Novel Role Of Lncrna H19 In Osteoarthritis Development Via Modulating Subchondral Bone Remodeling And Osteocyte Responses To Mechanical Stimulation
Invited conference paper presented and published in conference proceedings
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摘要Introduction:
Emerging evidence suggests that aberrant subchondral bone remodeling responding to excessive mechanical loading plays important role in earlyonset of OA [1]. Targeting the perturbed subchondral bone remodeling has been proposed to minimize uneven articular cartilage stress distribution, and thus anew therapeutic approach for OA. A long non-coding RNA (lncRNA) H19 has been reported to be associated with OA progression and regulate mechano-transduction at cellular level, however, its role in OA subchondral bone has not been reported [2]. This study aimed to examine the relationship between H19 andOA in clinical samples and animal models, and to determine its function in osteocyte upon mechanical stimulation.
Methods:
Subchondral bone were collected from patients with knee OA undergoing joint replacement surgery (n = 15). To verify the link between H19 and OAsubchondral bone, wild-type C57BL/6 mice and same background transgenic mice with osteocyte-specific deletion of H19 (cKO) by crossing Dmp1-Cre and H19floxed mice were used, and OA phenotype was induced by DMM (destabilization of the medial meniscus) surgery. Male mice at 12-week-old were used (n=7 pergroup). H19 expression was determined by fluorescence in situ hybridization (FISH) and qPCR, subchondral bone changes by microCT, and OA phenotypes byhistological analysis. To verify the effect of mechanical stimulation on osteocytes and to study the underlying mechanism, MLO-Y4 cells were subjected tounidirectional shear flow on ibidi flow chamber (Fig. D) [3]. H19 overexpression or knockdown was performed to verify the effect of H19 on MLO-Y4 cells. g.Paired t-test was used for comparing differences in human subchondral bone. One way ANOVA and Dunnett’s multiple comparisons test were used to comparedifferences in animal and cellular experiments.
Results:
Human subchondral bone of end-stage OA had higher level of H19 in both femoral condyle and tibial plateaus, which is associated with increased bonemass and more H19 positive osteocytes (Fig. A, B). In wild-type C57BL/6 mice, DMM surgery led to cartilage damage, subchondral sclerosis, and increased H19expression in subchondral bone. On the contrary, cKO mice with H19 ablation were less vulnerable to DMM induced OA phenotypic changes (Fig. C). In MLO-Y4cells, H19 overexpression resulted in higher response to shear stress as indicated by increased
Ptgs2, Tnfrsf11b, Opg/Rankl
and
Dmp1
mRNA levels (Fig. E). Thepromoting effect of H19 was abolished after H19 knockdown (Fig. F).
Discussion:
Our results provide new evidence that elevated H19 expression in osteocytes may contribute to the aberrant subchondral bone remodeling and thuscartilage damage in OA. H19 appears to be required for osteocytes’ response to mechanical stimulation. How does H19 mediate osteocyte activity and its role onOA onset/development is warranted for further investigation.
Significance/Clinical Relevance:
Further analysis on the biological function of H19 on osteocytes response to mechanical stimulation will shed light on thedevelopment of novel treatment for OA via targeting subchondral osteocytes.
Emerging evidence suggests that aberrant subchondral bone remodeling responding to excessive mechanical loading plays important role in earlyonset of OA [1]. Targeting the perturbed subchondral bone remodeling has been proposed to minimize uneven articular cartilage stress distribution, and thus anew therapeutic approach for OA. A long non-coding RNA (lncRNA) H19 has been reported to be associated with OA progression and regulate mechano-transduction at cellular level, however, its role in OA subchondral bone has not been reported [2]. This study aimed to examine the relationship between H19 andOA in clinical samples and animal models, and to determine its function in osteocyte upon mechanical stimulation.
Methods:
Subchondral bone were collected from patients with knee OA undergoing joint replacement surgery (n = 15). To verify the link between H19 and OAsubchondral bone, wild-type C57BL/6 mice and same background transgenic mice with osteocyte-specific deletion of H19 (cKO) by crossing Dmp1-Cre and H19floxed mice were used, and OA phenotype was induced by DMM (destabilization of the medial meniscus) surgery. Male mice at 12-week-old were used (n=7 pergroup). H19 expression was determined by fluorescence in situ hybridization (FISH) and qPCR, subchondral bone changes by microCT, and OA phenotypes byhistological analysis. To verify the effect of mechanical stimulation on osteocytes and to study the underlying mechanism, MLO-Y4 cells were subjected tounidirectional shear flow on ibidi flow chamber (Fig. D) [3]. H19 overexpression or knockdown was performed to verify the effect of H19 on MLO-Y4 cells. g.Paired t-test was used for comparing differences in human subchondral bone. One way ANOVA and Dunnett’s multiple comparisons test were used to comparedifferences in animal and cellular experiments.
Results:
Human subchondral bone of end-stage OA had higher level of H19 in both femoral condyle and tibial plateaus, which is associated with increased bonemass and more H19 positive osteocytes (Fig. A, B). In wild-type C57BL/6 mice, DMM surgery led to cartilage damage, subchondral sclerosis, and increased H19expression in subchondral bone. On the contrary, cKO mice with H19 ablation were less vulnerable to DMM induced OA phenotypic changes (Fig. C). In MLO-Y4cells, H19 overexpression resulted in higher response to shear stress as indicated by increased
Ptgs2, Tnfrsf11b, Opg/Rankl
and
Dmp1
mRNA levels (Fig. E). Thepromoting effect of H19 was abolished after H19 knockdown (Fig. F).
Discussion:
Our results provide new evidence that elevated H19 expression in osteocytes may contribute to the aberrant subchondral bone remodeling and thuscartilage damage in OA. H19 appears to be required for osteocytes’ response to mechanical stimulation. How does H19 mediate osteocyte activity and its role onOA onset/development is warranted for further investigation.
Significance/Clinical Relevance:
Further analysis on the biological function of H19 on osteocytes response to mechanical stimulation will shed light on thedevelopment of novel treatment for OA via targeting subchondral osteocytes.
著者Rongliang Wang, Michael Tim yun Ong, Vivas Hon Fai Chan, Gang Li, Wayne Yuk Wai Lee
會議名稱Orthopaedic Research Society 2023 Annual Meeting
會議開始日10.02.2023
會議完結日14.02.2023
會議地點Dallas, Texas
會議國家/地區美國
出版年份2023
語言美式英語
