Urinary Proteome Profile Research Articles

Urinary proteomic profiles of prostate cancer with different risk of progression and correlation with histopathological features

Prostate cancer (PCa) is the most common tumor in men with extremely variable outcome, varying from latent or indolent form to very aggressive behavior. High grade tumors, expansions exceeding the prostatic capsule into the surrounding soft tissues and spreading through lymph vascular channels, represent the most consistent unfavorable prognostic factors. However, accuracy in the prediction of the disease progression is sometimes difficult. Along with new molecular diagnostic techniques and more accurate histopathological approaches, proteomic studies challenge to identify potential biomarkers predictive of PCa progression. In our study we analyzed the urinary proteomes of 42 patients affected by PCa through two-dimensional electrophoresis associated with mass spectrometry. Proteomic profiles were correlated to histopathological features including pTNM stage and tumor differentiation in order to provide new promising markers able to define more accurately the PCa aggressiveness and driving new therapeutic approaches.

Urinary exosome proteomic profiling defines stage-specific rapid progression of autosomal dominant polycystic kidney disease and tolvaptan efficacy

ADPKD is the most common genetic disease of the kidney leading to end-stage renal disease necessitating renal replacement therapy at any time between the 1st and 8th decades of life due to widely variable rates of disease progression. This presents significant patient anxiety and a significant prognostic and therapeutic challenge. Tolvaptan is the only approved drug licensed to slow ADPKD progression by reducing renal cystic expansion but side-effects can limit its efficacy.To address the need to identify new biomarkers to monitor progression of ADPKD and to evaluate the therapeutic effects of Tolvaptan, proteomic analysis was conducted on defined (40-100nm) urinary exosomes isolated from ADPKD patients phenotyped and clinically monitored over a 10-year period. Comparative Gene Ontology analysis of Tandem Mass Tag labelled mass spectrometry-derived protein profiles from urinary exosomes from ADPKD patients with rapid (>10ml/min/5 years decline in estimated glomerular filtration rate) versus slow progression showed distinctive patterns of pathway up-regulation. Clear discrimination between rapid and slowly-progressive profiles were seen in all stages functional decline in ADPKD patients whether with mild (>70ml/min), moderate (50-69ml/min) or severe (<49ml/min) disease at onset. Discriminatory pathways and proteins included Notch-, integrin- and growth factor-signalling; microtubular kinase, vesicular proteins and epidermal growth factor substrates.Confocal microscopy of fluorescently-labelled normal versus ADPKD epithelial cell-derived exosomes in vitro also identified ADPKD-dependent abnormalities in intracellular vesicular trafficking and implicated changes in ADPKD-dependent exosome secretion and target cell uptake as factors underlying urinary exosome excretion biomarker properties.Comparative proteomic analysis of urinary exosomal proteins in individual patients before and after treatment with Tolvaptan for 4 years also identified distinct patterns of pathway modification dependent on the degree of effectiveness of the therapeutic response. Up-regulation of Wnt-pathway and vesicular proteins were characteristic of urinary exosomes from ADPKD patients with good responses to Tolvaptan while upregulation of angiogenesis pathways and additional molecular forms of vasopressin receptor AVPR2 were characteristic in urinary exosomes of ADPKD patients with poor responses.Taken together, these studies conclude that proteomic profiling of urinary exosome biomarkers provides a specific, sensitive and practical non-invasive method to identify and monitor the rate of disease progression and the effects of Tolvaptan therapy in individual ADPKD patients. This provides a means to identify those patients most likely to benefit maximally from therapy and to progress towards a personalization of ADPKD prognosis and management.

