Urinary Biomarkers of Acute Kidney Injury in the Ovation-65 trial: A Nested Analysis of the Urinary Proteome
CCCF ePoster library. Jabrane M. 11/12/19; 285171; EP96
Mehdi Jabrane
Mehdi Jabrane
Login now to access Regular content available to all registered users.

You may also access this content "anytime, anywhere" with the Free MULTILEARNING App for iOS and Android
Abstract
Rate & Comment (0)
ePoster
Topic: Basic or Translational Science

Sabrina Bouchard1,2, Mehdi Jabrane1,2, Gabrielle Côté1,2, Neill K. Adhikari3,4, Marie-Claude Battista1,2, Marie-Hélène Masse1,2, Marie-Pierre Garant2, Dominic Lévesque1,2, Michaël Chassé5,6, Martine Lebrasseur6, Irene Watpool7, Frédérick, D’Aragon1,2, Marc-André Leclair1, Yannick Poulin1, Brian Grondin Beaudoin1, Dominique Bérard1, Ruxandra Pinto3, Ron Wald4, Elizabeth Wilcox4, Élaine Carbonneau2, Francois-Michel Boisvert1,2 & François Lamontagne1,2.
1 Université de Sherbrooke, Sherbrooke, Canada
2 Centre de recherche du CHU de Sherbrooke, Sherbrooke, Canada
3 Sunnybrook Health Sciences Centre, Toronto, Canada
4 University of Toronto, Toronto, Canada
5 Université de Montréal, Montréal, Canada
6 Centre hospitalier universitaire de Montréal, Montréal, Canada
7 Ottawa Hospital Research Institute, Ottawa, Canada

Introduction: OVATION65 is a randomized controlled trial comparing permissive hypotension vs. usual care on organ injury biomarkers. Our overarching objective is to select a biomarker reflecting acute kidney injury (AKI). Herein, we report preliminary results of a proteomics study of candidate biomarkers of AKI in OVATION65.

Objectives: 1) Quantify peptides and proteins expressed in the urine of OVATION65 participants using the proposed methodology;
2) Determine whether these proteins include 6 prespecified candidate biomarkers of kidney injury;
3) Describe renal outcomes that will ultimately be associated with proteomics clusters.

Methods: This analysis includes 65 OVATION65 participants from 7 Canadian hospitals. Patients were 65 years old with vasodilatory hypotension, ≤12 hours since vasopressor initiation and anticipated to remain on vasopressors for ≥6 more hours. Exclusion criteria included acute spinal cord or brain injury; acute hemorrhage, ventricular failure or post-cardiopulmonary bypass vasoplegia; <1 year since solid organ transplantation; extracorporeal life support at baseline; lacking commitment to life-sustaining therapies; previous enrollment in OVATION65; and lack of physician equipoise. Patients were randomized to permissive hypotension (mean arterial pressure [MAP] target 60-65 mmHg) or usual care. Prespecified renal outcomes are incident KDIGO stage 3 AKI, incident renal replacement therapy (RRT), and renal SOFA score of 4. SOFA scores are measured on days 1 (baseline), 2, 3, 7, 14 and 28 while in ICU; other renal outcomes are screened daily.

Fresh urine samples were collected on study days 1, 3, and 7. Following centrifugation, supernatants were stored at -80C. After thawing, trypsin digestion of urinary proteins, peptides were quantified using a TimsTOF Pro mass spectrometer coupled to high-performance liquid chromatography (HPLC). Analyses were performed in duplicate and blinded to group allocation and renal outcomes. We ascertained if 6 candidate biomarkers (NGAL, FABPL, CYTC, KIM-1, combination of TIMP2 and IGFBP7) could be detected. Hierarchical clustering of urinary proteins was performed using complete-linkage methodology.

Results: The mean age of OVATION65 participants was 75 (standard deviation [SD] 7) years, and 24 (37%) were women. The mean APACHE II score was 24 (SD 6), 23 participants (35.6%) had chronic kidney disease, and 4 (6%) received chronic dialysis at baseline. Stage 3 AKI occurred in 12 (18.5%) participants at a median 0.5 (interquartile range 0-2) days after randomization. Six (9.2%) participants received new RRT and 22 (33.9%) participants achieved renal SOFA score of 4. Urine samples were collected from 62 (92%), 58 (89%), and 45 (69%) participants on days 1, 3, and 7, respectively. Following analysis of these 165 urine samples, over 2,600 hydrosoluble proteins were quantified (average of four peptides per proteins, Figure 1), and grouped in 3 distinct clusters. The 6 prespecified candidate biomarkers were among the proteins expressed in detectable levels.

Conclusion: Using mass spectrometer coupled to HPLC, we quantified over 2,600 hydrosoluble proteins in urine samples from OVATION-65 participants. While we remained blinded to their quantity, prespecified candidate biomarkers were expressed in detectable amounts and will be included in subsequent hierarchical clustering analyses linking urinary proteins to renal outcomes.

