Hepatocellular carcinoma (HCC) is an aggressive cancer with poor prognosis and high mortality rates. The increasing mortality associated with HCC highlights the urgent need for alternative therapeutic approaches. Natural products and their structural analogs have shown promise as novel treatment options. This study aimed to evaluate the potential therapeutic effects of the natural flavonoids apigenin and chrysin, administered individually and in combination, in an experimental HCC model. Forty-eight rats were divided into five groups: Control, DEN, DEN + Apigenin, DEN + Chrysin, and DEN + Combination. HCC was induced by intraperitoneal administration of diethylnitrosamine (DEN, 150 mg/kg), followed by oral 2-acetylaminofluorene (2-AAF, 20 mg/kg) according to a standardized protocol. Subsequently, Apigenin and chrysin were administered orally at 50 mg/kg for 8 weeks following tumor induction. At the end of the treatment, biochemical and histopathological analyses were performed on blood and liver tissue samples. Compared to controls, DEN-induced HCC was associated with activation of tumor-promoting signaling pathways, as evidenced by increased levels of survivin, annexin-V, smoothened, sonic hedgehog, jagged1, and notch1, alongside reduced caspase-3 expression. Histological examination revealed normal liver architecture in controls, while DEN-treated rats showed varying degrees of fibrosis (62.5% moderate to severe), inflammatory lesions, regenerative nodules, and 87.5% moderate to severe inflammation. Treatment groups displayed marked reductions in fibrosis and inflammation, with the most pronounced effects observed in DEN + Apigenin and combination groups. These findings suggest that apigenin, chrysin, and their combination may exert antitumor effects in experimental HCC, with apigenin demonstrating comparatively more pronounced activity based on biochemical and histopathological findings.

The salmon trout or rainbow trout (Oncorhynchus mykiss) is a teleost fish are characterized by the presence of lingual teeth. The objective of this study is to investigate their development, particularly in relation to the expression of genes associated with mammalian odontogenesis. Sixty fry were reared in a freshwater aquarium at 7°C. Specimens at developmental stages ranging from D0 (hatching) to D35 were collected and subjected to histological (Masson's trichrome) and immunohistochemical analysis, including the expression of proteins associated with the Pitx2, Shh, MSX1, Pax9, Bmp4 and β-catenin genes. The developmental patterns of the lingual teeth are similar to those of the maxillary and mandibular teeth. These are not giant papillae or pseudoteeth. The initiator gene Pitx2 is expressed in the epithelium during early stages but subsequently diminishes. Bmp4, Shh and β-Cat exhibit a pattern similar to that of mammals, except for the absence of the enamel node. Surprisingly, however, we observe expression of Msx1 and Pax9 in the dental mesenchyme as well as in the epithelium, a phenomenon unknown in mammals. The second week marks an increase in the expression of these two proteins and of Shh, while Bmp4 does not appear in large quantities until 10 days. Incidentally, the mucous glands (cement glands) express Shh, Pitx2, Msx1, Pax9 and β-catenin. In conclusion, tongue teeth are 'classical' teeth; their gene expression patterns are similar to those in mammals, except that Msx1 and Pax9 are expressed in both epithelial and mesenchymal tissues in this species.

GSK3β functions as a mechanosensitive regulator that links flow stress to Shh signaling.

