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Capturing the Kidne...
Capturing the Kidney Transcriptome by Urinary Extracellular Vesicles—From Pre-Analytical Obstacles to Biomarker Research
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- Barreiro, Karina (författare)
- University of Helsinki
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- Dwivedi, Om Prakash (författare)
- University of Helsinki
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- Rannikko, Antti (författare)
- University of Helsinki
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- Holthöfer, Harry (författare)
- University of Helsinki,University Medical Center Hamburg-Eppendorf
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- Tuomi, Tiinamaija (författare)
- Lund University,Lunds universitet,Molekylär endokrinologi,Forskargrupper vid Lunds universitet,Diabetiska komplikationer,Molecular Endocrinology,Lund University Research Groups,Diabetic Complications,Folkhälsan Research Center,Helsinki University Central Hospital,University of Helsinki
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- Groop, Per Henrik (författare)
- University of Helsinki,Folkhälsan Research Center,Monash University
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- Puhka, Maija (författare)
- University of Helsinki
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(creator_code:org_t)
- 2023
- 2023
- Engelska.
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Ingår i: Genes. - 2073-4425. ; 14:7
- Relaterad länk:
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http://dx.doi.org/10... (free)
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https://lup.lub.lu.s...
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https://doi.org/10.3...
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Abstract
Ämnesord
Stäng
- Urinary extracellular vesicles (uEV) hold non-invasive RNA biomarkers for genitourinary tract diseases. However, missing knowledge about reference genes and effects of preanalytical choices hinder biomarker studies. We aimed to assess how preanalytical variables (urine storage temperature, isolation workflow) affect diabetic kidney disease (DKD)—linked miRNAs or kidney—linked miRNAs and mRNAs (kidney-RNAs) in uEV isolates and to discover stable reference mRNAs across diverse uEV datasets. We studied nine raw and normalized sequencing datasets including healthy controls and individuals with prostate cancer or type 1 diabetes with or without albuminuria. We focused on kidney-RNAs reviewing literature for DKD-linked miRNAs from kidney tissue, cell culture and uEV/urine experiments. RNAs were analyzed by expression heatmaps, hierarchical clustering and selecting stable mRNAs with normalized counts (>200) and minimal coefficient of variation. Kidney-RNAs were decreased after urine storage at −20 °C vs. −80 °C. Isolation workflows captured kidney-RNAs with different efficiencies. Ultracentrifugation captured DKD -linked miRNAs that separated healthy and diabetic macroalbuminuria groups. Eleven mRNAs were stably expressed across the datasets. Hence, pre-analytical choices had variable effects on kidney-RNAs—analyzing kidney-RNAs complemented global correlation, which could fade differences in some relevant RNAs. Replicating prior DKD-marker results and discovery of candidate reference mRNAs encourages further uEV biomarker studies.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Urologi och njurmedicin (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Urology and Nephrology (hsv//eng)
Nyckelord
- diabetic kidney disease
- exosomes
- miRNA
- mRNA
- reference genes
- sequencing
- urinary extracellular vesicles
- urine
Publikations- och innehållstyp
- art (ämneskategori)
- ref (ämneskategori)
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