Osteoblastgenic and Osteogenic Effects of KY-273 with CDK8/19 Inhibitory Activity in Bone Marrow Mesenchymal Stem Cells and Female Rats

Uterine fibroids (UFs) are the most common non-cutaneous tumors in women worldwide. UFs arise from genetic alterations in myometrial stem cells (MM SCs) that trigger their transformation into tumor initiating cells (UF SCs). Mutations in the RNA polymerase II Mediator subunit MED12 are dominant drivers of UFs, accounting for 70% of these clinically significant lesions. Biochemically, UF driver mutations in MED12 disrupt CDK8/19 kinase activity in Mediator, but how Mediator kinase disruption triggers MM SC transformation remains unknown. Here, we show that pharmacologic inhibition of CDK8/19 in MM SCs removes a barrier to myogenic differentiation down an altered pathway characterized by molecular phenotypes characteristic of UFs, including oncogenic growth and extracellular matrix (ECM) production. These perturbations appear to be induced by transcriptomic changes, arising in part through epigenomic alteration and super-enhancer reprogramming, that broadly recapitulate those found in MED12-mutant UFs. Altogether these findings provide new insights concerning the biological role of CDK8/19 in MM SC biology and UF formation.

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A potent small molecule, AS2863619, enhances the osteogenic potential and ameliorates the inflammation-induced impairment of osteogenesis in periodontal ligament stem cells

He,

Li,

Huang

et al. 2024

Preprint

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Background Periodontal ligament stem cells (PDLSCs) represent a promising source for the regeneration of periodontal tissues. However, these cells may experience functional alterations during in vitro expansion and culture, as well as under inflammatory conditions. In this study, we investigated AS2863619 (AS), a small molecule drug with potential applications in immune disease research, to evaluate its effects on the osteogenic potential of PDLSCs and to elucidate the underlying mechanisms of action. Methods The effect of AS on the proliferation of PDLSCs was evaluated using a CCK-8 assay. The osteogenic potential of PDLSCs was assessed through alkaline phosphatase (ALP) activity staining or Alizarin Red S (ARS) staining to detect mineralized nodules. Inflammatory injury in PDLSCs was induced by treating them with recombinant human tumor necrosis factor-alpha (rhTNF-α) . Transcriptome sequencing was employed to investigate the potential target of AS for rescuing osteogenic differentiation from inflammatory injury. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were conducted to determine the expression levels of osteogenic gene markers and associated signaling pathways. Various small molecule inhibitors, agonists, and recombinant proteins were utilized to modulate relevant signaling pathways as specified in the text. Results The proliferation of PDLSCs was inhibited by AS at 250 nM after 1 day of culture, whereas AS at 125 nM inhibited their proliferation after 3 and 5 days of culture. Additionally, AS significantly enhanced the ALP activity of PDLSCs and accelerated the formation time of mineralized nodules, while suppressing their adipogenic differentiation ability. Furthermore, AS promoted the levels of bone morphogenetic protein (BMP)2 and phosphorylated (p)-SMAD1/5 without significant effects on p-ERK, p-P38, and p-JNK levels. Moreover, AS effectively augmented rhTNF-α-inhibited ALP activity in PDLSCs and attenuated rhTNF-α-induced components involved in retinoic acid-inducible gene I (RIG-I)-like receptor signaling pathway including RIG-I, MDA5, and ISG15. Conclusions AS is a potent small molecule that effectively enhances the osteogenic potential of PDLSCs, potentially by augmenting BMP2/SMAD signaling. Additionally, AS significantly improves the inflammation-induced impairment of osteogenic potential in PDLSCs and may contribute to osteogenic rescue through inhibition of rhTNF-α-induced RIG-I-like receptor signaling pathway.

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Osteoblastgenic and Osteogenic Effects of KY-273 with CDK8/19 Inhibitory Activity in Bone Marrow Mesenchymal Stem Cells and Female Rats

Cited by 3 publications

References 33 publications

Synthesis and Evaluation of a Novel Series of Diphenylamine and Diphenylether Derivatives with Osteoblastogenic and Osteogenic Effects <i>via</i> CDK8 Inhibition

Synthesis and Evaluation of a Novel Series of Diphenylamine and Diphenylether Derivatives with Osteoblastogenic and Osteogenic Effects <i>via</i> CDK8 Inhibition

Mediator kinase inhibition drives myometrial stem cell differentiation and the uterine fibroid phenotype through super-enhancer reprogramming

A potent small molecule, AS2863619, enhances the osteogenic potential and ameliorates the inflammation-induced impairment of osteogenesis in periodontal ligament stem cells

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