Human colostrum and mature milk contain oligosaccharides (Os), designated as human milk oligosaccharides (HMOs). Approximately 200 varieties of HMOs have been characterized. Although HMOs are not utilized as an energy source by infants, they have important protective functions, including pathogenic bacteria and viral infection inhibitors and immune modulators, among other functions, and HMOs stimulate brain-nerve development. The Os concentration is average 11 g/L in human milk but >100 mg/L in mature bovine milk, which is used to manufacture infant formula, suggesting that human-identical milk oligosaccharides (HiMOs) should be incorporated into milk substitutes. Some infant formulas incorporating 2'-fucosyllactose and lacto-N-neotetraose are now commercially available, and intervention trials have been concluded. We review basic HMO information, including their chemical structures and concentrations, attempts to synthesize HMOs at small and plant scale, studies that clarified HMO biological functions, and interventions with milk substitutes incorporating HiMOs in formula-fed infants.
Recent advancements in single-cell analysis have revolutionized our understanding of cellular heterogeneity, particularly in lipid metabolism. Single-cell lipidomics, enabled by ultra-sensitive mass spectrometry techniques such as Orbitrap and Fourier-transform ion cyclotron resonance (FT-ICR), provides unprecedented insights into lipid-mediated cellular functions. Unlike bulk analyses, single-cell approaches capture real-time metabolic changes, highlighting lipid species' roles in cell differentiation, signal transduction, and disease progression. Mass spectrometry imaging (MSI), including MALDI-MSI and SIMS, further delineates lipid distributions within tissues, revealing spatial heterogeneity critical to cellular function. Emerging evidence suggests that lipid alterations significantly impact developmental mechanisms, stem cell niches, and disease pathogenesis, challenging conventional bulk-level assumptions. However, a key challenge remains in deciphering how lipid networks coordinate cellular differentiation and transcriptional regulation. Future research must integrate lipidomic, proteomic, and genomic data to unravel lipid-mediated signaling and epigenetic modifications. Understanding these dynamics will advance regenerative medicine and therapeutic interventions, enabling precise targeting of lipid-driven pathways in disease contexts.
All eukaryotic cell surfaces are coated with various types of glycans, which are essential molecules in biological events. In this review, we summarize recent integrated glycomics studies using various biological samples. We introduce an improved sialic acid linkage-specific alkylamidation (SALSA) method for sialylated glycan analysis and an automated glycosphingolipid-glycan preparation system for large-scale glycomic analysis of human plasma/serum. Finally, we explain the importance of integrated glycomics of glycoconjugates through total glycomic analysis of human serum and mouse brain tissue, and discuss prospects for exploring glycans as effective biomarkers of biological phenomena.
In recent years, the concept of neuro-skeletal crosstalk, highlighting the reciprocal interactions between the nervous and skeletal systems, has opened new avenues for understanding the pathogenesis and intervention strategies of complex diseases. This review summarizes the roles of molecular networks such as neurotransmitters, endocrine factors, immune mediators, and extracellular vesicles in bone metabolism, repair, and neurodegenerative diseases, with an emphasis on recent advances regarding bone-derived signals-including the Piezo1 channel and osteocalcin-in neural regulation. Building on this foundation, we focus on advances in frontier materials such as nanomaterials and hydrogels for modulating the brain-bone microenvironment and facilitating coordinated tissue regeneration, as well as new strategies for targeted drug delivery and immune microenvironment modulation. Empowered by next-generation technologies-including multi-omics, artificial intelligence, and organ-on-a-chip systems-the investigation of the fundamental mechanisms and personalized interventions of the brain-bone axis is entering a new era of opportunity. We hope that this review will provide a theoretical basis and valuable reference for future mechanistic studies and innovation in this interdisciplinary field. By elucidating bidirectional regulatory networks, this review underscores the significant translational potential of targeting the brain-bone axis (BBA) for the treatment of skeletal disorders and neurodegenerative comorbidities. Therapeutic strategies harnessing neurotransmitters (e.g., norepinephrine, serotonin) and neuropeptides (e.g., CGRP) can directly modulate osteoblastic/osteoclastic activity and immune responses, thereby orchestrating fracture repair and metabolic homeostasis. The integration of functional materials-such as stimuli-responsive hydrogels, nanomaterials, and bioelectronic devices-enhances the spatiotemporal precision of signal modulation and facilitates drug delivery across biological barriers, including the blood-brain barrier (BBB). However, challenges regarding low cross-organ targeting efficiency, the complexity of dynamic pathological microenvironments, and physiological discrepancies between animal models and humans necessitate further optimization. Advances in multi-omics analysis, AI-driven network modeling, and intelligent biomimetic delivery systems hold promise for bridging these gaps, offering scalable solutions for clinical translation. This work highlights neuro-skeletal modulation as a transformative dual-targeting strategy for complex diseases, yet its implementation remains contingent upon the refinement of precise intervention technologies and rigorous clinical validation.
