Chaiyaboot Ariyachet

Fibrosis is a progressive pathological process characterized by excessive extracellular matrix (ECM) deposition and structural remodeling that ultimately leads to organ dysfunction. In organs such as the liver, lung, heart and kidney, sustained activation of fibroblasts and their differentiation into myofibroblasts are key drivers of fibrotic progression. Growing evidence suggests that these cellular transitions are regulated by intricate transcriptional networks that integrate inflammatory, metabolic and mechanical signals. Liver fibrosis provides a well‑established framework for studying the transcriptional regulation of fibroblast activation, largely driven by the differentiation of hepatic stellate cells (HSCs) into collagen‑secreting myofibroblasts. Various transcription factors coordinate major signaling pathways to regulate fibroblast proliferation, ECM production and cell survival. These transcriptional programs not only sustain fibrogenesis but also influence whether fibrotic responses resolve or progress to chronic tissue scarring. In the present review, current advances in understanding transcriptional regulatory networks governing fibroblast activation were summarized, with a primary focus on HSCs while highlighting shared mechanisms across multiple fibrotic organs. Emerging therapeutic strategies targeting transcription factors and their upstream regulatory pathways were further discussed. A deeper understanding of these transcriptional circuits may facilitate the development of novel antifibrotic therapies and enhance strategies for resolving fibrosis in various organs.

Organ fibrosis represents a terminal pathological consequence of chronic tissue injury and contributes substantially to global morbidity and mortality, yet effective therapeutic options remain limited. Characterized by the persistent activation of fibroblasts into myofibroblasts, fibrosis results in excessive extracellular matrix (ECM) deposition and progressive disruption of normal organ architecture, ultimately leading to functional failure. Increasing evidence places zinc finger proteins (ZNFs) as key mediators within the regulatory hierarchy of fibrotic diseases. As the largest and most diverse family of transcriptional regulators in the human genome, ZNFs utilize specialized zinc‐coordinating motifs to regulate gene expression, protein stability, and intracellular signaling pathways. Beyond their classical roles in DNA binding and transcriptional control, ZNFs are now recognized as key orchestrators of fibrotic programs through regulation of ubiquitin‐mediated protein turnover, cytoskeletal dynamics, and core pro‐fibrotic signaling pathways, including transforming growth factor‐β/Smad, mitogen‐activated protein kinase, and phosphoinositide 3‐kinase/protein kinase B cascades. Dysregulation of specific ZNFs has emerged as a critical driver of myofibroblast differentiation, ECM synthesis, and pathological tissue remodeling across multiple organ systems, including the liver, lung, kidney, skin, and heart. This review provides a comprehensive synthesis of the molecular roles of ZNFs in organ‐specific fibrosis, integrating their structural classification with their mechanistic influence on key signaling networks. Furthermore, we discuss the clinical potential of ZNFs as biomarkers of fibrotic progression and evaluate emerging therapeutic strategies aimed at targeting ZNF‐mediated pathways. Finally, we outline key knowledge gaps and future research directions. By consolidating recent advances, this review highlights ZNFs as promising molecular targets for the next generation of anti‐fibrotic interventions.

Liver fibrosis is characterized by excessive collagen accumulation, the principal component of the extracellular matrix (ECM), produced by activated hepatic stellate cells (HSCs). While the catabolic role of mannose receptor C‐type 2 (MRC2) in binding and degradation of collagen is established, its role in regulating pro‐fibrotic signaling remains undefined. We investigated the novel, non‐canonical signaling function of the collagen receptor MRC2 as a regulator of HSC activation. Using doxycycline‐inducible knockdown and overexpression systems in human HSCs, we show that MRC2 deficiency enhances activation phenotypes, including proliferation, migration, contraction, and collagen overproduction. Conversely, MRC2 overexpression suppressed these effects. Transcriptomic analysis of MRC2 knockdown cells revealed a pro‐fibrotic transcriptional shift, with enrichment of pathways related to ECM organization, collagen biosynthesis, and the PI3K/AKT signaling axis. Mechanistically, MRC2 depletion induced transcriptional upregulation of ECM genes and components of the integrin/PI3K/AKT axis, accompanied by increased AKT phosphorylation. Pharmacological inhibition of either PI3K or integrins reversed ECM gene expression and collagen phenotypes induced by MRC2 depletion, establishing these pathways as required mediators. Our findings identify a novel, non‐canonical signaling function of MRC2 as a negative regulator of HSC activation.

