TY - JOUR
T1 - MicroRNA-181a/b-1 enhances chondroprogenitor anabolism and downregulates aquaporin-9
AU - Bell-Hensley, Austin
AU - Brito, Victor Gustavo Balera
AU - Cai, Lei
AU - Liu, Jin
AU - Feeney, Kathryn
AU - Zheng, Hongjun
AU - McAlinden, Audrey
N1 - Publisher Copyright:
© 2024 The Authors
PY - 2025/3
Y1 - 2025/3
N2 - Objective: Effective osteoarthritis treatments that enhance the anabolic/regenerative capacity of chondrocytes are needed. Studying cartilage development processes may inform us of approaches to control chondrocyte differentiation and anabolism and, ultimately, how to effectively treat OA. MicroRNAs are broad-acting epigenetic regulators known to affect many skeletal processes. Previous reports from our group indicated that miR-181a-1 is upregulated during chondrocyte differentiation. The goal of this study was to determine how the entire miR-181a/b-1 cluster regulates in vitro chondrogenesis. Design: Precursor miR-181a/b-1 was over-expressed in cartilage progenitor cells using lentiviral technology Transduced cartilage progenitor cells were cultured as micromass pellets in hypoxic conditions and stimulated to undergo chondrogenic differentiation for five weeks. Bulk RNA-sequencing and immunostaining was applied to evaluate chondrogenic differentiation and matrix production. Results: Immunostaining of cartilage pellet sections showed that miR-181a/b-1 increased mature type II collagen and decreased expression of the chondroprogenitor type IIA collagen isoform. Bulk RNA-Seq at day 7 of chondrogenesis revealed upregulation of pro-anabolic genes such as COL2A1, COL9A2/3, COL11A2 and SNORC. Of the genes significantly downregulated by miR-181a/b-1, aquaporin 9 (AQP9) was the top hit which decreased in expression by over 14-fold. While a predicted target of miR-181a/b, our data showed that this miRNA cluster likely suppresses AQP9 via an indirect targeting mechanism. Conclusions: Our findings demonstrate a pro-differentiation/anabolic function for miR-181a/b-1 during in vitro chondrogenesis that may be due, in part, to suppression of AQP9. Future studies are needed to elucidate the role of this membrane channel protein in regulating chondrocyte differentiation and homeostasis.
AB - Objective: Effective osteoarthritis treatments that enhance the anabolic/regenerative capacity of chondrocytes are needed. Studying cartilage development processes may inform us of approaches to control chondrocyte differentiation and anabolism and, ultimately, how to effectively treat OA. MicroRNAs are broad-acting epigenetic regulators known to affect many skeletal processes. Previous reports from our group indicated that miR-181a-1 is upregulated during chondrocyte differentiation. The goal of this study was to determine how the entire miR-181a/b-1 cluster regulates in vitro chondrogenesis. Design: Precursor miR-181a/b-1 was over-expressed in cartilage progenitor cells using lentiviral technology Transduced cartilage progenitor cells were cultured as micromass pellets in hypoxic conditions and stimulated to undergo chondrogenic differentiation for five weeks. Bulk RNA-sequencing and immunostaining was applied to evaluate chondrogenic differentiation and matrix production. Results: Immunostaining of cartilage pellet sections showed that miR-181a/b-1 increased mature type II collagen and decreased expression of the chondroprogenitor type IIA collagen isoform. Bulk RNA-Seq at day 7 of chondrogenesis revealed upregulation of pro-anabolic genes such as COL2A1, COL9A2/3, COL11A2 and SNORC. Of the genes significantly downregulated by miR-181a/b-1, aquaporin 9 (AQP9) was the top hit which decreased in expression by over 14-fold. While a predicted target of miR-181a/b, our data showed that this miRNA cluster likely suppresses AQP9 via an indirect targeting mechanism. Conclusions: Our findings demonstrate a pro-differentiation/anabolic function for miR-181a/b-1 during in vitro chondrogenesis that may be due, in part, to suppression of AQP9. Future studies are needed to elucidate the role of this membrane channel protein in regulating chondrocyte differentiation and homeostasis.
KW - Anabolism
KW - Aquaporin-9
KW - Chondrocyte
KW - Chondrogenesis
KW - microRNA
KW - miR-181a/b
UR - http://www.scopus.com/inward/record.url?scp=85210361868&partnerID=8YFLogxK
U2 - 10.1016/j.ocarto.2024.100550
DO - 10.1016/j.ocarto.2024.100550
M3 - Article
C2 - 39691700
AN - SCOPUS:85210361868
SN - 2665-9131
VL - 7
JO - Osteoarthritis and Cartilage Open
JF - Osteoarthritis and Cartilage Open
IS - 1
M1 - 100550
ER -