Endometrioid endometrial cancer (EEC) is the most prevalent gynecological malignancy. While early-stage EEC typically carries a favorable prognosis, late-stage EEC shows significantly poorer outcomes, with a five-year survival rate of only 17-19%. Co-occurring PTEN and CTNNB1 variants are frequent in EEC and linked to poor prognosis. To investigate their functional impact, we generated uterine-specific Pten knockout mice and dominantly stabilized Ctnnb1 mutant mice (Pgrcre/+Ptenf/fCtnnb1f(ex3)/+; Ptend/dCtnnb1f(ex3)/+). These double-mutant mice exhibited significantly reduced survival compared to single Pten knockout (Pgrcre/+Ptenf/f; Ptend/d) mice. Histopathology revealed aggressive, metastatic endometrioid cancer in Ptend/dCtnnb1f(ex3)/+ mice; these mice developed myometrial invasion by four weeks of age - unlike Ptend/d mice. Transcriptomic analysis identified activation of multiple oncogenic pathways, including WNT/CTNNB1, PI3K/AKT, basal cell carcinoma, sonic hedgehog signaling and epithelial-mesenchymal transition (EMT). Immunohistochemistry confirmed hallmark features of EMT in the uterus of double-mutant mice, including strong downregulation of E-cadherin (CDH1) and upregulation of the EMT regulator SNAIL (Snai1). These findings demonstrate that synergistic mutations of PTEN and CTNNB1 promote early invasion, EMT activation, and metastatic progression, offering mechanistic insight into the aggressive behavior and poor clinical outcomes associated with this subtype of EEC.

Peer reviewed

Androgenetic alopecia (AGA) imposes a significant psychosocial burden, yet current treatments such as minoxidil and finasteride often yield suboptimal responses or adverse effects. Exosomes, nanoscale extracellular vesicles derived from mesenchymal stem cells and dermal papilla cells (DPCs), offer a promising regenerative alternative by modulating key hair-growth pathways. This systematic review evaluates the efficacy, mechanisms, and translational challenges of exosome-based therapies for hair restoration in AGA. A comprehensive search was conducted across Google Scholar, Embase, PubMed, Scopus, and Web of Science for studies published from 2019 to 2025. The search strategy prioritized AGA-related terminology, including androgenetic alopecia, male pattern hair loss, female pattern hair loss, baldness, exosomes, extracellular vesicles, and hair regrowth. Following duplicate removal, 39 studies meeting Population, Intervention, Comparison, Outcome, and Study design criteria were included for qualitative synthesis. Preclinical data demonstrate that exosomes promote hair regeneration through multiple synergistic mechanisms: activation of the Wnt/beta-catenin and Sonic Hedgehog pathways, suppression of transforming growth factor beta (TGF-beta)/SMAD3 signaling, delivery of anti-inflammatory cytokines (e.g., IL-10), and rejuvenation of senescent DPCs via microRNA cargo (e.g., miR-122-5p). Early clinical studies report improvements in hair density (8-20%) and shaft thickness; however, the evidence base remains limited by small sample sizes, retrospective designs, lack of control groups, and inconsistent outcome measures. Emerging delivery systems, such as thermoresponsive hydrogels and microneedle patches, show promise in enhancing follicular penetration but require further validation. Exosome therapy represents a multi-target regenerative approach for AGA with a favorable preliminary safety profile. However, widespread clinical adoption is hindered by critical gaps in manufacturing standardization, scalability, regulatory frameworks, and robust long-term efficacy data. Future research must prioritize large-scale randomized controlled trials with standardized endpoints and Good Manufacturing Practice (GMP)-compliant production protocols to validate these findings and establish exosomes as a mainstream therapeutic option for AGA.

Cytogenomics, including karyotyping, FISH, chromosomal microarrays, and optical genome mapping, has yielded significant results for clinical phenotypes in constitutional and cancer genetics, including intellectual disability, autism spectrum disorders, dysmorphic features, and hematological and solid-tissue neoplasia. However, some of these assays have yielded results of unclear significance because the abnormalities detected were often located in intergenic regions of the genome. Because these abnormalities are within the “dark matter” of the genome, their clinical significance has been a matter of speculation. However, functional genomics can explore the clinical implications of such abnormalities more robustly, whether the abnormalities disrupt topologically associating domains (TADs), delete regulatory regions, etc. Some human genetic diseases associated with these intergenic abnormalities and characterized by functional genomics include preaxial polydactyly (SHH gene), Pierre Robin syndrome (SOX9), and 5q14.3 microdeletion syndrome (MEF2C). While functional genomics is a broad research topic, this review focuses on prior and current efforts to leverage functional genomics within the intergenic regions for human health.