Urinary Proteomic Profile of Arterial Stiffness as a Predictor of Mortality and Cardiovascular Outcomes

Background: The underlying mechanisms of arterial stiffness has not been fully understood. This study aimed to illuminate its pathophysiology by using urinary proteomics and to determine the prognostic value of the urinary proteomic markers for adverse outcomes. Methods: Aortic stiffness was measured with carotid-femoral pulse wave velocity (PWV) in 669 randomly recruited Flemish (mean age, 50.5 years; 51.1% women). Urinary proteome was analyzed using capillary electrophoresis coupled with mass spectrometry. PWV-derived urinary proteomic score (PWV-UP) was developed by modelling PWV with proteomics data at baseline through orthogonal projections to latent structures. Adverse outcomes included all-cause mortality and fatal and non-fatal cardiovascular (CV) events. Linear regression and Cox proportional hazard regression were used. Pathway analysis was performed with the Reactome Pathway Database. Findings: PWV-UP that consisted of 2,336 peptides explained 65% variance of PWV, which was higher than 36% explained by clinical risk factors. After adjusting for potential confounders, PWV-UP was significantly associated with PWV (β=0.73 [95% confidence interval: 0.67–0.79], P<0.0001). Over a period of 9.2 years (median), 36 participants died and 75 experienced fatal and non-fatal CV events. The incidence of mortality and CV events was higher at the upper half of PWV-UP (P≤0.0002). The adjusted hazard ratios (+1 standard derivation) were 1.46 (1.08–1.97) for all-cause mortality, 2.04 (1.07–3.87) for CV mortality and 1.39 (1.11–1.74) for CV events (P≤0.031). For PWV, the corresponding estimates were 1.25 (0.97–1.60), 1.35 (0.85–2.15), and 1.22 (1.02–1.47), respectively (P≥0.033). The proteins in PWV-UP involved in multiple pathways, such as collagen turnover, cell adhesion, inflammation, and lipid metabolism. Interpretation: PWV-UP was independently associated with arterial stiffness and explained the variance of PWV better than clinical determinants. PWV-UP, but not PWV, independently predicted all-cause mortality and CV mortality, implying that PWV-UP-associated proteins may be multifaceted and involved in diverse pathological processes beyond arterial stiffness. Funding Information: European Union (HEALTH-F7-305507 HOMAGE), the European Research Council (Advanced Researcher Grant 2011-294713-EPLORE and Proof-of-Concept Grant 713601-uPROPHET), the European Research Area Net for Cardiovascular Diseases (JTC2017-046-PROACT), and KU Leuven (STG-18-00379) currently support the Studies Coordinating Centre in Leuven. Declaration of Interests: The authors declare no conflicts of interest. Ethics Approval Statement: The study was approved by the University of Leuven Ethics Committee. All participants provided written informed consent.

A multicenter prospective, randomized, placebo-controlled phase II/III trial for preemptive acute graft-versus-host disease therapy

Acute graft-versus-host disease (aGvHD) contributes to about 50% of transplant-related mortality (non-relapse mortality) after allogeneic hematopoietic stem cell transplantation (HSCT). Here the predictive value of a urinary proteomic profile (aGvHD_MS17) was tested together with preemptive prednisolone therapy. Two-hundred and fifty-nine of 267 patients were eligible for analysis. Ninety-two patients were randomized upon aGvHD_MS17 classification factor above 0.1 to receive either prednisolone (2–2.5 mg/kg, N = 44) or placebo (N = 47; N = 1 randomization failure) for 5 days followed by tapering. The remaining 167 patients formed the observation group. The primary endpoint of the randomized trial was incidence of aGvHD grade II between randomization and day +100 post HSCT. Analysis of the short-term preemptive prednisolone therapy in the randomized patients showed no significant difference in incidence or severity of acute GvHD (HR: 1.69, 95% CI: 0.66–4.32, P = 0.27). Prednisolone as preemptive treatment did not lead to an increase in relapse (20.2% in the placebo and 14.0% in the prednisolone group (P = 0.46)). The frequency of adverse events was slightly higher in the placebo group (64.4% versus 50%, respectively). Taken together, the results of the Pre-GvHD trial demonstrated the feasibility and safety of preemptive prednisolone treatment in the randomized patients.

