Ignite Creation Date:
2024-05-06 @ 1:19 PM
Last Modification Date:
2024-10-26 @ 1:12 PM
Study NCT ID:
NCT03991793
Status:
UNKNOWN
Last Update Posted:
2019-06-19
First Post:
2019-06-13
Brief Title:
Granzyme A in Patients With E Coli Bacteremic Urinary Tract Infections
Sponsor:
Instituto de Investigación Sanitaria Aragón
Organization:
Instituto de Investigación Sanitaria Aragón
Study Overview
Official Title:
Granzyme A in Patients With E Coli Bacteremic Urinary Tract Infections
Status:
UNKNOWN
Status Verified Date:
2019-06
Last Known Status:
NOT_YET_RECRUITING
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
GABEC
Brief Summary:
Background Survival in Granzyme A gene gzmA knocked-out mice was significantly longer than in wild-type mice in a murine peritonitis model cecal ligation puncture
Hypothesis GZM A has a pathogenic role in sepsis in humans and gzmA polymorphisms can help to predict the risk of sepsis among patients with systemic infections E coli bacteremic urinary tract infections
Objectives
1 To assess the correlation between GZM A serum levels and systemic inflammatory response in a human model of infectionsepsis E coli bacteremic UTI
2 To characterize gzmA polymorphisms among patients with E coli bacteremic UTI
3 To determine GZM A serum kinetics among patients with E coli bacteremic UTI
4 To characterize E coli strains causing bacteremic UTI antimicrobial phenotype and virulence factors virulome
Methods
Design and setting Prospective nested case-control study
Study population consecutive adult patients with bacteremic urinary tract infections UTIs caused by E coli
Exclusion criteria Patients with conditions that significantly compromise immune status or patients exposed to urologic procedures
Estimated sample size 50 patients with a sepsis non sepsis 11 ratio Septic and non septic patients will be matched on gender age - 10 years comorbidity Charlson score -1 time symptom onset to blood culture - 24h
Measurements GZM A serum levels will be determined on day 0 day 2-3 day 30 GZM A kinetics gzmA polymorphisms whole exome sequencingWhole genome sequencing of E coli isolates retrieved from blood cultures will be performed
Analysis Association between GZM A levels and gzmA polymorphisms and sepsis will be analyzed adjusting for patient infection and microorganism-related factors multivariate analysis
Hypothesis GZM A has a pathogenic role in sepsis in humans and gzmA polymorphisms can help to predict the risk of sepsis among patients with systemic infections E coli bacteremic urinary tract infections
Objectives
1 To assess the correlation between GZM A serum levels and systemic inflammatory response in a human model of infectionsepsis E coli bacteremic UTI
2 To characterize gzmA polymorphisms among patients with E coli bacteremic UTI
3 To determine GZM A serum kinetics among patients with E coli bacteremic UTI
4 To characterize E coli strains causing bacteremic UTI antimicrobial phenotype and virulence factors virulome
Methods
Design and setting Prospective nested case-control study
Study population consecutive adult patients with bacteremic urinary tract infections UTIs caused by E coli
Exclusion criteria Patients with conditions that significantly compromise immune status or patients exposed to urologic procedures
Estimated sample size 50 patients with a sepsis non sepsis 11 ratio Septic and non septic patients will be matched on gender age - 10 years comorbidity Charlson score -1 time symptom onset to blood culture - 24h
Measurements GZM A serum levels will be determined on day 0 day 2-3 day 30 GZM A kinetics gzmA polymorphisms whole exome sequencingWhole genome sequencing of E coli isolates retrieved from blood cultures will be performed
Analysis Association between GZM A levels and gzmA polymorphisms and sepsis will be analyzed adjusting for patient infection and microorganism-related factors multivariate analysis
Detailed Description:
1 Research hypothesis
The research team has explored the role of GZM A
1 Conceptual hypothesis
Granzyme A is a pathogenic sepsis mediator
Granzyme A gene polymorphisms determine the serum concentration of GZM A in patients with systemic infections
Granzyme A gene polymorphisms determine the risk of sepsis among patients with systemic infections
2 Operational hypothesis