Rare sugars are defined as monosaccharides and their derivatives that do not exist in nature at all or that exist in extremely limited amounts despite being theoretically possible. At present, no comprehensive dogma has been presented regarding how and why these rare sugars have deviated from the naturally selected monosaccharides. In this minireview, we adopt a hypothesis on the origin and evolution of elementary hexoses, previously presented by one of the authors (Hirabayashi, Q Rev Biol, 1996, 71:365-380). In this scenario, monosaccharides, which constitute various kinds of glycans in nature, are assumed to have been generated by formose reactions on the prebiotic Earth (chemical evolution era). Among them, the most stable hexoses, i.e., fructose, glucose, and mannose remained accumulated. After the birth of life, the "chemical origin" saccharides thus survived were transformed into a variety of "bricolage products", which include galactose and other recognition saccharides like fucose and sialic acid through the invention of diverse metabolic pathways (biological evolution era). The remaining monosaccharides that have deviated from this scenario are considered rare sugars. If we can produce rare sugars as we wish, it is expected that various more useful biomaterials will be created by using them as raw materials. Thanks to the pioneering research of the Izumori group in the 1990's, and to a few other investigations by other groups, almost all monosaccharides including l-sugars can now be produced by combining both chemical and enzymatic approaches. After briefly giving an overview of the origin of elementary hexoses and the current state of the rare sugar production, we will look ahead to the next generation of monosaccharide research which also targets glycosides including disaccharides.
OBJECTIVE: This study aimed to construct a perovskite quantum dot probe targeting isocitrate dehydrogenase 1 (IDH1) mutation. The probe is intended to enable rapid in vitro pathological diagnosis of IDH1-mutant glioma, guide surgeons in formulating scientific and rational resection strategies during surgery, and facilitate precise individualized intraoperative treatment for glioma patients. METHODS: Via surface ligand engineering, a perovskite nanocrystal (PNC) probe modified with bromobutyric acid (BBA) was developed in this study. This probe enhances water/oxygen stability and addresses the limitation that restricts the application of perovskite quantum dots as fluorescent probes. The probe was used to stain intraoperative tumor frozen sections to verify its specific recognition capability for IDH1-mutant glioma. RESULTS: Verification using 50 glioma frozen sections demonstrated that the probe could accomplish rapid pathological detection within 30 min (with a 5-minute incubation period). Fluorescence imaging showed specific green fluorescence in IDH1-mutant glioma. When compared with genetic testing results, the probe exhibited the following detection performance: sensitivity of 100%, specificity of 84%, positive predictive value of 86.2%, negative predictive value of 100%, and accuracy of 92% (κ = 0.84, P < 0.001). CONCLUSIONS: This BBA-stabilized perovskite probe enables rapid and specific in vitro detection of IDH1-mutant glioma. It provides real-time guidance for glioma surgery, is expected to assist surgeons in achieving precise resection of glioma, and thus advances the development of precision neuro-oncology.
Alpha-synuclein (α-Syn) is a neuronal protein implicated in the pathogenesis of several neurodegenerative disorders collectively known as synucleinopathies, including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. This article provides a comprehensive overview of the structural characteristics of α-Syn, emphasizing its fibrillar aggregation and the resulting polymorphic fibril forms. Using advances in cryo-electron microscopy, a diverse range of α-Syn fibril polymorphs has been elucidated, both from in vitro preparations and patient-derived brain samples. We further explore the impact of mutation and post-translational modifications-such as truncation and phosphorylation -on α-Syn structure and the unique polymorphs they induce. The article underscores the biological and pathological relevance of α-Syn polymorphism, highlighting how different structural strains may underlie the structural and pathological heterogeneity observed in synucleinopathies. Understanding the mechanisms driving polymorph formation is critical for deciphering disease progression and developing targeted therapeutic strategies.
