ABSTRACT
Gastric (GC) and breast (BrC) cancer are two of the most common and deadly tumours. Different lines of evidence suggest a possible causative role of viral infections for both GC and BrC. Wide genome sequencing (WGS) technologies allow searching for viral agents in tissues of patients with cancer. These technologies have already contributed to establish virus-cancer associations as well as to discovery new tumour viruses. The objective of this study was to document possible associations of viral infection with GC and BrC in Mexican patients. In order to gain idea about cost effective conditions of experimental sequencing, we first carried out an in silico simulation of WGS. The next-generation-platform IlluminaGallx was then used to sequence GC and BrC tumour samples. While we did not find viral sequences in tissues from BrC patients, multiple reads matching Epstein-Barr virus (EBV) sequences were found in GC tissues. An end-point polymerase chain reaction confirmed an enrichment of EBV sequences in one of the GC samples sequenced, validating the next-generation sequencing-bioinformatics pipeline.
Subject(s)
Female , Humans , Male , Breast Neoplasms/virology , DNA, Viral/isolation & purification , /genetics , High-Throughput Nucleotide Sequencing/methods , RNA, Viral/isolation & purification , Stomach Neoplasms/virology , Computers , Computational Biology/methods , Computer Simulation/economics , Cost-Benefit Analysis/methods , Mexico , Nucleic Acids/isolation & purification , Polymerase Chain Reaction/methods , Sequence Analysis, DNA/methods , Sequence Analysis, RNA/methodsABSTRACT
Abstract: Introduction: Survival of transplant patients and grafts depends largely on the use of immunosuppressive drugs. However, a balance remains to be established among immunosuppression, transplant rejection and cytomegalovirus (CMV) infection, which results in a high rate of morbidity and mortality. The aim of this study was to define a better strategy for monitoring transplanted patients based on the analysis of the blood concentration of sirolimus and tacrolimus and the burden of CMV. Methods: Fifty five post-transplant (kidney and liver) pediatric patients, nine treated with sirolimus and 46 treated with tacrolimus, were included. A total of 541 measurements were obtained. In each measurement the concentration of immunosuppressant in whole blood and CMV viral load in plasma and whole blood was quantified by real-time PCR. Pearson correlation coefficient (r) was estimated. Results: Values of r ≤ 0.0747 were found for the relationship between dose and concentration of immunosuppressant; r = 0.9406 for the relationship between viral load in whole blood and plasma, and r ≤ 0.4616 for the relationship between concentration of immunosuppressant and viral load. Conclusions: These data support that the doses of immunosuppressive drugs do not correlate with the levels of the same in whole blood. Therefore, systemic levels of immunosuppressant should be constantly monitored together with CMV load. Meanwhile, a high correlation between viral load measured in whole blood and plasma was found.
Resumen: Introducción: La supervivencia de pacientes trasplantados y de los injertos depende en gran medida del uso de fármacos inmunosupresores. Sin embargo, aún no se ha logrado establecer un balance entre la inmunosupresión, el rechazo al trasplante y la infección por citomegalovirus (CMV), lo cual deriva en una alta tasa de morbilidad y mortalidad. El objetivo de este trabajo fue definir una mejor estrategia de seguimiento de los pacientes trasplantados a partir del análisis de la concentración en sangre de sirolimus y tacrolimus y la carga de CMV. Métodos: Se incluyeron 55 pacientes pediátricos post-trasplante (riñón e hígado), nueve en tratamiento con sirolimus y 46 en tratamiento con tacrolimus. Se obtuvieron 541 mediciones en total. En cada medición se cuantificó la concentración de inmunosupresor en sangre total y la carga viral de CMV en plasma y sangre total mediante PCR en tiempo real. Se calculó el coeficiente de correlación de Pearson (r). Resultados: Se encontraron valores de r ≤ 0.0747 para la relación entre dosis y concentración del inmunosupresor; de r = 0.9406 para la relación de la carga viral entre suero y sangre total y de r ≤ 0.4616 para la relación entre concentración de inmunosupresor y carga viral. Conclusiones: Estos datos apoyan que la dosis de los fármacos inmunosupresores no correlaciona con los niveles de los mismos en sangre total. Por ello, deben ser constantemente monitoreados junto con la carga viral. Por su parte, se encontró alta correlación entre la carga viral medida en sangre total y plasma.