Glypican 2 (GPC2) is a potential immunological target in adult and childhood cancers

Identifying differentially expressed tumour-associated antigens is the major barrier to develop effective treatment against neuroendocrine tumours such as neuroblastoma (NB), lung cancer, and malignant melanoma (MM). Bearing this in mind, the aim of this study includes:  

I. Validate GPC2 protein expression and its dependence on MYC family MYCN/c-MYC expression in cancer cell lines of various origins including NB, lung cancer and melanoma by Western Blot (WB). 

II. Develop and characterise murine NB cell lines NXS2 and 9464D that stably express the human GPC2. 

In Experimental results section 1, different GPC2 Abs from various suppliers were tested by WB. GPC2 Ab1 demonstrated high sensitivity and specificity as compared to the other Abs. We found GPC2 and MYCN/c-MYC  expressions were detectable in all the cancer cell lines at various levels with no expression in negative controls: SHEP-TET-21N (MYCN-OFF) and HEK 293. We have also demonstrated that the tumour-specific GPC2-001 mRNA variant drives the full-size GPC2* (62 kDa) protein expression. Our RNA-seq data set analysis demonstrated no association between the expression levels of GPC2 and MYCN in NB and lung cancer cell lines.  

In Experimental results section 2, the modified murine NBcell line NXS2-hGPC2 was developed with human GPC2 using a liposomal-based approach. Our western blot results demonstrated a high expression of GPC2 in the NXS2-hGPC2 cell line compared to the parental NXS2 and 9464D cell lines. Cell viability and proliferation in NXS2-hGPC2 demonstrated that GPC2 overexpression increases cell proliferation. Additionally, the detection of human GPC2 by mouse monoclonal GPC2 Ab1 demonstrated high homology between human and mouse GPC2 sequences.