Multi-omics characterization of radiation-induced cerebellar remodeling and tumorigenic transcriptional programs.

Aberrant activation of the Sonic Hedgehog (SHH)-glioma-associated oncogene 1 (GLI1) signaling pathway is a determinant driver of basal cell carcinoma (BCC) and SHH-subgroup medulloblastoma (MBSHH). Here, we demonstrate the role of S-palmitoylation and depalmitoylation of GLI1 in the regulation of incidence and progression of MBSHH. High expression of acyltransferase ZDHHC13 in MBSHH induces S-palmitoylation of human GLI1 at Cys1034, stabilizes GLI1 by preventing its ubiquitin-dependent proteasomal degradation, and in turn activates this signaling pathway to promote the progression of MBSHH. Conversely, the deacylase ABHD17A depalmitoylates GLI1 at Cys1034, destabilizes GLI1, and in turn inactivates this signaling pathway. As a result, conditional knockout of Zdhhc13 in cerebellar granule neuron precursors (GNPs) significantly alleviates the severity of MBSHH induced by constitutively active Smoothened (Smo-M2) overexpression and markedly extends overall survival, whereas knockout of Abhd17a increases the incidence of MBSHH induced by Patched-1 ablation, rather than its progression. Together, these findings uncover the dynamic regulation of GLI1 palmitoylation and depalmitoylation by the ZDHHC13 and ABHD17A and subsequent GLI1 stabilization and destabilization as a hitherto uncharacterized mechanism controlling SHH/GLI1 signaling and may provide additional targets for therapeutic intervention of MBSHH.

RNF138 polyubiquitinates glioma-associated oncogene 1 on K815/K942 to suppress Sonic Hedgehog signaling in medulloblastoma.

Efficacy of Tinospora cordifolia bioactives as agonists of Smoothened (Smo) receptor to promote oligodendroglial lineage induction for remyelination-based therapy.

Introduction Diquat-induced toxic encephalopathy remains inadequately understood. Methods Data on patients with acute diquat poisoning were retrospectively collected from May 2017 to May 2023. The diagnosis was confirmed qualitatively by ultra-high performance liquid chromatography quadrupole-time of flight mass spectrometry. A follow-up study was designed to explore the long-term health effects of diquat poisoning. Results A total of 129 patients with acute diquat poisoning were included in this study. Thirty-four patients died (27 of whom developed toxic encephalopathy) following a median dose of diquat (200 g/L) 190 mL (IQR:100–200 mL). Forty-five patients were diagnosed with toxic encephalopathy, mainly involving the brainstem, bilateral cerebellar peduncles, and basal ganglia. The development of toxic encephalopathy was associated with a significantly higher risk of death (odds ratio = 16.5; 95%CI: 6.2–43.8; P <0.001). Twenty-five of the 95 survivors were enrolled in a follow-up study. The main persistent features were peripheral nerve injuries in 20%, suspected peripheral nerve injuries in 40%, and reduced compound muscle action potential in 84%; chronic-phase encephalopathy occurred in 28%. Transcriptomic analysis revealed significant changes in the expression profiles of neuroregulatory factors, including inhibin-βA and sonic hedgehog. Discussion Toxic encephalopathy represented an important organ injury caused by acute diquat poisoning, which was strongly associated with an increased risk of death. Peripheral nerve injuries were also common at follow-up. Several limitations should be noted, including the lack of diquat concentrations, the small sample size, the short follow-up duration, and the single-center design. Conclusions Thirty-four of 129 (26.4%) patients died from diquat poisoning. Toxic encephalopathy was diagnosed in 45 of 129 (34.9%) patients, and its development was associated with a significantly higher risk of death. Follow-up revealed peripheral nerve injuries in 20% of patients, and chronic-phase encephalopathy in 28%.

