20.500.12556/DiRROS-21562 Extracellular vesicle-bound DNA in urine is indicative of kidney allograft injury Extracellular vesicle-bound DNA (evDNA) is an understudied extracellular vesicle (EV) cargo, particularly in cancer-unrelated research. Although evDNA has been detected in urine, little is known about its characteristics, localization, and biomarker potential for kidney pathologies. To address this, we enriched EVs from urine of well-characterized kidney transplant recipients undergoing allograft biopsy, characterized their evDNA and its association to allograft injury. The SEC-based method enriched pure EVs from urine of kidney transplant recipients, regardless of the allograft injury. Urinary evDNA represented up to 29.2 ± 8% (mean ± SD) of cell-free DNA (cfDNA) and correlated with cfDNA in several characteristics but was less fragmented (P < 0.001). Importantly, using DNase treatment and immunogold labelling TEM, we demonstrated that evDNA was bound to the surface of urinary EVs. Normalised evDNA yield (P = 0.042) and evDNA copy number (P = 0.027) significantly differed between patients with normal histology, rejection injury and non-rejection injury, the later groups having significantly larger uEVs (mean diameter, P = 0.045) and more DNA bound per uEV. ddDNA is detectable in uEV samples of kidney allograft recipients, but its quantity is highly variable. In a proof-of-principle study, several evDNA characteristics correlated with clinical and histological parameters (P = 0.040), supporting that the potential of evDNA as a biomarker for kidney allograft injury should be further investigated. DNA donor-derived DNA extracellular vesicles DNK DNK pridobljena iz darovalca zunajcelični vezikli true false true Angleški jezik Slovenski jezik Neznano 2025-02-26 13:38:27 2025-02-26 13:38:27 2025-02-27 03:39:03 0000-00-00 00:00:00 2022 0 0 Soavtorji: Maja Štalekar, Magda Tušek Žnidarič, Katja Goričar, Nika Kojc, Polona Kogovšek, Vita Dolžan, Miha Arnol, Metka Lenassi; Nasl. z nasl. zaslona; Opis vira z dne 28. 9. 2022; Št. članka: e12268; str. 1-23 iss. 9 Vol. 11 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 2022-09-23 616.6 2001-3078 10.1002/jev2.12268 123229443 https://onlinelibrary.wiley.com/doi/10.1002/jev2.12268 1 94e2dd23-f43e-11ef-9f1e-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30844 RAZ_Sedej_Ivana_2022.pdf RAZ_Sedej_Ivana_2022.pdf 1 CFBC3A406854D04F5AF506E3F8A7D2A8 a20ed071515aea57e884c9d9284284d9eed1c528306b9e86734f22ae37170b13 ba773f8a-f43e-11ef-9f1e-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30845 https://doi.org/10.1002/jev2.12268 0 c3d08037-f43e-11ef-9f1e-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=30846 Nacionalni inštitut za biologijo 0 0 0