ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/089/SRR29760789/SRR29760789_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/093/SRR29760793/SRR29760793_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/087/SRR29760787/SRR29760787_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/091/SRR29760791/SRR29760791_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/092/SRR29760792/SRR29760792_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/087/SRR29760787/SRR29760787_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/093/SRR29760793/SRR29760793_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/092/SRR29760792/SRR29760792_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/090/SRR29760790/SRR29760790_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/095/SRR29760795/SRR29760795_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/094/SRR29760794/SRR29760794_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/090/SRR29760790/SRR29760790_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/091/SRR29760791/SRR29760791_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/089/SRR29760789/SRR29760789_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/088/SRR29760788/SRR29760788_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/095/SRR29760795/SRR29760795_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/094/SRR29760794/SRR29760794_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR297/088/SRR29760788/SRR29760788_2.fastq.gzprimary200OKGenomicsChonnam National Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA1133004Mus musculusxref:PubMed:39039245Bone is a highly dynamic tissue undergoing continuous formation and resorption. Here, we investigated differential but complementary roles of hypoxia-inducible factor (HIF)-1α and HIF-2α in regulating bone remodeling. Using RNA-seq analysis, we identified that specific genes involved in regulating osteoblast differentiation were similarly but slightly differently governed by HIF-1α and HIF-2α. We found that increased HIF-1α expression inhibited osteoblast differentiation via inhibiting RUNX2 function by upregulation of Twist2, confirmed using Hif1a conditional knockout (KO) mouse. Ectopic expression of HIF-1α via adenovirus transduction resulted in the increased expression and activity of RANKL, while knockdown of Hif1a expression via siRNA or osteoblast-specific depletion of Hif1a in conditional KO mice had no discernible effect on osteoblast-mediated osteoclast activation. The unexpected outcome was elucidated by the upregulation of HIF-2α upon Hif1a overexpression, providing evidence that Hif2a is a transcriptional target of HIF-1α in regulating RANKL expression, verified through an experiment of HIF-2α knockdown after HIF-1α overexpression. The above results were validated in an ovariectomized- and aging-induced osteoporosis model using Hif1a conditional KO mice. Our findings conclude that HIF-1α plays an important role in regulating bone homeostasis by controlling osteoblast differentiation, and in influencing osteoclast formation through the regulation of RANKL secretion via HIF-2α modulation. Overall design: RNA-seq analysis was performed to determine the gene expression profile caused by overexpression of HIF-1α and HIF-2α during primary culture calvarial preosteoblast differentiation.ENAfalseDifferential but complementary roles of HIF-1α and HIF-2α in the regulation of bone homeostasisDifferential but complementary roles of HIF-1α and HIF-2α in the regulation of bone homeostasis2025-09-242024-07-14PRJNA1133004GSE2717301009039039245