Charan Kumar V Devarakonda1, Emily R Shearier2, Chaoran Hu3, James Grady4, Jeremy L Balsbaugh5, John H Makari6, Fernando A Ferrer7, Linda H Shapiro8. 1. Center for Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA. Electronic address: Devarakonda@uchc.edu. 2. Center for Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA. Electronic address: Emily.Shearier@hhchealth.org. 3. Biostatistics Center, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA. Electronic address: chaoran.hu@uconn.edu. 4. Biostatistics Center, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA. Electronic address: jgrady@uchc.edu. 5. Proteomics and Metabolomics Facility, Center for Open Research Resources and Equipment, University of Connecticut, Storrs, CT, 06269, USA. Electronic address: jeremy.balsbaugh@uconn.edu. 6. Department of Surgery, Division of Urology, University of Nebraska, Omaha, NE, 68918, USA. Electronic address: jmakari@childrensomaha.org. 7. Center for Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA; Department of Urology, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA. Electronic address: Fernando.ferrer@mountsinai.org. 8. Center for Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA. Electronic address: lshapiro@uchc.edu.
Abstract
INTRODUCTION AND OBJECTIVE: Reliable urinary biomarker proteins would be invaluable in identifying children with ureteropelvic junction obstruction (UPJO) as the existing biomarker proteins are inconsistent in their predictive ability. Therefore, the aim of this study was to identify consistent and reliable urinary biomarker proteins in children with UPJO. METHODS: To identify candidate biomarker proteins, total protein from age-restricted (<2 years) and sex-matched (males) control (n = 22) and UPJO (n = 21) urine samples was analyzed by mass spectrometry. Proteins that were preferentially identified in UPJO samples were selected (2-step process) and ranked according to their diagnostic odds ratio value. The top ten proteins with highest odds ratio values were selected and tested individually by ELISA. The total amount of each protein was normalized to urine creatinine and the median with interquartile ranges for control and UPJO samples was determined. Additionally, fold change (UPJO/Control) of medians of the final panel of 5 proteins was also determined. Finally, we calculated the average + 3(SD) and average + 4(SD) values of each of the 5 proteins in the control samples and used it as an arbitrary cutoff to classify individual control and UPJO samples. RESULTS: In the first step of our selection process, we identified 171 proteins in UPJO samples that were not detected in the majority of the control samples (16/22 samples, or 72.7%). Of the 171 proteins, only 50 proteins were detected in at least 11/21 (52.4%) of the UPJO samples and hence were selected in the second step. Subsequently, these 50 proteins were ranked according to the odds ratio value and the top 10 ranked proteins were validated by ELISA. Five of the 10 proteins - prostaglandin-reductase-1, ficolin-2, nicotinate-nucleotide pyrophosphorylase [carboxylating], immunoglobulin superfamily-containing leucine-rich-repeat-protein and vascular cell adhesion molecule-1 were present at higher levels in the UPJO samples (fold-change of the median protein concentrations ranging from 2.9 to 9.4) and emerged as a panel of biomarkers to identify obstructive uropathy. Finally, the order of prevalence of the 5 proteins in UPJO samples is PTGR1>FCN2>QPRT>ISLR>VCAM1. CONCLUSION: In summary, this unique screening strategy led to the identification of previously unknown biomarker proteins that when screened collectively, may reliably distinguish between obstructed vs. non-obstructed infants and may prove useful in identifying informative biomarker panels for biological samples from many diseases.
INTRODUCTION AND OBJECTIVE: Reliable urinary biomarker proteins would be invaluable in identifying children with ureteropelvic junction obstruction (UPJO) as the existing biomarker proteins are inconsistent in their predictive ability. Therefore, the aim of this study was to identify consistent and reliable urinary biomarker proteins in children with UPJO. METHODS: To identify candidate biomarker proteins, total protein from age-restricted (<2 years) and sex-matched (males) control (n = 22) and UPJO (n = 21) urine samples was analyzed by mass spectrometry. Proteins that were preferentially identified in UPJO samples were selected (2-step process) and ranked according to their diagnostic odds ratio value. The top ten proteins with highest odds ratio values were selected and tested individually by ELISA. The total amount of each protein was normalized to urine creatinine and the median with interquartile ranges for control and UPJO samples was determined. Additionally, fold change (UPJO/Control) of medians of the final panel of 5 proteins was also determined. Finally, we calculated the average + 3(SD) and average + 4(SD) values of each of the 5 proteins in the control samples and used it as an arbitrary cutoff to classify individual control and UPJO samples. RESULTS: In the first step of our selection process, we identified 171 proteins in UPJO samples that were not detected in the majority of the control samples (16/22 samples, or 72.7%). Of the 171 proteins, only 50 proteins were detected in at least 11/21 (52.4%) of the UPJO samples and hence were selected in the second step. Subsequently, these 50 proteins were ranked according to the odds ratio value and the top 10 ranked proteins were validated by ELISA. Five of the 10 proteins - prostaglandin-reductase-1, ficolin-2, nicotinate-nucleotide pyrophosphorylase [carboxylating], immunoglobulin superfamily-containing leucine-rich-repeat-protein and vascular cell adhesion molecule-1 were present at higher levels in the UPJO samples (fold-change of the median protein concentrations ranging from 2.9 to 9.4) and emerged as a panel of biomarkers to identify obstructive uropathy. Finally, the order of prevalence of the 5 proteins in UPJO samples is PTGR1>FCN2>QPRT>ISLR>VCAM1. CONCLUSION: In summary, this unique screening strategy led to the identification of previously unknown biomarker proteins that when screened collectively, may reliably distinguish between obstructed vs. non-obstructed infants and may prove useful in identifying informative biomarker panels for biological samples from many diseases.
Authors: Michael S Forbes; Barbara A Thornhill; Jordan J Minor; Katherine A Gordon; Carolina I Galarreta; Robert L Chevalier Journal: Am J Physiol Renal Physiol Date: 2012-04-25
Authors: S Gupta; A R Jackson; D G DaJusta; D J McLeod; S A Alpert; V R Jayanthi; K McHugh; A R Schwaderer; B Becknell; C B Ching Journal: J Pediatr Urol Date: 2018-03-29 Impact factor: 1.830
Authors: Nicholas G Cost; Paul H Noh; Prasad Devarajan; Vesna Ivancic; Pramod P Reddy; Eugene Minevich; Michael Bennett; Christopher Haffner; Marion Schulte; W Robert DeFoor Journal: J Urol Date: 2013-06-20 Impact factor: 7.450
Authors: John W Froehlich; Stephen A Kostel; Patricia S Cho; Andrew C Briscoe; Hanno Steen; Ali R Vaezzadeh; Richard S Lee Journal: Mol Cell Proteomics Date: 2016-05-23 Impact factor: 5.911
Authors: Anna Wasilewska; Katarzyna Taranta-Janusz; Wojciech Dębek; Walentyna Zoch-Zwierz; Elżbieta Kuroczycka-Saniutycz Journal: Pediatr Nephrol Date: 2011-01-31 Impact factor: 3.714
Authors: Timothy D Cummins; David W Powell; Daniel W Wilkey; Makayla Brady; Fredrick W Benz; Michelle T Barati; Dawn J Caster; Jon B Klein; Michael L Merchant Journal: Glomerular Dis Date: 2022-01-27
Authors: Zuzana Seifriedova; Hana Flogelova; Jan Sarapatka; Oldrich Smakal; Vladimir Student Journal: Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub Date: 2022-02-03 Impact factor: 1.648