搜索结果：Biochimica et biophysica acta

Venlafaxine (VLX) and its active metabolite O-desmethylvenlafaxine (ODV) must partition into lipid membranes before reaching their membrane-embedded transporter targets. The principal active metabolite, ODV, differs from VLX by the removal of a methyl group and is itself marketed as an antidepressant. Here, we combined long-timescale all-atom molecular dynamics simulations with experimental membrane partitioning measurements to investigate how demethylation, stereochemistry, and drug concentration influence their interactions with phosphatidylcholine bilayers. All drug species spontaneously partitioned into the lipid-water interface and stabilized at ∼1.4 nm from the bilayer center while maintaining a conserved amphipathic orientation. Free-energy calculations revealed similar interfacial free-energy minima of approximately -20 kJ mol-1 for both compounds, whereas the energetic penalty for translocation toward the bilayer core exceeded 40 kJ mol-1. Demethylation of VLX lowered the energetic barrier for membrane insertion without altering the thermodynamically favored interfacial state, indicating that ODV accesses the membrane interface more efficiently while preserving comparable equilibrium membrane affinity. Experimental second-derivative UV-Vis spectroscopy further confirmed similar membrane partitioning for VLX and ODV. At low drug concentrations, membrane structure remained largely unperturbed following insertion. In contrast, elevated drug loading induced cooperative membrane perturbation characterized by lateral bilayer expansion, membrane thinning, and reduced acyl-chain order, with these effects consistently more pronounced for ODV. Together, these findings identify the lipid bilayer under studied conditions as a dynamically adaptive kinetic interface rather than a passive equilibrium reservoir, suggesting that functional differences between VLX and ODV arise primarily from differences in interfacial accessibility and subsequent protein-specific interactions rather than nonspecific membrane affinity alone.

Post-translational modifications (PTMs) play a key role in regulating protein/protein interactions and protein stability, thus influencing protein expression and function. HSPs and p53, including its wtp53 and mutp53 forms, make no exception to this rule, although the impact of PTMs on the regulation of these proteins has not yet been fully elucidated, particularly in the case of mutp53. These proteins, unlike wtp53, can behave as oncogenes, making their targeting an important step for successful anticancer therapy. We previously reported that mutp53 is degraded, preferentially via CMA, in colon cancer cells stressed by long-term TG treatment. Whether TG could induce PTMs and how they could contribute to mutp53 degradation has not yet been investigated and will be explored in this study. Acetylation of mutp53, as well as HSP90, has been reported to promote mutp53 degradation. However, we found that TG promoted deacetylation of mutp53 and HSP90, due to the sustained activity of HDAC6, a PTM that protected mutp53 from degradation. We then found that mutp53 was progressively demethylated by KDM1 at lysine K370 during TG treatment, which facilitated the interaction with HSC70 involved in mutp53 protein degradation via CMA. In conclusion, this study suggests that, in colon cancer cells subjected to stress by TG, mutp53 was degraded as a consequence of demethylation at lysine K370. Therefore, specific epigenetic drugs capable of reducing constitutive methylation and/or increasing acetylation could preemptively target mutp53 and improve the outcome of endoplasmic reticulum stress-inducing treatments in tumors harboring these proteins.

Immune checkpoint blockade (ICB) provides limited benefit in triple-negative breast cancer (TNBC), partly due to a lactate-rich, immunosuppressive tumor microenvironment (TME). Integrative bulk and single-cell transcriptomics, corroborated by tissue analyses, identify MCT4 as the predominant lactate transporter in TNBC, while supporting a context-dependent contribution of MCT1, and reveal an inverse association between MCT4 and intratumoral CD8+ T cells. We therefore tested whether inhibition of monocarboxylate transporters could recondition the TME. The repurposed agent syrosingopine, a dual MCT1/MCT4 inhibitor, consistently blocked lactate export across MCT1+ and MCT4+ models without altering transporter/LDH abundance, and reduced tumor cell PD-L1 while restoring T-cell effector function. Syrosingopine also sensitized tumor cells to NK-cell cytotoxicity without detectable toxicity in immune cells. In immunocompetent TNBC models, syrosingopine synergized with anti-PD-1 to suppress tumor growth, remodel the immune landscape, and prolong survival, while genetic perturbation of MCTs recapitulated key metabolic and immunologic phenotypes. These findings identify tumor lactate export as an important metabolic barrier to antitumor immunity in TNBC and support MCT-targeted lactate transport inhibition as a promising strategy to reprogram the TME and improve immunotherapeutic efficacy.

