Proteomic Pattern in Female Stress Urinary Incontinence: a Pilot Study
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|ClinicalTrials.gov Identifier: NCT02023502|
Recruitment Status : Unknown
Verified December 2013 by Univ.-Prof.Dr.Dr.h.c. Heinz Kölbl, Medical University of Vienna.
Recruitment status was: Recruiting
First Posted : December 30, 2013
Last Update Posted : December 30, 2013
Objective: The primary objective of the study is the comparison of protein concentrations between patients with stress urinary incontinence (SUI) and healthy controls.
Aim: This pilot study aims to determine a possible altered protein profile in women suffering from SUI, compared to healthy women and therefore to discriminate a potential protein biomarker for SUI.
Main outcome measure: mass spectrometric measuring of urinary proteomic secretome in diseased and healthy subjects (sequence coverage and number of identified proteins)
Trial design: Prospective case- control study
Setting: Department of Gynecology and Obstetrics of the Medical University of Vienna in co- operation with the Core Facilities Proteomics of the Medical University of Vienna
Population: Twenty female patients with SUI and twenty healthy patients who attend the outpatient clinic of the Department of Obstetrics and Gynaecology, Medical University of Vienna (sample size calculation FDR 0.05, power of 80%, assumed proportion of true H0 0.95, assumed standardized effect size of 1)
Methods: Examinations to be carried out: patient history, provocative stress test, ICIQ short form questionnaire, residual urine volume (ultrasound) and urine analysis (dipstick testing). A urine sample is obtained from the patient after spontaneous micturition, to which protease inhibitor will be added immediately. Two serum blood vials (each 9ml) are taken from a peripheral vein of the patient for routine laboratory and further research.
Proteomics analysis will be performed using chromatographic separation (LC) with mass spectrometric detection (MS).
|Condition or disease||Intervention/treatment|
|Female Stress Urinary Incontinence||Other: collection of urine and blood sample|
Sample preparation and analysis will include:
- Sample collection and immediate addition of protease inhibitor cocktail (Roche, Complete Protease Inhibitor Cocktail).
- Urine centrifugation and filtration for removal of cell debris.
- Protein precipitation by applying methanol/chloroform separation for removal of all possible non-proteinic substances.
- Protein digestion applying in-solution trypsin, pepsin, and Glu-C.
Peptide separation using nano HPLC and different chromatographic approaches.
- Reversed phase (RP) separation of peptides and MS detection
- Separation of peptides using HILIC (hydrophilic Interaction Liquid Chromatography) for discrimination of polar peptides.
- In addition to RP and HILIC directly coupled to MS, digested peptides will be separated using multidimensional approaches. Weak anion exchange columns operated under HILIC conditions will be used to separate peptides carrying posttranslational modifications such as phosphorylation or acetylation, thus, increasing the dynamic range of detection. During this separation, fractions will be collected and re-injected onto the RP and HILIC with MS detection.
- All nano HPLC separations will be performed using biocompatible separation system.
Mass spectrometric analysis of digested peptides will be performed using two different detection methods: ion-trap and the time-of-flight (qToF) MS. qToF mass spectrometric detection and analysis will also be used for the label-free quantitation of peptides and proteins detected in samples. All measurements will be performed in triplicate to provide corrections for technical variability of separation and ionization.
General database search will be performed using the Human SwissProt Database in its actual version at the time of analysis. Data search will be performed using Mascot (http://www.matrixscience.com/) and X!Tandem (http://www.thegpm.org/tandem/) search machines, and the final data allocation and filtering by using Scaffold (www.proteomsoftware.com).
Data analysis will be conducted with the bioconductor package limma. Before data analysis, data will be filtered by excluding proteins with measurements with a low interquartile range. Groups will be compared by moderated paired t-statistics adjusting for age. Adjustment for multiple testing will be done by Benjamini-Hochberg correction controlling the FDR at 0.05.
|Study Type :||Observational|
|Estimated Enrollment :||40 participants|
|Observational Model:||Case Control|
|Official Title:||Proteomic Pattern in Female Stress Urinary Incontinence: a Pilot Study|
|Study Start Date :||November 2013|
|Estimated Primary Completion Date :||April 2015|
stress urinary incontinence
Patients presenting with stress urinary incontinence according to the inclusion and exclusion criteria
Other: collection of urine and blood sample
healthy women with the same inclusion and exclusion criteria as the case group, except for stress urinary incontinence
Other: collection of urine and blood sample
- mass spectrometric measuring of urinary proteomic secretome in diseased and healthy subjects (sequence coverage and number of identified proteins) [ Time Frame: The primary outcome measure will be assessed for each participant in a time frame of 4-7 days after recruitment ]Comparison of protein concentrations based on number of identified proteins and mass spectrometric spectral count for patients with stress urinary incontinence and healthy controls.
To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT02023502
|Medical University of Vienna, Department of Obstetrics and Gynaecology||Recruiting|
|Vienna, Austria, 1090|
|Contact: Heinz Kölbl, Univ.-Prof.Dr.Dr.h.c. 0043 1 40400 2915 firstname.lastname@example.org|
|Principal Investigator: Heinz Kölbl, Univ.-Prof.Dr.Dr.h.c.|
|Sub-Investigator: Rosa Maria Laterza, Dr.med.univ.|
|Sub-Investigator: Marianne Koch, Dr.med.univ.|
|Sub-Investigator: Goran Mitulovic, Ass.-Prof.Mag.Dr.|
|Sub-Investigator: Engelbert Hanzal, Univ.-Prof.Dr.med.univ.|
|Sub-Investigator: Wolfgang Umek, Univ.-Prof.Dr.med.univ.|
|Sub-Investigator: Irene Steiner, Mag.|
|Principal Investigator:||Heinz Kölbl, Univ.-Prof.Dr.Dr.h.c.||Medical University of Vienna, Department of Obstetrics and Gynaecology|