Pathogenic variants in the GNAO1 gene, which encodes for Gαo, a major neuronal G protein, are associated with neurodevelopmental disorders, epilepsy, and movement disorders. We identified and characterized in detail a de novo heterozygous GNAO1 E246K pathogenic variant in an Israeli female infant with complex developmental delay and substantial motor difficulties. This variant has been reported in other cases as a recurrent pathogenic variant in patients with motor dysfunction and a broad range of neurological outcomes. To investigate the molecular and functional consequences of the Gαo E246K variant, we employed structural modeling and analysis, mass spectrometry-based proteomics, biochemical assays, and cellular functional assays. Our biochemical results show that this variant does not affect nucleotide binding, nor basal or RGS-accelerated GTP hydrolysis. Despite the E246 position location within a predicted effector binding region, mass spectrometry analysis did not identify any novel cellular partners. Instead, we demonstrate that the E246K variant disrupts the Gαo regulatory GTPase cycle by directly impairing Gβγ dissociation. This impairment overrides the function of wild-type Gαo, explaining the dominant effect and the severity of the neurogenetic phenotype despite a heterozygous background. These findings establish a new molecular mechanism for a GNAO1 variant with dominant-negative effects on the GTPase regulatory cycle. The insights gained from studying this mechanism of action provide a basis for developing specific and personalized therapeutic strategies based on the outcome of a missense mutation in GNAO1.
CYP2E1 expression is often negatively correlated with hepatocellular carcinoma (HCC) progression, and its overexpression induces apoptosis in HCC cells. However, the underlying mechanism of CYP2E1 in apoptosis induction remains unclear. Given the crucial role of lysosomal dysfunction through lysosomal membrane permeabilization (LMP) in promoting apoptosis, this study aimed to investigate whether and how lysosomal dysfunction and LMP contribute to CYP2E1-induced apoptosis in HCC. Here, we demonstrated that CYP2E1 overexpression induced LMP, leading to the translocation of cathepsin D and subsequent apoptosis in HCC cells. Notably, cathepsin D inhibitor attenuated CYP2E1 overexpression-induced LMP and apoptosis. In addition, through sialidase Neuraminidase 4 (NEU4), CYP2E1 overexpression resulted in the deglycosylation and degradation of lysosome-associated membrane protein 1 and 2 (LAMP1/2), contributing to LMP and apoptosis both in vitro and in vivo. Clinically, decreased CYP2E1 expression in the tumor tissues of HCC patients is strongly associated with upregulated LAMP1/2 expression and glycosylation. Collectively, our findings elucidate the role of LMP in CYP2E1-induced cell apoptosis with the involvement of NEU4, providing novel insight for CYP2E1 function in apoptosis.
Human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are a valuable tool for modeling cardiac diseases, drug testing, and regenerative applications. However, their application is limited by the immature phenotype of iPSC-CM. During maturation from fetal to adult phenotype cardiomyocytes undergo a transition from glycolysis to oxidative phosphorylation for energy production, which is supported by efficient tricarboxylic acid (TCA) cycle activity. Our metabolomics data suggest that the level of intermediates of TCA cycle including succinate, malate, fumarate, and α-ketoglutarate was very low in iPSC-CM. Therefore, we investigated the effect of supplementation with these metabolites on the maturation of iPSC-CM. We cultured iPSC-CM in glucose (Glu), galactose (Gal), or galactose plus TCA cycle intermediates (Gal+TCA) and evaluated the incremental effect of TCA cycle intermediates supplementation relative to Glu and Gal. The treatment with these TCA cycle intermediates led to improved calcium handling and cellular morphology of iPSC-CM relative to Glu and Gal. Furthermore, the treatment with TCA cycle metabolites enhanced electrical activity, improved mitochondrial health, and the cells were shifting toward oxidative phosphorylation relative to Glu only. This shift in the energy metabolism was associated with an upregulation in the expression of cardiomyocyte maturation genes and downregulation in the expression of fetal genes in Gal + TCA group relative to Glu. Overall, the benefits of Gal+TCA supplementation were quite evident compared to Glu alone but generally modest relative to Gal supplementation, supporting that TCA cycle intermediates supplementation can be used as an adjunct strategy to promote iPSC-CM maturation.