Abstract 1610: Development and characterization of a novel semi-spontaneous bladder cancer model by pathological evaluation, single cell sequencing and proteomic profiling of urine and serum

Abstract Bladder cancer continues to be a healthcare challenge and for many years there have been very few new therapies providing benefit for patients either with non-muscle or muscle invasive bladder cancer. The molecular profiling of bladder cancer has evolved over the last years and different classifications systems have arisen; however, there is still a lack of knowledge on the prognostic or predictive therapeutic value based on these stratification methods. The clinical heterogeneity of the disease impacts the use of the molecular tools; in addition to that, the heterogeneity in the tumor microenvironment may hamper therapeutic strategies based on monotherapy approaches. As check-point blocking antibodies have entered the clinic for the treatment of muscle-invasive bladder cancer, and as novel therapeutic approaches are gaining interest in the non-muscle invasive treatment schedule, we may have novel therapeutic options for these patients ahead of us. But to be successful we also need to understand how to combine these therapies to improve overall survival and to impact relapse and progression of the disease. To enable an improved understanding of the landscape of the bladder cancer therapies we have developed a novel model based on a transgenic mouse with overexpression of Hgf and a dysregulated cell cycle. When exposing this transgenic strain to OH-BBN in drinking water these mice develop a non-muscle invasive bladder cancer that differ from a wt C57BL/6 mouse exposed to OH-BBN. The tumors are of squamous differentiated Tis/T1 type after a course of 10 weeks OH-BBN exposure. After another 5-10 weeks there is a noted muscle invasive squamous-like bladder cancer. The tumors show a strong inflammatory reaction. The muscle-invasive stage also leads to a marked change in the urine proteomic profile (as measured by the Proximity Extension Assay [PEA 92]). We have profiled both the micro and macro-hematuria and the time-course proteomic changes along with the pathology in this model. In addition, we recently performed single-cell sequencing on tumors from the non-muscle and muscle invasive model to dissect the molecular profile, immune landscape along with the heterogeneity. Thus, we can compare this novel model to the frequently used syngeneic MB49 tumor model. We are also currently assessing anti-PD1 therapy in the novel semi-spontaneous tumor model to evaluate the responsiveness to check-point blocking therapy. Ahead we will process the single cell sequencing data and dissect out what type of targeted therapies along with immunotherapies that should be combined, to improve the understanding of how to resolve the progression from a non-muscle invasive to a muscle-invasive deadly disease, with a hope to translate this into a clinically relevant therapeutic strategy. Citation Format: Iliana Kerzeli, Ramy Elgendy, Ivan Stepanek, Milena Doroszko, Aikaterini Chourlia, Sven Nelander, Per-Uno Malmstrom, Ulrika Segersten, Anca Dragomir, Martin Lord, Sara Mangsbo. Development and characterization of a novel semi-spontaneous bladder cancer model by pathological evaluation, single cell sequencing and proteomic profiling of urine and serum [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1610.

Urinary Proteome of Newborn Calves-New Potential in Non-Invasive Neonatal Diagnostic.

