Ignite Creation Date:
2024-10-26 @ 3:41 PM
Last Modification Date:
2024-10-26 @ 3:41 PM
Study NCT ID:
NCT06611046
Status:
COMPLETED
Last Update Posted:
None
First Post:
2024-09-19
Brief Title:
MicroRNA Profile As a Biomarker of Liver Damage in Different Types of Liver Donors
Sponsor:
None
Organization:
None
Study Overview
Official Title:
Perfil De MicroRNA Como Biomarcador De Daño Hepático En Diferentes Tipos De Donantes De Hígado MicroRNA Profile As a Biomarker of Liver Damage in Different Types of Liver Donors
Status:
COMPLETED
Status Verified Date:
2024-09
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
No
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
The goal of this observational study is to analyze the expression of liver-derived miRNAs in different types of liver donors to gain in depth knowledge about distinctive physiologic features within donors and find potential biomarkers for graft quality assessment The main questions it aims to answer are
Is there a distinctive miRNA profile between donors after brain-stem death and donors after circulatory death
Is heparinase I treatment necessary to overcome miRNA quantification interference in heparinized liver donor samples
Researchers will compare miRNA expression in donors after brain-stem death and donors after circulatory death with I a subgroup of donors with unusable grafts due to significant steatosis 2 and with a subgroup of healthy controls undergoing elective cholecystectomy to set the reference miRNA profiles in extreme conditions
Is there a distinctive miRNA profile between donors after brain-stem death and donors after circulatory death
Is heparinase I treatment necessary to overcome miRNA quantification interference in heparinized liver donor samples
Researchers will compare miRNA expression in donors after brain-stem death and donors after circulatory death with I a subgroup of donors with unusable grafts due to significant steatosis 2 and with a subgroup of healthy controls undergoing elective cholecystectomy to set the reference miRNA profiles in extreme conditions
Detailed Description:
Between October 2019 and May 2021 perfusates liver biopsies and serum samples were collected prospectively from adult deceased organ donors in Vall d39Hebron University Hospital Barcelona Spain
Samples from adult human brain death donors n10 and donors after circulatory death n10 were analyzed for the presence of miR-122 miR-148 miR-155 miR-22 miR-222 and ratio miR122miR222 target biomarkers of liver injury selected from the literature
To set the reference miRNA profiles a subgroup of donors with unusable grafts due to significant steatosis n3 were analyzed
Donors for re-transplantation pediatric and split were excluded to avoid biopsy-related injury
As there were no living donor inclusions in the study period serum specimens from a subgroup of healthy controls undergoing elective cholecystectomy n7 were analyzed
This study is part of a PhD thesis and data collection was done by the main investigator based in organ donor availability in one tertiary hospital centre in Barcelona Spain Based on resource limitation constraints the sample size was limited to the consecutive samples obtained in a 2-year period
Written informed consent was obtained from patients and relatives of donors included in the study Samples and data from patients were provided by the Vall dHebron University Hospital Biobank PT2000107 integrated in the Spanish National Biobanks Network and they were processed following standard operating procedures The study protocol was approved by the medical ethics committee of the Vall dHebron University Hospital PRAG462019 and conforms to the principles outlined in the Declaration of Helsinki
Statistical analysis
To optimize the quality of data readings with extreme Ct values Ct less than 11 or Ct greater than 39 were discarded The mean of the Cts and the standard deviation for each of the miRNAs in the different samples analyzed technical triplicates were calculated To improve the quality of the data analyzed in triplicates with high standard deviation Ct SD greater than 03 the Ct value furthest from the mean was eliminated leaving a total of at least two values per miRNA per sample Likewise for each miRNA samples with less than two valid replicates were excluded To correct for potential RNA input or RT efficiency biases Ct values were normalized using the average Ct of endogenous references miR-103a miR-191 miR-30c miR-16 and let7a miRNA relative quantification RQ levels were analyzed using the Livak et al method 2-ΔΔCt calculated as follows ΔCt miRNA Ct target - averaged endogenous control Ct and the difference ΔΔCt between comparison groups ΔCt comparison - ΔCt reference sample Group comparisons were performed using T-test or U-Mann Whitney non-parametric for continuous data and Fisher test for qualitative P-values less than 005 were considered significant
Methodology substudy
A subgroup of adult human brain death donors n4 and donors after circulatory death n4 were analyzed for the presence of miR-122 and miR-148 Heparin is known to have an inhibiting effect in RNA qPCR analysis and liver donor samples are prone to have heparin traces as itamp39s used to avoid thrombus formation during organ procurement Heparinase-I has been used in the literature to overcome the heparin inhibiting effect
In this substudy the samples were treated with 0 IU 6 IU or 12 IU of heparinase-I to evaluate the counteracting effect of heparinase I on miRNA detection levels by RT-qPCR
Samples from adult human brain death donors n10 and donors after circulatory death n10 were analyzed for the presence of miR-122 miR-148 miR-155 miR-22 miR-222 and ratio miR122miR222 target biomarkers of liver injury selected from the literature
To set the reference miRNA profiles a subgroup of donors with unusable grafts due to significant steatosis n3 were analyzed
Donors for re-transplantation pediatric and split were excluded to avoid biopsy-related injury
As there were no living donor inclusions in the study period serum specimens from a subgroup of healthy controls undergoing elective cholecystectomy n7 were analyzed
This study is part of a PhD thesis and data collection was done by the main investigator based in organ donor availability in one tertiary hospital centre in Barcelona Spain Based on resource limitation constraints the sample size was limited to the consecutive samples obtained in a 2-year period
Written informed consent was obtained from patients and relatives of donors included in the study Samples and data from patients were provided by the Vall dHebron University Hospital Biobank PT2000107 integrated in the Spanish National Biobanks Network and they were processed following standard operating procedures The study protocol was approved by the medical ethics committee of the Vall dHebron University Hospital PRAG462019 and conforms to the principles outlined in the Declaration of Helsinki
Statistical analysis
To optimize the quality of data readings with extreme Ct values Ct less than 11 or Ct greater than 39 were discarded The mean of the Cts and the standard deviation for each of the miRNAs in the different samples analyzed technical triplicates were calculated To improve the quality of the data analyzed in triplicates with high standard deviation Ct SD greater than 03 the Ct value furthest from the mean was eliminated leaving a total of at least two values per miRNA per sample Likewise for each miRNA samples with less than two valid replicates were excluded To correct for potential RNA input or RT efficiency biases Ct values were normalized using the average Ct of endogenous references miR-103a miR-191 miR-30c miR-16 and let7a miRNA relative quantification RQ levels were analyzed using the Livak et al method 2-ΔΔCt calculated as follows ΔCt miRNA Ct target - averaged endogenous control Ct and the difference ΔΔCt between comparison groups ΔCt comparison - ΔCt reference sample Group comparisons were performed using T-test or U-Mann Whitney non-parametric for continuous data and Fisher test for qualitative P-values less than 005 were considered significant
Methodology substudy
A subgroup of adult human brain death donors n4 and donors after circulatory death n4 were analyzed for the presence of miR-122 and miR-148 Heparin is known to have an inhibiting effect in RNA qPCR analysis and liver donor samples are prone to have heparin traces as itamp39s used to avoid thrombus formation during organ procurement Heparinase-I has been used in the literature to overcome the heparin inhibiting effect
In this substudy the samples were treated with 0 IU 6 IU or 12 IU of heparinase-I to evaluate the counteracting effect of heparinase I on miRNA detection levels by RT-qPCR
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
None
Is a FDA Regulated Device?:
None
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None