Advanced glycation end products (AGEs) accumulate in the serum of people with diabetes, some chronic conditions, and aging and impair osseointegration by reshaping the protein corona and activating oxidative stress pathways at the biomaterial interface. Here, we introduce a pathology-adaptive surface engineering strategy in which plasma polymer coatings with defined functional chemistries are designed to selectively modulate AGE adsorption while preserving physiological protein interactions. Titanium was functionalized with carboxyl (-COOH), amine (-NH2), or hydrocarbon (-CH3) plasma polymers to precisely modulate interfacial chemistry. Carboxyl- and amine-functionalized surfaces significantly reduced AGE adsorption compared to hydrophobic controls while maintaining total serum protein adsorption, indicating selective exclusion of glycated species. This modulation of the AGE-modified protein corona attenuated the generation of reactive oxygen species, suppressed apoptosis, and restored osteogenic gene expression in bone marrow mesenchymal stem cells following exposure to AGEs. Functionalized coatings supported sustained proliferation and enhanced mineralization, demonstrating the recovery of regenerative capacity in a metabolically compromised environment. The AGE-RAGE axis was identified as a key driver of macrophage inflammatory activation. These findings establish pathology-adaptive surface chemistry as a materials strategy to selectively regulate disease-associated protein adsorption and restore regenerative performance under glycation stress.

ETS variant transcription factor 2 (ETV2) serves as a foundational transcription factor for endothelial lineage specification. However, the lineage-specific cofactors that orchestrate with ETV2 during endothelial fate commitment remain elusive. Here, we demonstrate that ETV2 drives the rapid forward programming of human pluripotent stem cells (hPSCs) into endothelial cells (ECs) by direct remodeling of endothelial-specific enhancers. Crucially, we identify T cell acute lymphocytic leukemia protein 1 (TAL1), which is traditionally characterized as a hematopoietic regulator, as an indispensable cofactor for ETV2-mediated endothelial commitment. Distinct from its role in murine development, TAL1 deficiency in hPSCs not only aborts the endothelial program by impairing H3K27ac deposition at key enhancers but also triggers a profound lineage redirection toward a mesenchymal fate. Mechanistically, TAL1 physically interacts with ETV2 to recruit the p300, thereby facilitating a permissive chromatin environment for endothelial identity. By leveraging an hPSC-based differentiation model, our findings establish TAL1 as a master gatekeeper of human EC specification and provide a molecular blueprint for how ETV2-centric complexes synergistically govern human cell fate.

Acute myeloid leukemia (AML) is an aggressive hematological malignancy arising from hematopoietic stem and progenitor cells (HSPCs). Current treatments often fail to eradicate AML; therefore, new therapeutic strategies are essential. Here, we reveal that RNA terminal uridylyl transferase enzymes 4 and 7 (TUT4/7) are druggable therapeutic targets, whose genetic deletion suppresses AML growth, induces apoptosis, and improves the survival in leukemic mouse models. Notably, a preclinical TUT4/7 inhibitor promotes cell death in samples from patients with AML and synergizes with venetoclax. Mechanistically, TUT4/7 inactivation suppresses mevalonate pathway gene expression, compromising the cholesterol synthesis pathway. Current AML therapies often cause severe hematopoietic toxicity. Although Tut4/7 deletion results in inflammatory activation throughout the hematopoietic system, this is permissive to a normal life span and Tut4/7 deficiency does not compromise HSPC function. Together, these findings identify TUT4/7 as druggable targets, whose inactivation suppresses AML while sparing normal hematopoiesis. In combination with venetoclax, this represents a promising therapeutic strategy.

