Search Results (1,950)

Autophagy was originally identified as a survival mechanism to allow cells to survive under nutrient-deprived or stressful conditions whereas cellular senescence was considered a tumor-suppressive mechanism. Both processes can be induced by similar stimuli and can influence each other. There have been continued debates about whether they are causally linked, whether autophagy promotes or prevents senescence or if they are independent of each other. Protein kinases play integral roles in cell fate decision and have a major influence on both autophagy and senescence. While mechanistic target of rapamycin complex 1 is considered the master regulator of autophagy, it also influences senescence. Mitogen-activated protein kinases originally associated with senescence can regulate autophagy. While there have been numerous review articles on the interplay between autophagy and senescence, a comprehensive review on how various kinases participate in this interplay is lacking. The purpose of this review is to learn lessons from some old and recent studies to understand how kinases contribute to this changing field. Since both autophagy and senescence can have beneficial and detrimental effects and kinases are important drug targets, insights regarding how kinases orchestrate these two processes should help develop therapeutic strategies to treat diseases, such as aging and cancer. Full article

Background/objective: Given the increasing complexity of the luminal breast cancer landscape, a proper characterization is required in everyday clinical practice, and the recurrence after the standard first-line treatment with CDK4/6 inhibitors with/without endocrine therapy should be managed. Method: The Hermione Exchange Educational Program was held in Milan, Italy, between September 2024 and January 2025. Two questionnaires were proposed regarding the use of targeted treatment or chemotherapy after progression from CDK4/6 inhibitors. The lecture and use cases enhanced the discussion during the workshops. Results: From the surveys, it emerged that most participants (69%) considered liver metastases at CDK4/6-inhibitor progression as a key reason to initiate chemotherapy, while lung progression influenced this choice for 50% of participants. Liver involvement guided the use of targeted therapy for 56%, and attitudes were divided on whether the duration of first-line CDK4/6 therapy should affect decisions (44% in agreement vs. 38% in disagreement). The willingness of patients to receive chemotherapy (88%) and comorbidities (81%) were significant drivers. Almost all participants agreed that both the duration of response and the molecular status were key aspects to consider when choosing a second line of therapy, along with the general clinical condition of the patient. In the lecture, tissue and liquid biopsy are considered powerful tools to describe tumor molecular features over time; such complexity should be harnessed by a close dialogue between oncologists, molecular biologists, and pathologists to optimize the therapeutic choice according to the mutational status of patients. The use cases illustrate three patients with visceral progression, non-visceral progression within 12 months, and non-visceral progression after 12 months following CDK4/6 inhibitors. Conclusion: Genomic testing should be considered at diagnosis and repeated during treatment to monitor the disease. The clinical experience acquired over the years must be integrated with new molecular knowledge. Full article

Yaks (Bos grunniens) are large mammals endemic to the Qinghai–Tibet Plateau. Efficient lung development is crucial for their adaptation to high-altitude hypoxia. As progenitor cells of the alveoli, type II alveolar epithelial (AT2) cells warrant further investigation into their physiological functions; however, relevant studies remain limited. In this study, primary AT2 cells were isolated from the lungs of yaks. Concurrently, lung tissues were collected from yaks at distinct developmental stages to investigate the role of the EGF/EGFR axis in regulating AT2 cell proliferation and apoptosis, as well as its essential contribution to yak lung development. Here, we demonstrate that the EGF/EGFR axis plays a beneficial role in yak alveolar development. Exogenous EGF supplementation or EGFR activation upregulated the downstream factors AKT and STAT3, enhanced AT2 cell proliferation, and reduced apoptosis. In contrast, EGFR inhibition promoted AT2 cell apoptosis and suppressed proliferation. Cell cycle analysis revealed that both exogenous EGF and EGFR activation increased the proportion of AT2 cells in the S and G2 phases, whereas EGFR inhibition caused cell cycle arrest at the G0/G1 phase. Moreover, the expression of cell cycle regulators cyclin D1, CDK4, and CDK6 was upregulated, while p16 and p21 expression was downregulated. Further comparative analyses indicated that the EGF/EGFR axis positively contributes to alveolar development in juvenile yaks. Collectively, these findings confirm that in plateau environments, activation of the EGF/EGFR axis promotes AT2 cell proliferation and inhibits apoptosis, thereby facilitating alveolar development in juvenile yaks. A key limitation is the lack of parallel comparisons with low-altitude cattle and other plateau-endemic species (e.g., Tibetan sheep), which precludes definitive assessment of the specificity of the EGFR/EGF axis in yak AT2 cell proliferation and lung development. Full article