Simple SummaryUrine is an underappreciated biological sample in the context of health monitoring of newborn calves. It can be collected noninvasively and used for diagnostic purposes, which is an attractive option in veterinary examination. The establishment of urinary protein maps of healthy newborn calves is important for diagnosing and monitoring the progression of various diseases. The aim of this study was to characterize the urinary proteome profile of healthy newborn calves. We applied 2-D gel electrophoresis combined with MALDI-TOF-TOF-MS/MS and 1-D gel electrophoresis coupled with LC-MS-MS analysis to separate and identify proteins present in the urine of newborn calves. Urinary proteomic analysis allowed us to detect proteins which are characteristic of embryonic and neonatal development and proteins involved in the cardiovascular and digestive system, and bone and kidney development. Moreover, we identified proteins that can be used to assess normal glomerular and tubular physiology. In calves, one of the main causes of mortality in the neonatal period is diarrhea, affecting fluid and electrolyte balance. We also identified proteins involved in the renal transport of water and electrolytes, which can be useful in the early diagnosis and prevention of these type of disorders. Urine is a biological diagnostic material suitable not only for the analysis of kidney and urinary tract functions but also the function of other tissues and organs. The urine proteome of adult mammals differs from the urine proteome of neonatal ones. The establishment of urinary protein maps of healthy newborn calves is important for diagnosing and monitoring the progression of various diseases. The experiment was carried out on a Polish-Friesian var. of Black-and-White male calves in the sixth day of postnatal life. The two proteomics approaches used for separation and identification of urinary proteins were: 2-DE with MALDI-TOF-TOF-MS/MS and 1-DE with LC-MS/MS. This resulted in the identification of 692 urinary proteins. The majority of them were classified as extracellular proteins (40.32%), as well as proteins involved in regulation of major cellular processes (31.07%). We have observed the presence of unique proteins associated with embryonic (ameloblastin, alpha-fetoprotein, Delta-like protein, embryo-specific fibronectin 1 transcript variant, Indian hedgehog homolog) and kidney development (angiotensin-converting enzyme, angiotensinogen, aquaporin-1, calbindin, glypican 3, nidogen 1, pro-cathepsin H). Additionally, proteins involved in the renal regulation of water and electrolyte balance (angiotensinogen, angiotensin-converting enzyme, aquaporin-1, ezrin, uromodulin) were detected. Presented in the current study 1-D and 2-D urinary proteomic maps are the basis for the identification and detection of prognostic biomarkers important for defining a calf’s health status.

MO041URINE PROTEOMICS FOR PREDICTION OF DISEASE PROGRESSION IN PATIENTS WITH IGA NEPHROPATHY

Abstract Background and Aims Although IgA nephropathy (IgAN) is the most common primary glomerulonephritis in many parts of the world, risk of progressive kidney function decline is significant. Furthermore, the effect of immunosuppressive treatment in IgAN currently remains uncertain. There is no validated tool to predict disease progression or response to treatment. To serve this unmet need, we aimed at developing a urinary biomarker-based algorithm that predicts fast disease progression in patients with IgAN, thus enabling a personalized risk stratification and potentially improving patient management. Method In this multicenter study in 7 centers in Europe and in Canada urine samples were collected as part of clinical routine at time of biopsy in n= 300 patients (63% male, age 42±14 years) with biopsy proven IgAN. The follow-up data were collected for at least one year. Progressive disease was defined as an annual loss of kidney function (estimated glomerular filtration rate, eGFR) of more than -5ml/min/1.73m2 per year or when end-stage kidney disease (ESKD) was reached. The institutional review boards of each of the participating centers approved this study. Urine samples were analyzed using capillary electrophoresis coupled mass spectrometry (CE-MS). Whole proteome/peptidome profile was obtained for each sample. This study received funding from the European Union´s ERA PerMed program. Results Urine proteome/peptidome profiles were obtained from n=294 patients. On average, 2383 peptides were detected per sample. The data were subsequently divided into a discovery (n=154) and validation cohort (n=140). The comparison of the progressors (n=35) and non-progressors (n=119) in the discovery cohort resulted in the definition of more than 100 significant peptides. These included mainly fragments from collagen, mostly type 1 (decreased in progressors) and from different blood derived proteins like alpha-1-antitrypsin, alpha-2-HS-glycoprotein and apolipoproteins (increased in progressors). The distribution of the 100 most significant peptides in the progressor and non-progressor group is shown in the figure. The peptides were combined into a classifier using support vector machine. After optimizing the classifier employing a take-one-out procedure combined with n-1 cross-validation, the urine-peptide based algorithm enabled separation of progressors versus no progressors with an accuracy of 90% in take-one-out cross-validation. This classifier was subsequently applied to the validation cohort and resulted in highly significant separation of progressive from non-progressive IgAN patients. Furthermore, this classifier will be further applied blinded in an independent well characterized multicenter cohort of 267 IgAN patients. Conclusion We identified a urinary proteome profile which was associated with progressive loss of GFR in patients with IgAN. Further validation of this profile in an independent cohort is ongoing. The data indicate that CE-MS-based urinary proteomics enables identifying IgAN patients at high risk of disease progression. These patients may benefit from aggressive immunosuppressive treatment. Upon validation of the classifier in an independent cohort, its value in predicting response to immunosuppression will be assessed, aiming at establishing an innovative strategy that could improve patient management and personalize treatment of IgAN patients.

