Combination of antibodies directed against different ErbB3 surface epitopes prevents the establishment of resistance to BRAF/MEK inhibitors in melanoma

Patients with metastatic melanoma bearing V600 mutations in BRAF oncogene clinically benefit from the treatment with BRAF inhibitors alone or in combination with MEK inhibitors. However, a limitation to such treatment is the occurrence of resistance. Tackling the adaptive changes helping cells survive from drug treatment may offer new therapeutic opportunities. Very recently the ErbB3 receptor has been shown to act as a central node promoting survival of BRAF mutated melanoma. In this paper we first demonstrate that ErbB3/AKT hyperphosphorylation occurs in BRAF mutated melanoma cell lines following exposure to BRAF and/or MEK inhibitors. This strongly correlates with increased transcriptional activation of its ligand neuregulin. Anti-ErbB3 antibodies impair the establishment of de novo cell resistance to BRAF inhibition in vitro. In order to more potently ablate ErbB3 activity we used a combination of two anti-ErbB3 antibodies directed against distinct epitopes of its extracellular domain. These two antibodies in combo with BRAF/MEK inhibitors potently inhibit in vitro cell growth and tumor regrowth after drug withdrawal in an in vivo xenograft model. Importantly, residual tumor masses from mice treated by the antibodies and BRAF/ERK inhibitors combo are characterized almost exclusively by large necrotic areas with limited residual areas of tumor growth. Taken together, our findings support the concept that triple therapy directed against BRAF/MEK/ErbB3 may be able to provide durable control of BRAF mutated metastatic melanoma

HDAC class I inhibitor domatinostat sensitizes pancreatic cancer to chemotherapy by targeting cancer stem cell compartment via FOXM1 modulation

Pancreatic ductal adenocarcinoma (PDAC) represents an unmet clinical need due to the very poor prognosis and the lack of effective therapy. Here we investigated the potential of domatinostat (4SC-202), a new class I histone deacetylase (HDAC) inhibitor, currently in clinical development, to sensitize PDAC to first line standard gemcitabine (G)/taxol (T) doublet chemotherapy treatment. Methods: Synergistic anti-tumor effect of the combined treatment was assessed in PANC1, ASPC1 and PANC28 PDAC cell lines in vitro as well as on tumor spheroids and microtissues, by evaluating combination index (CI), apoptosis, clonogenic capability. The data were confirmed in vivo xenograft models of PANC28 and PANC1 cells in athymic mice. Cancer stem cells (CSC) targeting was studied by mRNA and protein expression of CSC markers, by limiting dilution assay, and by flow cytometric and immunofluorescent evaluation of CSC mitochondrial and cellular oxidative stress. Mechanistic role of forkhead box M1 (FOXM1) and downstream targets was evaluated in FOXM1-overexpressing PDAC cells. Results: We showed that domatinostat sensitized in vitro and in vivo models of PDAC to chemotherapeutics commonly used in PDAC patients management and particularly to GT doublet, by targeting CSC compartment through the induction of mitochondrial and cellular oxidative stress. Mechanistically, we showed that domatinostat hampers the expression and function of FOXM1, a transcription factor playing a crucial role in stemness, oxidative stress modulation and DNA repair. Domatinostat reduced FOXM1 protein levels by downregulating mRNA expression and inducing proteasome-mediated protein degradation thus preventing nuclear translocation correlated with a reduction of FOXM1 target genes. Furthermore, by overexpressing FOXM1 in PDAC cells we significantly reduced domatinostatinducing oxidative mitochondrial and cellular stress and abolished GT sensitization, both in adherent and spheroid cells, confirming FOXM1 crucial role in the mechanisms described. Finally, we found a correlation of FOXM1 expression with poor progression free survival in PDAC chemotherapy-treated patients

Il Bilancio di Genere dell’Ateneo federiciano: dal rapporto di genere all’istituzionalizzazione del processo

[Italiano]:Le Università, in quanto enti di formazione e di ricerca, sono tenute a mettere in atto azioni e politiche volte a garantire la parità di genere e a rimuovere gli ostacoli che impediscono la piena realizzazione personale e professionale di uomini e donne. A tale scopo il Bilancio di Genere è lo strumento d’elezione per perseguire gli obiettivi di parità. Ma più che di un documento, si tratta di un vero e proprio articolato processo che richiede un significativo coinvolgimento della governance di Ateneo, attraverso un percorso di istituzionalizzazione dell’approccio alle questioni di genere. Il volume presenta gli esiti del secondo bilancio di genere della Federico II di Napoli, redatto secondo le Linee Guida elaborate dalla Conferenza dei Rettori delle Università Italiane. Oltre all’analisi di contesto, che consente di evidenziare i punti di forza e gli elementi di criticità dell’Ateneo federiciano rispetto alla parità di genere, il documento illustra il livello di integrazione della prospettiva di genere nei documenti strategici di Ateneo, la rete di organismi preposti alle tematiche di genere e il repertorio delle azioni messe in atto nel quinquennio precedente. L’analisi di genere degli impegni economico-finanziari, realizzata a partire dalla riclassificazione del bilancio, ha consentito inoltre di esplorare nel dettaglio le risorse espressamente destinate agli obiettivi di parità, identificando gli ambiti in cui è necessario investire maggiormente. Il documento include inoltre una prima analisi in ottica di genere dei questionari di rilevazione delle opinioni degli studenti e delle studentesse, che può essere adoperata come utile strumento per fornire spunti per la definizione di azioni positive destinate alla popolazione studentesca. Il quadro che emerge dall’analisi proposta è quello di un Ateneo che ha intrapreso un serio percorso di istituzionalizzazione dell’intero ciclo del Bilancio di Genere, attestato dall’impegno nella promozione di processi culturali ed organizzativi inclusivi volti a perseguire concretamente gli obiettivi di uguaglianza e di parità nella formazione, nella ricerca e nel lavoro. ./[English]:Universities, as training and research institutions, are required to implement actions and policies aimed at ensuring gender equality and at removing obstacles that prevent the full personal and professional fulfilment of men and women. To this end, the Gender Responsive Budgeting is the fundamental tool to pursue the objectives of equality. But more than a document, it is an articulated process that requires a significant involvement of the governance of the University, through a path of institutionalization of the approach to the gender issues. The volume presents the results of the second gender report of the University Federico II of Naples, drawn up according to the Conference of Rectors of Italian Universities Guidelines. In addition to the context analysis, which allows to highlight strengths and critical issues of the University with respect to gender equality, the document illustrates the level of integration of the gender perspective in the University's strategic documents, the network of bodies responsible for gender issues and the repertoire of actions implemented in the previous five years. The gender analysis of economic and financial commitments, carried out through the reclassification of the budget, has also made it possible to explore in detail the resources expressly allocated to the equality objectives, identifying the areas in which greatest investments are needed. The document also includes a preliminary gender analysis of student opinion questionnaires, which can be used as a useful tool to provide insights into the definition of positive actions for the student population. The picture that emerges from the proposed analysis is of a University that has embarked on a serious path of institutionalization of the gender budget process, attested by the commitment to promote inclusive cultural and organizational processes aimed at concretely pursuing equality objectives in training, research and work