S100A13, a calcium-binding protein containing two EF-hand motifs, contributes to intracellular calcium homeostasis, a key determinant of mitochondrial quality control. We previously showed that S100A13 modulates mitochondrial membrane potential (ΔΨm) in patient-derived skin fibroblasts carrying S100A13 (p.I80Gfs*13) and S100A3 (p.R77C) mutations. However, its role in regulating mitochondrial dynamics remains unclear. Here, we investigated whether S100A13 regulates mitochondrial fusion-fission balance in human bronchial epithelial cells (BEAS-2B) using Myc-tagged wild-type or p.I80Gfs*13 S100A13 constructs. The S100A13 p.I80Gfs*13 mutant markedly attenuated bradykinin- and ionophore-induced intracellular calcium transients and reduced ΔΨm compared with wild-type S100A13 (p < 0.05). These alterations were associated with severe mitochondrial ultrastructural abnormalities, disrupted cristae organization, and increased mitochondrial fragmentation (p < 0.05). Interestingly, both wild-type and p.I80Gfs*13 mutant S100A13 increased expression of the mitochondrial fusion-associated proteins MFN1/2 and OPA1 while reducing expression of the fission mediator MFF. However, despite these apparently pro-fusion molecular changes, the S100A13 p.I80Gfs*13 mutant failed to maintain mitochondrial fusion competency, suggesting a functional uncoupling between fusion protein abundance and mitochondrial fusion competency. Collectively, these findings identify S100A13 as an important regulator of intracellular calcium-dependent mitochondrial dynamics and demonstrate that C-terminal truncation disrupts calcium-dependent regulation of mitochondrial fusion and cristae integrity in lung epithelial cells.
A theoretical model of mitochondrial β-oxidation efficiency is presented, using ATP yield per oxygen atom (P:Oβ-ox) as a proxy for performance under oxygen-limited conditions. Expressed as a function of chain length and unsaturation, the model generates hyperbolic efficiency surfaces whose maxima mirrors the natural abundance of fatty acids in adipose tissues. It predicts a crossover near d ≈ 1.6, where efficiency becomes chain-length-independent-thus matching the dominance of monounsaturated FAs in mammals. The model aligns with empirical mobilization patterns and suggests an oxygen- and ATP-sensitive regulatory axis shaping lipid profiles in hypoxic or energy-stressed states.
Orofacial pain is difficult to manage. Treatments like nonsteroidal anti-inflammatory drugs (NSAIDs), and serotonin agonists provide limited relief or produce side effects. These limitations highlight the need for alternative strategies that reduce orofacial pain without adverse effects. One approach is polypharmacology, where a single compound modulates multiple targets involved in pain. Fatty acid amide hydrolase (FAAH) and soluble epoxide hydrolase (sEH) are regulators of pain that can be targeted simultaneously to enhance antinociceptive efficacy. We previously reported that the 4-phenylthiazole-based dual FAAH/sEH inhibitor SW-17 does not alleviate acute orofacial pain in rats, and it showed low stability in rat liver microsomes. We introduced electronic and steric modifications to the thiazole and phenyl rings of the 4-phenylthiazole scaffold to improve the stability and better understand structure-activity relationship of this set of analogs. An 11-compound library was synthesized by varying alkyl groups at position 5 of the thiazole ring and substituents on the phenyl ring. Several analogs exhibited nanomolar potency at both enzymes. The most potent compound, 4j, exhibited IC50 values of 18.7 nM and 25.1 nM for human FAAH and sEH, respectively. 4j was evaluated in rat liver microsomes where it exhibited relatively low metabolic stability but better than SW-17. ADMET studies indicated that 4j possesses promising safety features. At 3 mg/kg, 4j reversed acute orofacial inflammatory pain, whereas SW-17 was ineffective. This effect was comparable to sumatriptan and did not reduce voluntary wheel running. These findings imply that optimized dual FAAH/sEH inhibitors can alleviate orofacial pain without affecting normal behavior.
