Development and characterisation of monoclonal antibodies to tumour endothelial marker ‘melanoma cell adhesion molecule’ (MCAM) and approaches to targeting MCAM on tumour vessels

Čorić, Marta (2019). Development and characterisation of monoclonal antibodies to tumour endothelial marker ‘melanoma cell adhesion molecule’ (MCAM) and approaches to targeting MCAM on tumour vessels. University of Birmingham. Ph.D.

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Abstract

Renal cell carcinoma (RCC) patients are routinely treated with cytokines and VEGF targeted anti-angiogenics. Resistance to therapy due to the by-passing of VEGF pathway and severe systemic side effects induced by cytokines and VEGF targeting pose major drawbacks. Melanoma cell adhesion molecule (MCAM) is a marker of RCC blood vessels, with low or no expression on healthy vasculature. This thesis aimed to generate mouse MCAM targeted antibodies and recombinant human MCAM extracellular domain (hMCAMecd) receptor trap to investigate MCAM as a potential therapeutic target. Around 50 anti-mouse MCAM hybridoma clones were generated from immunized rats, with two of them sub-cloned to stable antibody expressing hybridomas (mMCAM10 and mMCAM66). The two antibodies recognize mMCAM by flow cytometry, immunohistochemistry, Western blot and immunoprecipitation. The mMCAM10, but not the mMCAM66, localises to tumour vessels within one hour of intravenous injection into the tumour bearing mice. This indicates MCAM’s potential as a target for tumour vascular disruption. The variable region of the mMCAM10 was sequenced for future antibody engineering. The hMCAMecd fused to an Fc tag caused significant reduction in endothelial cell migration, tube formation and transmigration, and had no effect on proliferation or cell cycle, highlighting MCAM’s potential for anti-angiogenic therapy.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):