Abstract 2994: In vivo targeting of mesothelioma’s molecular signature with nanoparticle functionalized with nucleolin-binding peptide
Authors: Fonseca Nuno A. , Moura Vera , Colelli Fabiana , Pesce Daniela , Signorino Giacomo , Focareta Laura , Fucci Alessandra , Cardile Francesco , Pisano Claudio , Simões Sérgio , Moreira João Nuno 
Published: August 2019

Abstract
Mesothelioma is a malignant tumor of the mesothelium, often associated with asbestos exposure, with an overall survival of only 8.8 months. A combination of cisplatin and pemetrexed has been used as an adjuvant treatment to surgical resection or in patients who have inoperable disease. This combination remains as the current standard of care, regardless the sub-type of mesothelioma, with only a 9% 5-year survival rate. Herein, it is hypothesized that a novel targeted treatment, based on a doxorubicin (DXR)-containing nanoparticle functionalized with the nucleolin-binding F3 peptide (named PEGASEMP), will have significant a therapeutic impact against the most aggressive subtype of mesothelioma (biphasic). The hypothesis relies on nucleolin deregulated overexpression in cancer and endothelial cells from tumor blood vessels. Human biphasic mesothelioma cells were stably transduced with luciferase-reporter gene, and orthotopically injected intrapleurally into female immunocompromised mice. Three weeks (Early Stage) and 4 weeks (Advanced Stage) after cell injection, animals were randomly allocated to different treatment groups: vehicle, PEGASEMP at 5.6 or 7 mg of DXR/kg alone (q7dx5w); cisplatin at 4.0 mg/kg alone or combined with PEGASEMP at 5.6 mg of DXR/kg (q7dx5w); and the standard of care, a combination of cisplatin at 4.0 mg/kg (q7dx5w) plus pemetrexed at 100.0 mg/kg (q2dx3x5w). Bioluminescence was monitored weekly with IVIS Spectrum In Vivo Imaging system. Differential gene expression upon PEGASEMP treatment of advanced stage tumors was also performed in comparison with non-treated animals using the GeneChip Human Transcriptome Array 2.0. In the early stage of development, PEGASEMP, at 7.0 mg/kg, inhibited tumor growth by 2,713-fold relative to standard of care, which translated into a 1223-fold decrease in tumor burden. At an advanced stage of tumor development, the combination of PEGASEMP and cisplatin enabled 107-fold tumor growth inhibition, enabling a 55-fold reduction in tumor burden, relative to standard of care. In both cases, no decrease of the body weight was observed. Furthermore, 75% (146) gene transcripts were found to be deregulated (while the remaining 25% were upregulated), involving the downregulation of cell division, including ontologies such as cell cycle or chromatin organization and assembly. Specifically, downregulation of topoisomerase 2, cyclin B1 or Ki67 transcripts, part of the described molecular signature of mesothelioma, was observed and was consistent with the mechanism of action of doxorubicin. Overall, the novel mechanism of action associated with PEGASEMP, enables a significant benefit in terms of efficacy and safety in the treatment of biphasic mesothelioma, as compared with the current standard of care, affecting the molecular signature of the disease and thus supporting future clinical evaluation.
Citation Format: Nuno A. Fonseca, Vera Moura, Fabiana Colelli, Daniela Pesce, Giacomo Signorino, Laura Focareta, Alessandra Fucci, Francesco Cardile, Claudio Pisano, Sérgio Simões, João Nuno Moreira. In vivo targeting of mesothelioma’s molecular signature with nanoparticle functionalized with nucleolin-binding peptide [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2994.