Among patients with bacteremic E coli urinary tract infections UTIs GZM A levels are significantly higher in those patients who develop sepsis as compared with those who do not develop sepsis
There are significant differences in the GZM A gene polymorphism profile of patients with bacteremic E coli UTIs who develop sepsis as compared with those who do not develop sepsis
2 Aims and objectives
Aims
1 To assess the pathogenic role of GZM A in sepsis in patients with bacteremic E coli UTIs
2 To explore the capability of GZM A polymorphisms to anticipate the risk of developing sepsis in patients with bacteremic E coli UTIs
3 To evaluate the potential usefulness of GZM A as a diagnostic biomarker of sepsis in patients with bacteremic E coli UTIs
4 To characterize E coli virulome among circulating uropathogenic strains
Objectives
1 To evaluate the correlation between serum levels of GZM A and systemic inflammatory response in patients with bacteremic E coli UTIs
2 To characterize GZM A gene polymorphisms among patients with bacteremic E coli UTIs
3 To assess GZM A serum kinetics among patients with bacteremic E coli UTIs
4 To phenotypically and molecularly characterize E coli strains causing bacteremic UTIs including their virulence factors virulome
3 Expected outcomes
Characterization of the pathogenic role of GZM A in sepsis in patients with systemic infections
Characterization of GZM A as a sepsis biomarker in a human model of infection-sepsis
Phenotypical and molecular characterization of uropathogenic E coli causing bloodstream infections
4 Methods
41 Design and project scope
- Prospective exploratory nested case-control study to be conducted at one academic hospital Hospital Clinico Universitario Lozano Blesa affiliated with the Instituto de Investigacion Sanitaria Aragon IIS Aragon
42 Study period June 2019 - December 2020
43 Patients and sample size
Inclusion criteria To meet all
Age 18 years old
E coli bloodstream infection
Urinary source Urinary source should be considered if any of the following
1 Urinary source is clinically suspected and both E coli isolates in blood and in urine culture share the same phenotype antibiogram
2 Urinary sourceorigin is clinically suspected and the urine culture is negative but the patient had received at least one dose of any systemic antibiotic with antimicrobial activity against the E coli strain causing the BSI before blood cultures were obtained
3 Both isolates in blood and in urine culture share the same phenotype antibiogram and there is no alternative source
Exclusion criteria
1 Use of systemic antibiotics for 48h in the two months preeeding the episode
2 Immunocompromised hosts - Patients receiving systemic steroid use 10 mg prednisoneday during 10 or more days in the previous 2 months
Patients receiving biological therapy previous 2 months
Active solid or hematological cancer
HIV
Neutropenia 500 PMNmicrol
3 Basal urinary tract abnormalities or locally modified vesical microbiome any
- Ureteroileostomy ureterosigmoidostomy ureterostomy Bricker or nephrostomy
Indwelling urinary catheter in the last two months
Urological surgery in the last two months
Intravesical chemotherapy in the last two months
Intravesical BCG instillation in the last two months
Potential candidates will be detected daily by the microbiologists on the research team Inclusion criteria will be verified in the multidisciplinary meeting that antimicrobial stewardship teams AST conduct on a daily basis at the participating hospital
Estimated size of the study population Matching
- 50 patients with a sepsis non sepsis 11 ratio will be included
- Septic and non-septic patients will be matched on gender age - 10 years comorbidity Charlson score -1 time symptom onset to blood culture - 24h
44 Definitions
Cases sepsis septic shock
- Sepsis or shock septic are defined as life-threatening organ dysfunction caused by a dysregulated host response to infection according to the 2001 SCCMESICMACCPATSSIS International Sepsis Definition Conference
Controls
- Absence of sepsis or septic shock according to the 2001 SCCMESICMACCPATSSIS International Sepsis Definition Conference
45 Variables
451 Patient-related variables
- Demographical gender age
Comorbidity Modified Charlson Index
Baseline serum creatinine
452 Infection-related variables
