Cell Biology and Toxicology最新文献

Age-related decreases in follicle numbers and oocyte quality are major contributors to the decline in female fertility, which is associated with increased infertility rates. Emerging evidence suggests that targeting granulosa cell senescence could delay ovarian aging and depletion of the ovarian reserve, highlighting the potential for therapeutic interventions focused on granulosa cells. Advanced glycation end products (AGEs) accumulate with age and result in oxidative stress in the follicular microenvironment, but their direct impact on human granulosa cell (hGC) senescence and the fundamental processes are still mostly unknown. In this study, we found that AGEs treatment significantly exacerbated hGC senescence, impaired mitochondrial function, and suppressed mitophagy in a concentration-dependent manner. Importantly, these deficits were lessened by urolithin A-induced mitophagy activation, whereas Cyclosporine A-induced mitophagy inhibition had the reverse consequences. In addition, silencing Sirtuin 3 (SIRT3) or PINK1 further aggravated these adverse effects, while SIRT3 overexpression attenuated senescence and restored mitochondrial function by enhancing mitophagy. Furthermore, SIRT3 overexpression promoted the synthesis of estradiol-17β and progesterone, key hormones for ovarian function. Our findings demonstrated that AGEs induced hGC senescence by disrupting mitochondrial function and inhibiting mitophagy, with SIRT3 playing a protective role. Enhancing mitophagy by targeting SIRT3 may be a promising treatment approach to counteract age-related declines in female fertility.

Diabetic patients face delayed wound healing due to angiogenesis dysfunction. This study aims to investigate the function of endothelial progenitor cell (EPC)-derived extracellular vesicles (EVs) in wound healing of diabetic mice, providing a theoretical basis for treating difficult-to-heal diabetic wounds. The full-thickness skin wound model was used as an animal model. After treatment with EPC-EVs, wound healing and histopathological structures were evaluated. Peripheral blood was collected to analyze circulating EPCs. In cell models, EV endocytosis, cell viability, angiogenic capacity, and cell migration were detected. miR-204-5p, lncRNA SNHG1, EIF4A3, and HDAC6 were detected. EVs derived from EPCs with miR-204-5p overexpression were extracted to investigate their effects on wound healing. The bindings between miR-204-5p and SNHG1, SNHG1 and EIF4A3, and EIF4A3 and HDAC6 mRNA were validated. EPC-EVs promoted wound healing in diabetic mice. EPC-EVs enhanced angiogenesis and migration in cell models. EPC-EVs with miR-204-5p overexpression exhibited better therapeutic effects. EPC-EVs delivered miR-204-5p into tissues/cells to lower SNHG1 expression. SNHG1 bound to EIF4A3 to increase HDAC6 expression. SNHG1/HDAC6 overexpression partly reversed the pro-angiogenic effects of EPC-EVs on diabetic wound healing and HG-impaired endothelial cells. In conclusion, EPC-EVs enhance EPC mobilization and angiogenesis to accelerate wound repair in diabetic mice via the miR-204-5p/SNHG1/HDAC6 axis.

CSF3 exerts a significant function in the progression of colorectal cancer (CRC). N6-methyladenosine (m6A) modification is now considered the main driving factor of RNA influence for maintaining homeostasis in cancer cells. Nevertheless, how m6A mediates the role of CSF3 and its influence in pathogenesis of CRC is still elusive. After neutrophil isolation from bone marrow, the purity and survival rate of neutrophils were assessed. Azoxymethane (AOM)/Dextran Sodium Sulfate (DSS) was employed to construct the CRC mice model. Both loss-of-function and gain-of-function experiments were conducted to explore the influence of CSF3 on NETosis and tumorigenesis of CRC in vitro and in vivo. The purity and survival rate of neutrophils were 88.07% and 94.84%, respectively. Overexpression of CSF3 (oe-CSF3) markedly enhanced NETosis, while CSF3 knockdown (sh-CSF3) suppressed it. Intriguingly, CSF3 expression positively correlated with relaxin-2 (RLN2) levels in CRC cells, and RLN2 supplementation rescued tumorigenesis and NETosis after sh-CSF3 treatment. Mechanistically, fat mass and obesity-associated protein (FTO)-mediated m6A demethylation of CSF3 mRNA suppressed CRC tumorigenesis in vivo. CSF3 upregulation counteracted the tumor-suppressive effects of FTO overexpression, restoring NETosis and tumor growth. Consistent with this, FTO overexpression in CRC mice alleviated disease severity, as evidenced by improved body weight, reduced tumor burden, and diminished NETosis. Collectively, our findings establish a novel regulatory axis in which FTO-dependent m6A demethylation of CSF3 suppresses NETosis by inhibiting RLN2 expression, offering new insights into therapeutic targeting of the m6A-CSF3-RLN2 pathway in CRC.

Background: Programmed cell death (PCD) patterns play important roles in lung adenocarcinoma (LUAD) development as well as treatment resistance, and in-depth study of PCD is beneficial for improving the therapeutic paradigm for LUAD.