Given that extracellular vesicles (EVs) secreted from stem cells contain angiogenic microRNAs (miRNAs), these EVs show equivalent angiogenic therapeutic effect to cell transplantation. This study aimed to identify the angiogenic miRNAs from several miRNAs involved in EVs secreted from stem cells. In human dental pulp stem cells (hDPSCs) under hypoxic culture, vascular endothelial growth factor (VEGF) involved in EVs increased. We hypothesized that angiogenic miRNAs increase in EVs that are secreted from hDPSCs under hypoxic culture compared with those under normoxic culture. In the first screening, the expression levels of miRNAs involved in EVs that were secreted from hDPSCs cultured under hypoxic and normoxic conditions were analyzed using miRNA array. In the second screening, 12 miRNAs were individually involved in EVs, and the growth of human aortic endothelial cells was assayed. In the third screening, miRNA-encapsulated EVs were injected in BALB/c mouse hindlimb ischemia model, followed by angiogenesis evaluation by blood flow analysis. Results showed that miR-765 is an angiogenic miRNA, targeting the 3' untranslated region of dipeptidyl peptidase 4 (DPP4) and upregulating fibroblast growth factor 2 (FGF2) in vitro and in vivo. In conclusion, as an angiogenesis mechanism, miR-765 increased FGF2 expression levels by inhibiting DPP4.
Storage of biological materials underpins medical, research, and biotechnological applications. Although cold-chain preservation is effective, it is costly, infrastructure-dependent, and vulnerable to disruption. Room-temperature dry storage, inspired by desiccation-tolerant organisms, offers an alternative by stabilizing biomolecules in vitrified matrices that limit molecular motion and degradation. Trehalose is widely used as a vitrifying agent, but its protective capacity depends on glassy properties shaped by drying conditions, environment, storage duration, and biomolecule type. However, systematic links between these factors and stability remain poorly defined. Here, we examine how drying conditions and storage duration influence the stability of DNA, RNA, and enzymes in trehalose-based vitrified systems. DNA remained stable across all conditions, independent of trehalose or drying parameters, reflecting intrinsic resistance to desiccation damage. RNA exhibited moderate sensitivity to drying without trehalose but was stabilized in its presence, although RNA integrity did not consistently correlate with measured glassy properties. In contrast, enzymes were highly sensitive to drying in the absence of trehalose and strongly protected under conditions that promoted favorable vitrified properties. Short-term enzyme protection (30 min) positively correlated with higher glass transition temperature (Tg). However, during prolonged dry storage, higher Tg was inversely correlated with enzyme stability and instead tracked detrimental physical aging of the vitrified matrix. These findings demonstrate that optimal glass properties depend on both biomolecule class and timescale, providing a framework for rationally designing room-temperature preservation strategies.
Resistance of tumor cells to chemotherapy remains a critical obstacle to effective cancer treatment. Although paclitaxel is one of the most commonly used chemotherapeutic agents for treating triple-negative breast cancer (TNBC), the mechanisms underlying paclitaxel resistance are not fully understood. We previously found that phosphodiesterase 1C (PDE1C) was substantially upregulated in a paclitaxel-resistant T50RN cell clone established from the human TNBC cell line MDA-MD-231. In this study, we aimed to explore whether and how PDE1C modulates resistance to paclitaxel in T50RN cells. Our results showed that depletion of PDE1C enhanced paclitaxel cytotoxicity, and that pharmacological inhibition of PDE1 potentiated paclitaxel-induced antiproliferative and antimitotic effects in T50RN cells. Additionally, intracellular cyclic adenosine monophosphate (cAMP) levels were lower in T50RN cells than in parental MDA-MB-231 cells. PDE1 inhibition restored the cAMP level, suggesting that cAMP-degrading activity of PDE1 is elevated in the T50RN cells. Similar to PDE1 inhibitors, the cell permeable cAMP analog 8‑bromo-cAMP or the adenylate cyclase activator forskolin increased cAMP levels and concurrently augmented paclitaxel-induced cytotoxicity and spindle abnormalities in T50RN cells. Furthermore, PDE1 inhibitors, forskolin, and an agonist of the cAMP downstream effector EPAC enhanced paclitaxel-mediated microtubule (MT) stabilization. Thus, PDE1 inhibition may act through cAMP/EPAC signaling to facilitate MT stabilization and potentiate the antiproliferative and antimitotic effects of paclitaxel in T50RN cells. Upon PDE1 inhibition, paclitaxel-treated T50RN cells exhibited signs of endoplasmic reticulum (ER) stress and apoptosis. Together, our in vitro findings indicate that PDE1C overexpression contributes to paclitaxel resistance.
