Di Somma Sarah, Iannuzzi Carmelina Antonella, Passaro Carmela, Forte Iris Maria, Iannone Raffaella, Gigantino Vincenzo, Indovina Paola, Botti Gerardo, Giordano Antonio, Formisano Pietro, Portella Giuseppe, Malfitano Anna Maria, Pentimalli Francesca
Dipartimento Scienze Mediche Traslazionali, Università di Napoli "Federico II", Naples, Italy.
Cell Biology and Biotherapy Unit, Istituto Nazionale Tumori IRCCS, Fondazione G. Pascale, Naples, Italy.
Front Oncol. 2019 Jul 12;9:564. doi: 10.3389/fonc.2019.00564. eCollection 2019.
Malignant pleural mesothelioma (MPM) is an aggressive cancer associated with asbestos exposure that urgently requires effective therapeutic strategies. Current treatments are unable to increase significantly patient survival, which is often limited to <1 year from diagnosis. Virotherapy, based on the use of oncolytic viruses that exert anti-cancer effects by direct cell lysis and through the induction of anti-tumor immune response, represents an alternative therapeutic option for rare tumors with limited life expectancy. In this study, we propose the use of the adenovirus 922-947, engineered to allow selective replication in cancer cells, to counteract MPM. We performed a thorough preclinical assessment of 922-947 effects in a set of MPM cell lines and xenografts. Cytotoxicity of 922-947 alone and in combination assays was evaluated by sulforhodamine B assay. Cell cycle, calreticulin expression, and high mobility group box protein 1 (HMGB1) secretion were determined by flow cytometry, whereas ATP content was determined by a luminescence-based bioassay. The modulation of angiogenic factors in MPM-infected cells was evaluated through ELISA. We found that 922-947 infection exhibits cytotoxic effects in MPM cell lines, affecting cell viability, cell cycle progression, and regulating main hallmarks of immunogenic cell death inducing calreticulin surface exposure, HMGB1 and ATP release. Our results also suggest that 922-947 may affect angiogenic signals by regulation of VEGF-A and IL-8 secretion. Furthermore, 922-947 shows anti-tumor efficacy in murine xenograft models reducing tumor growth and enhancing survival. Finally, the combination with cisplatin potentiated the cytotoxic effect of 922-947. Overall our data identify virotherapy, based on the use of 922-947, as a new possible therapeutic strategy against MPM, which could be used alone, in combination with standard chemotherapy drugs, as shown here, or other approaches also aimed at enhancing the antitumoral immune response elicited by the virus.
恶性胸膜间皮瘤（MPM）是一种与接触石棉相关的侵袭性癌症，迫切需要有效的治疗策略。目前的治疗方法无法显著提高患者生存率，患者从诊断起的生存期通常限于不到1年。病毒疗法基于使用溶瘤病毒，通过直接细胞裂解和诱导抗肿瘤免疫反应发挥抗癌作用，是预期寿命有限的罕见肿瘤的一种替代治疗选择。在本研究中，我们提议使用经过基因工程改造以允许在癌细胞中选择性复制的腺病毒922 - 947来对抗MPM。我们在一组MPM细胞系和异种移植模型中对922 - 947的作用进行了全面的临床前评估。通过磺酰罗丹明B测定法评估922 - 947单独及联合测定的细胞毒性。通过流式细胞术测定细胞周期、钙网蛋白表达和高迁移率族蛋白B1（HMGB1）分泌，而通过基于发光的生物测定法测定ATP含量。通过酶联免疫吸附测定法评估MPM感染细胞中血管生成因子的调节。我们发现922 - 947感染在MPM细胞系中表现出细胞毒性作用，影响细胞活力、细胞周期进程，并调节免疫原性细胞死亡的主要特征，诱导钙网蛋白表面暴露、HMGB1和ATP释放。我们的结果还表明，922 - 947可能通过调节血管内皮生长因子 - A（VEGF - A）和白细胞介素 - 8（IL - 8）分泌来影响血管生成信号。此外，922 - 947在小鼠异种移植模型中显示出抗肿瘤功效，可减少肿瘤生长并提高生存率。最后，与顺铂联合增强了922 - 947的细胞毒性作用。总体而言，我们的数据确定基于使用922 - 947的病毒疗法是一种针对MPM的新的可能治疗策略，它可以单独使用，如本研究所示与标准化疗药物联合使用，或与其他旨在增强病毒引发的抗肿瘤免疫反应的方法联合使用。
