MetaboLightsapplication/xmlftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/m_MTBLS15834_LC-MS_alternating_hilic_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/s_MTBLS15834.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/a_MTBLS15834_LC-MS_alternating_hilic.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/i_Investigation.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siSTK39-5.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siNC-2.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siSTK39-4.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siNC-5.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siSTK39-3.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siNC-1.mzMLftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834/FILES/DERIVED_FILES/AngII-siSTK39-2.mzMLprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS15834<p>Identified metabolites were annotated using KEGG compound database (http://www.kegg.jp/kegg/compound/), annotated metabolites were then mapped to KEGG Pathway database (http://www.kegg.jp/kegg/pathway.html). Pathways with significantly regulated metabolites mapped to were then fed into MSEA (metabolite sets enrichment analysis), their significance was determined by hypergeometric test's P-Values.</p>MetaboLightsPublicLiquid Chromatography MS - alternating - hilic<p>The sample extracts were analyzed using an LC-ESI-MS/MS system (Waters ACQUITY H-ClassD, https://www.waters.com/nextgen/cn/zh.html; MS, QTRAP 6500+ System, https://sciex.com/). The analytical conditions were as follows.</p><p><br></p><p>Amide method: HPLC: column, ACQUITY UPLC BEH Amide (i.d. 2.1x100 mm, 1.7 um); solvent system, water with 10 mM Ammonium acetate and 0.3% Ammonium hydroxide (A), 90% acetonitrile/water (V/V) (B); The gradient was started at 95% B (0-1.2 min), decreased to 70% B (8 min), 50% B (9-11 min), finally ramped back to 95% B (11.1-15 min); flow rate, 0.4 mL/min; temperature, 40 degC; injection volume: 2 uL.</p>Targeting STK39 ameliorates cardiac remodeling by Improving Mitochondrial function through degradation of IFIT3.The Second Affiliated Hospital of Harbin Medical UniversityHuibin Liuheartmass spectrometry assay<p>The sample was thawed on ice, 100 uL of ultrapure water extract was added to resuspend the cell pellet. Divide 50 uL cell suspension and add 200 uL of methanol (precooled at -20 degC) and vortexed for 2 min under the condition of 2500 r/min. The sample was frozen in liquid nitrogen for 5 min, removed on ice for 5 min, after that, the sample was vortexed for 2 min. The previous step was repeated for 3 times. The sample was centrifuged at 12000 r/min for 10 min at 4 degC. Take 200 uL of supernatant into a new centrifuge tube and place the supernatant in -20 degC refrigerator for 30 min. Then the supernatant was centrifuged at 12000 r/min for 10 min at 4 degC. After centrifugation, transfer 180 uL of supernatant through Protein Precipitation Plate for further LC-MS analysis. The left 50 uL cell suspension was frozen and thawed for 3 times, centrifuged at 12,000 r/min for 10 min, and the supernatant was taken to determine the protein concentration by BCA Protein Assay kit.</p>Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS15834jie zhang. The Second Affiliated Hospital of Harbin Medical University. jiezhang202411@163.com.Huibin Liu. The Second Affiliated Hospital of Harbin Medical University. liuhuibin66@126.com.<p>Mass spectrometer parameters including the declustering potentials (DP) and collision energies (CE) for individual MRM transitions were done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period.</p>Treatmentliuhuibin66@126.com<p>Ventricular cardiomyocytes were isolated from neonatal mice. The neonatal ventricular cardiomyocytes were maintained in DMEM medium containing 10% fetal bovine serum (FBS) and antibiotics, placed in a humid environment with 5% CO2 at 37 degC. On the second day after the cells attached (marked as day 0), siSTK39 was transfected, and the cells were kept silent for 12 hours, then treated with AngII (MCE) for 48 hours to induce cardiac hypertrophy. After the experimental treatment, the culture medium was aspirated to completely remove it. The cells were quickly washed 3 times with pre-chilled PBS to eliminate residual medium and serum components. A small volume of pre-chilled PBS was then added, and the cells were gently scraped off using a cell scraper and transferred to a centrifuge tube. The suspension was centrifuged at 4 degC and 500g for 5 min, after which the supernatant was discarded. The cells were resuspended and washed with pre-chilled PBS, centrifuged at 4 degC and 1000g for 5 min at low speed, and the supernatant was discarded again. After an additional wash with PBS, the cells in the PBS suspension were counted, and an aliquot of 1x10^7 cells per sample was transferred into a 2 mL sterile centrifuge tube. The aliquot was centrifuged at 4 degC and 1000g for 10 min at low speed, and the supernatant was discarded to retain the cell pellet. The cell pellet was immediately snap-frozen in liquid nitrogen and subsequently stored at -80 degC, with all samples shipped on dry ice to ensure they remained frozen during transport.