Background/Objectives: Colorectal cancer (CRC) remains one of the leading causes of cancer-related morbidity and mortality worldwide. Despite advances in treatment, outcomes for advanced CRC remain unsatisfactory due to uncontrolled proliferation, metastasis, and recurrence. This study investigated the anti-cancer effects of KMU-11342, an indolin-2-one-based multi-protein kinase inhibitor with previously reported anti-inflammatory properties, in human colorectal cancer models. Methods: The anti-cancer effects of KMU-11342 were evaluated in colorectal cancer cells and further investigated in three-dimensional (3D) spheroid and patient-derived organoid models. Cell proliferation, migration, apoptosis, and cell cycle progression were assessed. Kinase activity profiling and molecular docking analyses were performed to identify potential targets and characterize the underlying signaling pathways. Results: KMU-11342 significantly inhibited the proliferation and migration of CRC cells. It reduced CRC cell density by 58.9% and 83.3% at 0.5 and 1 μM, respectively. These effects were accompanied by G2/M cell cycle arrest and apoptotic cell death. In 3D models, spheroid formation was markedly reduced and stemness-related characteristics were diminished. Patient-derived CRC organoids also showed decreased viability, exhibiting 38.6% and 77.4% reductions at 1 and 2 μM, respectively. These effects were observed in a dose-dependent manner in both two-dimensional (2D) and 3D colorectal cancer models. Kinase activity profiling and molecular docking analyses identified glycogen synthase kinase 3 beta (GSK3β) and cyclin-dependent kinase 1 (CDK1) as potential mediators of the anti-cancer effects of KMU-11342 through the p53/nuclear factor kappa B (NF-κB) and FoxO1 signaling axes, respectively. Conclusions: KMU-11342 exhibits potent anti-tumor activity against CRC through suppressing proliferation, migration, and stemness in both 2D and 3D models, including patient-derived organoids. Its effects may be mediated, at least in part, through modulation of GSK3β and CDK1 via the p53/NF-κB and FoxO1 signaling pathways. Full article

WEE2, an oocyte-specific kinase of the WEE family, is a core regulator of oocyte meiosis. It maintains germinal vesicle (GV) arrest and prevents premature meiotic resumption by phosphorylating cyclin-dependent kinase 1 (CDK1), thereby inhibiting maturation-promoting factor (MPF) activity. WEE2 also regulates exit from metaphase II (MII), ensuring orderly meiotic progression. Consequently, the functional integrity of WEE2 is essential for female reproduction. Homozygous or compound heterozygous mutations in the WEE2 gene represent a major genetic cause of total fertilization failure and primary infertility, as these mutations lead to reduced or abolished kinase activity, impair meiotic control, and disrupt oocyte maturation and embryonic development. This review systematically summarizes the protein structure, core functions, and mutation types of WEE2, along with its association with total fertilization failure and female primary infertility. It also highlights research advances in WEE2-targeted inhibitors and discusses the potential applications and future directions of WEE2 in the diagnosis and management of reproductive disorders. Full article