Tumor heterogeneity is a fundamental feature of malignant tumors and a major driver of treatment resistance, ultimately contributing to treatment resistance, therapeutic failure, disease recurrence, and poor outcome. Rather than being static, heterogeneity arises early during tumorigenesis and continues to evolve during progression, metastasis, and therapeutic selection. Advances in single-cell and spatial omics, integrative multi-omics profiling, liquid biopsy, and computational modeling have markedly improved the characterization of heterogeneity across genetic, epigenetic, transcriptomic, phenotypic, and microenvironmental dimensions. However, translating heterogeneity profiling into routine clinical decision-making remains challenging due to sampling bias, technical variability, data-integration complexity, and the need for prospectively validated, actionable metrics. In this review, we summarize the biological origins and major classifications of tumor heterogeneity, discuss current approaches for its detection and longitudinal monitoring, examine its implications for established and emerging therapies, and highlight precision-oncology strategies aimed at anticipating, tracking, and ultimately exploiting heterogeneity to achieve durable cancer control.

Innate immune activation is tightly coupled to metabolic remodeling, yet how STING-associated signaling intersects with macrophage metabolism during bacterial infection remains incompletely understood. Here, using Listeria monocytogenes infection models, we show that infection is accompanied by increased glycolytic gene expression, glucose consumption, and lactate production in macrophages, and that STING deficiency attenuates these metabolic changes, whereas cGAS deficiency has a comparatively limited effect under our experimental conditions. Reduced glucose availability, 2-deoxy-d-glucose, and metformin enhanced AMPK activation, TBK1 phosphorylation, and IFN/ISG transcription, whereas Compound C treatment or Prkaa1 knockdown attenuated this enhancement. STING deficiency or acute STING inhibition reduced infection-associated glucose metabolic responses and limited the amplification of type I interferon-response gene expression under metabolic stress. Although the upstream mechanism by which STING regulates glucose metabolism remains to be fully defined, our findings support a model in which STING-associated metabolic changes and AMPK activity cooperate to enhance infection-induced interferon signaling during Listeria monocytogenes infection.

Lipid droplets (LDs) are dynamic organelles that coordinate lipid storage, trafficking, and metabolic adaptation under physiological and stress conditions. Despite their emerging role in cellular homeostasis, the molecular basis of treatment-induced lipid droplet remodeling remains insufficiently defined. Here, we combine lipid droplet isolation with label-free Raman spectroscopy to characterize biochemical and spectroscopic signatures associated with structural remodeling of isolated lipid droplets (iLDs) derived from normal Schwann cells and malignant peripheral nerve sheath tumor (MPNST) cells exposed to cannabidiol (CBD), ionizing radiation, and their combination. Our analysis reveals pronounced intrinsic spectral heterogeneity within iLD fractions and identifies treatment- and cell type-specific alterations in lipid composition, Raman spectral markers associated with acyl chain packing, and conformational order. Notably, stress-induced remodeling involves coordinated changes in lipid chain organization, highlighting lipid droplets as dynamic regulators of cellular metabolic adaptation. These findings provide molecular insight into lipid droplet-mediated stress responses and establish Raman-based profiling of isolated LDs as a powerful approach for investigating lipid remodeling mechanisms within isolated lipid droplet-enriched fractions. We further propose the Raman intensity ratio I₁₁₆₇/I₁₂₉₂ as a semiquantitative Raman-derived spectral index associated with stress-induced lipid remodeling and CBD-mediated radiosensitization.

Novel therapeutic strategies for Parkinson's disease (PD) are urgently needed. Neuroinflammation is a critical driver of disease progression and represents a promising target for intervention. Emerging evidence highlights lactate as a signaling metabolite that regulates inflammatory responses through protein lactylation. Given the involvement of 14-3-3 proteins in PD pathogenesis, we investigated whether lactate confers neuroprotection by promoting 14-3-3 lactylation and modulating neuroinflammatory signaling in PD. A rat model of PD was induced by subcutaneous injection of Rotenone (ROT) into the dorsal cervical region. Lactate was administered intracerebroventricularly. Motor function was assessed using open field, grid, and suspension tests. TH-positive neurons in the substantia nigra were evaluated by immunohistochemistry. The lactylation of 14-3-3 proteins and their interaction with NLRP3 were examined by co-immunoprecipitation (Co-IP). Mitochondrial localization of GSDMD was visualized by immunoelectron microscopy. The cytosolic mtDNA was assessed using qPCR. NLRP3 inflammasome components, the cGAS-STING pathway, and mitochondrial GSDMD were analyzed by western blotting. Levels of inflammatory cytokines and cGAMP were quantified by ELISA. Lactate ameliorated motor deficits and dopaminergic neuron loss in ROT-treated rats. Lactate increased 14-3-3 lactylation and enhanced 14-3-3 binding to NLRP3, accompanied by reduced NLRP3 inflammasome activation, attenuated GSDMD-associated mitochondrial injury, decreased cytosolic mtDNA levels, and suppressed cGAS-STING pathway activation. Lactate exerts neuroprotective effects in PD through a mechanism associated with enhanced 14-3-3 lactylation, reduced NLRP3/GSDMD pathway activation, attenuated GSDMD-associated mitochondrial injury, decreased cytosolic mtDNA levels, and suppression of cGAS-STING signaling.

Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease, with proximal tubule fibrosis being a key pathological feature. Our previous study identified significant upregulation of cytosolic nonspecific dipeptidase 2 (CNDP2) in the renal tubules of a DKD mouse model, yet its functional role remains unclear. This study aimed to investigate the role of CNDP2 in renal tubular fibrosis during DKD, focusing specifically on the "sulfur-containing amino acids (SAAs) metabolism-mammalian target of rapamycin (mTOR)" signaling axis. We employed integrated in vivo and in vitro models. Kidney- specific Cndp2 knockdown was achieved using an adeno-associated virus vector, and the CNDP2 inhibitor bestatin was used for pharmacological intervention. Renal function, fibrosis, the Ragulator-Rag GTPase-mTOR signaling pathway, amino acid profiles, and ultrastructural changes were assessed. A dietary intervention restricting SAAs was also applied. CNDP2 was specifically upregulated in renal tubules under DKD conditions. Both genetic knockdown and pharmacological inhibition of CNDP2 significantly improved renal function and attenuated fibrosis. Mechanistically, CNDP2 hydrolyzes dipeptides, leading to elevated levels of SAAs (cysteine/cystine). This promotes the activation of the Ragulator-Ras-related GTPase (Rag) complex, resulting in subsequent hyperactivation of mTOR signaling and driving tubular fibrosis. Notably, dietary restriction of SAAs similarly ameliorated DKD pathology. CNDP2 drives renal tubular fibrosis in DKD by activating mTOR signaling through disruption of SAAs metabolism. Our findings reveal a novel "CNDP2-SAAs-mTOR" pathway, identifying CNDP2 as a promising therapeutic target for DKD intervention.

Komagataella phaffii, formerly known as Pichia pastoris, is a methylotrophic yeast employed as a recombinant protein factory for academic and industrial purposes. Despite its advantages, some recombinant proteins, which are engineered to be secreted, are retained intracellularly and subject to degradation. Serendipitously, we discovered that fetal bovine serum (FBS) improved the secretion of several proteins. When a P. pastoris strain expressing enhanced green fluorescent protein (eGFP) was grown with FBS, western analysis revealed a 2-3 fold increased level of eGFP secretion as well as the induction of an upper molecular band (30 kD) in addition to the expected band (26.8 kD). FBS protected the C-terminus of eGFP from degradation, and the alterations to this upper molecular weight variant were triggered intracellularly by an FBS component that was most likely not a protein. To elucidate the mechanisms behind the production of the novel recombinant protein variant, we utilized site-directed mutagenesis and mass spectrometry. Through these strategies, we were able to localize posttranslational modifications to the C-terminus associated with FBS treatment. Analysis of the intracellular lysate revealed proteomic alterations, suggesting that genes involved in translation, trafficking, stress response and energetics were key players in FBS's effect on the P. pastoris secretion mechanism through interaction with eGFP's C-terminus. Our study suggests that C-terminal degradation may affect other recombinant proteins produced in P. pastoris, a problem that may be resolved by FBS.