Methods: Fourteen PCD-related patterns were integrated and multiple datasets from TCGA and GEO were collected to develop a PCD signature using 101 machine learning algorithm combinations. Prognosis, immune cell infiltration, and sensitivity to chemotherapy and immunotherapy were compared between different risk groups and validated by multiple bulk RNA-seq and scRNA-seq datasets of patients receiving immunotherapy. CellChat was used to analyze the cellular interactions between patients with different PCD groups. Immune cell infiltration in the tumor tissues of 38 LUAD patients treated with anti-PD-1 therapy was validated by multiplex immunohistochemistry (mIHC).

Results: A PCD signature containing 7 genes was constructed using 101 machine learning algorithm combinations and validated across multiple datasets. High PCD scores in patients are associated with poorer prognosis, lower immune cell infiltration, and reduced responsiveness to immunotherapy. In addition, the PCD signature were comprehensively analyzed by scRNA-seq, and the results showed that the high PCD signature was concentrated mainly in advanced LUAD. Moreover, pathways associated with tumor progression and immune resistance were more strongly promoted in the high PCD signature group. The expression of the key gene NAPSA correlated with immune cell infiltration and immunotherapy response, as confirmed by IHC and mIHC.

Conclusion: The PCD signature confers significant potential to predict prognosis of LUAD in patients, and NAPSA is promising as a new marker for predicting the efficacy of immunotherapy.

Although the effects of noise-induced hearing loss (NIHL) on cognitive functions have been widely investigated, the cognitive effects of noise-induced hidden hearing loss (NIHHL), particularly its impact on memory, remain poorly understood. The Dnah11 gene, which encodes a dynein motor protein involved in synaptic development, may play a role in NIHHL-related cognitive impairment. We aimed to investigate whether NIHHL induces memory impairment and explore the role of Dnah11 expression in this process. Behavioral experiments identified the peak of memory impairment at 1 month following noise exposure. To elucidate molecular changes, hippocampal gene expression was analyzed using transcriptomic sequencing, reverse transcription quantitative polymerase chain reaction (RT-qPCR), and immunofluorescence. RNA sequencing revealed significant Dnah11 upregulation, with immunofluorescence confirming DNAH11 overexpression in hyperactivated CaMKIIα-positive excitatory neurons. Stereotaxic injection of recombinant adeno-associated virus (rAAV) vectors to knock down hippocampal Dnah11 expression improved memory performance in NIHHL mice without improving hearing loss. This cognitive improvement was accompanied by partial restoration of synaptic plasticity-related proteins, including SYN and PSD95. These findings indicate that Dnah11 upregulation in hippocampal excitatory neurons contributes to NIHHL-induced memory impairment, and targeting Dnah11 may offer a therapeutic strategy for memory impairment associated with hidden hearing loss.

The role of platelets in blood coagulation and vascular repair is well known. In recent years, extensive attention has been given to the fact that the impact of tumors on peripheral blood platelets plays a key role in cancer progression. This review systematically summarizes the latest research progress on how tumors regulate the quantity, volume, composition, and activation status of peripheral blood platelets through multiple mechanisms. First, tumor cells can induce excessive platelet production by activating AhR-RUNX1 signaling through paracrine pathways and the release of kynurenine, thereby leading to thrombocytosis, which is associated with advanced tumor stages, metastasis, and poor prognosis. Second, tumor progression may trigger disseminated intravascular coagulation (DIC) or chemotherapy-related bone marrow suppression, which in turn results in consumptive thrombocytopenia. In addition, dynamic changes in the mean platelet volume (MPV) are related to tumor type and progression stage, which may reflect abnormal megakaryocyte differentiation or inflammatory status. In terms of platelet composition, tumor cells can remodel the proteome and transcriptome of platelets by secreting ADP, IgG, and functional RNA. Changes in RNA profiles have been confirmed to have potential for tumor diagnosis. In terms of activation status, tumor cells can induce platelet activation and aggregation (TCIPA) by releasing procoagulant factors such as tissue factors and exosomes (EVs), accelerating thrombosis and promoting angiogenesis. In clinical applications, platelet-related biomarkers have become a research hotspot for early cancer diagnosis and prognostic evaluation. Moreover, targeting platelets affected by tumors provides new strategies for tumor treatment. On the basis of the scientific findings of numerous existing studies, it is speculated that there seems to be a "dynamic balance" among platelets, which also provides a new direction for future research.