Background/Objectives: Advanced hormone receptor-positive (HR+), epidermal growth factor 2-negative (HER2−) breast carcinoma (BC) patients receive frontline therapy with cyclin-dependent tyrosine kinase 4/6 inhibitors + endocrine therapy (ET). At progression, the best management includes mutational analysis for ESR-1, allowing second-line therapy with elacestrant. The aim of this study was to evaluate the efficacy and safety of elacestrant in an Italian real-world setting. Methods: A multicenter, observational study with a mixed retrospective and prospective design was conducted in 13 medical oncology units across Italy. The study population included adult patients with HR+/HER2− locally advanced or metastatic breast cancer with an activating ESR1 mutation documented by liquid biopsy and progressing after at least one line of endocrine therapy containing a CDK4/6 inhibitor. Mutational analysis of plasma was performed using next-generation sequencing with a multigene panel that included ESR1, PIK3CA, AKT, and PTEN. The sample size was calculated according to the two-stage Simon design. Toxicity was classified according to CTCAE version 5.0 criteria. Survival analyses were conducted using the Kaplan–Meier method. Results: At the time of analysis, 39 evaluable patients were enrolled, all female and Caucasian, with a median age of 67 years (range 41–89). The efficacy analysis documented an overall ORR of 28% and a disease control rate of 56%. The median duration of response was 6+ months (95% CL: 3.5–10.6 m). Median overall survival was not reached with a median follow-up of 10 months. The toxicity profile was overall favorable: grade ≥2 asthenia was the most frequent adverse event (23%), followed by gastrointestinal toxicity, which was generally mild. No treatment-related toxicity was reported in 64% of patients. Dose reductions were necessary in 15% of cases, while permanent treatment discontinuation due to toxicity occurred in only 4%. Conclusions: The results of this Italian multicenter observational study confirm the efficacy and tolerability of elacestrant in HR+/HER2− metastatic breast cancer with ESR1 mutation, in a real-world context consistent with the data from the pivotal EMERALD study and with real-world data present in the literature. Full article

Background: Hematopoietic stem cell (HSC) ex vivo culture causes severe loss of repopulation and regenerative capacity without compromising multilineage differentiation, which greatly limits the efficacy of HSC transplantation. The molecular mechanisms underlying culture-triggered HSC dysfunction remain poorly understood. Methods: Human CD34⁺ HSCs were cultured ex vivo for 96 h to establish a culture-induced HSC dysfunction model. Single-cell RNA sequencing was applied to screen key regulatory genes. TSC22D3 function was verified via overexpression assays, and immunodeficient mice were used to assess HSC engraftment. Transcriptomic profiling were performed to explore downstream molecular mechanisms. Results: Ex vivo culture induced G0 quiescence exit, elevated early apoptosis and impaired in vivo repopulation in human CD34⁺ HSCs. TSC22D3 was highly enriched in freshly isolated quiescent HSCs and gradually downregulated during culture. TSC22D3 overexpression restored HSC G0 arrest and improved hematopoietic engraftment in mice. Mechanically, TSC22D3 upregulated HSC self-renewal genes, suppressed cell cycle-related genes (CDK2/4), and activated the P53-P21-P27 pathway. Conclusions: This study demonstrates that TSC22D3 preserves HSC function during ex vivo culture by maintaining stem cell quiescence and restricting excessive proliferation. These findings reveal a novel transcriptional mechanism regulating HSC homeostasis and provide a promising target for improving functional HSC ex vivo expansion for clinical transplantation. Full article