Urine metabolomics and proteomics for bladder cancer prediction by LC/MS based strategy

Early diagnosis and life‐long surveillance are clinically important to improve the long‐term survival of bladder cancer (BCa) patients. Cystoscopy is considered the gold standard for the clinical diagnosis of human BCa. However, it is expensive and invasive. The objective of this study was to discover novel biomarkers to distinguish BCa from healthy controls, urothelium carcinoma and benign bladder lesions by LC‐MS based metabolomics and proteomics. We also tried to differentiate high‐grade and low‐grade BCa. A total of 234 subjects were enrolled, including 80 healthy adults, 22 benign patients, 26 urothelium carcinoma patients and 106 BCa patients (63 high‐grade BCa, 43 low‐grade BCa). The results showed that both urine metabolites and urine proteins could distinguish the BCa group from the healthy group, urothelium carcinoma group and benign bladder lesions group. And the combination of metabolites and proteins could achieve higher accuracy. Similarly, BCa clinical stages (high‐ vs. low‐grade BCa) could be better separated with a panel consisting of metabolites and proteins than either the metabolite panel or the protein panel alone. In conclusion, this study showed that urine proteomic and metabolic profiling were able to assist in the detection of BCa and grading distinguish.Support or Funding InformationNational Basic Research Program of China (No. 2013CB530805, 2014CBA02005)

Identification of urinary proteomic profile of patients with chronic allograft nephropathy

ABSTRACT Background and objectives Chronic allograft nephropathy (CAN) is a major complication that occurs post-transplantation. At present, the diagnosis of CAN is based on renal biopsy. Therefore, there is an ultimate need to identify more specific and sensitive noninvasive methods for the early diagnosis of CAN. Recently, proteomic-based modalities have been developed to discover biomarkers of CAN. Methods Urine samples from 75 participants were collected. Participants were divided into three groups: Group I: 25 patients with CAN, Group II: 25 transplanted patients with stable renal functions, and Group III: 25 healthy control subjects matched for age and sex. Each group was divided into training set and test set. Specimens were purified with magnetic beads-based weak cation exchange chromatography and analyzed using MALDI-TOF MS. Results A Genetic Algorithm (GA) was used to set up the classification models. Five peaks represented the proteomic profile that differentiates between the CAN patients and the control group with sensitivity of 100%, specificity of 100%, recognition capability of 100%, and cross-validation of 91.7% and five peaks differentiate between the transplant patients with normal renal functions and the control groups with sensitivity of 96.8%, specificity of 95.5%, recognition capability of 98%, and cross-validation of 100%. Interpretation and conclusions We identified a pattern for CAN and transplant patients with normal renal functions by proteomic profiling using MALDI-TOF-MS and magnetic beads.

Re: Urine Proteomic Profiling in Patients with Nephrolithiasis and Cystinuria.

No AccessJournal of UrologyUrological Survey1 Jul 2019Re: Urine Proteomic Profiling in Patients with Nephrolithiasis and Cystinuria Dean G. AssimosMD Dean G. AssimosDean G. Assimos More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000265AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "Re: Urine Proteomic Profiling in Patients with Nephrolithiasis and Cystinuria." The Journal of Urology, 202(1), p. 25 Suggested Reading : Do urinary cystine parameters predict clinical stone activity?J Urol 2018; 199: 495. Link, Google Scholar : The interaction of thiol drugs and urine pH in the treatment of cystinuria. J Urol 2013; 189: 2147. Link, Google Scholar : Penicillamine therapy for pediatric cystinuria: experience from a cohort of American children. J Urol 2008; 180: 2620. Link, Google Scholar © 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 202Issue 1July 2019Page: 25-25 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Dean G. Assimos More articles by this author Expand All Advertisement Loading ...

