Multi-target regulation by artemether in MAFLD through EGFR/HSP90 pathways.

Background and aims The escalating prevalence of metabolic dysfunction-associated fatty liver disease (MAFLD) poses a significant global health burden. MAFLD is characterized by abnormal lipid accumulation in liver parenchymal cells and activation of endoplasmic reticulum stress. However, the effect of Krüppel-like factor 16(KLF16) on glycolipid metabolism has been limited. We investigated whether KLF16 could alleviate MAFLD through endoplasmic reticulum stress. Methods HepG2 and mouse primary hepatocytes were treated with oleic acid (OA) to model MAFLD in vitro. siRNA was used to downregulate KLF16 expression in cells. C57/BL6J mice were fed a high-fat diet to model MAFLD in vivo, and AAV8 was used to regulate KLF16 and ATF4 expression. Western-blot, RT-qPCR, oil red O staining, and dual-luciferase reporter assays were used to explore the underlying mechanisms in MAFLD. T-test and ANOVA were used to compare the differences among groups. Results KLF16 expression was upregulated in MAFLD models. KLF16 downregulation aggravates lipid deposition in liver cells. The expression of ATF4 protein was downregulated in MAFLD models with KLF16 knockdown. KLF16 transcriptionally regulates ATF4 expression through two binding sites. ATF4 overexpression alleviates lipid deposition exacerbated by KLF16 knockdown in mice. Conclusion In the face of abnormal lipid deposition in MAFLD, liver cells can play a spontaneous protective role by upregulating KLF16, which can promote the expression of ATF4 at the transcriptional level, further affecting downstream lipid metabolism-related genes. KLF16 may serve as a promising target gene to improve the progression or prognosis of MAFLD.

Gastric cancer (GC) remains the fifth most common cancer worldwide. Heat-shock protein 90 (HSP90) has become an attractive therapeutic target in treating cancers, because of its abnormally high expression in cancers. Several successful cases of HSP90 inhibitors capable of inhibiting GC inspired us to try ganetespib, a clinically promising and actively investigated second-generation HSP90 inhibitor in GC treatment. In our study, we show that ganetespib markedly reduced the growth of MGC-803 and also significantly inhibited the growth of SGC-7901 and MKN-28 in a dose-dependent manner. It induced G2/M cell-cycle arrest and apoptosis in all three cell lines, together with the related markers affected significantly. Mechanistically, ganetespib caused pronounced decrease of expression of classic HSP90 client proteins. Specifically, it greatly affected epidermal growth factor receptor (EGFR) signaling cascades by markedly decreasing the levels of total EGFR and EGFR on cell membranes. EGFR knockdown also induced cell-cycle arrest and apoptosis accompanied with a decrease of several EGFR downstream proteins. These results strongly support that EGFR signaling greatly contributes to the ganetespib inhibitory effects. Besides, we found that the responses of GC cell lines to ganetespib correlated well with their EGFR expression levels: MGC-803, as well as AGS and BGC-803, with higher EGFR expression responded to ganetespib better, whereas SGC-7901 and MKN-28 with lower EGFR levels were much less sensitive to ganetespib. Although SGC-7901 and MKN-28 were not very sensitive to ganetespib, ganetespib worked synergistically with radiation and cisplatin in killing them. Importantly, ganetespib significantly inhibited the growth of xenograft tumors in vivo as a single agent or in combination with cisplatin. Results of hematoxylin/eosin staining, TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) assays, and immunohistochemistry staining of phosphorylated cyclin-dependent kinase 1 (pCDK1), EGFR and Ki-67 revealed significant differences in ganetespib-treated tumors. Collectively, our data suggest that ganetespib, as a new potent treatment option, can be used for the molecularly targeted therapy of GC patients according to their expression profiles of EGFR.

