RT Journal Article
T1 p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins
A1 Montero Calle, Ana
A1 Garranzo Asensio, María
A1 Torrente Rodríguez, Rebeca Magnolia
A1 Ruiz Valdepeñas Montiel, Víctor
A1 Poves, Carmen
A1 Dziakova, Jana
A1 Sanz, Rodrigo
A1 Díaz Del Arco, Cristina
A1 Pingarrón Carrazón, José Manuel
A1 Fernández Aceñero, María Jesús
A1 Campuzano Ruiz, Susana
A1 Barderas Manchado, Rodrigo
AB Colorectal cancer (CRC) is the third most common cancer and the second most frequent cause of cancer-related death worldwide. The detection in plasma samples of autoantibodies against specific tumor-associated antigens has been demonstrated to be useful for the early diagnosis of CRC by liquid biopsy. However, new studies related to the humoral immune response in cancer are needed to enable blood-based diagnosis of the disease. Here, our aim was to characterize the humoral immune response associated with the different p53 and p63 proteoforms derived from alternative splicing and previously described as aberrantly expressed in CRC. Thus, here we investigated the diagnostic ability of the twelve p53 proteoforms and the eight p63 proteoforms described to date, and their specific N-terminal and C-terminal end peptides, by means of luminescence HaloTag beads immunoassays. Full-length proteoforms or specific peptides were cloned as HaloTag fusion proteins and their seroreactivity analyzed using plasma from CRC patients at stages I-IV (n = 31), individuals with premalignant lesions (n = 31), and healthy individuals (n = 48). p53γ, Δ40p53β, Δ40p53γ, Δ133p53γ, Δ160p53γ, TAp63α, TAp63δ, ΔNp63α, and ΔNp63δ, together with the specific C-terminal end α and δ p63 peptides, were found to be more seroreactive against plasma from CRC patients and/or individuals with premalignant lesions than from healthy individuals. In addition, ROC (receiver operating characteristic) curves revealed a high diagnostic ability of those p53 and p63 proteoforms to detect CRC and premalignant individuals (AUC higher than 85%). Finally, electrochemical biosensing platforms were employed in POC-like devices to investigate their usefulness for CRC detection using selected p53 and p63 proteoforms. Our results demonstrate not only the potential of these biosensors for the simultaneous analysis of proteoforms’ seroreactivity, but also their convenience and versatility for the clinical detection of CRC by liquid biopsy. In conclusion, we here show that p53 and p63 proteoforms possess differential seroreactivity in CRC patients in comparison to controls, distinctive from canonical proteins, which should improve the diagnostic panels for obtaining a blood-based biomarker signature for CRC detection.
PB MDPI
SN 2072-6694
YR 2023
FD 2023-03-31
LK https://hdl.handle.net/20.500.14352/103634
UL https://hdl.handle.net/20.500.14352/103634
LA eng
NO Montero-Calle, A.; Garranzo-Asensio, M.; Torrente-Rodríguez, R.M.; Ruiz-Valdepeñas Montiel, V.; Poves, C.; Dziaková, J.; Sanz, R.; Díaz del Arco, C.; Pingarrón, J.M.; Fernández-Aceñero, M.J.; et al. p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins. Cancers 2023, 15, 2102. https://doi.org/10.3390/ cancers15072102
NO The humoral immune response in cancer has been demonstrated to be useful for distinguishing patients from healthy individuals using serum or plasma. Our study aimed to assess whether p53 and p63 proteoforms derived from alternative splicing could have a differential seroreactivity and higher diagnostic value than canonical proteins in colorectal cancer. Using luminescence assays and biosensing approaches with the proteoforms expressed in vitro fused to HaloTag, we demonstrate the appearance of a differential seroreactivity among the proteoforms in colorectal cancer patients. Our findings reveal increased complexity of the humoral immune response in cancer against specific autoantigens, since specific seroreactivity and different diagnostic values were observed among the p53 and p63 proteoforms and the canonical proteins.
NO European Commission
NO Ministerio de Ciencia e Innovación (España)
NO Comunidad de Madrid
NO Instituto de Salud Carlos III (España)
DS Docta Complutense
RD 20 jul 2024