Endocrine resistance is a leading cause of mortality in oestrogen receptor-positive and human epidermal growth factor receptor 2-negative (ER+HER2−) breast cancer (BC), highlighting the urgent need to understand its underlying molecular mechanisms and identify potentially resistant patients for effective management. In this study, we constructed endocrine-resistant cell lines through long-term oestrogen deprivation and identified differentially expressed genes (DEGs) via transcriptome analysis. Key endocrine-resistant genes were defined through Cox regression analysis. Our findings revealed that the genes CLEC3A, PCDH10, and ST3GAL1 were significantly upregulated in endocrine-resistant cells and serve as independent prognostic factors for ER+HER2− BC patients. We developed an endocrine resistance score (ERS), and a nomogram model incorporating ERS demonstrated robust predictive capabilities for patient prognosis. Single-cell RNA sequencing analysis demonstrated that the ERS and the three core genes constituting the ERS were significantly upregulated in tissue specimens from patients with resistance to endocrine neoadjuvant therapy. Additionally, knocking down CLEC3A, PCDH10, and ST3GAL1 led to reduced malignancy progression in endocrine-resistant BC cells. Mechanistic studies revealed that CLEC3A promotes endocrine resistance by upregulating the PI3K-AKT pathway. This study suggests that CLEC3A, PCDH10, and ST3GAL1 are associated with endocrine resistance and can reflect the prognosis of ER+HER2− BC patients receiving endocrine therapy, providing potential therapeutic targets and a valuable prognostic indicator for clinicians.

Cuproptosis, a copper-dependent cell death mechanism driven by tricarboxylic acid (TCA) cycle collapse, shows limited efficacy in hypoxic or glycolytic renal cell carcinoma (RCC). Here, through systematic screening of 688 glycolysis inhibitors combined with elesclomol (ES), we identified PIK-III as a potent cuproptosis sensitiser. Multi-omics analysis revealed that PIK-III restores sensitivity by rewiring thiamine metabolism. Mechanistically, PIK-III induces macropinocytosis, enabling thiamine uptake to replenish thiamine pyrophosphate (TPP), which activates pyruvate dehydrogenase E1-alpha 1 (PDHA1) and redirects pyruvate into the TCA cycle. Concurrently, ES-induced DLAT oligomerisation disrupts TCA flux, creating a metabolic crisis. In vivo, PIK-III synergises with ES to suppress tumour growth in xenograft and patient-derived models without systemic toxicity. Our work uncovers a metabolic vulnerability in cuproptosis-resistant RCC and positions PIK-III as a therapeutic candidate to overcome resistance via dual targeting of thiamine transport and mitochondrial dysfunction.

Differences in gene expression, which arise from divergence in cis-regulatory elements or alterations in transcription factors (TFs) binding specificity, are one of the most important causes of phenotypic diversity during evolution. On one hand, changes in the cis-elements located in the vicinity of target genes affect TF binding and/or local chromatin environment, thereby modulating gene expression in cis. On the other hand, alterations in trans-factors influence the expression of their target genes in a more pleiotropic fashion. Although the evolution of amino acid sequences is much slower than that of non-coding regulatory elements, particularly for the TF DNA binding domains (DBDs), it is still possible that changes in TF-DBD might have the potential to drive large phenotypic changes if the resulting effects have a net positive effect on the organism's fitness. If so, species-specific changes in TF-DBD might be positively selected. So far, however, this possibility has been largely unexplored. By protein sequence analysis, we observed high sequence conservation in the DBD of the TF caudal-type homeobox 2 across many vertebrates, whereas three amino acid changes were exclusively found in mouse Cdx2 (mCdx2), suggesting potential positive selection in the mouse lineage. Multi-omics analyses were then carried out to investigate the effects of these changes. Surprisingly, there were no significant functional differences between mCdx2 and its rat homologue (rCdx2), and none of the three amino acid changes had any impact on its function. Finally, we used rat-mouse allodiploid embryonic stem cells to study the cis effects of Cdx2-mediated gene regulation between the two rodents. Interestingly, whereas Cdx2 binding is largely divergent between mouse and rat, the transcriptional effect induced by Cdx2 is conserved to a much larger extent. There were no significant functional differences between mCdx2 and its rat homologue (rCdx2), and none of the three amino acid changes had any impact on its function. Moreover, Cdx2 binding is largely divergent between mouse and rat; the transcriptional effect induced by Cdx2 is conserved to a much larger extent.