Gorlin-Goltz syndrome, also known as nevoid basal cell carcinoma syndrome, is a hereditary disorder characterized by a predisposition to the development of multiple neoplasms and developmental anomalies. While rare in our clinical setting, it is crucial to recognize this entity for accurate early diagnosis, multidisciplinary follow-up, and effective treatment, given the high recurrence rate of its lesions. The principal clinical manifestations include basal cell carcinomas, palmoplantar pits, and odontogenic cysts. Diagnosis is based on specific clinical criteria and confirmed through analysis of pathogenic variants in the PTCH1 or SUFU genes, which are components of the Sonic Hedgehog molecular pathway. This study presents a series of 10 patients meeting the clinical criteria for Gorlin syndrome, who underwent genetic testing to confirm the diagnosis. The frequency and therapeutic management of the main dermatological manifestations are analyzed, and an algorithm is proposed for managing patients diagnosed with Gorlin syndrome from dermatology or maxillofacial surgery consultations.

Deregulation of the Sonic Hedgehog (SHH)/GLI signaling pathway has been strongly implicated in carcinogenesis, particularly in brain and skin tumors, through mutations affecting various pathway components. Given its central role in tumor development, the SHH/GLI axis has emerged as a key target for therapeutic intervention, underscoring the need to identify GLI-specific targets in cancer cells. Netrin-1, a multifunctional signaling molecule with pro-oncogenic properties, is overexpressed in diverse tumor types. While its tumor-promoting functions are well documented, the regulatory mechanisms governing its expression remain poorly understood. Here, we provide through gene expression profiling and in vitro and ex vivo analyses evidence demonstrating that Netrin-1 (NTN1) is a direct transcriptional target of canonical SHH signaling. We further show that in conditions of SHH pathway hyperactivation-such as in SHH-driven medulloblastoma (MB) NTN1 is significantly upregulated. This overexpression promotes the self-renewal of MB stem-like cells through the modulation of ERK signaling, thereby contributing to enhanced cancer stemness. Our findings identify NTN1 as a novel downstream effector of the SHH/GLI pathway and demonstrate that quantifying NTN1 levels can refine molecular classification of SHH-subtype medulloblastomas-distinguishing tumors with high SHH pathway activation and enhanced stem-like properties, with potential implications for prognosis and targeted therapy selection.

Astrocyte-mediated angiogenesis in CNS diseases: Mechanisms and therapeutic implications.

The dentition of mice comprises only two tooth types, molars and incisors. Despite similar epithelial-mesenchymal signalling interactions during development, their final morphologies differ dramatically. Both molars and incisors grow from a similarly symmetric epithelial organ bud starting at embryonic (E) day 12.5. By E13.5, they develop outgrowths known as cervical loops (CLs), which grow symmetrically in the molar but highly asymmetrically in the incisor. Prior work has shown epithelial Shh is required for the symmetric growth of molar CLs, but the cellular basis of incisor asymmetry is not known. Here, we report that differential proliferation in the epithelium and asymmetric distal Shh expression at E12.5 underlie asymmetric incisor morphogenesis. Inhibiting proliferation or Shh expression at this stage ex vivo resulted in a loss of asymmetry. Furthermore, symmetrizing the source of Shh at this stage by adding ectopic SHH protein ex vivo or expressing ectopic Shh in vivo similarly caused a loss of asymmetry. Together, these results demonstrate that spatial asymmetry of differential proliferation dependent on Shh expression plays a crucial role in the developing murine incisor.