Characterization of low molecular weight urinary proteins at varying time intervals in type 2 diabetes mellitus and diabetic nephropathy patients

BackgroundTo identify low molecular weight urinary proteins capable of detecting diabetic nephropathy patients which may predict renal alterations at early stages and prevent it from worsening further.MethodThree hundred ninety (390) age-matched subjects were divided into 8 groups depending upon duration of diabetes and the severity of renal damage. Urinary proteome profile of all subjects was determined with the help of microfluidic array. Participants with similar profile were further selected to study proteome map of urinary low molecular weight proteins with the help of 2 dimensional gel electrophoresis.ResultsOut of 390 total patients 268 patients showed a similar one dimensional proteomic pattern. Further, two-dimensional urinary proteomic pattern of these patients with molecular weight < 50 kDa was studied. Eight proteins with molecular weight 11, 15, 17, 23, 34, 38 and 46 kDa were identified with MALDI-QTOF. These low molecular weight proteins showed gradual increase in urinary excretion along with the duration of diabetes and severity of renal damage.ConclusionThe study concludes that proteomic analysis might be a useful tool for detecting some novel markers capable of detecting patients susceptible to diabetic nephropathy in the early phase.

Urine proteomic profiling in patients with nephrolithiasis and cystinuria.

The purpose of the study was to assess the differences in the concentration and function of urinary proteins between patients with cystine stones (CYS) and healthy controls (HC). We postulated that CYS and HC groups would demonstrate different proteomic profiles. A pilot study was performed comparing urinary proteomes of 10 patients with CYS and 10 age- and gender-matched HC, using liquid chromatography-mass spectrometry. Proteins which met the selection criteria (i) ≥ 2 unique peptide identifications; (ii) ≥ twofold difference in protein abundance; and (iii) ≤ 0.05 p value for the Fisher's Exact Test were analyzed using Gene Ontology classifications. Of the 2097 proteins identified by proteomic analysis, 398 proteins were significantly different between CYS and HC. Of those, 191 were involved in transport processes and 61 in inflammatory responses. The majority were vesicle-mediated transport proteins (78.5%), and 1/3 of them were down-regulated; of those, 12 proteins were involved in endosomal transport (including 6 charged multivesicular body proteins (CHMP) and 3 vacuolar sorting-associated proteins) and 9 in transmembrane transport. Myosin-2 and two actin-related proteins were significantly up-regulated in the vesicle-mediated transport group. We provide proteomic evidence of impaired endocytosis, dysregulation of actin and myosin cytoskeleton, and inflammation in CYS. Endosomal transport proteins were down-regulated mainly through defective CHMP. These findings may contribute to further understanding of the pathogenesis of CYS, potentially affecting its management.

Fast quantitative urinary proteomic profiling workflow for biomarker discovery in kidney cancer

BackgroundUrine has evolved as a promising body fluids in clinical proteomics because it can be easily and noninvasively obtained and can reflect physiological and pathological status of the human body. Many efforts have been made to characterize more urinary proteins in recent years, but few have focused on the analysis throughput and detection reproducibility. Increasing the urine proteomic profiling throughput and reproducibility is urgently needed for discovering potential biomarker in large cohorts.MethodsIn this study, we developed a fast and robust workflow for streamlined urinary proteome analysis. The workflow integrate highly efficient sample preparation technique and urinary specific data-independent acquisition (DIA) approach. The performance of the workflow was systematically evaluated and the workflow was subsequently applied in a proof-of-concept urine proteome study of 21 kidney cancer (KC) patients and 22 healthy controls.ResultsWith this workflow, the entire sample preparation process takes less than 3 h and allows multiplexing on standard centrifuges. Without pre-fractionation, our newly developed DIA method allows quantitative analysis of ~ 1000 proteins within 80 min of MS time (~ 15 samples/day). The quantitation accuracy of the whole workflow was excellent with median CV of 9.1%. The preliminary study on KC identified 125 significantly changed proteins.ConclusionsThe result suggested the feasibility of applying the high throughput workflow in extensive urinary proteome profiling and clinical relevant biomarker discovery.

