Acute cellular rejection occurs frequently during the first few weeks following liver transplantation. During this period, its molecular phenotype is confounded by peri- and postoperative proinflammatory events. To unambiguously define the molecular profile associated with rejection, we collected sequential biological specimens from 55 patients at least 3 years after liver transplantation who developed rejection during trials of intentional immunosuppression withdrawal. We analyzed liver tissue and blood samples obtained before initiation of drug withdrawal and at rejection, alongside blood samples collected during the weaning process. Gene expression profiling was conducted using whole-genome microarrays and real-time polymerase chain reaction. Rejection resulted in distinct blood and liver tissue transcriptional changes in patients who were either positive or negative for hepatitis C virus (HCV). Gene expression changes were mostly independent from pharmacological immunosuppression, and their magnitude correlated with severity of histological damage. Differential expression of a subset of genes overlapped across all conditions. These were used to define a blood predictive model that accurately identified rejection in HCV-negative, but not HCV-positive, patients. Changes were detectable 1-2 mo before rejection was diagnosed. Our results provide insight into the molecular processes underlying acute cellular rejection in liver transplantation and help clarify the potential utility and limitations of transcriptional biomarkers in this setting.
Biomarkers/metabolism , Gene Expression Profiling , Graft Rejection/diagnosis , Immune Tolerance/genetics , Liver Transplantation , Postoperative Complications , Withholding Treatment , Female , Follow-Up Studies , Gene Expression Regulation , Graft Rejection/etiology , Graft Rejection/metabolism , Graft Survival , Humans , Immunosuppression Therapy , Immunosuppressive Agents/therapeutic use , Liver Diseases/surgery , Male , Middle Aged , Prospective Studies
Objetivo: Analizar los perfiles de expresión génica del cáncer de próstata (CaP) e identificar los genes diferencialmente expresados. Determinar si la expresión diferencial en tejido se mantiene en muestras de orina-posmasaje prostático (PMP). Material y métodos: Un total de 46 muestras de tejido prostático (36 de pacientes con CaP y 10 controles) y 158 orinas-PMP (113 de pacientes con CaP y 45 controles) se recogieron entre diciembre de 2003 y mayo de 2007. Se utilizaron microarrays de ADN para identificar los genes diferencialmente expresados entre las muestras de tejido tumorales y las controles. Diez genes fueron seleccionados para la validación técnica de los microarrays en las mismas muestras tisulares mediante PCR cuantitativa (RT-qPCR). Se seleccionaron 42 genes para ser validados en muestras de orina-PMP mediante RT-qPCR. Resultados: El gráfico de escalado multidimensional mostró una clara separación entre las muestras de tejido tumorales y las controles. Se han identificado 1.047 genes diferencialmente expresados (FDR ≤ 0,1) entre los 2 grupos. La correlación entre los datos de microarrays y RT-qPCR fue alta (r = 0,928, p < 0,001). Trece genes mantuvieron el mismo sentido de expresión diferencial al ser analizados en orinas-PMP y 4 de ellos (HOXC6, PCA3, PDK4 y TMPRSS2-ERG) mostraron diferencias de expresión estadísticamente significativas entre orinas-PMP tumorales y controles (p < 0,05). Conclusión: Existe un perfil de expresión génica diferencial en el CaP. Aunque la extrapolación de la expresión génica obtenida en tejido prostático a orina-PMP se debe realizar con precaución, el análisis del tejido prostático permite la identificación de nuevos biomarcadores para diagnóstico no invasivo del CaP
Objective: To analyze gene expression profiles of prostate cancer (PCa) with the aim of determining the relevant differentially expressed genes and subsequently ascertain whether this differential expression is maintained in post-prostatic massage (PPM) urine samples. Material and methods: Forty-six tissue specimens (36 from PCa patients and 10 controls) and158 urine PPM-urines (113 from PCa patients and 45 controls) were collected between December 2003 and May 2007. DNA microarrays were used to identify genes differentially expressed between tumour and control samples. Ten genes were technically validated in the same tissue samples by quantitative RT-PCR (RT-qPCR). Forty two selected differentially expressed genes were validated in an independent set of PPM-urines by qRT-PCR. Results: Multidimensional scaling plot according to the expression of all the microarray genes showed a clear distinction between control and tumour samples. A total of 1047 differentially expressed genes (FDR≤0.1) were indentified between both groups of samples. We found a high correlation in the comparison of microarray and RT-qPCR gene expression levels (r = 0.928,P < 0.001). Thirteen genes maintained the same fold change direction when analyzed in PPM urine samples and in four of them (HOXC6, PCA3, PDK4 and TMPRSS2-ERG), these differences were statistically significant (P < 0.05). Conclusion: The analysis of PCa by DNA microarrays provides new putative mRNA markers for PCa diagnosis that, with caution, can be extrapolated to PPM-urines
Humans , Male , Gene Expression , Prostatic Neoplasms/genetics , Oligonucleotide Array Sequence Analysis/methods , Genetic Markers , Genetic Predisposition to Disease , Case-Control Studies , Real-Time Polymerase Chain Reaction
Due to its low level of nephrotoxicity and capacity to harness tolerogenic pathways, sirolimus (SRL) has been proposed as an alternative to calcineurin inhibitors in transplantation. The exact mechanisms underlying its unique immunosuppressive profile in humans, however, are still not well understood. In the current study, we aimed to depict the in vivo effects of SRL in comparison with cyclosporin A (CSA) by employing gene expression profiling and multiparameter flow cytometry on blood cells collected from stable kidney recipients under immunosuppressant monotherapy. SRL recipients displayed an increased frequency of CD4 + CD25highFoxp3 + T cells. However, this was accompanied by an increased number of effector memory T cells and by enrichment in NFkB-related pro-inflammatory expression pathways and monocyte and NK cell lineage-specific transcripts. Furthermore, measurement of a transcriptional signature characteristic of operationally tolerant kidney recipients failed to detect differences between SRL and CSA-treated recipients. In conclusion, we show here that the blood transcriptional profile induced by SRL monotherapy in vivo does not resemble that of operationally tolerant recipients and is dominated by innate immune cells and NFkB-related pro-inflammatory events. These data provide novel insights on the complex effects of SLR on the immune system in clinical transplantation.