KL4, a 21-residue mimetic of the C-terminus of surfactant protein B, is effective in treating infant respiratory distress syndrome (RDS). It works by reducing alveolar surface tension and improving oxygen exchange. Here, we use steady-state fluorescence measurements to probe how KL4 modulates bilayer order and organization in pulmonary surfactant-like lipid mixtures with varying acyl chain saturation. We show how fluorescence-based methods can distinguish between a more surface-associated and a more deeply inserted mode of interaction of KL4 in phospholipid membranes with varying acyl chain saturation. Specifically, in saturated liposomes with a tail-labeled pyrene analog, KL4 addition decreases excimer-to-monomer (Ie/Im) ratios and produces larger changes in the steady-state anisotropy of rhodamine-labeled lipids, consistent with reduced lipid mobility and peptide-induced changes in bilayer packing, and is qualitatively consistent with previous ssNMR/EPR studies of KL4. Conversely, fluorescence results for unsaturated liposomes show smaller changes in Ie/Im and anisotropy, suggesting greater interfacial interaction between KL4 and a more modest impact on bilayer order. Increasing concentrations of KL4 in unsaturated lipids result in a notable decrease in rhodamine headgroup emission intensity, suggesting local environmental shifts or collisional quenching, consistent with a more surface-proximal peptide. Overall, these fluorescence readouts are sensitive to KL4-induced changes in bilayer order, lipid dynamics, and lateral organization, but do not, by themselves, define peptide orientation or insertion depth. These assays require minimal sample and simple equipment, making them practical for monitoring how KL4 and similar surfactant mimics influence bilayer behavior across different lipid compositions and for screening lipid and peptide combinations in future models of healthy and diseased surfactant relevant to neonatal RDS and adult ARDS, including COVID-19.

Most deaths from pancreatic ductal adenocarcinoma (PDAC) result from metastatic organ failure rather than primary tumor burden. Across the metastatic cascade, tumor cells encounter hypoxia, nutrient limitation, oxidative stress, immune pressure, and changing stromal conditions, and only those capable of dynamic metabolic adaptation successfully disseminate and colonize distant sites. In this review, we summarize how metabolic plasticity shapes PDAC dissemination, organ-specific colonization, and metastatic outgrowth. We further discuss how this framework may shift therapeutic strategy from broad metabolic blockade toward precision interventions that target metastatic fitness.

The intratumoral microbiota is now recognized as a key component of the tumor microenvironment, where it influences tumor metabolism, shapes immune activity, and modulates treatment responses. Microbial metabolites such as short-chain fatty acids regulate pathways that control energy use in cancer cells and modify immune signaling within tumors. Microbial imbalance disrupts metabolic homeostasis and promotes immune escape, contributing to cancer progression and resistance to therapy. Specific taxa including Fusobacterium nucleatum drive distinct protumorigenic effects through metabolic and immunologic routes. Spatial heterogeneity of microbial colonization further defines metabolic gradients and immune niches that influence treatment efficacy. Advances in sequencing, multi-omics, and spatial profiling have clarified these interactions and identified microbial signatures with diagnostic and prognostic potential. Therapeutic strategies such as precision probiotics, engineered bacteria, and nanotechnology-based delivery systems offer avenues to target microbial metabolic pathways and enhance treatment response. Continued integration of microbiology, oncology, and bioinformatics will support translation of these findings into personalized cancer therapies.

Trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) can affect lipid metabolism, leading to weight loss and attracting widespread attention. However, its suitability for use in lactating mothers remains unclear. A transgenic mouse, named Pai (homozygous) and Pai/wt (heterozygous), capable of producing endogenous t10c12-CLA was successfully established. In this study, embryo transfer technology was utilized to transplant embryos of wild-type (wt) mice into the transgenic mice, in order to investigate the effects of t10c12-CLA on biometric and reproductive parameters. Embryo transfer efficiency remained unchanged following the procedure. The male-to-female ratio of offspring in Pai/wt group was significantly higher than that in wt and Pai groups. Additionally, the birth weight, 21-day body weight, serum glucose level and triglyceride level of pups in Pai group were significantly lower than those in wt group. Simultaneously, the Pai pups exhibited modulated gut microbiota composition and regulated metabolic profiles in white adipose tissue and liver. Further investigations in liver tissue and HepG2 cells revealed that these effects featured suppressed lipogenesis (FASN, ChREBP), enhanced fatty acid oxidation (CPT1A) through activation of AMPK pathway in liver. Maternal t10c12-CLA could suppress lipogenesis and lower lipid levels by activating the AMPK pathway, and alter the composition of gut microbiota, thereby impacting offspring lipid metabolism.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by the pathological accumulation of amyloid-β (Aβ) plaques and neurofibrillary tangles composed of hyperphosphorylated Tau protein. While microRNAs (miRNAs) have emerged as critical regulators of gene expression and potential biomarkers for AD, their role in Tau pathology remains incompletely understood. In this study, we investigated the role of the miR-92 family and its downstream targets in modulating Tau-induced neurodegeneration using the Drosophila model. Overexpression of miR-311, miR-312, and miR-313, members of the miR-92 family, exacerbated Tau-induced phenotypes. We identified deltex (dx) and engrailed (en) as miR-92 target genes with genetic screening with in silico target prediction. Overexpression of en or EN2, a human ortholog of en, reduced Tau protein levels in Drosophila and human neuroblastoma cells, respectively. Furthermore, overexpression of en attenuates Tau-associated memory deficits in Drosophila. These findings suggest that the engrailed gene is an evolutionarily conserved regulator of Tauopathy and highlight the utility of the Drosophila AD model for identifying genetic modulators with therapeutic potential in AD.

