Journal Article

Afatinib and Temozolomide combination inhibits tumorigenesis by targeting EGFRvIII-cMet signaling in glioblastoma cells

Gorantla, Santhi, Batra, Surinder K., Shonka, Nicole, Siddiqui, Jawed A., Rachagani, Satyanarayana, Mallya, Kavita, Ponnusamy, Moorthy P., Jain, Maneesh, Macha, Muzafar A., Nimmakayala, Rama Krishna, Vengoji, Raghupathy

2019

Overview

Background Glioblastoma (GBM) is an aggressive brain tumor with universal recurrence and poor prognosis. The recurrence is largely driven by chemoradiation resistant cancer stem cells (CSCs). Epidermal growth factor receptor (EGFR) and its mutant EGFRvIII are amplified in ~ 60% and ~ 30% of GBM patients, respectively; however, therapies targeting EGFR have failed to improve disease outcome. EGFRvIII-mediated cross-activation of tyrosine kinase receptor, cMET, regulates GBM CSC maintenance and promote tumor recurrence. Here, we evaluated the efficacy of pan-EGFR inhibitor afatinib and Temozolomide (TMZ) combination on GBM in vitro and in vivo. Methods We analyzed the effect of afatinib and temozolomide (TMZ) combination on GBM cells U87MG and U251 engineered to express wild type (WT) EGFR, EGFRvIII or EGFRvIII dead kinase, CSCs isolated from U87 and U87EGFRvIII in vitro . The therapeutic utility of the drug combination was investigated on tumor growth and progression using intracranially injected U87EGFRvIII GBM xenografts. Results Afatinib and TMZ combination synergistically inhibited the proliferation, clonogenic survival, motility, invasion and induced senescence of GBM cells compared to monotherapy. Mechanistically, afatinib decreased U87EGFRvIII GBM cell proliferation and motility/invasion by inhibiting EGFRvIII/AKT, EGFRvIII/JAK2/STAT3, and focal adhesion kinase (FAK) signaling pathways respectively. Interestingly, afatinib specifically inhibited EGFRvIII-cMET crosstalk in CSCs, resulting in decreased expression of Nanog and Oct3/4, and in combination with TMZ significantly decreased their self-renewal property in vitro. More interestingly, afatinib and TMZ combination significantly decreased the xenograft growth and progression compared to single drug alone. Conclusion Our study demonstrated significant inhibition of GBM tumorigenicity, CSC maintenance in vitro , and delayed tumor growth and progression in vivo by combination of afatinib and TMZ. Our results warrant evaluation of this drug combination in EGFR and EGFRvIII amplified GBM patients.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Afatinib / Afatinib - administration & dosage / Afatinib - pharmacology / Analysis / Animals / Antineoplastic agents / Antineoplastic Combined Chemotherapy Protocols - administration & dosage / Antineoplastic Combined Chemotherapy Protocols - pharmacology / Apoptosis / Binding sites / Biomedical and Life Sciences / Biomedicine / Brain cancer / Brain Neoplasms - drug therapy / Brain Neoplasms - genetics / Brain Neoplasms - metabolism / Brain tumors / Cancer / Cancer Research / Cancer stem cells / Cancer therapies / Cancer treatment / Care and treatment / Cell culture / Cell growth / Cell Line, Tumor / Cell Proliferation - drug effects / Cell Survival - drug effects / Cellular signal transduction / Combination drug therapy / Cytotoxicity / Epidermal growth factor / Epidermal growth factor receptor / Epidermal growth factors / ErbB Receptors - genetics / Gene Expression Regulation, Neoplastic - drug effects / Glioblastoma / Glioblastoma - drug therapy / Glioblastoma - genetics / Glioblastoma - metabolism / Glioblastomas / Gliomas / Humans / Immunology / Kinases / Lung cancer / Metastasis / Mice / Mutation / Neoplastic Stem Cells - drug effects / Neoplastic Stem Cells - metabolism / Oncology / Patients / Pemetrexed / Phenols (Class of compounds) / Phosphorylation / Proto-Oncogene Proteins c-met - metabolism / Recurrence (Disease) / Signal Transduction - drug effects / Stem cells / Studies / Temozolomide / Temozolomide - administration & dosage / Temozolomide - pharmacology / Tetracyclines / Tumors / Tyrosine / Xenograft Model Antitumor Assays