- Time from the onset of symptoms to the start of antimicrobial treatment
- Time from the onset of symptoms to the start of appropriate antimicrobial treatment
- Time form the onset of symptoms to the surgical therapy if needed
Clinical severity at time 0 blood culture and day 2- 3 and day 30 sepsis score
453 Inflammation sepsis mediators and biomarkers - The following biomarkers will be determined during patient enrollment - White blood cell count and differential - Platelet count - Fibrinogen - Prothrombin activity
- C reactive protein
Procalcitonin
GZM A
GZM A serum levels will be obtained in all patients during the enrollment visit the 2-3 day and the 30 day visits GZM A kinetics GZM A levels will be determined by an ELISA commercial assay Human Granzyme A ELISA development kitHRP Mabtech
GzmA gene polymorphisms as well as other potentially associated mutations will be screened by Whole Exome Sequencing WES To this aim we will use DNA isolated from peripheral blood cells and the AmpliSeq technology kit on the Ion Torrent platform following the manufacturers instructions This platform is available at the Genomics Central Research Unit CRU at CIBA from University of ZaragozaIIS Aragon All kits for isolating and analyzing DNA samples are commercially available and optimized by Thermo Scientific Bioninformatic analysis will be performed by an agreement established between Genomics CRU and Micromics SL led by Pedro Gonzalez at CRG in Barcelona
454 Microbiological variables
Minimum inhibitory concentrations MIC of the E coli isolates retrieved from urine and blood cultures will be determined through automated microdilution panels as routinely performed by the Microbiology Laboratory of participating hospitals
Whole genome sequencing WGS of the E coli strains isolated in blood and urine cultures The presence of E coli known virulence factors will be analyzed and a virulence score for each strain will be calculated Mora-Rillo et al 2015
The research team has explored the role of GZM A
1 Conceptual hypothesis
Granzyme A is a pathogenic sepsis mediator
Granzyme A gene polymorphisms determine the serum concentration of GZM A in patients with systemic infections
Granzyme A gene polymorphisms determine the risk of sepsis among patients with systemic infections
2 Operational hypothesis
Among patients with bacteremic E coli urinary tract infections UTIs GZM A levels are significantly higher in those patients who develop sepsis as compared with those who do not develop sepsis
There are significant differences in the GZM A gene polymorphism profile of patients with bacteremic E coli UTIs who develop sepsis as compared with those who do not develop sepsis
2 Aims and objectives
Aims
1 To assess the pathogenic role of GZM A in sepsis in patients with bacteremic E coli UTIs
2 To explore the capability of GZM A polymorphisms to anticipate the risk of developing sepsis in patients with bacteremic E coli UTIs
3 To evaluate the potential usefulness of GZM A as a diagnostic biomarker of sepsis in patients with bacteremic E coli UTIs
4 To characterize E coli virulome among circulating uropathogenic strains
Objectives
1 To evaluate the correlation between serum levels of GZM A and systemic inflammatory response in patients with bacteremic E coli UTIs
2 To characterize GZM A gene polymorphisms among patients with bacteremic E coli UTIs
3 To assess GZM A serum kinetics among patients with bacteremic E coli UTIs
4 To phenotypically and molecularly characterize E coli strains causing bacteremic UTIs including their virulence factors virulome
3 Expected outcomes
Characterization of the pathogenic role of GZM A in sepsis in patients with systemic infections
Characterization of GZM A as a sepsis biomarker in a human model of infection-sepsis
Phenotypical and molecular characterization of uropathogenic E coli causing bloodstream infections
4 Methods
41 Design and project scope
- Prospective exploratory nested case-control study to be conducted at one academic hospital Hospital Clinico Universitario Lozano Blesa affiliated with the Instituto de Investigacion Sanitaria Aragon IIS Aragon
42 Study period June 2019 - December 2020
43 Patients and sample size
Inclusion criteria To meet all
Age 18 years old
E coli bloodstream infection
Urinary source Urinary source should be considered if any of the following
