Aptamer targeting EGFRvIII mutant hampers its constitutive autophosphorylation and affects migration, invasion and proliferation of glioblastoma cells

Glioblastoma Multiforme (GBM) is the most common and aggressive human brain tumor, associated with very poor survival despite surgery, radiotherapy and chemotherapy. The epidermal growth factor receptor (EGFR) and the platelet-derived growth factor receptor beta (PDGFR beta) are hallmarks in GBM with driving roles in tumor progression. In approximately half of the tumors with amplified EGFR, the EGFRvIII truncated extracellular mutant is detected. EGFRvIII does not bind ligands, is highly oncogenic and its expression confers resistance to EGFR tyrosine kinase inhibitors (TKIs). It has been demonstrated that EGFRvIII-dependent cancers may escape targeted therapy by developing dependence on PDGFR beta signaling, thus providing a strong rationale for combination therapy aimed at blocking both EGFRvIII and PDGFR beta signaling. We have recently generated two nuclease resistant RNA aptamers, CL4 and Gint4. T, as high affinity ligands and inhibitors of the human wild-type EGFR (EGFRwt) and PDGFR beta, respectively. Herein, by different approaches, we demonstrate that CL4 aptamer binds to the EGFRvIII mutant even though it lacks most of the extracellular domain. As a consequence of binding, the aptamer inhibits EGFRvIII autophosphorylation and downstream signaling pathways, thus affecting migration, invasion and proliferation of EGFRvIII-expressing GBM cell lines. Further, we show that targeting EGFRvIII by CL4, as well as by EGFR-TKIs, erlotinib and gefitinib, causes upregulation of PDGFR beta. Importantly, CL4 and gefitinib cooperate with the anti-PDGFR beta Gint4. T aptamer in inhibiting cell proliferation. The proposed aptamer-based strategy could have impact on targeted molecular cancer therapies and may result in progresses against GBMs.