Protein misfolding in multidomain assemblies emerges from a complex interplay between intra- and interdomain interactions. Here, we dissect how neighboring domains shape the folding and misfolding of the PDZ4 domain from the scaffold protein MAGI1. Exploiting the single intrinsic tryptophan in PDZ4, we monitored its behavior in the isolated domain, in the PDZ3-4 tandem, and within the five-domain PDZ2-6 supramodule by equilibrium and kinetic (un)folding experiments over a broad pH range. Equilibrium denaturation reveals that PDZ4 displays a cooperative, two-state-like transition with similar thermodynamic stability in all constructs, indicating that domain adjacency does not appreciably affect its native state. In contrast, kinetic measurements uncover pronounced deviations from two-state behavior. PDZ4 alone populates a transient intermediate, whose population and associated kinetic slowdown are markedly amplified in the tandem and even more in the supramodular context, consistent with the formation of a misfolded ensembles stabilized by non-native interdomain contacts. Acidic pH selectively enhances the kinetic trap in PDZ3-4 and PDZ2-6, but not in PDZ4 alone, pointing to a key role of electrostatics at interdomain interfaces, potentially involving specific salt bridges. Structural inspection suggests the presence of potential specific salt bridges (Asp871/Glu901 with Arg873/Lys905), although further investigation is required. Our data show that misfolding of MAGI1 PDZ4 is a context-dependent, cooperative property of adjacent domains rather than a simple by-product of folding, and illustrate how supramodular organization encodes kinetic plasticity in scaffold proteins.
Glycoinformatics has entered the artificial intelligence era. Stymied by a lack of big data, high sequence complexity, and biosynthetic dependencies, the application of machine learning to glycomics data has largely emerged this decade. In this mini-review, we explore the latest groundbreaking computational approaches applied to glycan sequencing, classifying disease risk from glycan biomarkers, and predicting protein-glycan interactions. We detail advancements in the architectures of these models, concluding that they have matured to the stage of extracting predictive information from glycan sequences. We also discuss their challenges and limitations, and how to fully reap their rewards in the future.
Lipids are fundamental to the biology of all organisms, including the functioning of subcellular organelles. The plant chloroplast is the site of photosynthesis and serves as a hub of metabolic events that are dynamically regulated between the light and dark periods. The regulation of chloroplast events, like most other biological processes, is often controlled by protein phosphorylation. Here, we explored the lipidome of Arabidopsis thaliana under both light and dark conditions, examining lipid changes upon overexpression or knockout of the chloroplast-specific phosphatase Shewanella-like protein phosphatase 1 (SLP1). To date, no substrates have been identified for the phosphatase SLP1. Results indicate that Arabidopsis thaliana possesses a dynamic lipidome that undergoes significant changes between light and dark conditions, and that protein phosphorylation, in part controlled by SLP1, plays a crucial role in regulating chloroplast metabolism and its associated lipidome. Alterations due to loss or over-expression of SLP1 in the chloroplast are transmitted to other cellular compartments, leading to altered lipids throughout the cell. We report that although both SLP1 mutant lines display perturbed glycerolipid metabolism, knockout and overexpression affect distinct pathways. In the three Arabidopsis lines studied, we also identify dramatic diurnal changes in the abundance of oxidized lipids. Together, these findings establish SLP1-dependent dephosphorylation as an upstream regulator of diel lipid remodelling, linking chloroplast activity to broader cellular lipid organization.
Glaucoma is the second leading cause of irreversible blindness globally, with elevated intraocular pressure (IOP) being its primary risk factor. Current therapeutic approaches, such as beta-blockers, alpha-adrenergic agonists, Rho-kinase inhibitors, etc., aim to reduce IOP levels. However, the molecular mechanisms underlying altered IOP remain poorly understood. In this study, we have treated primary human trabecular meshwork cells (HTM) with exogenous dexamethasone (dex) or transforming growth factor beta-2 (TGF-β2) to investigate their effects on glaucoma candidate genes. Interestingly, our findings reveal that FOXC1 acts as a repressor of CYP1B1, and optineurin (OPTN) facilitates the ubiquitination of FOXC1, thereby inducing the expression of CYP1B1. Furthermore, we found that the miR-200 family and other miRNAs regulate these glaucoma candidate genes. Furthermore, TGF-β2 downregulates the miR-200 family, whereas the miR-200 family targets FOXC1, exerting reversible effects by altering the extracellular matrix. Thus, modulating the TGF-β2/OPTN/FOXC1/miR-200 axis appears critical in regulating actin dynamics in the anterior eye segment.