The copy-number events in skull base chordoma stratify tumours into four biologically coherent groups

Hedgehog (HH) signaling in vertebrates is dependent on the primary cilium, an organelle that scaffolds signal transduction. HH signals induce ciliary enrichment of Smoothened (SMO) and ciliary departure of the G protein-coupled receptor (GPCR) GPR161 to trigger GLI activation of the HH transcriptional program. Recently, SMO has been shown to inhibit protein kinase A (PKA). To test the hypothesis that SMO inhibits PKA at cilia to activate the HH signal transduction pathway, we developed a ciliary PKA reporter. Ciliary PKA activity was graded during zebrafish development. Activation of the HH signal transduction pathway by either Sonic hedgehog (SHH) or SMO agonist (SAG) inhibited ciliary PKA activity. Blocking SMO phosphorylation by GRK2/3 prevented ciliary SMO from inhibiting ciliary PKA activity. Converting the SMO C-terminal PKA pseudosubstrate site into a consensus PKA substrate blocked SMO-mediated inhibition of ciliary PKA activity. A ciliary GPCR, SSTR3, activated ciliary PKA and induced HH transcriptional responses in NIH/3T3 cells via a different mechanism: activation of Gαi/o. A different ciliary GPCR, GPR161, possesses an A-Kinase Anchoring Protein (AKAP), which we found was critical for the ciliary localization of the catalytic subunit of PKA (PKA-C) to promote ciliary PKA activity. We propose that HH signal transduction is inhibited by GPR161-mediated ciliary enrichment of PKA-C, and activated by GRK2/3-phosphorylated SMO inhibition of ciliary PKA activity.

Current treatment strategies for medulloblastoma remain ineffective owing to extensive tumor heterogeneity. We generated five platforms of omics data including liquid chromatography and mass spectrometry-based proteome and performed integrated multi-omic characterization to improve the conventional molecular classification of medulloblastoma. We identified seven refined distinct subtypes. The sonic hedgehog (SHH) group was reclassified into two subgroups, SHHα and SHHβ, whereas group 4 was divided into three subgroups, G4α, G4β, and G4γ. SHH and group 4 subtypes exhibit two distinct neuronal differentiation trajectories: granular neuron and unipolar brush cell differentiation (SHHβ and G4γ, respectively), both of which associated with more favorable clinical outcomes. Furthermore, we uncovered unique proteomic and kinomic properties that conferred increased treatment vulnerabilities to targeted therapeutic interventions against each of the three medulloblastoma subtypes associated with poor clinical outcomes. We demonstrated the therapeutic potential of exploiting these vulnerabilities by utilizing a proteasome inhibitor and subtype-specific agents, including CDK1/2, PARP, CLK1, and MET inhibitors. Mechanistic insights were further elucidated through in-depth proteome analyses. Our study qualifies the use of proteomic signatures and activation of neuronal differentiation trajectories to tailor selective therapeutic opportunities for distinct subgroups of patients with medulloblastoma.

The evolutionarily conserved RNA exosome complex modulates gene expression during development. Mutations in RNA exosome complex subunits have been implicated in various human brain disorders, suggesting that defects in RNA decay are linked to impaired neural development. In our study, we identified de novo variants of EXOSC10 in microcephalic individuals. The patient's phenotype can be replicated by heterozygous conditional knockout of Exosc10 in the developing mouse forebrain. The heterozygous loss of Exosc10 in the developing mouse cortex leads to increased cell cycle exit, with a premature differentiation of radial glial cells to intermediate progenitors and neurons in the embryonic cortex. This premature neurogenesis at the expense of neural stem cell proliferation coincides with a smaller cortical size. RNA sequencing and RNA immunoprecipitation sequencing revealed upregulation of many Sonic hedgehog (Shh) signalling genes. We further show the direct degradation of Shh pathway transcripts such as Scube1 and Scube3 by Exosc10. In Exosc10 mouse mutants, the reduced cortical size could be largely rescued by reducing Shh activity. We propose that increased Shh activity due to Exosc10 deficiency leads to premature neurogenesis and ultimately to microcephaly. These observations offer new insights into the neurodevelopmental role of Exosc10 and highlight the dosage-dependent regulation of Shh signalling by Exosc10 in cortical development.