Mutations in the transcription factor gene Wilms Tumor 1 (WT1) are one of the leading causes of congenital glomerular disease, characterized by severe urinary protein loss and glomerular scarring. No disease-modifying therapies exist for WT1 glomerulopathies, and affected children rely on dialysis or kidney transplantation. We evaluated a previously uncharacterized treatment in a mouse model with an orthologous human mutation in Wt1 (Wt1+/R394W) that replicates the pathology of WT1 glomerulopathy. Lipid nanocomplexes were engineered to target integrin αvβ3 for efficient delivery of mRNA to primary podocytes and glomerular endothelial cells in vitro. A minimally invasive ultrasound-guided renal artery injection enabled precise and specific kidney localization in vivo, a finding not replicated by systemic administration. Nanocomplex-derived protein localized in glomeruli for up to 7 days in healthy and diseased mice. This platform was then used to perform an interventional preclinical trial in Wt1+/R394W mice, delivering angiopoietin-1 (Angpt1) mRNA, a vascular growth factor critical for glomerular health that is reduced in Wt1+/R394W podocytes. Angpt1 nanocomplex therapy reduced albuminuria, preserved glomerular endothelial integrity, prevented podocyte loss, and alleviated glomerulosclerosis in Wt1+/R394W mice. These findings demonstrate the therapeutic potential of this targeted approach for WT1 glomerulopathy and provide a foundation for clinical translation to improve outcomes in children with glomerular disease.

Embryonic stem cells (ESCs) are refractory to inflammatory stimuli but acquire inflammatory competence upon differentiation, yet the underlying mechanisms remain unclear. Here, we report that H2BK5ac regulates the acquisition of inflammatory potential in mouse ESCs during differentiation. H2BK5ac levels increased at the regulatory regions of inflammatory genes upon differentiation, and this modification was required for their transcriptional upregulation. An acetylation-deficient H2BK5A mutant impaired the upregulation of inflammatory genes, whereas an acetylation-mimicking H2BK5Q mutant had the opposite effect. We identified p300 and HDAC3 as the acetyltransferase and deacetylase, respectively, that control H2BK5ac: p300 protein levels increased upon differentiation and its depletion reduced H2BK5ac levels, whereas loss of HDAC3 increased H2BK5ac levels despite its unchanged protein levels. H2BK5Q overexpression in mouse embryos increased embryonic mortality and inflammatory cytokine secretion, consistent with its pro-inflammatory effect. Thus, our study reveals H2BK5ac as a regulator that establishes inflammatory potential in mouse ESCs during differentiation.

Mesenchymal glioma stem cells (MES GSCs) are closely associated with glioblastoma radioresistance, yet the mechanisms linking MES-state maintenance to ferroptosis resistance remain incompletely defined. Here, we show that MES GSCs exhibit enhanced glycolytic activity and lactate production, driven in part by MES-associated transcriptional regulators that promote LDHA expression. LDHA-derived lactate induces p300-dependent H3K18 lactylation, which enhances USP30 transcription. USP30 subsequently stabilizes the lipid-remodeling enzyme MBOAT2 by limiting its ubiquitination, leading to phosphatidylethanolamine remodeling toward ferroptosis-resistant PE-MUFA species. Disruption of this LDHA-H3K18la-USP30-MBOAT2 axis by LDHA inhibition, impaired H3K18 lactylation, USP30 inhibition, or MBOAT2 depletion increases lipid peroxidation, promotes ferroptosis, and sensitizes MES GSCs to irradiation. Lipid rescue experiments further identify PE-MUFA remodeling as a functional mediator of MBOAT2-dependent ferroptosis resistance. In intracranial xenografts, combined targeting of glycolysis and USP30 enhances radiotherapeutic efficacy without obvious treatment-associated body weight loss under the tested conditions. These findings reveal a metabolic-epigenetic-lipid remodeling circuit that protects MES GSCs from ferroptosis and promotes radioresistance.

Androgenetic alopecia (AGA) is a common form of hair loss with limited treatment options, driven by poorly understood molecular mechanisms. Using single-cell RNA sequencing of human scalp follicles from balding and non-balding regions, we constructed a high-resolution molecular anatomy atlas of the hair follicle and its microenvironment. We identified significant silencing of Wingless-related integration site 5A (WNT5A) signaling, which was widely known as the trigger of non-canonical WNT signaling pathway, in hair follicle stem cells (HFSCs) from balding areas, leading to downregulation of the transcription factor ATF3 and its target FOSB. Therapeutically, functional validation showed that activating the WNT5A-ATF3-FOSB axis maintained HFSC competence in AGA murine models and significantly inhibited the progression of AGA. Our findings reveal a critical molecular axis in AGA pathogenesis, highlighting its potential as a therapeutic target for hair loss disorders.

