p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins
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 biops...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/15/7/2102 |
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| author | Ana Montero-Calle María Garranzo-Asensio Rebeca M. Torrente-Rodríguez Víctor Ruiz-Valdepeñas Montiel Carmen Poves Jana Dziaková Rodrigo Sanz Cristina Díaz del Arco José Manuel Pingarrón María Jesús Fernández-Aceñero Susana Campuzano Rodrigo Barderas |
| author_facet | Ana Montero-Calle María Garranzo-Asensio Rebeca M. Torrente-Rodríguez Víctor Ruiz-Valdepeñas Montiel Carmen Poves Jana Dziaková Rodrigo Sanz Cristina Díaz del Arco José Manuel Pingarrón María Jesús Fernández-Aceñero Susana Campuzano Rodrigo Barderas |
| author_sort | Ana Montero-Calle |
| collection | DOAJ |
| description | 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 (<i>n</i> = 31), individuals with premalignant lesions (<i>n</i> = 31), and healthy individuals (<i>n</i> = 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. |
| first_indexed | 2024-03-11T05:40:34Z |
| format | Article |
| id | doaj.art-2c64b4d8f4c24eddb953597d5d21ab1b |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-11T05:40:34Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-2c64b4d8f4c24eddb953597d5d21ab1b2023-11-17T16:26:12ZengMDPI AGCancers2072-66942023-03-01157210210.3390/cancers15072102p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical ProteinsAna Montero-Calle0María Garranzo-Asensio1Rebeca M. Torrente-Rodríguez2Víctor Ruiz-Valdepeñas Montiel3Carmen Poves4Jana Dziaková5Rodrigo Sanz6Cristina Díaz del Arco7José Manuel Pingarrón8María Jesús Fernández-Aceñero9Susana Campuzano10Rodrigo Barderas11Chronic Disease Programme (UFIEC), Instituto de Salud Carlos III, 28220 Madrid, SpainChronic Disease Programme (UFIEC), Instituto de Salud Carlos III, 28220 Madrid, SpainDepartment of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, 28014 Madrid, SpainDepartment of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, 28014 Madrid, SpainGastroenterology Unit, Hospital Universitario Clínico San Carlos, 28040 Madrid, SpainSurgical Digestive Department, Hospital Universitario Clínico San Carlos, 28040 Madrid, SpainSurgical Digestive Department, Hospital Universitario Clínico San Carlos, 28040 Madrid, SpainSurgical Pathology Department, Hospital Universitario Clínico San Carlos, 28040 Madrid, SpainDepartment of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, 28014 Madrid, SpainSurgical Pathology Department, Hospital Universitario Clínico San Carlos, 28040 Madrid, SpainDepartment of Analytical Chemistry, Faculty of Chemistry, Complutense University of Madrid, 28014 Madrid, SpainChronic Disease Programme (UFIEC), Instituto de Salud Carlos III, 28220 Madrid, SpainColorectal 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 (<i>n</i> = 31), individuals with premalignant lesions (<i>n</i> = 31), and healthy individuals (<i>n</i> = 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.https://www.mdpi.com/2072-6694/15/7/2102colorectal cancerimmunomicsautoantibodiesp53 familyp53 and p63alternative splicing |
| spellingShingle | Ana Montero-Calle María Garranzo-Asensio Rebeca M. Torrente-Rodríguez Víctor Ruiz-Valdepeñas Montiel Carmen Poves Jana Dziaková Rodrigo Sanz Cristina Díaz del Arco José Manuel Pingarrón María Jesús Fernández-Aceñero Susana Campuzano Rodrigo Barderas p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins Cancers colorectal cancer immunomics autoantibodies p53 family p53 and p63 alternative splicing |
| title | p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins |
| title_full | p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins |
| title_fullStr | p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins |
| title_full_unstemmed | p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins |
| title_short | p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins |
| title_sort | p53 and p63 proteoforms derived from alternative splicing possess differential seroreactivity in colorectal cancer with distinct diagnostic ability from the canonical proteins |
| topic | colorectal cancer immunomics autoantibodies p53 family p53 and p63 alternative splicing |
| url | https://www.mdpi.com/2072-6694/15/7/2102 |
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