(484.13) Pharmacological reactivation of RBL2/p130 could be an effective antitumoral strategy for malignant pleural mesothelioma

Poster Board Number: A25

Aurora Costa (University of Siena, University of Siena, Temple University), Iris Maria Forte (Istituto Nazionale Tumori, IRCCS, Fondazione G. Pascale), Elisa Ventura (Temple University), Francesca Pentimalli (Libera Università Mediterranea), Antonio Giordano (University of Siena, Temple University)

Malignant mesothelioma (MM) is an aggressive tumor developing from the mesothelium covering the body cavities. The most common MM type affects the pleura surrounding the lungs. MM is mainly associated with asbestos exposure. At present, no curative modalities are effective against MM exist. The molecular mechanisms underlying MM development seem to depend mostly on the inactivation of tumor suppressors, among which also the dysfunction of the retinoblastoma (RB) protein pathway that is a hallmark of cancers. We previously demonstrated that RBL2/p130 is a direct AKT target and its reactivation following AKT inhibition mediates apoptosis in MM.
So, we hypothesize that concomitant inhibition of CDK and AKT activity, both converging on the reactivation of RBL2/p130 tumor suppressor role, synergize in counteracting MM.

We first assessed, through an MTS assay, the effect of three new generation CDK inhibitors (CDKi), palbociclib, ribociclib, abemaciclib, on the cell viability of MM (NCI-H28, IST-MES TWO, NCI-H2452, MMB, MSTO-211H, NCI-H2052) and immortalized human normal mesothelial (MET-5A) cells at 72h. Then, to verify whether CDKi exerted long-term cell growth inhibition, we performed clonogenic assays upon treatment of all the MM All the MM and mesothelial cell lines. Finally, we also examined, by MTS, the possible synergistic effects of abemaciclib in combination with an AKT inhibitor (AKT1/2 VIII).

Our preliminary data show that ribociclib and palbociclib only moderately reduce viability of the MPM cells, whereas abemaciclib shows cytotoxic effects at very low doses in all MM cell lines. Abemaciclib treatment induces a dose-dependent cytotoxic effect in all the MM cell lines without meaningfully affecting the normal mesothelial cells. Interestingly, abemaciclib also induced a cytotoxic effect in the most aggressive histotype, the sarcomatoid NCI-H2052 cells. To verify whether abemaciclib exerted long-term cell growth inhibition, we performed clonogenic assays upon treatment with IC50 of abemaciclib and found that the CDKi dramatically reduced colony formation in all the MM cell lines. We also examined, by MTS, the possible synergistic effects of abemaciclib in combination with a kinase AKT inhibitor (iAKT1/2 VIII), so we treated the MM cell lines for 72 h with the two agents, both alone and in combination, at five different concentrations, in a constant ratio. The cell viability data were evaluated through the Chou–Talalay method and showed a strong synergism revealing a combination index (CI) values lt; 1 for all cell.

Although reactivating oncosoppressor genes is difficult to achieve because they are hardly ‘druggable’, our preliminary findings will provide the rationale for pharmacological strategies able to unleash RBL2/p130 tumor suppressor potential. The pharmacological reactivation of RBL2/p130 could be an effective strategy against MM, which can be rapidly implemented in the clinical practice.

the Commonwealth of Pennsylvania: DoH-17 PA Dept of Health Title: 2020amp;ndash;2021 BT Bio-Technology Sbarro Grant (2021amp;ndash;2022) PA DoH Grant Number: SAP#4100089059



Figure 1: MTS assay of cell viability (y-axis) of NCI-H28, IST-MES-TWO, NCI-H2452,MMB,MSTO-211H, NCI-H2052 AND MET5A cells treated for 72h with different concentrations of abemaciclib (x-axis). The results showed that abemaciclib inhibited the growth of NCI-H28, IST-MES-TWO, NCI-H2452,MMB,MSTO-211H, NCI-H2052 AND MET5A cells in a dose-dependent manner