Transcriptomics studies have identified integrin receptor β2 subunit (ITGB2) as a core gene in osteoarthritis (OA), strongly linked to osteoclast function in the subchondral bone. However, the mechanism through which ITGB2 regulates osteoclast function in OA remains unclear. In this study, we found that ITGB2 was negatively correlated with ITGB1 in the human subchondral bone. Proteomic analysis indicated that integrin binding is crucial in OA subchondral bone, with ITGB2 identified as a significantly upregulated protein in OA. In vitro experiments using immunoprecipitation and bimolecular fluorescence complementation revealed that ITGB2, but not ITGB1, directly interacts with Rac1 during osteoclast differentiation. Activated Rac1 promotes osteoclast differentiation and bone resorption through several mechanisms. ITGB2 knockdown reduced Rac1-GTP levels and increased ITGB1 expression. ITGB2 inhibition reduced actin ring formation and microtubule migration to the cell edge during osteoclast differentiation. Additionally, overexpression of ITGB1 in ITGB2-knockdown cells not only further suppressed ITGB2 expression but also exacerbated the inhibition of osteoclast differentiation. In a DMM mouse model, ITGB2 was associated with osteoclast activity in the subchondral bone. ITGB2 knockdown significantly reduced bone resorption and slowed OA progression by inhibiting osteoclastogenesis. In conclusion, our study identified a novel mechanism for the reciprocal regulation of integrin subunits. Moreover, inhibition of the ITGB2 signalling pathway slows subchondral bone remodelling in osteoarthritis by inhibiting osteoclast differentiation, offering a potential strategy for targeted therapeutic interventions.

Keratinocyte differentiation factor 1 (Kdf1) reportedly plays a significant role in enamel formation. In terms of tooth morphogenesis, human KDF1 variants are associated with crown morphological abnormalities, suggesting that Kdf1 may also be essential for tooth morphogenesis. However, the involvement of Kdf1 in tooth morphogenesis and its underlying mechanisms remains unclear. In this study, we observed that mice lacking epithelial Kdf1 (K14-Cre;Kdf1^fl/fl) displayed rounded and blunt molar cusps, resembling the morphological anomalies observed in patients with Kdf1 variants. 5-Ethynyl-2′-deoxyuridine assays revealed increased proliferative activity of the inner enamel epithelial (IEE) cells in the cusp region of K14-Cre;Kdf1^fl/fl mice during the bell stage. RNA sequencing and western blot analysis confirmed the overactivation of PI3K/AKT/mTOR signalling in the molar IEE cells of K14-Cre;Kdf1^fl/fl mice. Furthermore, in utero microcapillary injection of the PI3K/AKT/mTOR pathway inhibitor LY294002 partially rescued the molar cusp defects in K14-Cre;Kdf1^fl/fl mice. Collectively, our findings provide in vivo evidence supporting the regulatory role of Kdf1 in molar cusp morphogenesis, highlighting its function in modulating dental epithelial cell proliferation via the PI3K/AKT/mTOR signalling pathway.

Loss of function mutations of NDUFS4 resulted in Leigh syndrome, which is a progressive neurodegenerative disease and characterized by mitochondrial oxidative stress, inflammation and aberrant mitochondrial dynamics. However, there is currently no effective treatment. Here, we demonstrate that pioglitazone significantly mitigates mitochondrial reactive oxygen species (ROS) generation, lowers cyclooxygenase-2 (COX-2) mRNA levels, and rescues aberrant mitochondrial dynamics in vitro (increasing Opa-1 expression while decreasing Drp-1 expression). Furthermore, similar effects were observed with the selective Drp-1 inhibitor mdivi-1, suggesting that inhibiting mitochondrial fission mediates the therapeutic effects of pioglitazone. Pioglitazone administration activated AMPK phosphorylation, but these effects, along with pioglitazone's ability to reverse oxidative stress, inflammation, and mitochondrial fission, were abolished by the AMPK inhibitor compound C. In vivo, pioglitazone alleviated motor dysfunction, prolonged lifespan, and promoted weight gain in Ndufs4 KO mice. This was accompanied by enhanced mitochondrial fusion and increased levels of mitochondrial complex subunits. Consistently, pioglitazone attenuated neuroinflammation and oxidative stress in vivo. Collectively, our findings indicate that pioglitazone alleviates mitochondrial oxidative stress and inflammation through an AMPK-dependent inhibition of Drp-1-mediated mitochondrial fission. Therefore, suppression of mitochondrial fission may represent a novel therapeutic strategy for Leigh syndrome (LS).

The maternal gene products stored in oocytes control the initial development of multicellular animals. Alteration within the dual allelic variants of transcription factor TCF12 causes female infertility; however, its impact on female reproduction is still unknown. In this study, we provide evidence that TCF12 is abundantly expressed within the nucleus of oocytes during growth at the germinal vesicle (GV) stage, recognising and binding to the functional domain of target genes to moderate transcriptional activity. The absence of Tcf12 in oocytes during the primordial follicular phase causes female sterility. Tcf12 does not participate in meiotic maturation; however, unlike Tcf3, it is essential for fertilisation and preimplantation development. Tcf12 maintains fertilisation competence by controlling the proper expression and location of cortical granules and protease ovastacin (encoded by Astl). In contrast, zygotes without TCF12 have a prolonged mitotic cell cycle upon a decrease in protein phosphatase 2A (PP2A) activity inhibition, resulting in zygotic genome activation (ZGA) failure during the 2-cell stage. Maternal knockout embryos gradually lose their developmental potential in subsequent developmental processes. These observations indicate that the maternal effect induced by Tcf12 ensures preimplantation development.