Human induced pluripotent stem cells (hiPSCs) represent a powerful platform for disease modeling, especially in monogenic diseases as they preserve the donor's genetic background while enabling directed differentiation into disease-relevant cell types. This makes them highly suitable for studying disease mechanisms in a patient-specific and physiologically relevant context. Although CRISPR/Cas9 is widely applied for genome editing, precise correction of pathogenic variants in hiPSCs remains challenging due to the lack of standardized CRISPR component selection and experimental design. Here, we describe an optimized CRISPR-based strategy for correcting a heterozygous HNF1A frameshift mutation (c.235_236insG; p.Glu79Glyfs*16) in HNF1A-MODY patient-derived hiPSCs. Using electroporation, we efficiently delivered CRISPR components, including a ribonucleoprotein complex of Cas9 and single-guide RNA, along with a single-stranded oligodeoxynucleotide repair template. Corrected hiPSC lines were validated for pluripotency, absence of exogenous reprogramming factors, and off-target effects. Additionally, we discuss key technical challenges encountered during the editing process and provide practical recommendations that may improve the generation of mutation-corrected hiPSC lines. These guidelines could serve as a useful reference for researchers employing CRISPR-based strategies for generation of reliable disease modelling tools.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold significant promise for cardiac regeneration therapies. However, the efficacy of such treatments depends on the ability of transplanted cells to migrate and integrate into the damaged myocardium, a process that remains poorly understood. In this study, we investigated the migratory behaviour of hiPSC-CMs using homogenised rat MI tissue to simulate myocardial infarction (MI) in vitro. Transwell migration assays demonstrated a concentration-dependent chemotactic response, with hiPSC-CM migration increasing up to threefold towards MI tissue homogenate. Wound-healing assays further confirmed enhanced migration under MI-mimetic conditions. Bulk RNA sequencing revealed activation of the TGF-β signalling pathway as a key regulator of this response. Inhibition of TGF-β signalling, both pharmacologically and through antibody neutralisation, significantly reduced hiPSC-CM migration. These findings uncover a previously underappreciated chemotactic capability of hiPSC-CMs and identify TGF-β signalling as a central mediator, offering new mechanistic insights and potential therapeutic targets to improve the integration and efficacy of hiPSC-CM-based cardiac regeneration strategies.

Diabetic foot ulcer (DFU) is a clinically challenging complication characterized by poor healing outcomes, and conventional therapies provide limited benefit. Mesenchymal stem cell (MSC) transplantation offers a promising strategy for DFU repair. However, the low survival of transplanted MSCs in the hostile wound microenvironment, coupled with the lack of real-time, non-invasive methods to track these cells in vivo, severely hampers their therapeutic efficacy and clinical translation.

The isolation efficiency of extracellular vesicles (EVs) is essential to ensure their purity and biological properties. Ultracentrifugation is a traditional method which utilizes high centrifugal forces to separate EVs based on their density. The polyethylene glycol (PEG)-based precipitation method is a scalable and cost-efficient alternative to ultracentrifugation. In this study, two popular techniques used for the isolation of EVs from umbilical cord tissue-derived mesenchymal stem cells (UCT-MSCs): ultracentrifugation and polyethylene glycol PEG-based precipitation methods are examined.

Type 2 diabetes mellitus (T2DM) is one of the most common metabolic disorders arising from decreased insulin secretion and insulin sensitivity. Patients with T2DM take statins to prevent diabetic complications and mitigate mortality risk associated with this disease. Statins primarily function by blocking HMG-CoA, resulting in reduced cholesterol levels. Statins have been suggested to possess certain antidiabetic properties. However, the overall effects of statins remain controversial. The aim of this study was to investigate the mechanisms and triggering conditions for the development of statin-induced T2DM, despite their reported pleiotropic antidiabetic effects.

