Ignite Creation Date:
2024-05-06 @ 6:48 PM
Last Modification Date:
2024-10-26 @ 2:55 PM
Study NCT ID:
NCT05795257
Status:
RECRUITING
Last Update Posted:
2024-01-24
First Post:
2023-03-21
Brief Title:
Pulmonary Immune Cell-microbiome Interactions in ARDS
Sponsor:
Hvidovre University Hospital
Organization:
Hvidovre University Hospital
Study Overview
Official Title:
Pulmonary Immune Cell-microbiome Interactions in Acute Respiratory Distress Syndrome The ILLUMINA-1 Study
Status:
RECRUITING
Status Verified Date:
2024-01
Last Known Status:
None
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:
ILLUMINA-1
Brief Summary:
The overall aim is to compare the composition and spatial heterogeneity of the following in critically ill intensive care unit ICU patients i immune cell populations and their activation patterns ii the surrounding cytokine-chemokine milieu including trans-compartmental fluxes of these mediators between the lung and bloodstream and iii the lung microbiome
Main hypotheses
The immune cell population in bronchoalveolar lavage fluid BALF from patients with ARDS is dominated by neutrocytes while T cells are depleted and show evidence of hyper-activation and exhaustion
T cell hyper-activation and exhaustion is specifically compartmentalised to the lungs and much more pronounced in moderate-to-severe than none-to-mild ARDS
Cyto- and chemokines derived from pulmonary immune cells are higher in moderate-to-severe than none-to-mild ARDS with a greater release from lungs to the bloodstream notably of IL-6 and IL-8
The differences in T cell profile in BALF notably the ratio between regulatory T cells and T helper 17 cells will change with disease severity over time and can be explained by the presence of tI-IFN antibodies andor a low microbial diversity of the respiratory tract with low enrichment from the oral cavity
Main hypotheses
The immune cell population in bronchoalveolar lavage fluid BALF from patients with ARDS is dominated by neutrocytes while T cells are depleted and show evidence of hyper-activation and exhaustion
T cell hyper-activation and exhaustion is specifically compartmentalised to the lungs and much more pronounced in moderate-to-severe than none-to-mild ARDS
Cyto- and chemokines derived from pulmonary immune cells are higher in moderate-to-severe than none-to-mild ARDS with a greater release from lungs to the bloodstream notably of IL-6 and IL-8
The differences in T cell profile in BALF notably the ratio between regulatory T cells and T helper 17 cells will change with disease severity over time and can be explained by the presence of tI-IFN antibodies andor a low microbial diversity of the respiratory tract with low enrichment from the oral cavity
Detailed Description:
Background Pulmonary hyperinflammation with neutrocyte and macrophage invasion and T cell depletion are common features of the acute respiratory distress syndrome ARDS but it remains to be elucidated whether the T cells in the lungs of patients with ARDS are hyperactivated andor exhausted and to which extent this contributes to neutrocyte invasion and thus lung tissue destruction Furthermore the pulmonary microbiome has shown a reduction in diversity and interactions in ARDS which may be important for normal T cell function At present immune cell-microbiome interactions and their relation to disease severity and progression have not yet been studied in ARDS
Overall design In 20 mechanically ventilated patients with none-to-mild and 20 with moderate-to-severe non-COVID-19 ARDS according to the Berlin definition an endotracheal aspirate and BAL fluid BALF from separate lung segments will be obtained Furthermore an oral and nasal swab and blood samples will be collected This will be done within 72 hours after intubation and again after 7-10 days if the patient is still intubated
Patients electronic health record The following is obtained after inclusion diagnosis codes and medication type and dosage including vasopressors and sedatives and smoking history currentpreviousnever smoker pack years admission time blood pressure heart rhythm and heart rate temperature ventilator settings supportive care dialysis ECMO blood tests results at admission and on the study days blood cell counts coagulation parameters renal liver and electrolyte panel arterial and mixed venous blood gases clinical scores at admission and on the study days SAPS3 APACHE II SOFA death within 30 days
Blood sampling Arterial blood samples are drawn from the patients invasive arterial catheter inserted at ICU admission for clinical purposes for continuous invasive blood pressure monitoring and repeated arterial blood gas collection immediately before BALF collection