The Janus kinase (JAK)/STAT signaling pathway plays a pivotal role in cancer biology as well as in inflammatory and autoimmune disorders such as psoriasis. Recent advances in biomedical research and targeted therapies have highlighted the importance of computational approaches for accelerating the discovery of selective kinase inhibitors. This study aimed to develop a robust computational framework for predicting the inhibitory potency of JAK2 ligands and for analyzing their binding interactions using structure-based methods. A curated dataset of 1869 chemically valid JAK2 ligands with experimentally reported Ki values was compiled, standardized, and converted to pKi. Using this dataset, a bond-aware graph neural network (GNN) was trained and evaluated for pKi prediction. Top-ranked predicted ligands were further examined via molecular docking, pharmacophore modeling, and molecular dynamics simulations to assess their interactions within the JAK2 ATP-binding site. The proposed model achieved strong predictive performance, yielding an average test-set R2 of 0.91 ± 0.01, MAE of 0.14 ± 0.01, and RMSE of 0.26 ± 0.02 across repeated data splits. Structure-based analyses supported the predicted binding poses and identified key stabilizing interactions within the JAK2 ATP-binding site. Overall, this integrative computational framework provides a reliable approach for predicting JAK2 inhibitory potency and offers mechanistic insights that may support the computational prioritization of candidate molecules for future experimental evaluation.

Dysregulated lipid metabolism drives atherosclerosis (AS). Yacon, an Andean lipid-modulating tuber, exerts anti-AS potential, but mechanisms remain unclear. We integrated network pharmacology, machine learning, single-cell RNA sequencing (scRNA-seq), and in vivo validation to explore its anti-AS effects and targets. Active constituents and targets were curated from literature, TCMSP, and SwissTargetPrediction; lipid/AS genes from GeneCards, OMIM, and GEO were filtered via limma, WGCNA, LASSO, randomForest, and SVM-RFE. Immune infiltration and external validation confirmed hub gene relevance. scRNA-seq prioritized FABP5; docking and dynamics quantified compound-FABP5 interactions. In vivo efficacy was tested in high-fat diet (HFD)-fed ApoE-/- mice via histology (Oil Red O, H&E, and Masson) and molecular assays (RT-qPCR, Western blot, and immunofluorescence). We identified 12 constituents, 384 targets, and seven core targets (AURKA, MMP9, FABP5, etc.), with FABP5 top-ranked. Docking and dynamics identified Enhydrin as the strongest FABP5 binder. Enhydrin administration was associated with reduced hepatic lipid accumulation, decreased serum triacylglycerol (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels, and increased high-density lipoprotein cholesterol (HDLC) levels. Histopathological analysis of arterial tissues revealed attenuated vascular lipid deposition and delayed atherosclerotic lesion progression. Across assays, Enhydrin downregulated FABP5, reduced abnormal fatty acid trafficking, and upregulated PPARγ and ABCA1, with markedly reduced vascular lipid deposition and improved serum lipid profiles, reflecting enhanced cholesterol efflux. Conclusion: Our integrative multi-omics analysis pinpointed FABP5 as a promising novel target for yacon-derived Enhydrin in atherosclerosis. In vivo, Enhydrin markedly downregulated FABP5 and upregulated PPARγ and ABCA1, suggesting this axis mediates its anti-atherosclerotic activity. SIGNIFICANCE STATEMENT: 1: This study identified FABP5 as a candidate target for atherosclerosis through integrative multi-omics, and its association with the anti-atherosclerotic effects of Enhydrin suggests therapeutic potential. 2: The anti-atherosclerotic effects of yacon's active components and their underlying molecular pathways were systematically screened and preliminarily characterized by integrating bioinformatic prediction with in vivo validation, laying a preliminary theoretical foundation for further pharmacological investigation and clinical translation.

