Vpletenost s paratiroidnim hormonom uravnavanih miRNA v signalni poti Wnt in RANK/RANKL/OPG

Emkić, Erma (author), Ostanek, Barbara (mentor), Vrščaj, Lucija Ana (comentor)

Kostna prenova je usklajeni proces kostne razgradnje in izgradnje. V njej igrata pomembno vlogo signalna pot Wnt in signalna pot RANK/RANKL/OPG. Parathormon (PTH) vpliva tako na kostno razgradnjo kot na izgradnjo. V primeru intermitentne izpostavljenosti pride do povečanja kostne mase, zato v terapiji osteoporoze uporabljamo njegovo rekombinantno obliko (teriparatid) kot osteoanabolno zdravilo. Eden od mehanizmov vpliva PTH na kosti je preko sprememb izražanja mikro RNA (miRNA). Le-te predstavljajo kratke, nekodirajoče RNA, ki zavirajo izražanje genov po transkripciji. Namen magistrske naloge je bil ugotoviti, katerim miRNA, ki uravnavajo izražanje genov v signalni poti Wnt ali RANK/RANKL/OPG, se spremeni izražanje pod vplivom PTH. V elektronski knjižnici PubMed smo pregledali literaturo, nato s pomočjo bioinformatičnih orodij DIANA tools, miRWalk, TargetScan ter miRDB naredili napovedi tarčnih genov za izbrane miRNA, z bioinformatičnim orodjem miRTarBase preverili eksperimentalno potrjene interakcije ter izrisali interaktome z orodjem Cytoscape. S pregledom literature smo zbrali 17 relevantnih člankov, iz katerih smo pridobili nabor 69 miRNA, za katere smo naredili napovedi. Skupno število vseh potencialnih tarčnih genov je bilo 12782, eksperimentalno potrjenih pa 477. V nadaljevanju smo analizirali miRNA, ki vplivajo na signalno pot Wnt in signalno pot RANK/RANKL/OPG, ločeno ali na obe hkrati. MiRNA, ki vpliva na obe signalni poti, lahko predstavlja obetavno potencialno zdravilo, v kolikor so njeni učinki tišanja ustrezni, torej da deluje hkrati osteanabolno in antiresorptivno. Kot potencialno zdravilo za zdravljenje osteoporoze bi lahko uporabili miRNA hsa-miR-24-2-5p, katere izražanje v kostnih celicah bi bilo potrebno v nadaljnjih študijah izmeriti. Naši rezultati potrjujejo pomemben vpliv miRNA na obe signalni poti in predstavljajo izhodišče za nove zdravilne učinkovine v terapiji osteoporoze.

• Language: Slovenian
• Keywords: osteoporoza, PTH, signalna pot Wnt, signalna pot RANK/RANKL/OPG, miRNA
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2022
• PID: 20.500.12556/RUL-140843
• Publication date in RUL: 20.09.2022

Secondary language

• Language: English
• Title: Involvement of parathyroid hormone-regulated miRNAs in the Wnt and RANK/RANKL/OPG signalling

Abstract

Bone remodeling is a coordinated process of coupled bone resorption and formation in which Wnt signaling pathway and the RANK/RANK/OPG signaling pathway play an important role. Parathyroid hormone (PTH) is a hormone that affects bone formation and resorption. In the case of intermittent exposure, there is an increase in bone mass, so its recombinant form (teriparatide) is used in the treatment of osteoporosis as an osteoanabolic drug. Changes of micro RNA (miRNA) expression represent one of the mechanisms of PTH action on bone. MiRNAs are small non-coding RNA molecules that inhibit gene expression after transcription. The aim of the master's thesis was to find miRNAs regulated by PTH, which are involved in the Wnt signaling pathway or RANK/RANKL/OPG pathway. Literature search was performed in the electronic library PubMed, followed by target predictions for the identified miRNAs with bioinformatics tools DIANA tools, miRWalk, TargetScan and miRDB. Experimentally confirmed interactions were identified with the bioinformatics tool miRTarBase and interactomes drawn with Cytoscape. Literature review resulted in 17 relevant articles, with a set of 69 miRNAs for which predictions were made. The total number of all target genes was 12782 and 477 were experimentally confirmed. Next, we undertook the analysis of miRNAs, which have the potential to influence the Wnt signaling pathway, RANK/RANKL/OPG signaling pathway or both. MiRNA that affects both signaling pathways can represent a promising potential therapeutic, as long as its silencing effects are appropriate, so it acts as an osteoanabolic and antiresorptive. As a potential therapeutic for the treatment of osteoporosis, miRNA hsa-miR-24-2-5p could be used, the expression of which in bone cells should be measured in the following studies. Our results confirm the significant influence of miRNA on both signaling pathways and represent a starting point for development of new active substances in the treatment of osteoporosis.

• Keywords: osteoporosis, PTH, Wnt signaling pathway, RANK/RANKL/OPG signaling pathway, miRNA