Research Projects

Epidermal Growth Factor Receptor (EGFR) variant-III (EGFR-vIII) is a constitutively active deletion mutant of EGFR expressed specifically on cancer cells with poor prognosis due to refractoriness to chemo and anti-EGFR therapy. Importantly, 50% or more cancers express EGFR-vIII. Thus, EGFR-vIII targeted therapeutics development has huge clinical significance. Therefore, we developed EGFR-vIII specific humanized single chain fragment variable (hscFv) and validated it’s in vitro efficacy regarding EGFR-vIII specific binding, suppression of EGFR-vIII signalling and down regulation of metabolic fuelling component GLUT3. The results demonstrated significant growth suppression of EGFR-vIII expressing cancer cells. Despite EGFR-vIII’s cancer specificity and the poor prognosis, no EGFR-vIII specific therapy is yet available. This indicates the immense translational potential of targeted anti-EGFR-vIII hscFv therapy. Based on the promising in vitro data on hscFv (see preliminary results and publications 4,5,6), we hypothesize that in vivo treatment of EGFR vIII expressing tumors in orthotopic xenograft models will confirm its therapeutical significance in an in vivo setting. Sicentific objectives: Validation of hscFv’s biotherapeutic potential in vivo using EGFR-vIII expressing glioblastoma orthotopic xenograft models in swiss albino nude mice towards tumor targeting and growth suppression for translation. Experiments to be carried out: (a) hscFv expression and purification (b) orthotopic xenograft tumor model development using EGFR-vIII expressing U87 glioma cell line and (c) identify the therapeutic hscFv dose (d) evaluate hscFv binding/accumulation in tumor tissue (e) quantification of EGFR-vIII phosphorylation levels and GLUT3 expression followed by (f) biodistribution, pharmacokinetics and toxicology analysis of hscFv, (g) tumor proteome analysis before and after treatment with hscFv. The significance of anti-EGFR vIII biotherapeutic lies in the fact that being humanized scFv(hscFv) with specificity and functionality to EGFR-vIII, translatability related to growth suppression and targeted delivery of chemodrugs is high. The purification strategy developed can be scaled up for hscFv production. Considering the fact that 50% or more cancers express EGFR vIII, development of targeted therapy has immense clinical potential.