Although generally acknowledged as a plasma membrane protein, the epidermal growth factor (EGF) receptor has been found in the nucleus and subcellular organelles. Recently, the mitochondrial localization of the EGF receptor (EGFR) was reported; nevertheless, the molecular mechanism underlying EGFR localization in mitochondria is largely unknown. Using immunofluorescence and immunoelectron microscopy, we observed that EGFR did localize within mitochondria. Moreover, EGFR mitochondrial translocation can be increased by rapamycin treatment in A431 cells and greatly reduced by the presence of 3-methyladenine (3-MA), an inhibitor of autophagy. The reduction of mitochondrial EGFR via autophagy inhibition is further confirmed by small interference RNA (siRNA), through which the essential protein Beclin 1 was depleted. Knocking down Beclin 1 markedly decreased the mitochondrial translocation of EGFR that was induced by rapamycin. We also noticed that the content of mitochondrial EGFR transfer is decreased when the cells are exposed to the apoptotic inducer etoposide. Additionally, either EGF treatment or EGFR knockdown by siRNA results in a greater decline of cell viability in cells possessing more mitochondrial EGFRs. Taken together, we conclude that EGFR mitochondrial localization is regulated by either autophagy or programmed cell death and is correlated with cell survival.

Hepatocellular carcinoma (HCC) represents a highly heterogeneous malignancy whose pathogenesis involves multiple etiological factors, including hepatitis viruses, metabolic disorders, and environmental carcinogens. Nevertheless, clinical management continues to face significant challenges due to high rates of postoperative recurrence, metastasis, and drug resistance. Emerging evidence indicates that Annexin A2 (ANXA2), a calcium-dependent phospholipid-binding protein, plays a pivotal role throughout HCC progression by integrating multiple oncogenic pathways. This review systematically elucidates ANXA2’s multifaceted regulatory mechanisms in HCC pathogenesis. Functioning as a critical molecular nexus, ANXA2 is dynamically modulated in its expression and activity by hepatic insults, particularly HBV/HCV infection, Alcoholic Liver Disease (ALD), Metabolic Dysfunction-associated Fatty Liver Disease (MAFLD), and tobacco exposure, thereby bridging chronic liver injury and malignant transformation. Mechanistically, ANXA2 promotes tumor progression through multiple pathways, including ubiquitination regulation, stem cell characteristic regulation, cytoskeletal remodeling, Epithelial–Mesenchymal Transition (EMT) induction, and autophagy modulation. Furthermore, it confers chemotherapy resistance specifically through regulating apoptosis evasion, ferroptosis suppression, and autophagy homeostasis, while its effects on radiotherapy and immunotherapy responses involve distinct mechanisms. Promisingly, emerging ANXA2-targeted strategies demonstrate therapeutic potential, including exosome and microenvironment modulation, molecular targeted therapy, regulation of the ANXA2-NF-κB axis, Nanotechnology, and radionuclide theranostics. Collectively, this review establishes an integrated “etiology-mechanism-intervention” framework while providing novel molecular targets and a theoretical foundation for precision medicine approaches in HCC.

Taohong Siwu decoction ameliorates hepatic lipid accumulation in female mice with heart failure and preserved ejection fraction via AMPK/SREBP1 pathway.

The rising prevalence of chronic hepatitis B (CHB) and metabolic dysfunction-associated fatty liver disease (MAFLD) comorbidity necessitates a deeper understanding of their clinical interplay. Although miR-122 is a known liver-specific regulator of chronic liver diseases, its specific role in the context of CHB-MAFLD remains elucidated. In this study, serum analysis revealed that CHB-MAFLD patients exhibit significantly lower levels of HBV DNA, pgRNA, and HBsAg compared to patients with CHB alone, which correlated with elevated miR-122 expression. Through bioinformatics and molecular assays, SOX4 was identified as a direct downstream target of miR-122-5p. Utilizing an HBV-infected steatotic cell model, we observed that the comorbid state suppresses HBV replication markers-including DNA, pgRNA, HBsAg, and HBcrAg-while concurrently upregulating the miR-122-5p/SOX4 axis. Functional experiments demonstrated that miR-122-5p and SOX4 both act as inhibitors of HBV activity, as their knockdown enhanced viral replication, while their overexpression led to significant suppression. These findings suggest that the metabolic environment in CHB-MAFLD may naturally suppress HBV replication through the activation of a novel miR-122/SOX4 regulatory axis. This study highlights miR-122 as a potential therapeutic target and provides a mechanistic basis for the altered viral kinetics observed in patients with dual liver pathology. Trial Registration: Chinese Clinical Trial Registry identifier: ChiCTR2200063555.

