RESUMEN
Exposure to ionizing radiation induces a complex cascade of systemic and tissue-specific responses that lead to functional impairment over time in the surviving population. However, due to the lack of predictive biomarkers of tissue injury, current methods for the management of survivors of radiation exposure episodes involve monitoring of individuals over time for the development of adverse clinical symptoms and death. Herein, we report on changes in metabolomic and lipidomic profiles in multiple tissues of nonhuman primates (NHPs) that were exposed to a single dose of 7.2 Gy whole-body 60Co γ-radiation that either survived or succumbed to radiation toxicities over a 60-day period. This study involved the delineation of the radiation effects in the liver, kidney, jejunum, heart, lung, and spleen. We found robust metabolic changes in the kidney and liver and modest changes in other tissue types at the 60-day time point in a cohort of NHPs. Remarkably, we found significant elevation of long-chain acylcarnitines in animals that were exposed to radiation across multiple tissue types underscoring the role of this class of metabolites as a generic indicator of radiation-induced normal tissue injury. These studies underscore the utility of a metabolomics approach for delineating anticipatory biomarkers of exposure to ionizing radiation.
Asunto(s)
Rayos gamma/efectos adversos , Radiación Ionizante , Animales , Biomarcadores/metabolismo , Carnitina/análogos & derivados , Carnitina/metabolismo , Femenino , Expresión Génica/efectos de los fármacos , Técnicas In Vitro , Riñón/efectos de los fármacos , Riñón/metabolismo , Lipidómica , Macaca mulatta , Masculino , Metabolómica/métodos , PrimatesRESUMEN
The aims of this study were to demonstrate the feasibility of centrally collecting and processing high-quality cerebrospinal fluid (CSF) samples for proteomic studies within a multi-center consortium and to identify putative biomarkers for medulloblastoma in CSF. We used 2-DE to investigate the CSF proteome from 33 children with medulloblastoma and compared it against the CSF proteome from 25 age-matched controls. Protein spots were subsequently identified by a combination of in-gel tryptic digestion and MALDI-TOF TOF MS analysis. On average, 160 protein spots were detected by 2-DE and 76 protein spots corresponding to 25 unique proteins were identified using MALDI-TOF. Levels of prostaglandin D2 synthase (PGD2S) were found to be six-fold decreased in the tumor samples versus control samples (p<0.00001). These data were further validated using ELISA. Close examination of PGD2S spots revealed the presence of complex sialylated carbohydrates at residues Asn(78) and Asn(87) . Total PGD2S levels are reduced six-fold in the CSF of children with medulloblastoma most likely representing a host response to the presence of the tumor. In addition, our results demonstrate the feasibility of performing proteomic studies on CSF samples collected from patients at multiple institutions within the consortium setting.
Asunto(s)
Biomarcadores/metabolismo , Neoplasias Encefálicas/enzimología , Perfilación de la Expresión Génica , Oxidorreductasas Intramoleculares/metabolismo , Lipocalinas/metabolismo , Meduloblastoma/enzimología , Isoformas de Proteínas/metabolismo , Secuencia de Aminoácidos , Asparagina/metabolismo , Biomarcadores/líquido cefalorraquídeo , Neoplasias Encefálicas/líquido cefalorraquídeo , Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/genética , Secuencia de Carbohidratos , Estudios de Casos y Controles , Niño , Electroforesis en Gel Bidimensional , Ensayo de Inmunoadsorción Enzimática , Humanos , Oxidorreductasas Intramoleculares/líquido cefalorraquídeo , Oxidorreductasas Intramoleculares/genética , Lipocalinas/líquido cefalorraquídeo , Lipocalinas/genética , Meduloblastoma/líquido cefalorraquídeo , Meduloblastoma/diagnóstico , Meduloblastoma/genética , Datos de Secuencia Molecular , Pediatría , Isoformas de Proteínas/líquido cefalorraquídeo , Isoformas de Proteínas/genética , Proteoma/análisis , Proteómica , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Tripsina/metabolismoRESUMEN
Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy wherein a majority of patients present metastatic disease at diagnosis. Although the role of epithelial to mesenchymal transition (EMT), mediated by transforming growth factor beta (TGFß), in imparting an aggressive phenotype to PDAC is well documented, the underlying biochemical pathway perturbations driving this behaviour have not been elucidated. We used high-resolution mass spectrometry (HRMS) based molecular phenotyping approach in order to delineate metabolic changes concomitant to TGFß-induced EMT in pancreatic cancer cells. Strikingly, we observed robust changes in amino acid and energy metabolism that may contribute to tumor invasion and metastasis. Somewhat unexpectedly, TGFß treatment resulted in an increase in intracellular levels of retinoic acid (RA) that in turn resulted in increased levels of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (COL1). These findings were further validated in plasma samples obtained from patients with resectable pancreatic cancer. Taken together, these observations provide novel insights into small molecule dysregulation that triggers a molecular cascade resulting in increased EMT-like changes in pancreatic cancer cells, a paradigm that can be potentially targeted for better clinical outcomes.