Novel Urinary Peptidomic Classifier Predicts Incident Heart Failure.

BackgroundDetection of preclinical cardiac dysfunction and prognosis of left ventricular heart failure (HF) would allow targeted intervention, and appears to be the most promising approach in its management. Novel biomarker panels may support this approach and provide new insights into the pathophysiology.Methods and ResultsA retrospective comparison of urinary proteomic profiles generated by mass spectrometric analysis from 49 HF patients, 36 patients who progressed to HF within 2.6±1.6 years, and 192 sex‐ and age‐matched controls who did not progress to HF enabled identification of 96 potentially HF‐specific peptide biomarkers. Based on these 96 peptides, the classifier called Heart Failure Predictor (HFP) was established by support vector machine modeling. The incremental prognostic value of HFP was subsequently evaluated in urine samples from 175 individuals with asymptomatic diastolic dysfunction from an independent population cohort. Within 4.8 years, 17 of these individuals progressed to overt HF. The area under receiver‐operating characteristic curve was 0.70 (95% CI, 0.56–0.82); P=0.0047 for HFP and 0.57 (0.42–0.72; P=0.62) for N‐terminal pro b‐type natriuretic peptide. Hazard ratios were 1.63 (CI, 1.04–2.55; P=0.032) per 1‐SD increment in HFP and 0.70 (CI, 0.35–1.41; P=0.32) for a doubling of the logarithmically transformed N‐terminal pro b‐type natriuretic peptide.Conclusions HFP is a novel biomarker derived from the urinary proteome and might serve as a sensitive tool to improve risk stratification, patient management, and understanding of the pathophysiology of HF.

Proteomic profile‑based screening of potential protein biomarkers in the urine of patients with nephrotic syndrome.

Nephrotic syndrome is not a single disease; rather, it is a term for numerous diseases and pathological types. Renal biopsy is of use in determining the diagnosis and prognosis, and for guiding treatment; however, the use of this intervention is limited due to its invasive nature. Abnormal kidney‑derived proteins in the urine of patients provide useful information regarding numerous pathological processes that occur in the kidneys, and may be considered a potential non‑invasive biomarker for kidney disease. Proteomic analysis exhibits the advantage of being high‑throughput and has previously been used to identify biomarkers of disease. The present study aimed to identify abnormal kidney‑derived proteins in the urine of patients with nephrotic syndrome using a novel proteomic strategy. Urine samples from 5patients with nephrotic syndrome were subjected to acetone precipitation and albumin/immunoglobulin G depletion prior to analysis by two‑dimensional liquid chromatography tandem mass spectrometry. The resulting data were compared to a publicly available proteomic database of normal human plasma/urine and normal human kidney in PeptideAtlas, and of normal human kidney in the Human Protein Atlas. Candidate biomarkers were validated using ELISA analysis in 60patients with nephrotic syndrome: 30with focal segmental glomerulosclerosis (FSGS) and 30with minimal change disease (MCD), as well as in 30healthy controls. The initial screening identified 809 proteins in the urine of patients with nephrotic syndrome. A total of 13/809 proteins were additionally present in the kidney proteome of PeptideAtlas and the Human Protein Atlas, although not in normal human urine and normal human plasma according to PeptideAtlas; these were referred to as 'kidney‑derived disease‑associated proteins'. One of the kidney‑derived disease‑associated proteins, ubiquitin‑60S ribosomal protein L40 (UBA52) was observed to be increased in the urine of patients compared with normal controls [Creatinine, 637ng/mg (216‑1,851) vs.1.89ng/mg (1.37‑3.33), P<0.001; and 18.58ng/mg (11.11‑46.25) vs. 1.89ng/mg (1.37‑3.33), P<0.001)], and the urinary UBA52 levels were significantly increased in patients with FSGS compared with in patients with MCD (P<0.001). In conclusion, the present study identified potential novel urinary protein biomarkers for nephrotic syndrome, in addition to an extensive urinary proteomic profile of patients with nephrotic syndrome.