Background/Objectives: First-generation CDK4/6 inhibitors (palbociclib, ribociclib, abemaciclib) target the conserved ATP-binding pocket of CDK4 and, despite clinical success, are limited by acquired resistance and insufficient exploration of alternative regulatory sites. This study aimed to identify a putative allosteric small-molecule candidate at the CDK4 αE-helix–Cyclin D1 α1-helix protein–protein interaction (PPI) interface within the CDK4/Cyclin D1/p21 ternary complex using RapidFunnel-AI, a decision-interpretable virtual-screening pipeline. Methods: Starting from 50,000 ChEMBL 33 molecules, the pipeline sequentially applied a Q-Fold/RapidFunnel topological Tanimoto scan based on clinical CDK4/6 inhibitor motifs, fragment-level electronic-property enrichment, ADMET/PAINS filtering, dry Vina-GPU docking, hydration-mediated AutoDock-GPU (Version 1.6) docking, explicit-solvent molecular dynamics, contact-retention analysis, and MM-GBSA energy decomposition. The Q-Fold Thermo-Core surrogate model provided fragment-level enrichment, predicting the HOMO–LUMO gap (R² = 0.93) and isotropic polarizability (R² = 0.98) on QM9. Candidate selection did not rely on the lowest docking or MM-GBSA score alone, but on pose persistence, contact continuity, and energy-component consistency. Results: The workflow reduced the initial library to 43 topologically prioritized candidates, 25 ADMET/PAINS-filtered ligands, and 9 docking-derived complexes for MD validation. Ligand_020 emerged as the only candidate that preserved a persistent binding mode at Site 2 during a 500 ns simulation—an interface engagement reproduced across three independent 500 ns replicates with no full dissociation in any replicate—with a protein Cα RMSD of 2.88 ± 0.32 Å, a ligand heavy-atom RMSD of 3.56 ± 0.28 Å, and a van der Waals-dominated MM-GBSA profile (ΔGbind = −28.23 ± 3.57 kcal/mol). In contrast, palbociclib and ribociclib, forcibly placed at Site 2 as negative controls, lost most initial contacts within 5 ns and tended to detach despite more favorable MM-GBSA values. Conclusions: These results suggest that single-score docking or MM-GBSA ranking can generate false positives at shallow PPI interfaces. By integrating AI-assisted prioritization, multipocket docking, explicit-solvent MD, contact-retention analysis, and energy-component consistency, RapidFunnel-AI nominated Ligand_020 as an experimentally testable putative allosteric hit targeting the CDK4/Cyclin D1 interface, offering a reusable platform for PPI-focused oncological drug discovery. Full article

Fisetin is a bioactive flavanol with reported anticancer activity, although its mechanisms in leukaemia and potential for combination therapy remain incompletely understood. This study investigated the cytotoxic and mechanistic effects of fisetin, alone and combined with pinocembrin, in human leukaemia cells. Cell viability, apoptosis, and cell cycle progression were assessed by flow cytometry; protein expression in Jurkat cells was assessed by Western blotting; and molecular docking was used to evaluate interactions with the Fas receptor. Drug interactions were quantified using ZIP synergy analysis, and cytotoxicity and clonogenic survival were evaluated using soft-agar colony formation assays in K562 cells. Fisetin significantly reduced cell viability and induced apoptosis, accompanied by caspase-8 cleavage, p62 accumulation, and CDK4 downregulation, consistent with activation of extrinsic apoptosis, impaired autophagic flux, and cell cycle inhibition in Jurkat cells. Docking analysis supported a potential interaction with the Fas receptor, which was confirmed using the Fas receptor antagonist Met-12. Co-treatment with pinocembrin enhanced fisetin-mediated cytotoxicity and produced synergistic effects, particularly in Jurkat cells (ZIP score > 10), while synergistic interactions at specific sub-IC₅₀ concentrations were also observed in K562 cells. Combination treatment further enhanced caspase-8 activation, reduced CDK4 expression in Jurkat cells, and significantly suppressed clonogenic survival in K562 cells compared with single-agent treatments. These findings suggest that fisetin promotes caspase-8-dependent apoptosis, potentially involving Fas-associated signalling, and highlight fisetin–pinocembrin combination therapy as a promising strategy for leukaemia treatment. Full article

Interleukin-13 receptor α2 (IL-13Rα2) has traditionally been considered a decoy receptor; however, its cellular functions beyond the immune system remain unclear. We aimed to investigate the role of IL-13Rα2 in C2C12 myoblast proliferation and differentiation. IL-13Rα2 expression was knocked down in C2C12 cells using siRNA. Myogenic differentiation was evaluated by myosin heavy chain (MyHC) immunostaining and by quantifying the expression of myogenic regulatory and fusion-related genes. Myoblast proliferation was assessed using BrdU incorporation and cell number analyses, and signaling events induced by IL-13Rα2 knockdown were analyzed via immunoblotting and immunocytochemical analysis. IL-13Rα2 knockdown did not alter myogenic differentiation or the expression of fusion-associated genes. In contrast, IL-13Rα2 knockdown significantly increased BrdU incorporation and cell number, accompanied by increased Akt phosphorylation and decreased ERK phosphorylation. Cyclin D1 and cyclin-dependent kinase 4 (CDK4) levels were also increased. Akt inhibition abolished the enhanced proliferation and normalized Cyclin D1/CDK4 levels, whereas ERK activation did not further modify the knockdown-associated phenotype. These findings demonstrate that IL-13Rα2 negatively regulates myoblast proliferation by modulating the Akt–Cyclin D1–CDK4 signaling pathway, while being dispensable for myogenic differentiation. Full article