Cyclosporine/therapeutic use , Immunosuppressive Agents/therapeutic use , Kidney Transplantation/immunology , Sirolimus/therapeutic use , T-Lymphocytes/immunology , CD4 Lymphocyte Count , Flow Cytometry , Gene Expression Profiling , Humans , Immunity, Innate/drug effects , Phenotype , T-Lymphocytes/drug effects , T-Lymphocytes/metabolism
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Humans , Voice Disorders/etiology , Neuromuscular Diseases/diagnosis , Electromyography
Introducción. La terapia convencional basada en fármacos dopaminérgicosno frena ni ralentiza de modo significativo el cursoprogresivo de la enfermedad de Parkinson (EP). La fase final de la EPse caracteriza por la presencia de síntomas y signos resistentes a laterapia dopaminérgica (depresión, demencia, disartria, caídas, etc.).Es urgente desarrollar terapias que eviten llegar a estas fases deteniendoo retardando la progresión de la enfermedad. Sin embargo,no se dispone de estrategias neuroprotectoras efectivas.Método. Se realizó un estudio de informadores clave en el queexpertos en EP que cumplimentaron un cuestionario de 10 preguntassobre la problemática más importante en el área de la neuroprotecciónen la EP. Tras ello se estableció un consenso sobre la situaciónactual y se sugirieron nuevas direcciones de investigación.Resultados. La mayoría de respuestas coincidieron en la necesidadde nuevos conceptos, en las limitaciones de los actuales modelosanimales o las dificultades de demostrar un efecto protector en humanospor la falta de biomarcadores. Algunos participantes opinanque ya se está ejerciendo un cierto efecto modificador del curso dela enfermedad.Conclusiones. El concepto de neuroprotección debe ser ampliado,los modelos animales deben mejorarse y urge encontrar un biomarcadorfiable para planificar la terapia en fases más precoces ypara determinar el efecto neuroprotector (AU)
Introduction. Currently used antiparkinsonian drugs neitherstop nor slow-down the progressive nature of the disease. The finalphase of PD is characterized by the presence of symptomsand signs resistant to dopaminergic agents, such as depression,dementia, freezing and falls. Therefore, it is urgent to develop therapies able to positively modify this outcome. Despite neuroprotectionis a research priority in PD, no effective strategieshave been found so far.Method. A key informants study was conducted. A group ofexperts in PD fulfilled a questionnaire of 10 questions to explorethe most important topics related to neuroprotection. Afterwardsa consensus about the cur-rent situation of neuroprotection inPD was established and future directions of development weresuggested.Results. Most of the answers emphasized the need of newconcepts, the limitations of animal models and the difficulties inthe difficulties in demonstrating a neuroprotective effects in humansowing to a lack of biomarkers. Some of the experts believethat we are already exerting a disease modifying effect.Conclusions. The concept of neuroprotection should be widened.Animal models should be improved. A reliable biomarkerto start neuroprotective therapies long before the appearance ofmotor symptoms and to evaluate the neuroprotective effect ofany therapy should be urgently developed (AU)
Humans , Animals , Consensus , Antiparkinson Agents/therapeutic use , Neuroprotective Agents/therapeutic use , Parkinson Disease/drug therapy , Parkinson Disease/prevention & control , Biomarkers/metabolism , Disease Models, Animal , Disease Progression , Parkinson Disease/physiopathology , Practice Guidelines as Topic , Surveys and Questionnaires , Treatment Outcome