Titanium implants are widely used in prosthodontics, but their bioinert surfaces can limit early osseointegration. This study examined whether electrolyte-tuned anodization can tailor TiO2 nanotube (TNT) coatings to improve in vitro osteogenesis, angiogenesis, and inflammation-related responses.

Mammals have four tropomyosin (TPM) genes (TPM1, TPM2, TPM3, and TPM4) that produce various isoforms through alternative splicing. TPM1 generates both myofibrillar (Tpm1.1, Tpm1.2) and non-myofibrillar isoforms, including those with an exon 9d peptide or with an exon 9a peptide. These non-myofibrillar isoforms are involved in regulating actin-based structures and processes in non-muscle cells. This study focused on the expression of the non-myofibrillar HMW Tpm1 isoforms (Tpm1.3, Tpm1.4, Tpm1.5, Tpm1.6, Tpm1.7, and Tpm1.14) during human inducible pluripotent stem cells (hiPSC) differentiation into cardiomyocytes (CMs) at different time points (Days 0, 5, 10, 15, 20). We have determined the expression of various Tpm1 transcripts by qRT-PCR using isoform-specific primer-pairs. Western blotting with Tpm1-exon 6a and Tpm1-exon 9d antibodies and 2D Western blotting with Tpm1-exon 6a antibody followed by mass spectra analyses were used to evaluate protein expression. Transcripts of non-myofibrillar Tpm1 isoforms peaked at Day 15 and continued at a slightly lower level in mature hiPSC-CM until Day 20. However, no expression of Tpm1.3 or Tpm1.14 has been observed. Protein expression of Tpm1.4, Tpm1.5, Tpm1.6, and Tpm1.7 increases up to Day 15 but practically disappears by Day 20 hiPSC-CM, suggesting their production is decreased, or more likely they are degraded intracellularly. The results suggest that the proteins have the potential to be transiently involved in the early stages of CM differentiation. This study also demonstrates that Tpm1.5, when fused with YFP in an expression construct and transfected into embryonic chicken CM and Day 20 mature hiPSC-CMs, can be organized into cardiac myofibrils despite being previously characterized as a non-muscle isoform. Our observation is further substantiated by the fact that YFP-Tpm1.5 fusion protein can be ectopically expressed and incorporated into the myofibrils of chicken myotube skeletal muscle, which is known to be more stringent than cardiac muscle with regards to myofibril remodeling. Paradoxically, anti-6a antibody fails to recognize the organized YFP-Tpm1.5 fusion protein in embryonic chicken CMs, embryonic chicken skeletal muscle myotubes, or in Day 20 mature hiPSC-CMs. Interestingly, the antibody recognizes the denatured YFP-Tpm1 fusion protein in Western blot analyses. It is well-documented in the literature that an antibody epitope may fail to recognize its antigen when the antigen is in its native state in living cells, often because the epitope is buried, altered conformationally, or inaccessible. Conversely, while the Tpm1.Ex6a antibody stains Tpm1.5 in cell nuclei, the YFP-Tpm1.5 fusion protein does not localize to the cell nuclei. This suggests conformational differences between nuclear and cytoplasmic Tpm1 protein(s) and differential access of the fusion protein to different cellular locations. This also suggests that it is endogenous Tpm1 6a-containing protein identified in the nuclei.

Post mortem proteomic analysis of Alzheimer's disease (AD) brain tissue has identified novel target genes and proteins for potential therapeutic development. Moesin (MSN) and CD44 were identified as candidate targets. Using human induced pluripotent stem cells (hiPSCs)-derived neurons, we assayed how reducing gene expression of MSN and CD44 affected amyloid beta secretion, tau phosphorylation, and transcriptional state.

Isocitrate dehydrogenase (IDH)-mutant gliomas are infiltrative brain tumors with heterogeneous clinical courses. Predicting individual responses to chemotherapy and long-term outcomes remains challenging. We aimed to characterize distinct patterns of tumor evolution during first-line chemotherapy using longitudinal tumor volume dynamics and clinical outcomes.