H1N1, a globally pervasive subtype of influenza A virus (IAV), poses an ongoing threat to human health and occasionally leads to multi-organ dysfunction in severe cases. Evidence confirms that the H1N1 virus is enabled to penetrate the placental barrier; however, the underlying mechanisms by which maternal infection contributes to detrimental fetal outcomes remain elusive. In this study, a systematic literature review and meta-analysis demonstrated a strong association between maternal H1N1 infection during pregnancy and adverse fetal outcomes. Using a chicken embryo model, we found that the H1N1 virus specifically targets the developing liver and lung tissues, activates immune and stromal cells, and induces localised inflammatory responses, thereby triggering excessive oxidative stress. The resulting imbalance in oxidative stress disrupts antioxidant defence systems and promotes ferroptosis in parenchymal cells. Persistent ferroptosis subsequently initiates tissue repair processes, activates fibroblasts, and leads to aberrant extracellular matrix deposition, ultimately contributing to early fibrosis in the liver and lung tissues. Collectively, this study elucidates the molecular mechanisms by which H1N1 selectively infects fetal liver and lung, inducing ferroptosis-mediated parenchymal cell death and tissue fibrosis, thereby impairing fetal development. These findings provide novel theoretical insights for the clinical management and prevention of H1N1-associated maternal-fetal infections and adverse pregnancy outcomes.

Atherosclerosis remains a significant global health challenge, arising from the complex interactions among dysregulated lipid metabolism, chronic inflammation and immune activation. Ferroptosis, marked by lipid peroxide buildup dependent on iron, is gaining recognition as a modulator of macrophage activity in atherosclerosis. Macrophages are the pivotal orchestrators of chronic inflammation and atherosclerotic plaque formation. The marked heterogeneity and plasticity of macrophages within plaques dynamically shape the local microenvironment, contributing to phenomena such as lipid overload, cytokine overactivation, hypoxia, and programmed cell death. This review examines how dysregulated iron handling, lipid metabolism, and redox imbalances synergise to induce macrophage ferroptosis in atherosclerosis. Moreover, ferroptosis contributes to the development and progression of atherosclerosis by causing dysfunction in vascular smooth muscle cells (VSMCs), vascular endothelial cells (VECs), and macrophages, thereby promoting plaque formation and instability. Furthermore, macrophages are intricately linked to ferroptosis, with this iron-dependent cell death enhancing oxidative stress and inflammatory pathways. Macrophage ferroptosis drives plaque progression and destabilisation, ultimately heightening the risk of rupture and cardiovascular events. By inhibiting macrophage ferroptosis, it may be possible to reduce oxidative stress and inflammation, stabilise atherosclerotic plaques, and ultimately lower the risk of cardiovascular events. This review highlights the therapeutic potential of targeting macrophage ferroptosis for the treatment of atherosclerosis.

Brain organoids have become an essential platform for studying human neural development and neurological disorders. Yet, one major limitation of conventional brain organoids is their lack of vascular structures. This deficiency restricts organoid size, contributes to necrotic core formation, and hampers their functional maturation. Introducing vascularization offers a compelling solution—it enhances nutrient delivery, supports neurogenesis, and fosters the development of interfaces that resemble the blood–brain barrier (BBB). In this review, we explore how vascularization enhances the structural and physiological relevance of brain organoids and its growing significance in disease modelling and therapeutic screening. We examine current methodologies for engineering vascularized brain organoids (vBOs), including co-culturing with endothelial cells (ECs), transcriptional programming, tissue fusion techniques, microfluidic perfusion systems, and 3D bioprinting. These strategies vary in complexity, scalability, and the extent to which they achieve vascular integration. Functionally, vBOs demonstrate improved oxygen diffusion, enhanced synaptic development, and more robust barrier properties. Such advances enable modelling of complex neurovascular conditions like stroke, glioblastoma, and BBB dysfunction. Moreover, vBOs are emerging as valuable tools in developmental studies and personalised medicine, supporting patient-derived modelling and large-scale drug testing in BBB-relevant contexts. Despite these advances, replicating the structural complexity, functionality, and long-term stability of native vasculature remains challenging. We discuss current limitations and highlight innovative approaches, including the use of next-generation biomaterials and dynamic perfusion technologies. Ultimately, vBOs mark a significant step towards creating physiologically accurate in vitro models of the human brain—offering new opportunities for neuroscience research, drug development, and regenerative medicine.

X. Zhou, R. Yu, Y. Long, et al., “BMAL1 Deficiency Promotes Skeletal Mandibular Hypoplasia via OPG Downregulation,” Cell Proliferation 51 (2018): e12470, https://doi.org/10.1111/cpr.12470.

(1) The images in Figure 5A of RAW+BMSCs OPG (0 ng/mL) and RAW+(MC3T3-shRNA#1) OPG (50 ng/mL) appeared to overlap.

(2) The TRAP Staining image appeared incorrectly in Figure 6C (Wild Type twice a week, 6 weeks, OPG 0 ng/mL).

Corrected Figures 5A and 6C are provided below. The correction does not alter any findings and conclusions reported in this article.

We apologise for these errors.