Immune dysregulation by aberrant chemokine production underlies many diseases. Targeting chemokine receptors with small molecule inverse agonists, antagonists, or neutralizing antibodies has proven challenging due to non-specific effects and receptor upregulation. Locked dimers of chemokines, generated via cysteine substitutions to produce constitutively homodimeric molecules, offer a promising alternative for receptor-specific inhibition. This study evaluates the in vivo safety and dosing of an engineered CCL20 locked dimer (CCL20LD), which selectively binds CCR6 without inducing chemotaxis. The antagonist-like properties of CCL20LD make it a potential therapeutic for CCL20-CCR6 driven diseases. Daily 14-day subcutaneous administration of CCL20LD at doses previously shown to be therapeutically effective in preclinical models of psoriasis or psoriatic arthritis did not result in weight loss or immune suppression. CCL20LD administration had little to no effects on the complete blood count with differential, comprehensive metabolic panel, urinalysis, organ weights, or bone marrow progenitors. At single cell resolution, doses near 7.5mg/kg/day modestly disrupted T cell dependent B cell activation. While splenomegaly due to extramedullary hematopoiesis was observed at the highest tested dose, serum cytokine levels were largely unchanged. Combined, these findings indicate that selective targeting of CCR6 with an engineered CCL20 dimer is broadly safe in vivo, exhibiting a wide therapeutic window with minimal adverse or immunomodulatory effects.

Exosomes play a crucial role in the transmission of drug resistance in tumors. However, the mechanism of exosomes-mediated transmission in non-small cell lung cancer (NSCLC) under gefitinib treatment remains limited. In this work, we demonstrated that exosomes derived from HCC827/GR cells (drug-resistant) enhanced the survivability of HCC827 cells (drug-sensitive) under treatment with gefitinib. A total of 157 shared upregulated proteins between exosomes and their parent cells were identified in the comparison of the gefitinib-resistant groups versus the gefitinib-sensitive groups. Notably, 69 of these shared proteins are enzymes, and many of them were enriched in pathways related to fatty acid metabolism. Among these enzymes involved in fatty acid metabolism, ACC1 exhibited the highest fold change in upregulated expression in both drug-resistant groups (exosomes and cells). Moreover, the expression of ACC1 was upregulated in gefitinib-sensitive cells after uptake of exosomes from gefitinib-resistant cells. The role of ACC1 in enhancing the survival of HCC827/GR cells under gefitinib treatment was demonstrated using an inhibitor and siRNA-mediated knockdown. Specifically, the upregulated ACC1 stabilized fatty acid oxidation and reactive oxygen species levels in HCC827/GR cells, thereby maintaining cellular metabolic homeostasis. Collectively, this work reveals the transmission of drug resistance in NSCLC via exosomes that carry the ACC1 protein.

COVID-19 has caused millions of deaths worldwide since 2019. Vaccination has reduced both transmission and disease severity. However, emerging viral variants have weakened vaccine effectiveness, highlighting the need for new antiviral therapies. This study examines how the SARS-CoV-2-Spike protein (SARS-2-S) induces the VSIR-ISX signaling pathway, leading to metabolic disturbances that may worsen disease progression. Using RNA sequencing, we found that SARS-2-S expression in pulmonary cells activates genes involved in tryptophan and arachidonic acid (AA) metabolism, altering bioactive mediators like kynurenine and prostanoids, which are crucial for inflammation and immune responses. Mechanistically, the ACE2-MYD88 pathway, activated by SARS-2-S, enhances the VSIR-ISX axis through NF-κB signaling, driving these metabolic disruptions. Chromatin immunoprecipitation and genome sequencing revealed that ISX, activated via VSIR-MAPK signaling, upregulates enzymes involved in AA metabolism by binding directly to their gene promoters. Notably, disrupting the VSIR-ISX axis using shRNA interference or NF-κB inhibitors effectively mitigated these metabolic disturbances. Our findings suggest that the VSIR-ISX pathway could be a promising therapeutic target for treating COVID-19 by addressing virus-induced metabolic disruptions.

Paclitaxel (PTX), a commonly utilized chemotherapy drug, is linked to peripheral neuropathy, which limits dosing and significantly affects patients' quality of life. C-terminal binding protein 1 (CtBP1) is a transcriptional coregulator that participates in epigenetic gene regulation, but its role in PTX-induced neuropathic pain remains unclear. In this study, the role of CtBP1 in PTX-induced neuropathic pain is examined, with a focus on its epigenetic regulation in the dorsal root ganglia (DRGs). PTX administration markedly increased CtBP1 protein levels in DRG neurons, which coincided with the development and continuation of mechanical allodynia and thermal hyperalgesia in rat models. Our findings also revealed that CtBP1 interacts with the histone demethylase LSD1-a regulator of H3K9me2-at ErbB2 promoter sites in DRG neurons. PTX treatment increased CtBP1 protein levels, which subsequently induced LSD1 expression and decreased H3K9me2 protein levels at the ErbB2 promoter, indicating epigenetic activation of ErbB2 signaling in DRG neurons implicated in neuropathic pain. Reducing either CtBP1 or LSD1 expression reversed ErbB2 upregulation and attenuated PTX-induced pain sensitivity. These results suggest that the CtBP1-LSD1 complex epigenetically increases ErbB2 expression in DRG neurons, contributing to PTX-induced neuropathy. Targeting the CtBP1-LSD1 pathway could represent a promising therapeutic strategy for the treatment of chemotherapy-induced neuropathic pain.