Cholesterol, an indispensable structural and signaling lipid, is fundamental to cellular membrane integrity, steroidogenesis, and developmental morphogen pathways. Its homeostasis hinges on the precise coordination of four interdependent metabolic modules: de novo biosynthesis, intestinal absorption, enzymatic conversion, and systemic clearance. This review delineates the molecular machinery governing these processes—from the Bloch/Kandutsch-Russell synthesis pathways and niemann-pick C1-like 1 (NPC1L1)-mediated cholesterol uptake to cholesterol 7α-hydroxylase (CYP7A1)-driven bile acid synthesis and HDL-dependent reverse transport. We further elucidate cholesterol’s multifaceted roles in lipid raft assembly, Hedgehog signal transduction, and vitamin D/hormone production. Critically, dysregulation of cholesterol flux underpins pathogenesis in atherosclerosis, metabolic dysfunction-associated fatty liver disease (MAFLD), neurodegenerative disorders, and oncogenesis, with disrupted synthesis, efflux, or esterification cascades serving as key drivers. Emerging therapeutic strategies extend beyond conventional statins and proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors to include transformative modalities: CRISPR-based in vivo gene editing (e.g., VERVE-101 targeting PCSK9), small interfering RNA (siRNA) therapeutics (inclisiran), and microbiota-directed interventions. Pioneering approaches against targets Such as angiopoietin-like 3 (ANGPTL3), lipoprotein(a) [Lp(a)], and asialoglycoprotein receptor 1 (ASGR1)—alongside repurposed natural agents (berberine, probiotics)—offer promise for mitigating residual cardiovascular risk and advancing precision cardiometabolic medicine. By integrating mechanistic insights with clinical advancements, this review underscores the transition from broad-spectrum therapies to personalized, multi-target regimens, offering a roadmap for mitigating cholesterol-related diseases in the era of genomic and metabolic medicine.

Procyanidin C1 from hawthorn modulates metabolic-inflammatory axis to combat vascular senescence in atherosclerosis.

IA36: Treatment strategies to overcome EGFR TKI resistance.