Bronchoscopy with BALF collection This procedure is performed in a standardized fashion according to current clinical guidelines Immediately prior to the procedure an oral swab nasal swab and an endotracheal aspirate ETA are obtained FIO2 is then increased to 10 and the bronchoscopy procedure is performed using a disposable videoscope with an outer diameter of 50 mm Three successive 50-ml aliquots of prewarmed 37C isotonic saline are instilled in the medial segment of the right middle lobe aspirated immediately with low negative suction pressure 100 cm H2O and pooled into a sterile glass container on ice to obtain a BALF specimen
Afterwards a mini-BAL is performed in the upper and lower lobe of the right lung with a single installation of 20 ml isotonic saline in each lobe with immediately aspiration into a sterile container
Measurements The composition of the immune cell population as well as the function and differentiation of various cell lines will be investigated by single-cell RNA sequencing scRNA-seq on selected immune cells from BALF ETA and blood and this will be supplemented by bulk RNA sequencing with sample barcoding and multiplexing giving a detailed expression pattern of all samples
The composition microbiome in BALF ETA and oral swabs will be assessed by targeted amplicon sequencing of the hyper-variable regions 1 through 3 of the 16S subunit of ribosomal RNA gene for bacteria
Statistical analyses will be performed using R statistical software version 411 R Project for Statistical Computing within RStudio statistical software version 141717 RStudio and p005 considered statistically significant Inspection of normality and variance homogeneity will be done by creating qq-plots and histograms The statistical inference tools SPIEC-EASI and HeatMaps will be used and based on correlational analyses principal component analyses including non-hierarchal cluster analysis will be applied to identify traits in the two groups
Overall design In 20 mechanically ventilated patients with none-to-mild and 20 with moderate-to-severe non-COVID-19 ARDS according to the Berlin definition an endotracheal aspirate and BAL fluid BALF from separate lung segments will be obtained Furthermore an oral and nasal swab and blood samples will be collected This will be done within 72 hours after intubation and again after 7-10 days if the patient is still intubated
Patients electronic health record The following is obtained after inclusion diagnosis codes and medication type and dosage including vasopressors and sedatives and smoking history currentpreviousnever smoker pack years admission time blood pressure heart rhythm and heart rate temperature ventilator settings supportive care dialysis ECMO blood tests results at admission and on the study days blood cell counts coagulation parameters renal liver and electrolyte panel arterial and mixed venous blood gases clinical scores at admission and on the study days SAPS3 APACHE II SOFA death within 30 days
Blood sampling Arterial blood samples are drawn from the patients invasive arterial catheter inserted at ICU admission for clinical purposes for continuous invasive blood pressure monitoring and repeated arterial blood gas collection immediately before BALF collection
Bronchoscopy with BALF collection This procedure is performed in a standardized fashion according to current clinical guidelines Immediately prior to the procedure an oral swab nasal swab and an endotracheal aspirate ETA are obtained FIO2 is then increased to 10 and the bronchoscopy procedure is performed using a disposable videoscope with an outer diameter of 50 mm Three successive 50-ml aliquots of prewarmed 37C isotonic saline are instilled in the medial segment of the right middle lobe aspirated immediately with low negative suction pressure 100 cm H2O and pooled into a sterile glass container on ice to obtain a BALF specimen
Afterwards a mini-BAL is performed in the upper and lower lobe of the right lung with a single installation of 20 ml isotonic saline in each lobe with immediately aspiration into a sterile container
Measurements The composition of the immune cell population as well as the function and differentiation of various cell lines will be investigated by single-cell RNA sequencing scRNA-seq on selected immune cells from BALF ETA and blood and this will be supplemented by bulk RNA sequencing with sample barcoding and multiplexing giving a detailed expression pattern of all samples
The composition microbiome in BALF ETA and oral swabs will be assessed by targeted amplicon sequencing of the hyper-variable regions 1 through 3 of the 16S subunit of ribosomal RNA gene for bacteria
Statistical analyses will be performed using R statistical software version 411 R Project for Statistical Computing within RStudio statistical software version 141717 RStudio and p005 considered statistically significant Inspection of normality and variance homogeneity will be done by creating qq-plots and histograms The statistical inference tools SPIEC-EASI and HeatMaps will be used and based on correlational analyses principal component analyses including non-hierarchal cluster analysis will be applied to identify traits in the two groups
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