Izražanje izbranih mikro RNA v serumu in napoved odziva na zdravljenje pomenopavzne osteoporoze s teriparatidom

Sedej, Neža (author), Ostanek, Barbara (mentor), Vrščaj, Lucija Ana (comentor)

Pomenopavzna osteoporoza je kronična presnovna bolezen kosti, za katero je značilno postopno zmanjševanje mineralne kostne gostote, poslabšanje mikroarhitekture kostnega tkiva ter povečano tveganje za zlome. Glavni vzrok za nastanek bolezni je znižana raven estrogena po menopavzi, kar povzroči neravnovesje med kostno resorpcijo in izgradnjo. Ena izmed terapevtskih možnosti je zdravljenje z rekombinantno obliko analoga parathormona – teriparatidom, ki deluje osteoanabolno. Kljub temu, da je pri večini bolnic zdravljenje uspešno, obstaja manjši delež, pri katerih ne pride do pričakovanega zvišanja mineralne kostne gostote. Namen naše naloge je bil ugotoviti, ali se izražanje izbranih mikro RNA, za katere je znano, da so vključene v osteogenezo (hsa miR 20b 5p, hsa miR 31 3p, hsa miR 133a 3p in hsa miR 375-3p), spreminja tekom zdravljenja, ali se razlikuje med odzivnimi in neodzivnimi bolnicami, ter ali bi lahko te miRNA služile kot napovedni označevalci terapevtskega odziva. V raziskavo je bilo vključenih 51 bolnic s pomenopavzno osteoporozo, ki so prejemale teriparatid eno leto. Glede na relativno spremembo mineralne kostne gostote v ledvenem delu hrbtenice v obdobju enega leta smo jih razvrstili v odzivno (sprememba večja ali enaka 3 %) in neodzivno skupino (sprememba manjša od 3 %). Izražanje izbranih miRNA v serumskih vzorcih pred začetkom in po šestih mesecih zdravljenja smo določili z verižno reakcijo s polimerazo v realnem času (RT-qPCR). Najpomembnejše razlike v izražanju smo zaznali pri dveh negativnih regulatorjih osteogeneze, to sta hsa‑miR‑133a‑3p in hsa‑miR‑375‑3p, pri katerih je bilo relativno izražanje pri neodzivnih preiskovankah že izhodiščno statistično značilno višje, ta razlika pa se je ohranila tudi po šestih mesecih zdravljenja. Pri hsa‑miR‑20b‑5p smo po šestih mesecih v primerjavi z odzivno skupino zaznali višje relativno izražanje v neodzivni skupini, kar je nekoliko nepričakovano, glede na njeno znano proosteogeno delovanje. Pri hsa‑miR‑31‑3p spremembe niso dosegle statistične značilnosti, viden pa je bil trend povečanega izražanja pri neodzivnih. Rezultati kažejo, da imata hsa‑miR‑133a‑3p in hsa‑miR‑375‑3p potencial za klinično uporabnost kot napovedna označevalca odziva na zdravljenje s teriparatidom. To odpira možnost bolj individualiziranega zdravljenja osteoporoze, za potrditev pa bodo potrebne še nadaljnje raziskave.

• Language: Slovenian
• Keywords: pomenopavzna osteoporoza, mineralna kostna gostota, teriparatid, miRNA, RT-qPCR
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2025
• PID: 20.500.12556/RUL-174223
• Publication date in RUL: 30.09.2025

Secondary language

• Language: English
• Title: Expression of selected microRNAs in serum and prediction of response to teriparatide treatment in postmenopausal osteoporosis

Abstract

Postmenopausal osteoporosis is a chronic metabolic bone disease characterized by a gradual decrease in bone mineral density, deterioration of bone microarchitecture, and an increased risk of fractures. The primary cause is reduced estrogen levels after menopause, disrupting the balance between bone resorption and formation. One treatment option is teriparatide, a recombinant analogue of parathyroid hormone with an anabolic effect on bone. Although treatment is effective in most patients, there is a smaller proportion in whom the expected increase in bone mineral density does not occur. The aim of our study was to determine whether the expression of selected microRNAs known to be involved in osteogenesis (hsa‑miR‑20b‑5p, hsa‑miR‑31‑3p, hsa‑miR‑133a‑3p, hsa‑miR‑375‑3p) changes during treatment, whether it differs between responsive and non-responsive patients, and whether these microRNAs could serve as predictive biomarkers of therapeutic response. The study included 51 women with postmenopausal osteoporosis who received teriparatide treatment for one year. Based on the relative change in bone mineral density in the lumbar spine, they were classified into responder and non-responder groups (responders had a BMD increase of three percent or more, non-responders had an increase of less than three percent). Expression levels of selected microRNAs in serum samples, collected before treatment and after six months, were analysed using real-time polymerase chain reaction (RT-qPCR). The most prominent differences were observed in hsa‑miR‑133a‑3p and hsa‑miR‑375‑3p, where relative expression levels were already significantly higher in non-responsive patients at baseline and remained elevated after six months of treatment. For hsa‑miR‑20b‑5p, a higher relative expression was observed in the non-responsive group after six months, which was unexpected given its known pro-osteogenic role. No statistically significant differences were found for hsa‑miR‑31‑3p, although a trend toward increased expression in the non-responsive group was noted. Our results suggest that hsa‑miR‑133a‑3p and hsa‑miR‑375‑3p have potential clinical value as predictive biomarkers of response to teriparatide. This highlights the possibility of more personalized approaches to osteoporosis treatment, but confirmation with further research is needed.

• Keywords: postmenopausal osteoporosis, bone mineral density, teriparatide, miRNA, RT qPCR