Acknowledgement: This project was funded by the Lotte & John Hecht Memorial Foundation.

ePoster
Topic: Basic or Translational Science

Sabrina Bouchard1,2, Mehdi Jabrane1,2, Gabrielle Côté1,2, Neill K. Adhikari3,4, Marie-Claude Battista1,2, Marie-Hélène Masse1,2, Marie-Pierre Garant2, Dominic Lévesque1,2, Michaël Chassé5,6, Martine Lebrasseur6, Irene Watpool7, Frédérick, D’Aragon1,2, Marc-André Leclair1, Yannick Poulin1, Brian Grondin Beaudoin1, Dominique Bérard1, Ruxandra Pinto3, Ron Wald4, Elizabeth Wilcox4, Élaine Carbonneau2, Francois-Michel Boisvert1,2 & François Lamontagne1,2.
1 Université de Sherbrooke, Sherbrooke, Canada
2 Centre de recherche du CHU de Sherbrooke, Sherbrooke, Canada
3 Sunnybrook Health Sciences Centre, Toronto, Canada
4 University of Toronto, Toronto, Canada
5 Université de Montréal, Montréal, Canada
6 Centre hospitalier universitaire de Montréal, Montréal, Canada
7 Ottawa Hospital Research Institute, Ottawa, Canada

Introduction: OVATION65 is a randomized controlled trial comparing permissive hypotension vs. usual care on organ injury biomarkers. Our overarching objective is to select a biomarker reflecting acute kidney injury (AKI). Herein, we report preliminary results of a proteomics study of candidate biomarkers of AKI in OVATION65.

Objectives: 1) Quantify peptides and proteins expressed in the urine of OVATION65 participants using the proposed methodology;
2) Determine whether these proteins include 6 prespecified candidate biomarkers of kidney injury;
3) Describe renal outcomes that will ultimately be associated with proteomics clusters.

Methods: This analysis includes 65 OVATION65 participants from 7 Canadian hospitals. Patients were 65 years old with vasodilatory hypotension, ≤12 hours since vasopressor initiation and anticipated to remain on vasopressors for ≥6 more hours. Exclusion criteria included acute spinal cord or brain injury; acute hemorrhage, ventricular failure or post-cardiopulmonary bypass vasoplegia; <1 year since solid organ transplantation; extracorporeal life support at baseline; lacking commitment to life-sustaining therapies; previous enrollment in OVATION65; and lack of physician equipoise. Patients were randomized to permissive hypotension (mean arterial pressure [MAP] target 60-65 mmHg) or usual care. Prespecified renal outcomes are incident KDIGO stage 3 AKI, incident renal replacement therapy (RRT), and renal SOFA score of 4. SOFA scores are measured on days 1 (baseline), 2, 3, 7, 14 and 28 while in ICU; other renal outcomes are screened daily.

Fresh urine samples were collected on study days 1, 3, and 7. Following centrifugation, supernatants were stored at -80C. After thawing, trypsin digestion of urinary proteins, peptides were quantified using a TimsTOF Pro mass spectrometer coupled to high-performance liquid chromatography (HPLC). Analyses were performed in duplicate and blinded to group allocation and renal outcomes. We ascertained if 6 candidate biomarkers (NGAL, FABPL, CYTC, KIM-1, combination of TIMP2 and IGFBP7) could be detected. Hierarchical clustering of urinary proteins was performed using complete-linkage methodology.

Results: The mean age of OVATION65 participants was 75 (standard deviation [SD] 7) years, and 24 (37%) were women. The mean APACHE II score was 24 (SD 6), 23 participants (35.6%) had chronic kidney disease, and 4 (6%) received chronic dialysis at baseline. Stage 3 AKI occurred in 12 (18.5%) participants at a median 0.5 (interquartile range 0-2) days after randomization. Six (9.2%) participants received new RRT and 22 (33.9%) participants achieved renal SOFA score of 4. Urine samples were collected from 62 (92%), 58 (89%), and 45 (69%) participants on days 1, 3, and 7, respectively. Following analysis of these 165 urine samples, over 2,600 hydrosoluble proteins were quantified (average of four peptides per proteins, Figure 1), and grouped in 3 distinct clusters. The 6 prespecified candidate biomarkers were among the proteins expressed in detectable levels.

Conclusion: Using mass spectrometer coupled to HPLC, we quantified over 2,600 hydrosoluble proteins in urine samples from OVATION-65 participants. While we remained blinded to their quantity, prespecified candidate biomarkers were expressed in detectable amounts and will be included in subsequent hierarchical clustering analyses linking urinary proteins to renal outcomes.

Acknowledgement: This project was funded by the Lotte & John Hecht Memorial Foundation.

    This eLearning portal is powered by:
    This eLearning portal is powered by MULTIEPORTAL
Anonymous User Privacy Preferences

Strictly Necessary Cookies (Always Active)

MULTILEARNING platforms and tools hereinafter referred as “MLG SOFTWARE” are provided to you as pure educational platforms/services requiring cookies to operate. In the case of the MLG SOFTWARE, cookies are essential for the Platform to function properly for the provision of education. If these cookies are disabled, a large subset of the functionality provided by the Platform will either be unavailable or cease to work as expected. The MLG SOFTWARE do not capture non-essential activities such as menu items and listings you click on or pages viewed.


Performance Cookies

Performance cookies are used to analyse how visitors use a website in order to provide a better user experience.


Save Settings