Fine particulate matter (PM_2.5) exposure increases the risk of neurodevelopmental abnormalities by disrupting neural stem cell (NSC) proliferation and cell cycle homeostasis, which are critical for normal neurodevelopment. This study investigated the impact of fine particulate matter (PM_2.5) on NSC proliferation and cell cycle using a three-dimensional (3D) graphene oxide (GO) scaffold that mimics the NSC microenvironment. PM_2.5 exposure led to concentration-dependent decreases in NSC viability and induced G0/G1 phase arrest via the marked downregulation of Cyclin D1-Cdk4 and Cyclin E-Cdk2, which critically impact G1/S transition. NSCs in 3D GO scaffolds maintained higher expression of key cell cycle regulators (Cyclin A, Cdk1/Cdk2, APC, and Cdc20) and superior cell viability when suffering PM_2.5 exposure, demonstrating the 3D culture environment was beneficial for NSC proliferation. We speculate that the 3D culture environment is more favorable and protective for cell proliferation. Therefore, these findings highlight the utility of the 3D GO scaffold for studying PM_2.5 effects on growing neural stem cells. This work provides a physiologically relevant in vitro platform that captures microenvironment-dependent neurotoxic responses, consequently offering valuable mechanistic insights into PM_2.5-induced developmental neurotoxicity. Full article

Tousled-like kinases 1 and 2 (TLK1 and TLK2) are paralogous serine/threonine kinases that share high sequence similarity yet exhibit functional divergence in cellular processes such as DNA replication, damage response, and chromatin organization. This study elucidates the paralog-specific co-phosphoregulatory networks underlying this divergence through a comprehensive analysis of 3825 human phosphoproteomic articles. Predominant phosphosites were identified as S134 and T38 for TLK1 and S73, S99, and S111 for TLK2, revealing context-dependent regulation across cancers and perturbations. Co-phosphoregulation analyses uncovered distinct networks: TLK1 associates with DNA damage signaling via proteins like ABRAXAS1, PML, and RAD9A, while TLK2 integrates with chromatin remodeling and replication through CHD4, DOT1L, NASP, and RNF20. Upstream kinases for TLK2, predominantly CDKs, link it to cell-cycle progression, whereas downstream substrates and binary interactors converge on genome stability pathways with paralog-specific nuances. These findings highlight the potential role of TLK1 on checkpoint activation and TLK2 on replication-coupled chromatin maintenance, providing insights into their roles in cancer amplification and therapeutic resistance, as well as neurodevelopmental disorders, where emerging evidence also support the involvement of TLK1 alongside TLK2. Full article

Invertebrates rely exclusively on innate immunity but exhibit memory-like responses termed immune priming or trained immunity. In the commercially vital whiteleg shrimp (Litopenaeus vannamei), infection by Vibrio parahaemolyticus causes severe economic losses, yet the molecular networks driving secondary immune recall remain poorly understood. In this study, we established a two-step immune challenge model in L. vannamei using formaldehyde-inactivated V. parahaemolyticus and performed transcriptomic analysis on hemocytes to compare primary and secondary immune responses. Differentially expressed gene (DEG) screening and enrichment analyses (GO, KEGG, and GSEA) suggest that shrimp hemocytes undergo a broad and coordinated transcriptional reprogramming rather than uniform upregulation of immune genes. Transcriptomic data show potential associations between secondary immune priming and the modulation of cell fate processes: genes related to cell cycle progression (e.g., CDK1, CCNB3) and spindle assembly (e.g., MPS1) were significantly upregulated alongside apoptosis inhibition (CASP6 downregulation). Concurrently, metabolic remodeling was observed through the upregulation of lipid synthesis (SREBF1, FASN) and carbohydrate uptake pathways, potentially providing anabolic support for hemocyte growth and immune activation. Furthermore, the humoral effector responses appear to be strengthened, characterized by upregulated antimicrobial peptides (PEN, ALF) and the proPO melanization cascade (PPAF3, PPO3), whereas the expression of intracellular NLR was relatively suppressed, which might help mitigate excessive immune inflammation and immunopathological damage. Collectively, these transcriptomic findings identify a putative coordinated transcriptional signature of hemocyte recall responses in L. vannamei. This study expands our understanding of innate immune memory in invertebrates and provides candidate molecular markers for further study in disease-resistant breeding research in shrimp aquaculture. Full article

