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Multiplexed Biosensing Diagnostic Platforms Detecting Autoantibodies to Tumor-Associated Antigens from Exosomes Released by CRC Cells and Tissue Samples Showed High Diagnostic Ability for Colorectal Cancer

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Engineering 2021年第10期7卷 1393-1412页

作者： Ana Montero-Calle Itziar Aranguren-Abeigon María Garranzo-Asensio Carmen Poves María Jesús Fernández-Aceñero Javier Martínez-Useros Rodrigo Sanz Jana Dziaková Javier Rodriguez-Cobos Guillermo Solís-Fernández Eloy Povedano Maria Gamella Rebeca Magnolia Torrente-Rodríguez Miren Alonso-Navarro Vivian de los Ríos J.Ignacio Casal gemma domínguez Ana Guzman-Aranguez Alberto Peláez-García JoséManuel Pingarrón Susana Campuzano Rodrigo Barderas Chronic Disease Program(UFIEC) Carlos III Health InstituteMadrid E-28220Spain Department of Biochemistry and Molecular Biology Faculty of Optics and OptometryComplutense University of MadridMadrid E-28040Spain Gastroenterology Unit San Carlos Clinical HospitalMadrid E-28040Spain Surgical Pathology Department San Carlos Clinical HospitalMadrid E-28040Spain Translational Oncology Division OncoHealth InstituteJimenez Diaz Foundation University HospitalMadrid E-28040Spain Department of Biochemistry Faculty of MedicineAlberto Sols Institute of Biomedical ResearchCSIC-UAMMadrid E-28029Spain Department of Analytical Chemistry Faculty of Chemical SciencesComplutense University of MadridMadrid E-28040Spain Center for Biological Research Spanish National Research Council(CSIC)MadridE-28040Spain Molecular Pathology and Therapeutic Targets Group La Paz University Hospital(IdiPAZ)Madrid E-28046Spain

Colorectal cancer(CRC)is the second leading cause of cancer-related death worldwide.The five-year survival rate of CRC patients depends on the stage at diagnosis,being higher than 80%when CRC is diagnosed in the early stages but lower than 10%when CRC is diagnosed in advanced stages.Autoantibodies against specific CRC autoantigens(tumor-associated antigens(TAAs))in the sera of patients have been widely demonstrated to aid in early diagnosis.Thus,we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy.To that end,we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis.A total of 103 proteins were identified as potential autoantigens specific to CRC.After bioinformatics and meta-analysis,we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot(WB)and immunohistochemistry(IHC).We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients,along with an association of nine of these proteins with CRC prognosis.After validation,all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals,either by luminescence Halotag-based beads,or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection.Taken together,our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases;they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care(POC)de

关键词： Autoantibodies Diagnosis Colorectal cancer Exosomes Tumor microenviroment Humoral immune response Point of care Biosensors

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