Glycan affinity magnetic nanoplatforms for urinary glycobiomarkers discovery in bladder cancer

Bladder Cancer (BC) presents one of the highest recurrence rates amongst solid tumours and constitutes the second deadliest disease of the genitourinary track. Non-invasive identification of patients facing disease recurrence and/or progression remains one of the most critical and challenging aspects in disease management. To contribute to this goal, we demonstrate the potential of glycan-affinity glycoproteomics nanoplatforms for urinary biomarkers discovery in bladder cancer. Briefly, magnetic nanoprobes (MNP) coated with three broad-spectrum lectins, namely Concanavalin A (ConA; MNP@ConA), Wheat Germ Agglutinin (WGA; MNP@WGA), and Sambucus nigra (SNA; MNP@SNA), were used to selectively capture glycoproteins from the urine of low-grade and high-grade non-muscle invasive as well as muscle-invasive BC patients. Proteins were identified by nano-LC MALDI-TOF/TOF and data was curated using bioinformatics tools (UniProt, NetOGlyc, NetNGlyc, ClueGO app for Cytoscape and Oncomine) to highlight clinically relevant species. Accordingly, 63 glycoproteins were exclusively identified in cancer samples compared with healthy controls matching in age and gender. Specific glycoprotein sets exclusively found in low-grade non-muscle invasive bladder tumours may aid early diagnosis, while those only found in high-grade non-invasive and muscle-invasive tumours hold potential for accessing progression. Amongst these proteins is bladder cancer stem-cell marker CD44, which has been associated with poor prognosis. Orthogonal validation studies by slot-blotting demonstrated an elevation in urine CD44 levels of high-grade patients, which became more pronounced upon muscle-invasion, in mimicry of the primary tumour. These observations demonstrate the potential of MNP@lectins for identification of clinically relevant glycoproteomics signatures in bladder cancer. Future clinical validation in a larger and well characterized patient subset is required envisaging clinical translation of the results.

Talanta. 2018 Mar 12 [Epub]

Rita Azevedo, Janine Soares, Cristiana Gaiteiro, Andreia Peixoto, Luís Lima, Dylan Ferreira, Marta Relvas-Santos, Elisabete Fernandes, Ana Tavares, Sofia Cotton, Ana Luísa Daniel-da-Silva, Lúcio Lara Santos, Rui Vitorino, Francisco Amado, José Alexandre Ferreira

Experimental Pathology and Therapeutics Group, Research Centre, Portuguese Oncology Institute of Porto (IPO-Porto), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal; Institute of Biomedical Sciences Abel Salazar, University of Porto, R. Jorge de Viterbo Ferreira 228, 4050-013 Porto, Portugal., Experimental Pathology and Therapeutics Group, Research Centre, Portuguese Oncology Institute of Porto (IPO-Porto), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal., Experimental Pathology and Therapeutics Group, Research Centre, Portuguese Oncology Institute of Porto (IPO-Porto), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal; Institute of Biomedical Sciences Abel Salazar, University of Porto, R. Jorge de Viterbo Ferreira 228, 4050-013 Porto, Portugal; Institute for Research and Innovation in Health (i3S), University of Porto, R. Alfredo Allen, 4200-135 Porto, Portugal., Experimental Pathology and Therapeutics Group, Research Centre, Portuguese Oncology Institute of Porto (IPO-Porto), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal; Institute for Research and Innovation in Health (i3S), University of Porto, R. Alfredo Allen, 4200-135 Porto, Portugal; Glycobiology in Cancer, Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), R. Júlio Amaral de Carvalho 45, 4200-135 Porto, Portugal; Porto Comprehensive Cancer Centre(P.ccc), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal., CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal., iBiMED - Institute of Biomedicine, Department of Medical Sciences, University of Aveiro, Campus Universitario de Santiago, Agra do Crasto - Edificio 30, 3810-193 Aveiro, Portugal; Research Unit of the Physiology and Cardiothoracic Surgery Department of the Faculty of Medicine, University of Porto, Alameda Prof. Hernâni Monteiro, 4200-319 Porto, Portugal., Mass Spectrometry Centre, Organic Chemistry and Natural Products Unit, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal., Experimental Pathology and Therapeutics Group, Research Centre, Portuguese Oncology Institute of Porto (IPO-Porto), R. Dr. António Bernardino de Almeida 62, 4200-162 Porto, Portugal; Institute of Biomedical Sciences Abel Salazar, University of Porto, R. Jorge de Viterbo Ferreira 228, 4050-013 Porto, Portugal; Institute for Research and Innovation in Health (i3S), University of Porto, R. Alfredo Allen, 4200-135 Porto, Portugal; Glycobiology in Cancer, Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), R. Júlio Amaral de Carvalho 45, 4200-135 Porto, Portugal; International Iberian Nanotechnology Laboratory (INL), Avda. Mestre José Veiga, 4715 Braga, Portugal. Electronic address: .