In a report accompanying our article, Spigel et al present their findings on combining erlotinib, an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), with onartuzumab (MetMAb; Genentech, South San Francisco, CA), a humanized monovalent monoclonal antibody that blocks ligand activation of MET receptor tyrosine kinase (RTK). MET-positive patients (defined by immunohistochemistry) treated with erlotinib and onartuzumab demonstrated improved progression-free and overall survival, whereas MET-negative patients had worse outcomes. This article provides an overview of signaling control by EGFR and MET and the biologic rationale for cotargeting strategies. EGFR and MET are membrane-bound RTKs consisting of extracellular domains that bind to growth factors and intracellular domains that contain a kinase domain (Fig 1). MET is activated by its cognate ligand, hepatocyte growth factor (HGF), which is produced by stromal cells, also in an autocrine fashion. EGFR is activated by several ligands, including EGF, transforming growth factor alpha (TGF ), and heparin-binding EGF. On receptor binding of the ligands, the kinase domain becomes activated, leading to transautophosphorylation of tyrosine residues on the cytoplasmic tail. Phosphorylation of these tyrosine residues creates intramolecular docking sites for proteins containing Src homology 2 domains that serve as readers for tyrosine phosphorylation. This includes adaptor proteins such as growth factor receptor–bound protein 2 (GRB2), GAB1 (GRB2-associated binding protein 1), and Src homology 2–containing transforming protein 1 (SHC1). These proteins serve as scaffolds to support signal transduction to the mitogen-activated protein kinase (MAPK), phosphoinositide 3-kinase (PI3K), phospholipase C (PLC), and signal transducer and activator of transcription (STAT) pathways. EGFR and MET use a highly overlapping repertoire of signaling adaptors and downstream effector pathways, highlighting their combined ability to codrive oncogenic signaling. Three general approaches are being employed in clinic to target EGFR and MET. The first uses small-molecule TKIs, such as erlotinib for EGFR and crizotinib for MET, that bind the ATP-binding pocket in the kinase domain and inhibit the kinase function. The second approach uses antibody therapy directed against the extracellular domain of the MET receptor (onartuzumab) or the HGF ligand (ficlatuzumab). The third approach targets downstream proteins involved in signaling, such as inhibitors of PI3K, or molecules regulating receptor trafficking and degradation, such as heat shock proteins. The EGFR pathway can be activated in lung cancer through genomic events, such as activating EGFR mutations or gene amplification, or through tumor environmental events, such as autocrine production of EGFR-activating ligands. Likewise, MET activation occurs through HGF produced in the tumor microenvironment and through MET gene amplification. MET amplification by fluorescent in situ hybridization occurs in approximately 5% to 15% of lung adenocarcinomas. Increased MET is associated with advanced stage of disease, poor outcome, brain metastasis, and high levels of EGFR. Coactivation of EGFR and MET occurs in subsets of lung cancer cell lines and tumor tissues and is associated with aggressive disease. In cells lacking EGFR mutation or high levels of MET, activation of either receptor can produce extensive signaling crosstalk to the opposing RTK and downstream substrates. EGFR activation can induce accumulation of activated MET protein through a c-SRC– dependent mechanism, and dual MET/EGFR inhibition has shown increased activity in xenograft models. Phosphoproteomic analysis of lung tumors and cell lines can identify tumors with concomitant EGFR and MET activation. EGFR, either through mutation or ligand-mediated activation, can also regulate MET levels through hypoxia-inducible factor-1 . Importantly, MET signaling is one actionable mechanism driving resistance to EGFR TKIs. MET activation can drive intrinsic as well as acquired resistance to EGFR through genomic and tumor microenvironment–mediated mechanisms. Primary resistance to anti-EGFR antibody therapy has been associated with MET activation in primary explant models of lung cancer. Approximately 5% of EGFR mutation–positive tumors with acquired resistance to EGFR TKIs are found to have MET gene amplification. Mechanistically, amplified MET co-opts use of ERBB3 to maintain the PI3K pathway activation. Combining MET and EGFR TKIs or EGFR/HSP90 inhibition has been effective in mouse models of lung cancer with T790M EGFR and MET amplification. High levels of HGF are associated with intrinsic or acquired resistance to EGFR TKIs in patients with lung cancer with activating EGFR mutations and in those with other solid tumors. HGF production by stromal fibroblasts leads to fibroblast-induced JOURNAL OF CLINICAL ONCOLOGY U N D E R S T A N D I N G T H E P A T H W A Y VOLUME 31 NUMBER 32 NOVEMBER 1

The identification of somatic mutations in the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) in patients with non-small cell lung cancer (NSCLC) and the association of such mutations with the clinical response to EGFR tyrosine kinase inhibitors (TKI), such as gefitinib and erlotinib, have had a substantial effect on the treatment of this disease. EGFR gene amplification has also been associated with an increased therapeutic response to EGFR-TKIs. The effects of these two types of EGFR alteration on EGFR function have remained unclear, however. We have now examined 16 NSCLC cell lines, including eight newly established lines from Japanese NSCLC patients, for the presence of EGFR mutations and amplification. Four of the six cell lines that harbor EGFR mutations were found to be positive for EGFR amplification, whereas none of the 10 cell lines negative for EGFR mutation manifested EGFR amplification, suggesting that these two types of EGFR alteration are closely associated. Endogenous EGFRs expressed in NSCLC cell lines positive for both EGFR mutation and amplification were found to be constitutively activated as a result of ligand-independent dimerization. Furthermore, the patterns of both EGFR amplification and EGFR autophosphorylation were shown to differ between cell lines harboring the two most common types of EGFR mutation (exon 19 deletion and L858R point mutation in exon 21). These results reveal distinct biochemical properties of endogenous mutant forms of EGFR expressed in NSCLC cell lines and may have implications for treatment of this condition.