Transforming growth factor-β (TGF-β) signaling plays a crucial role in maintaining the quiescence and self-renewal potential of hematopoietic stem cells (HSCs). Despite decades of research, the underlying mechanisms of TGF-β signaling in HSCs remain elusive due to conflicting phenotypes of various knockout (KO) mouse models. Here, we show that HSCs co-express Tgfbr2, Tgfbr3, and Endoglin (Eng) but rarely express Tgfbr1, whereas lymphocytes co-express Tgfbr1 and Tgfbr2 but rarely express Tgfbr3 or Eng. We also demonstrate that Tgfbr3 is dispensable for the maintenance of immune homeostasis, in contrast to Tgfbr1 and Tgfbr2, either of which is essential for lymphocyte homeostasis. Serial transplantation assays revealed that deletion of Tgfbr3 in HSCs had little effect on short-term reconstitution but impaired long-term self-renewal potential, a similar phenotype observed in Eng conditional KO mice. Therefore, we propose that lymphocytes rely on the Tgfbr1/Tgfbr2 complex as suggested by the classical model, whereas HSCs require the unique Tgfbr2/Tgfbr3/Eng complex to orchestrate TGF-β signaling. Collectively, this study reveals Tgfbr3 as a critical regulator in the maintenance of HSC self-renewal potential and suggests a novel TGF-β receptor complex specific to HSCs.

The gut microbiota and its metabolites change dynamically with age, but their trajectories and links to healthy aging are unclear, hindering the development of related interventions.

Human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) exhibits exceptional sensitivity to chemoradiotherapy, with 5-year overall survival exceeding 80%, thereby providing a compelling rationale for treatment de-escalation. However, 15%-20% of patients develop therapeutic resistance and locoregional recurrence, resulting in markedly inferior outcomes. This Review systematically dissects the molecular architecture of treatment resistance in HPV-positive OPSCC, with particular emphasis on mechanisms conferring resistance to radiotherapy, platinum-based chemotherapy, and EGFR-targeted therapy (cetuximab). We delineate core drivers - aberrant DNA damage response (DDR) signaling, epigenetic reprogramming, cancer stem cell (CSC) plasticity, and non-coding RNA networks-and pinpoint actionable vulnerabilities. The contributions and limitations of patient-derived organoids, genetically engineered mouse models, and pivotal clinical trials (notably RTOG 1016) are critically appraised. We address persistent controversies, including the complex relationship between de-escalation and resistance, and highlight the lack of integrative resistance biomarkers. Finally, we propose a roadmap to bridge translational gaps and accelerate the development of precision strategies that circumvent resistance.

Ovarian cancer remains a leading cause of gynecologic cancer death worldwide, largely due to late diagnosis, frequent recurrence, and metastatic tendencies. Thrombosis is a common and life-threatening complication in these patients, contributing to poor prognosis and therapy resistance. Emerging evidence highlights a mechanistic link between tumor-derived extracellular vesicles and thrombotic events - particularly podoplanin-positive small extracellular vesicles (PDPN⁺ sEVs). Secreted by PDPN-expressing ovarian tumor cells, these vesicles act as biologically active messengers that circulate systemically. A central mechanism involves the binding of PDPN⁺ sEVs to C-type lectin-like receptor 2 (CLEC-2) on platelets, inducing platelet activation, aggregation, and the release of pro-inflammatory mediators. This interaction creates a hypercoagulable and pro-inflammatory microenvironment. Beyond coagulation, PDPN⁺ sEVs promote cancer aggressiveness by enhancing cancer stem cell plasticity, driving epithelial-to-mesenchymal transition, and facilitating immune evasion - hallmarks of metastasis and chemoresistance. This dual activity establishes a thromboinflammatory tumor niche that accelerates disease progression while undermining treatment efficacy. The ability of PDPN⁺ sEVs to circulate in ascitic fluid and peripheral blood positions them as promising candidates for liquid biopsy-based diagnostics. Furthermore, targeting the PDPN-CLEC‑2 axis or disrupting sEV biogenesis offers a novel therapeutic strategy to curb both thrombosis and metastatic spread. In conclusion, PDPN⁺ sEVs represent a critical molecular link between coagulation and cancer progression, offering valuable diagnostic, prognostic, and therapeutic potential for improving outcomes in ovarian cancer.