Bone remodeling relies on a balance between osteoclast-mediated resorption and osteoblast-mediated formation. Roxadustat, a hypoxia-inducible factor prolyl hydroxylase inhibitor, promotes osteoblast differentiation but its effects on osteoclasts remain unclear. This study investigated roxadustat’s impact on osteoclast differentiation and function in vitro using primary murine bone marrow-derived mononuclear cells differentiated with M-CSF and RANKL. Cell viability, TRAP staining, bone resorption assays, RNA-seq, flow cytometry, immunofluorescence, Western blot for p27, and rescue experiments with the cyclin-dependent kinases 4 and 6 (CDK4/6) inhibitor abemaciclib were performed. Roxadustat suppressed osteoclast differentiation and resorption without cytotoxicity in a concentration-dependent manner. RNA-seq revealed enrichment of cell cycle pathways; although differentiation was inhibited, roxadustat paradoxically promoted osteoclast precursor proliferation, evidenced by increased Ki67 and decreased p27 expression. The inhibitory effects on osteoclastogenesis and resorption were partially reversed by abemaciclib. Given that terminal differentiation typically requires cell cycle exit, these findings suggest that roxadustat may inhibit osteoclast differentiation at least in part by disrupting this process, promoting precursor proliferation, and downregulating p27. Together with its known anabolic effects on osteoblasts, roxadustat might have dual therapeutic potential for bone disorders with renal anemia, such as osteoporosis in chronic kidney disease. Full article

Germline BRCA1/2-mutated (gBRCAm) hormone receptor-positive/HER2-negative (HR+/HER2-) metastatic breast cancer (MBC) is a biologically distinct subset in which the efficacy of cyclin-dependent kinase 4/6 (CDK4/6) inhibitors remains incompletely characterized. We evaluated real-world outcomes and prognostic factors in a multicenter retrospective Turkish cohort treated with a CDK4/6 inhibitor plus endocrine therapy (June 2020–September 2025). Progression-free survival (PFS) and overall survival (OS) were estimated using Kaplan–Meier and Cox methods. Among 121 patients, 30 (24.8%) had BRCA1, 88 (72.7%) had BRCA2, and three (2.5%) had dual mutations; 66.9% received first-line therapy, with ribociclib in 69.4% and palbociclib in 29.8%. Objective response rate was 69.4% and the clinical benefit rate was 82.6%. Median PFS was 17.0 months and OS 47.0 months. PFS was numerically longer in BRCA1 than in BRCA2 carriers (25.0 vs. 14.0 months), although the difference was not statistically significant in the pairwise comparison (HR 1.50, 95% CI 0.88–2.56; log-rank p = 0.135); the dual BRCA1/2 subgroup (n = 3) had the poorest outcomes and was assessed descriptively. OS did not differ significantly between BRCA1 and BRCA2 carriers (57.0 vs. 49.0 months; log-rank p = 0.520). PFS did not differ between ribociclib and palbociclib (p = 0.192); OS favored ribociclib at borderline significance (p = 0.050), but this was not confirmed in Cox regression. In multivariable analysis, ECOG ≥ 1 (HR 1.85; p = 0.010) and fulvestrant-based therapy (HR 1.74; p = 0.041) predicted shorter PFS; fulvestrant also predicted worse OS (HR 2.39; p = 0.008). CDK4/6 inhibitor-based therapy shows meaningful activity in gBRCAm HR+/HER2- MBC; the numerically poorer outcomes observed in BRCA2 carriers are hypothesis-generating and warrant validation in larger cohorts. Full article