Characterized by cytoplasmic lipid accumulation and fatty-acid metabolic reprogramming, clear cell renal cell carcinoma (ccRCC) is closely related to tumor progression and invasion. The impact of SPDEF, an ETS transcription factor, in ccRCC and its involvement in lipid metabolism remain unclear. SPDEF expression and prognostic relevance were analyzed using TCGA-KIRC data and validated in ccRCC tissues and cell lines. Functional experiments were conducted to evaluate influence on cell proliferation, migration, lipid metabolism, and apoptosis. Regulatory interactions with ELOVL2 were examined via transcriptomic analysis, dual-luciferase reporter assays, ChIP-qPCR, and site-directed promoter mutagenesis. SPDEF was significantly overexpressed and closed to higher TNM stage and unfavorable survival. The higher expression of SPDEF enhanced malignant phenotypes, increased lipid accumulation, and impeded apoptotic processes. Mechanistically, SPDEF bound to the -141 bp site of the ELOVL2 promoter, thereby activating its transcription; mutation of this site abolished activation. Altering ELOVL2 expression partially rescued or mimicked SPDEF-driven phenotypes. SPDEF acts as an oncogenic transcriptional activator in ccRCC by directly upregulating ELOVL2, thereby driving lipid metabolic reprogramming and tumor progression. These findings provide mechanistic insight into the SPDEF-ELOVL2 axis in lipid-associated ccRCC progression, while its clinical and translational relevance requires further validation in larger patient cohorts and clinically relevant models.

Resistance to molecular-targeted agents, represented by sorafenib (SOR), poses a major challenge in the treatment of hepatocellular carcinoma (HCC). Huaier (HUA), a traditional Chinese medicine used as an adjuvant cancer therapy, has demonstrated favorable efficacy in clinical practice for advanced HCC. This study demonstrates that HUA reverses SOR resistance in HCC and exhibits synergistic effects with SOR by modulating NCOA4-mediated ferritinophagy, thereby influencing iron and lipid metabolic pathways associated with ferroptotic vulnerability. In SOR-resistant Huh7R cells and mouse xenograft models, HUA alone or combined with SOR disrupted iron homeostasis through ferritinophagy induction, promoted Fe3+ reduction to Fe2+, enhanced ROS generation, induced lipid peroxidation, and concurrently reduced lipid droplet storage and triglyceride accumulation. Mechanistically, HUA upregulated NCOA4 and increased the LC3B-II/I ratio, leading to FTH1 degradation. Accumulated Fe2+ drove robust ROS production via the Fenton reaction, exacerbated lipid peroxidation, upregulated ACSL4, and downregulated SCD1 and GPX4, collectively inducing ferroptosis, a form of regulated cell death specifically rescued by ferrostatin-1 or deferoxamine but not by Z-VAD-FMK or necrostatin-1, with minimal caspase activation. Furthermore, this cascade markedly downregulated PLIN2, resulting in energy depletion in Huh7R cells and tumor tissues, thereby impairing adaptive survival and ultimately reversing SOR resistance. Notably, NCOA4 silencing substantially attenuated HUA's efficacy in overcoming SOR resistance. These findings highlight the role of HUA in promoting NCOA4-dependent ferritinophagy and modulating iron and lipid metabolism to enhance ferroptotic sensitivity, providing a rationale for the clinical development of HUA + SOR combination therapy in HCC.

Cellular organelles are uniquely specialized membrane-bound structures that enable cells to organize and coordinate biochemical processes. Specifically, mitochondria are essential organelles for cellular metabolism, coordinating energy production, and connecting signaling networks for cellular homeostasis. 99% of mitochondrial proteins are encoded by nuclear genes that require precise and efficient translation and import into mitochondria for biological processes. This process is mediated by coordinated pathways involving the mitochondrial specific translocation complexes, chaperones, and specialized targeting routes. Tight regulation of these import mechanisms allows for proper protein localization, folding, and assembly. Disruptions in the mitochondrial protein import pathway compromise organelle homeostasis and activate proteostatic stress and quality control pathways. Such defects have been observed in a wide range of pathophysiological conditions, including cardiovascular disease, neurodegeneration, and cancer. The import defects destabilizing mitochondrial proteins can impair oxidative phosphorylation and metabolic signaling. In sum, defects to mitochondrial function can highlight a central role of mitochondrial protein import beyond maintaining cellular function and how defects at distinct stages of import contribute to disease, underscoring opportunities for therapeutic intervention targeting mitochondrial proteostasis.