly associated with worse patient prognosis, suggesting that they are potentially interesting therapeutic targets. Evaluating the FGFRs expression in ACC cell lines we found that each cell model tested, namely NCI-H295R, MUC-1, JIL-2266, CU-ACC2 and TVBF-7 cells, showed a particular pattern of FGFRs expression. Exposure of each cell line to increasing concentrations of erdafitinib, rogaratinib and fisogatinib induced different grades of cytotoxicity depending on the cell line. These and other previous results strengthen the role of Pg in ACC, and this hypothesis is now under study in the ongoing randomized phase II clinical trial PESETA. The hope is that the ongoing projects regarding WEE1 inhibitor adavosertib and FGFRs-inhibitors generate robust results that will lead to potential clinical applications for this rare disease with otherwise dismal outcomes.File | Dimensione | Formato | |
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