1 Urinary source is clinically suspected and both E coli isolates in blood and in urine culture share the same phenotype antibiogram
2 Urinary sourceorigin is clinically suspected and the urine culture is negative but the patient had received at least one dose of any systemic antibiotic with antimicrobial activity against the E coli strain causing the BSI before blood cultures were obtained
3 Both isolates in blood and in urine culture share the same phenotype antibiogram and there is no alternative source
Exclusion criteria
1 Use of systemic antibiotics for 48h in the two months preeeding the episode
2 Immunocompromised hosts - Patients receiving systemic steroid use 10 mg prednisoneday during 10 or more days in the previous 2 months
Patients receiving biological therapy previous 2 months
Active solid or hematological cancer
HIV
Neutropenia 500 PMNmicrol
3 Basal urinary tract abnormalities or locally modified vesical microbiome any
- Ureteroileostomy ureterosigmoidostomy ureterostomy Bricker or nephrostomy
Indwelling urinary catheter in the last two months
Urological surgery in the last two months
Intravesical chemotherapy in the last two months
Intravesical BCG instillation in the last two months
Potential candidates will be detected daily by the microbiologists on the research team Inclusion criteria will be verified in the multidisciplinary meeting that antimicrobial stewardship teams AST conduct on a daily basis at the participating hospital
Estimated size of the study population Matching
- 50 patients with a sepsis non sepsis 11 ratio will be included
- Septic and non-septic patients will be matched on gender age - 10 years comorbidity Charlson score -1 time symptom onset to blood culture - 24h
44 Definitions
Cases sepsis septic shock
- Sepsis or shock septic are defined as life-threatening organ dysfunction caused by a dysregulated host response to infection according to the 2001 SCCMESICMACCPATSSIS International Sepsis Definition Conference
Controls
- Absence of sepsis or septic shock according to the 2001 SCCMESICMACCPATSSIS International Sepsis Definition Conference
45 Variables
451 Patient-related variables
- Demographical gender age
Comorbidity Modified Charlson Index
Baseline serum creatinine
452 Infection-related variables
- Time from the onset of symptoms to the start of antimicrobial treatment
- Time from the onset of symptoms to the start of appropriate antimicrobial treatment
- Time form the onset of symptoms to the surgical therapy if needed
Clinical severity at time 0 blood culture and day 2- 3 and day 30 sepsis score
453 Inflammation sepsis mediators and biomarkers - The following biomarkers will be determined during patient enrollment - White blood cell count and differential - Platelet count - Fibrinogen - Prothrombin activity
- C reactive protein
Procalcitonin
GZM A
GZM A serum levels will be obtained in all patients during the enrollment visit the 2-3 day and the 30 day visits GZM A kinetics GZM A levels will be determined by an ELISA commercial assay Human Granzyme A ELISA development kitHRP Mabtech
GzmA gene polymorphisms as well as other potentially associated mutations will be screened by Whole Exome Sequencing WES To this aim we will use DNA isolated from peripheral blood cells and the AmpliSeq technology kit on the Ion Torrent platform following the manufacturers instructions This platform is available at the Genomics Central Research Unit CRU at CIBA from University of ZaragozaIIS Aragon All kits for isolating and analyzing DNA samples are commercially available and optimized by Thermo Scientific Bioninformatic analysis will be performed by an agreement established between Genomics CRU and Micromics SL led by Pedro Gonzalez at CRG in Barcelona
454 Microbiological variables
Minimum inhibitory concentrations MIC of the E coli isolates retrieved from urine and blood cultures will be determined through automated microdilution panels as routinely performed by the Microbiology Laboratory of participating hospitals
Whole genome sequencing WGS of the E coli strains isolated in blood and urine cultures The presence of E coli known virulence factors will be analyzed and a virulence score for each strain will be calculated Mora-Rillo et al 2015
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None