</p>MetabolomicsMetabolomicsTandem Mass Spectrometry£¬QTRAP? 6500+Mus musculusUbiquitinationtargeted analysismitochondrial homeostasisSTK39cardiac hypertrophyheartIFIT3Ultra Performance Liquid Chromatography£¬Waters ACQUITY H-ClassDMetabolomicsTandem Mass Spectrometry£¬QTRAP? 6500+UbiquitinationMus musculustargeted analysismitochondrial homeostasisSTK39cardiac hypertrophyIFIT3heartUltra Performance Liquid Chromatography£¬Waters ACQUITY H-ClassD<p>Linear ion trap (LIT) and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (QTRAP), QTRAP 6500+ LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in both positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: ion source, ESI+/-; source temperature 550 degC; ion spray voltage (IS) 5500 V (Positive), -4500 V (Negative); curtain gas (CUR) was set at 35 psi, respectively. Energy and its metabolites were analyzed using scheduled multiple reaction monitoring (MRM). Data acquisitions were performed using Analyst 1.6.3 software (Sciex). Multiquant 3.0.3 software (Sciex) was used to quantify all metabolites. Mass spectrometer parameters including the declustering potentials (DP) and collision energies (CE) for individual MRM transitions were done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period.</p>6-Phosphogluconic-acidSuccinyl-CoASuccinic-AcidLactateGuanosineFructose-1,6-bisphosphatec-di-AMPOxaloacetateDL-Glyceric-AcidGlucuronic-acidADPNicotinaMide-adenine-dinucleotide(NAD)GluconatePhosphoenolpyruvic-acidL-AlanineD-Ribose-5-phosphate-disodiumItaconic-acid2-Phospho-D-glycerateAlpha-Ketoglutaric-Acid2-Oxoadipic-acidL-LeucineTyrosineL-CystineAMPThreonineFlavin-mononucleotideD-Erythrose-4-phosphateD-Fructose-6-phosphate3-phenyllactic-acidGlyceraldehyde-3-phosphateIMPD-Ribulose-5-phosphateArgininosuccinic-acidPhosphorylethanolamineLysineL-Glutamic-acidFumaric-acidUMPD-Glucose-1-phosphateOrnithinedTMPTrehalose-6-phosphateCitric-acidD-Glucose-6-phosphateSerineL-2-Hydroxyglutaric-acid-disodiumCyclic-AMPL-AsparagineUDP-GlcNAcDihydroxyacetone-phosphateMalic-acidD(+)-GlucoseL-citrullineGlycerol-3-phosphateArginineD-ribulose-1,5-bisphosphate2,3-DPGIsocitric-acidGlycolic-acidPyruvic-acidSedoheptulose-7-phosphatecis-Aconitic-acidAdenineAcetyl-CoAdCMPD-Mannose-6-phosphatedAMPCysteic-acidXylulose-5-phosphateTriphosphate-guanosine3-phosphoglycerateInosinedUMPDihydronicotinamide-adenine-dinucleotide-phosphate(NADPH)ATPUracilUreidopropionateL-AspartateGuanosine-diphosphateGlutaminefalseTargeting STK39 ameliorates cardiac remodeling by Improving Mitochondrial function through degradation of IFIT3<p>Pathological cardiac remodeling in response to sustained pressure overload is a pivotal process in the progression of heart failure (HF), yet the underlying molecular mechanisms remain incompletely understood, limiting therapeutic development. Here, we identify serine/threonine kinase 39 (STK39) as a critical mediator across this remodeling progression, showing its marked upregulation in both hypertrophic and failing human hearts, as well as in mouse hearts after transverse aortic constriction. Cardiomyocyte-specific Stk39 knockdown ameliorates cardiac hypertrophy, improves ventricular function, and attenuates myocardial fibrosis in vivo, while STK39 silencing suppresses angiotensin II (Ang II)-induced hypertrophic growth, fibrosis, and mitochondrial dysfunction in mice cardiomyocytes in vitro. Integrated multi-omics and experimental validation identify interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) as a key downstream target. Mechanistically, STK39 physically interacts with IFIT3 and competes with the E2/E3 ligase UBE2O for binding, thereby protecting IFIT3 from UBE2O-mediated ubiquitination and degradation. Stabilized of IFIT3 impairs mitochondrial homeostasis via VDAC2, promoting mitochondrial reactive oxygen species (ROS) production, which triggers Ca2+ dysregulation and subsequent Calcineurin/NFATc3 activation, ultimately driving pathological gene transcription and cardiac remodeling. Collectively, our findings uncover a reveal a previously unrecognized novel function of STK39 and establish the STK39-IFIT3 axis that links mitochondrial dysregulation to classical hypertrophic signaling, positioning this axis as a potential therapeutic target for pressure overload-induced HF.</p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