In vitro and In Vivo Evidences Propound Potential of Lipocalin 2 as a Therapeutic Target in Cervical Carcinoma
Author(s): Nehanjali Dwivedi, Tahmina Mazumder, Shivani Tihara, Gayathri Veeraraghavan, Ramanujam Siva, Smitha P K, Rohit Ranade, Anil K M, Sujan K Dhar, Manjula Das
Aim: This study assessed efficacy of a novel α-Lipocalin 2 (LCN2) monoclonal antibody (MAb) employing in vitro and in vivo xenograft models of cervical cancer (CC).
Main methods: Anti-proliferative and anti-invasive effects of α-LCN2 MAb was assessed in vitro on STR-authenticated-HeLa cells. Gelatin zymography and qPCR were used to decipher molecular mechanism of the observed effects. To assess its anti-tumorigenic potential in vivo, Hela cell tumors grown as mouse model xenografts were treated with α-LCN2 MAb. Treatment efficacy was assessed by tumor palpability and volume regression followed by histopathological and image texture analysis of tumor. Molecular pathways were deciphered by analyzing differential RNASeq data separately for the injected human and recruited mouse cell-components in both MAb-treated and mock-treated xenografts.
Key findings: α-LCN2 MAb caused tumor regression and softening through necrosis of both tumor and stromal cells. Differential gene expression analysis of human component indicated LCN2 inhibition abolished tumorigenic pathways while significantly increasing necrosis and cell death pathways in tumor via reduction in LCN2 and a downfall of TNFα-IL17 axis leading to further decline in LCN2 expression, suggestive of a negative feedback loop. In mouse transcriptome component, T-cell activation and decline in M2 macrophage population upon α-LCN2 treatment underlies its potential as an immune sensitizer that can devour tumor cells through T-cell mediated cytotoxicity leading to an anti-tumorigenic milieu.
Significance: Results of this study strongly suggests LCN2 to be a potential therapeutic target for CC. α-LCN2 MAb used in this study can be further developed for clinical usage in CC using intra-tumoral delivery.