Urinary proteome profile of renal papillary necrosis in experimental model rats

Urinary proteome profile of renal papillary necrosis in experimental model rats

The Urine Proteome Profile Is Different in Neuromyelitis Optica Compared to Multiple Sclerosis (P5.304)

The Urine Proteome Profile Is Different in Neuromyelitis Optica Compared to Multiple Sclerosis (P5.304)

Proteomic identification of potential cancer markers in human urine using subtractive analysis.

Urine is an ideal medium in which to focus diagnostic cancer research due to the non-invasive nature and ease of sampling. Many large-scale proteomic studies have shown that urine is unexpectedly complex. We hypothesised that novel diagnostic cancer biomarkers could be discovered using a comparative proteomic analysis of pre-existing data. We assembled a database of 100 published datasets of 5,620 urinary proteins, as well as 46 datasets of 8,620 non-redundant proteins derived from kidney and blood proteome analyses. The data were then used to either subtract or compare molecules from a novel urinary proteome profiling dataset that we generated. We identified 1,161 unique proteins in samples from either cancer-bearing or healthy subjects. Subtractive analysis yielded a subset of 44 proteins that were found uniquely in urine from cancer patients, 30 of which were linked previously to cancer. In conclusion, this approach is useful in discovering novel biomarkers in tissues where unrelated profiling data is available. Only a limited disease-specific novel dataset is required to define new targets or substantiate previous findings. We have shared this discovery platform in the form of our Large Scale Screening Resource database, accessible through the Proteomic Analysis DataBase portal (www.PADB.org).

G.O.22 - Serum and urine proteomic profiling reveals biomarkers suitable for monitoring the outcome of therapeutic interventions in muscular dystrophies

G.O.22 - Serum and urine proteomic profiling reveals biomarkers suitable for monitoring the outcome of therapeutic interventions in muscular dystrophies

Differentially expressed urinary biomarkers in children with idiopathic nephrotic syndrome.

We performed a discovery phase of urinary proteomic profile in children with idiopathic nephrotic syndrome and validated selected biomarkers. Urinary proteomic profile was performed using isobaric tags for relative and absolute quantitation labeling, coupled with liquid chromatography-matrix assisted laser desorption and ionization analysis. Validation of biomarkers apolipoprotein A1, alpha 2 macroglobulin, orosomucoid 2, retinol binding protein 4 and leucine-rich alpha 2-glycoprotein 1 was done by enzyme-linked immunosorbent assay. Apolipoprotein A1 levels of <0.48 µg/mg of creatinine-differentiated steroid-resistant nephrotic syndrome (SRNS) from first episode nephrotic syndrome, area under curve (AUC) [0.99 (CI 0.9-1.0), 100 % sensitivity and 100 % specificity] and a value of <0.24 µg/mg of creatinine could differentiate SRNS from frequently relapsing nephrotic syndrome/steroid dependent nephrotic syndrome [AUC 0.99 (CI 0.9-1.0), 100 % sensitivity and 100 % specificity]. Alpha 2 macroglobulin could differentiate children with SRNS-focal segmental glomerulosclerosis (FSGS) from SRNS-minimal change disease (MCD) at values >3.3 µg/mg of creatinine [AUC 0.84 (CI 0.62-1.0), 90 % sensitivity and 85 % specificity]. Orosomucoid 2 >1.81 µg/mg of creatinine could distinguish SRNS-FSGS from SRNS-MCD [AUC 0.84 (CI 0.62-1.0), sensitivity 90 % and specificity 85.5 %]. RBP 4 value of >1.54 µg/mg of creatinine differentiated SRNS-FSGS from SRNS-MCD [AUC 0.87 (CI 0.68-1.0), sensitivity 90 % and specificity 85.7 %]. Lower level of apolipoprotein A1 in urine is suggestive of SRNS. Alpha 2 macroglobulin, retinol binding protein 4 and orosomucoid 2 are markers associated with FSGS, with alpha 2 macroglobulin being most predictive.