The epidermal growth factor receptor (EGFR) located in dorsal root ganglion has been found as a new target for chronic pain treatment. However, it is not clear whether the change of EGFR expression in the dorsal root ganglion contributes to neuropathic pain development. In this study, we used a chronic compression of unilateral lumbar dorsal root ganglions (CCD)-induced rat neuropathic pain model and found that CCD caused the upregulation of both phosphorylated EGFR and total EGFR expression in compressed lumbar 4/5 (L4/L5) dorsal root ganglions by western blotting and immunohistochemistry methods. Either inhibition of EGFR activation by EGFR inhibitor or knockdown of EGFR expression by EGFR small interference RNA (siRNA) relieved CCD-induced pain hypersensitivities to mechanical, thermal, and cold stimuli in rats. Moreover, EGFR knockdown reversed CCD-induced the increase of intracellular mammalian target of rapamycin (mTOR) expression as well as the activation of the satellite glial cells in the ipsilateral compressed L4/L5 dorsal root ganglions. These findings suggest that not only activated EGFR but also total EGFR contribute to CCD-induced neuropathic pain by enhancing intracellular mTOR signaling.

Free Energies of Interaction of Lipids with Regulatory Binding Sites on the Transmembrane Domain of the EGF Receptor

Growth factor receptor-signaling pathways are potentially important targets for anticancer therapy. The interaction of anticancer agents with specific molecular targets can be identified by correlating target expression patterns with cytotoxicity patterns. We sought to identify new agents that target and inhibit the activity of the epidermal growth factor (EGF) receptor and of c-erbB2 (also called HER2 or neu), by correlating EGF receptor, transforming growth factor (TGF)-alpha (a ligand for EGF receptor), and c-erbB2 messenger RNA (mRNA) expression levels with the results of cytotoxicity assays of the 49000 compounds in the National Cancer Institute (NCI) drug screen database. The levels of mRNAs were measured and used to generate a molecular target database for the 60 cell lines of the NCI anticancer drug screen. The computer analysis program, COMPARE, was used to search for cytotoxicity patterns in the NCI drug screen database that were highly correlated with EGF receptor, TGF-alpha, or c-erbB2 mRNA expression patterns. The putative EGF receptor-inhibiting compounds were tested for effects on basal tyrosine phosphorylation, in vitro EGF receptor tyrosine kinase activity, and EGF-dependent growth. Putative ErbB2-inhibiting compounds were tested for effects on antibody-induced ErbB2 tyrosine kinase activity. EGF receptor mRNA and TGF-alpha mRNA levels were highest in cell lines derived from renal cancers, and c-erbB2 mRNA levels were highest in cells derived from breast, ovarian, and colon cancers. Twenty-five compounds with high correlation coefficients (for cytotoxicity and levels of the measured mRNAs) were tested as inhibitors of the EGF receptor or c-erbB2 signaling pathways; 14 compounds were identified as inhibitors of these pathways. The most potent compound, B4, inhibited autophosphorylation (which occurs following activation) of ErbB2 by 50% in whole cells at 7.7 microM. Novel EGF receptor or c-erbB2 pathway inhibitors can be identified in the NCI drug screen by correlation of cytotoxicity patterns with EGF receptor or c-erbB2 mRNA expression levels.

Berbamine attenuates acetaminophen-induced liver injury by engaging GCLC and enhancing ferroptosis-regulatory antioxidant pathways.