Investigation and Evaluation of Interferon Inducer Alone and in Combination with Chemotherapeutic Agents as Targeted Anti-Cancer Therapy

Abstract

Breast cancer has been denoted as the second most common cause of cancer-related deaths globally. Amongst it, triple negative breast cancer which is defined as the absence of ER, PR and HER2 receptors, accounts for 15-20% of all breast cancers. Currently, chemotherapy and radiation therapy is the mainstay for TNBC and the current research pertains to novel targets such as VEGF, EGFR, PARP and angiogenesis inhibitors. Immunotherapy has also shown advancement in cancer therapy mimicking the body’s natural defense system. DEAE-Dextran has shown interferon inducing property and interferons have reported to have anti-angiogenic potential. Therefore, we have investigated the role of DEAEDextran in TNBC. We carried out various in vitro studies of DEAE-Dextran such as MTT assay and trypan blue exclusion assay and it has shown the cytotoxic effect of DEAE-Dextran as well as β-interferon inducing property of DEAE-Dextran. Further, various angiogenic models were evaluated for the anti-angiogenic potential of DEAE-Dextran and it was observed that DEAE-Dextran portrayed promising anti-angiogenic activity in cell migration assay, aortic ring assay, chick yolk sac membrane assay as well as matrigel plug xenograft model. In depth molecular mechanism was evaluated through gene expression studies and found that DEAE-Dextran significantly upregulated β-interferon and downregulated VEGF and NOTCH1 expressions. Flow cytometry studies also showed the apoptotic nature of DEAE-Dextran with cell targeting and cellular uptake within one hour of treatment. In vivo dose ranging study was performed to determine the dose of DEAE-Dextran for release of β- interferon levels in the body and it was found that 100 mg/kg was sufficient dose for the increased β-interferon levels. DMBA induced mammary cancer model was initiated and it showed potent anti-cancer activity by virtue of DEAE-Dextran through β-interferon induction. Acute and sub-acute toxicity studies for DEAE-Dextran showed the LD50 as 1000 mg/kg with mortality above the LD50 due to renal toxicity and doses administered in the 28 day repeated dose study at 400 mg/kg showed no significant toxicity in various morphological and histopathological determinations. We further prepared paclitaxel nanoparticles and DEAE-Dextran coated paclitaxel nanoparticles which showed potent anti-cancer activity in the xenograft model. We therefore conclude that DEAE-Dextran exhibits excellent anticancer activity due to its β-interferon inducing property via its anti-angiogenic, apoptotic and cytotoxic mechanism.