ALKBH5-mediated m6A demethylation of Runx2 mRNA promotes extracellular matrix degradation and intervertebral disc degeneration.

BACKGROUND: N6-methyladenosine (m6A) methylation is a prevalent RNA modification implicated in various diseases. However, its role in intervertebral disc degeneration (IDD), a common cause of low back pain, remains unclear. RESULTS: In this investigation, we explored the involvement of m6A demethylation in the pathogenesis of IDD. Our findings revealed that ALKBH5 (alkylated DNA repair protein AlkB homolog 5), an m6A demethylase, exhibited upregulation in degenerative discs upon mild inflammatory stimulation. ALKBH5 facilitated m6A demethylation within the three prime untranslated region (3'-UTR) of Runx2 mRNA, consequently enhancing its mRNA stability in a YTHDF1 (YTH N6-methyladenosine RNA binding protein F1)-dependent manner. The subsequent elevation in Runx2 expression instigated the upregulation of ADAMTSs and MMPs, pivotal proteases implicated in extracellular matrix (ECM) degradation and IDD progression. In murine models, subcutaneous administration of recombinant Runx2 protein proximal to the lumbar disc in mice elicited complete degradation of intervertebral discs (IVDs). Injection of recombinant MMP1a and ADAMTS10 proteins individually induced mild to moderate degeneration of the IVDs, while co-administration of MMP1a and ADAMTS10 resulted in moderate to severe degeneration. Notably, concurrent injection of the Runx2 inhibitor CADD522 with recombinant Runx2 protein did not result in IVD degeneration in mice. Furthermore, genetic knockout of ALKBH5 and overexpression of YTHDF1 in mice, along with lipopolysaccharide (LPS) treatment to induce inflammation, did not alter the expression of Runx2, MMPs, and ADAMTSs, and no degeneration of the IVDs was observed. CONCLUSION: Our study elucidates the role of ALKBH5-mediated m6A demethylation of Runx2 mRNA in activating MMPs and ADAMTSs, thereby facilitating ECM degradation and promoting the occurrence of IDD. Our findings suggest that targeting the ALKBH5/Runx2/MMPs/ADAMTSs axis may represent a promising therapeutic strategy for preventing IDD.

USP24-dependent stabilization of Runx2 recruits a p300/NCOA3 complex to transactivate ADAMTS genes and promote degeneration of intervertebral disc in chronic inflammation mice.

BACKGROUND: Intervertebral disc degeneration (IDD) naturally occurs during the aging process. Its occurrence is closely related to chronic inflammation; however, the causal relationship between them is controversial. This study aimed to investigate if inflammation would promote IDD incidence and explore the underlying mechanism. METHODS: A chronic inflammation mouse model was established by intraperitoneal injection of lipopolysaccharide (LPS). Enzyme-linked immunosorbent assay was performed to determine proinflammatory cytokines in serum. Histological staining was used to evaluate the degeneration of IVDs. Immunoblots and RT-qPCR analyses were performed to measure protein and mRNA expression levels. Immunoprecipitation, mass spectrometry, and co-immunoprecipitation assays were used to determine the assembly of protein complex. RESULTS: We found that an inflammatory microenvironment activated p38 kinase, which phosphorylated the Runx2 transcription factor at the Ser28 site. The phosphorylated Runx2 (pRunx2) then recruited a deubiquitinase, ubiquitin-specific peptidase 24 (USP24), which stabilized pRunx2 and protected it from ubiquitin-dependent proteasomal degradation. The stabilized pRunx2 recruited histone acetyltransferase p300 and nuclear receptor coactivator 3 (NCOA3) to assemble a complex. This NCOA3-p300-pRunx2 complex then transactivated the expression of 13 ADAMTS (a disintegrin and metalloproteinase with thrombospondin motif) genes, thereby promoting the degradation of extracellular matrix (ECM) in intervertebral discs (IVDs) and causing IDD. Administration of either a p38 inhibitor (doramapimod), a NCOA3 inhibitor (bufalin), or a p300 inhibitor (EML425) significantly decreased the expression of the 13 ADAMTS genes and slowed the degeneration of IVDs. CONCLUSION: In summary, our results demonstrate that USP24 protects pRunx2 from proteasomal degradation under chronic inflammation conditions, enabling pRunx2 to transactivate ADAMTS genes and degrade ECM. Our findings provide direct evidence that chronic inflammation triggers IDD and offer a therapeutic strategy for retarding IDD in patients with chronic inflammation.

A Novel Method for Notable Reducing Phase Transition Temperature of VO2 Films for Smart Energy Efficient Windows.

Although Vanadium dioxide (VO2) has a potential application value for smart energy efficient windows because of its unique phase transition characteristic, there are still many obstacles that need to be overcome. One challenge is to reduce its high transition temperature (ζc = 68 °C) to near room temperature without causing its phase transition performance degradation. In this paper, a novel method was employed that covered a 3 nm ultra-thin heavy Cr-doped VO2 layer on the pure VO2 films. Compared with the as-grown pure VO2, obviously, phase transition temperature decreasing from 59.5 °C to 48.0 °C was observed. Different from previous doping techniques, almost no phase transition performance weakening occurred. Based on the microstructure and electrical parameters measurement results, the mechanism of ζc reducing was discussed. The upper ultra-thin heavy Cr-doped layer may act as the induced role of phase transition. With temperature increasing, carrier concentration increased from the upper heavy Cr-doped layer to the bottom pure VO2 layer by diffusion, and induced the carrier concentration reach to phase transition critical value from top to bottom gradually. The present method is not only a simpler technique, but also avoids expensive alloy targets.

Notable Enhancement of Phase Transition Performance and Luminous Transmittance in VO2 Films via Simple Method of Ar/O Plasma Post-Treatment.

Ar/O plasma irradiation is proposed for post-treatment of vanadium dioxide (VO2) films. Oxidation and surface migration were observed in the VO2 films following irradiation. This combined effect leads to an effective stoichiometry refinement and microstructure reconstruction in the interfacial area. A notable improvement in luminous transmittance and an enhancement in phase transition performance of the treated VO2 films were achieved. Compared with that of as-deposited VO2 films, the electrical phase transition amplitude of treated films increased more than two-fold. The relative improvement in luminous transmittance (380⁻780 nm) is 47.4% (from 25.1% to 37%) and the increase in solar transmittance is 66.9% (from 29.9% to 49.9%), which is comparable to or better than the previous work using anti-reflection (AR) coatings or doping methods. The interfacial boundary state proved to be crucial and Ar/O plasma irradiation offers an effective approach for further refinement of thermochromic VO2 films.