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ISSN 2007-6800 (Print)

2022, Number 3

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Rev Mex Traspl 2022; 11 (3)

Protein expression analysis in patients with renal transplant rejection

Olivares-del MJI, Ocharán-Hernández ME, Reyes-Grajeda JP, Charco-Cruz M, Florez-Zorrilla C, Sánchez-Cedillo IA, Cano-Cervantes JH, Núñez-Venzor A, Lina-López L, Fonseca-González GT

Language: Spanish
References: 38
Page: 81-90
PDF size: 562.22 Kb.


ABSTRACT

Introduction: renal transplantation is the only treatment that improves the quality of life of patients with chronic kidney disease; at the end of 2021, 1,971 transplants were performed in Mexico. Among the main complications observed after transplantation are infections, alterations associated with immunosuppression, recurrence of the underlying disease and rejection. The incidence of rejection reported worldwide ranges between 12 and 14%. The diagnosis of rejection is histopathological; however, no molecular markers have been reported that allow us to suspect the development of this entity before the onset of structural deterioration of the graft. Current methods to monitor graft rejection are creatinine elevation, the presence of proteinuria and the development of specific anti-donor antibodies (ADES). A renal biopsy implies a risk of hematuria and perirenal hematoma (2 to 4%). Background: there are a large number of new, non-invasive biomarkers in urine and peripheral blood samples under investigation in recent years, most of them with prognostic or diagnostic value, as well as varying degrees of clinical and preclinical success. Some of them, in relation to acute rejection have been investigated miR146a-5p in recipient serum and urine, three-gene panel (CTOT 03 study for T-cell mediated rejection (TCMR), the KALIBRE study (7-gene panel for TCMR), GoCAR study (17-gene panel for TCMR), kSORT (AART study), for T cell-mediated rejection and antibody-mediated rejection, complement fragments for antibody-mediated rejection, CXCL 9 for TCMR and ABMR, CSCL 10, for ABMR to mention some of the work that has been done so far. The development of new technologies to identify proteins associated with a metabolic process has demonstrated the possibility of having a favorable impact in identifying processes such as transplant graft rejection. Proteomics consists of a series of biochemical and analytical techniques to identify differentially expressed proteins that could be used as biomarkers for this purpose. Material and methods: 23 kidney transplanted patients from the Centro Médico Nacional 20 de Noviembre, who underwent renal graft biopsy in a period of four months, were classified in two groups, one group with stable graft function n = 13 and one group with active rejection mediated by antibodies n = 13. Demographic analysis was performed with mode, median and ranges without finding significant differences in both groups. The patients' serum samples were analyzed for integrity, once they were quantified by the 2D-Quant colorimetric method, the most representative samples were chosen for their protein content, i.e., those whose protein concentration was above 20 mg/dL. Subsequently, these were subjected to electrophoretic separation analysis of two-dimensional gels in order to identify differential proteins in both study groups. Results: we systematized the collection, preservation and transport of samples of kidney transplanted patients from the 20 de Noviembre hospital. Protein quantification was standardized for analysis with two-dimensional gels and a preliminary analysis of differential expression patterns was performed, setting the precedents for subsequent analysis and identification of differential proteins by mass spectrometry. Comparing both gels, we found the representation of high abundance proteins such as immunoglobulin, haptoglobin, complement factors, alpha 1 antitrypsin, to mention some that are of greater representation in the gels and denote the adequate preservation and management of tissues, molecules such as CXC 7 and ABMP protein, with greater representation in the master gel of patients with rejection. Conclusions: classical proteomics in the study of differential proteins for the development of protocols for the discovery of proteins in patients receiving a transplant, with the aim of identifying potential molecular markers that could be useful for the diagnosis of rejection is a useful tool that is still under development and requires time and analysis of specific data to achieve objective and transcendental results. Proteins such as CXC 7 and ABMP protein are representative for this study, and validation studies should be performed in addition to mass spectrometry identification to assess the possibility of identifying proteins different from those reported in the literature.


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