Prohibitin is a prognostic marker and therapeutic target to block chemotherapy resistance in Wilms' tumor.

Wilms' tumor is the most common type of childhood kidney cancer. To improve risk stratification and identify novel therapeutic targets for patients with Wilms' tumor, we used high-resolution mass spectrometry proteomics to identify urine tumor markers associated with Wilms' tumor relapse. We determined the urine proteomes at diagnosis of 49 patients with Wilms' tumor, non-Wilms' tumor renal tumors, and age-matched controls, leading to the quantitation of 6520 urine proteins. Supervised analysis revealed specific urine markers of renal rhabdoid tumors, kidney clear cell sarcomas, renal cell carcinomas as well as those detected in patients with cured and relapsed Wilms' tumor. In particular, urine prohibitin was significantly elevated at diagnosis in patients with relapsed as compared with cured Wilms' tumor. In a validation cohort of 139 patients, a specific urine prohibitin ELISA demonstrated that prohibitin concentrations greater than 998 ng/mL at diagnosis were significantly associated with ultimate Wilms' tumor relapse. Immunohistochemical analysis revealed that prohibitin was highly expressed in primary Wilms' tumor specimens and associated with disease stage. Using functional genetic experiments, we found that prohibitin was required for the growth and survival of Wilms' tumor cells. Overexpression of prohibitin was sufficient to block intrinsic mitochondrial apoptosis and to cause resistance to diverse chemotherapy drugs, at least in part by dysregulating factors that control apoptotic cytochrome c release from mitochondrial cristae. Thus, urine prohibitin may improve therapy stratification, noninvasive monitoring of treatment response, and early disease detection. In addition, therapeutic targeting of chemotherapy resistance induced by prohibitin dysregulation may offer improved therapies for patients with Wilms' and other relapsed or refractory tumors.

Prenatal smoking exposure and cardio-metabolic risk factors in adulthood: a general population study and a meta-analysis.

OBJECTIVE: Prenatal smoking exposure is associated with obesity and other cardio-metabolic risk factors in children, but no previous meta-analysis has been conducted in adults. METHODS: We investigated the association of prenatal smoking exposure in the Danish General Suburban Population Study (GESUS) with BMI, waist circumference, total cholesterol, type 2 diabetes, gestational type 2 diabetes, and hypertension in adulthood. We subsequently performed a meta-analysis, adding published studies investigating the association between prenatal smoking and the risk of cardio-metabolic outcomes among individuals at least 18 years of age. RESULTS: We included 19 eligible observational studies with various cardio-metabolic outcomes (N = 24,201-308,981 adults). In individuals exposed to prenatal smoking, the pooled random effects adjusted odds ratio were 1.35 (95% CI: 1.16-1.56) for being overweight, 1.46 (1.39-1.54) for being obese, 1.07 (0.89-1.29) for type 2 diabetes, 1.17 (0.92-1.48) for hypertension, and 1.38 (1.19-1.61) for gestational diabetes mellitus (GDM), compared with no exposure. The standardized means in waist circumference, total cholesterol, diastolic, and systolic blood pressure were not different in individuals exposed vs. not exposed to prenatal smoking. Heterogeneity was moderate to high (51% < I2 < 99%). However, removal of the high heterogeneity removed the associated uncertainty in the point estimate and revealed that prenatal smoking is associated with increased BMI in adulthood. There was also no evidence of publication bias in the meta-analyses. CONCLUSIONS: The findings from the meta-analyses suggested that prenatal smoking exposure is associated with an increased odds ratio of overweight, obesity, and GDM in adulthood, but not with type 2 diabetes, hypertension, waist circumference, or total cholesterol. These findings highlight the importance of abstaining from smoking by pregnant women.

Improved separation and analysis of plasma amino acids by modification of the MassTrak™ AAA Solution Ultraperformance® liquid chromatography method.

BACKGROUND: Ultraperformance® Liquid Chromatography (UPLC) is increasingly used for quantitative amino acid screening. The Waters MassTrak™ UPLC Amino Acid Analysis (AAA) Solution kit offers rapid analysis with minimal sample preparation. We describe a simple modification of this method enabling enhanced chromatographic separation of previously problematic analytes with improvements in quantification. METHODS: The commercial UPLC method was compared with our modified version of the same method. The modification incorporates eluent buffer of increased organic content run at reduced column temperature. UPLC methods were compared by analyzing amino acids from 57 plasma samples. A comparison (n=131) between the modified UPLC method and ion-exchange chromatography was also carried out. RESULTS: The commercial method produced a large negative bias for Tyrosine (-22.72%±14.10) and ornithine (-15.02%±10.07). Assay imprecision of Tyrosine using the commercial method (mean Tyrosine: 72.58 and 31.17µmol/l) produced values of 10.86% and 21.12% respectively, compared with the modified method (3.39% and 4.47%). The comparison of modified UPLC and ion-exchange methods was favorable, validating the improvements observed in amino acid quantification. CONCLUSION: The modified UPLC method has eliminated significant bias associated with the commercially available method. The modification is simple, robust and readily adaptable to the current MassTrak™ AAA Solution kit for clinical applications.