Liver fibrosis, the precursor to cirrhosis and hepatocellular carcinoma (HCC), represents a global health crisis with millions affected and few effective treatments available. This progressive pathology is characterized by excessive extracellular matrix (ECM) deposition, primarily driven by the activation of hepatic stellate cells (HSCs) into profibrogenic myofibroblasts. Elucidating the molecular control of HSC activation is paramount for therapeutic development. Protein-tyrosine phosphatase 1B (PTP1B), an established target in metabolic diseases, has emerging pro-fibrotic roles, as evidenced by protection against liver fibrosis in PTP1B-deficient mice. However, the regulatory function of PTP1B in human-relevant models remains poorly understood. Here, we define the role and mechanistic significance of PTP1B in primary human HSCs and investigate its clinical relevance in human fibrotic liver. Using loss- and gain-of-function approaches in primary human HSCs, we demonstrate that PTP1B significantly promotes HSC activation, evidenced by enhanced proliferation, migration, and increased expression of key ECM genes and collagen production. Mechanistically, PTP1B acts as a positive regulator of the pro-fibrotic SRC kinase, directly dephosphorylating the inhibitory tyrosine 527 (Y527) residue to induce its activation. Pharmacological inhibition of SRC effectively reversed the PTP1B-driven pro-fibrotic phenotypes. Consistent with our findings, re-analysis of human fibrotic liver tissues revealed that PTP1B and SRC expressions are significantly upregulated and positively correlated with ECM genes. Collectively, these findings establish a novel PTP1B-SRC signaling axis that critically drives human HSC activation and hepatic fibrogenesis, positioning PTP1B as a high-potential therapeutic target for liver fibrosis.

Pulmonary fibrosis is characterized by persistent activation of lung fibroblasts and excessive extracellular matrix (ECM) deposition; however, the transcriptional mechanisms sustaining this activated state remain poorly understood. Given the established profibrotic role of zinc finger protein 469 (ZNF469) in other mesenchymal lineages, we investigated its function in lung fibroblasts using an integrated functional and transcriptomic approach. We found that ZNF469 depletion attenuated key features of fibroblast activation, including proliferation, migration, and contractile capacity, concomitantly reducing collagen production and deposition. Mechanistically, combined transcriptomic profiling and Cleavage Under Targets and Release Using Nuclease (CUT&RUN) assays revealed that ZNF469 modulates ECM-associated transcriptional regulation, demonstrating preferential occupancy at the promoters of key collagen genes, such as collagen type I alpha 1 chain (COL1A1) and collagen type III alpha 1 chain (COL3A1). Furthermore, we demonstrated that transforming growth factor-β1 (TGF-β1) stimulated ZNF469 expression in a SMAD family member 3 (SMAD3)-dependent manner, with CUT&RUN further revealing SMAD3 occupancy at the ZNF469 promoter. Importantly, ZNF469 depletion blunted TGF-β1-induced collagen production, indicating that ZNF469 contributes to the profibrotic effects of the TGF-β1/SMAD3 axis. Finally, we underscored the clinical relevance of ZNF469 by analyzing bulk and single-cell RNA sequencing data from patients with idiopathic pulmonary fibrosis and systemic sclerosis. ZNF469 was found to be upregulated in fibrotic lung tissues, particularly within the activated fibroblast lineage, and correlated with key ECM gene signatures. Our findings collectively support a model wherein the TGF-β1/SMAD3/ZNF469 axis contributes significantly to ECM gene regulation and fibroblast activation, thereby positioning ZNF469 as a critical profibrotic factor and a promising therapeutic target in pulmonary fibrosis.

Adipogenesis is a critical factor in causing obesity, which is a global health problem associated with metabolic disorders, such as insulin resistance and cardiovascular diseases. Natural compounds with anti-adipogenic activity may represent potential approaches for modulating adipocyte function. However, despite increasing interest in natural products, the anti-adipogenic potential of acridone alkaloids, particularly prenylated derivatives, remains largely unexplored. This study examined the effects of N-methylatalaphylline (NMA), a prenylated acridone alkaloid, on adipocyte differentiation, lipid accumulation, and glucose uptake. NMA exhibited anti-adipogenesis, particularly toward preadipocytes, and significantly reduced lipid accumulation in murine 3T3-L1 and human PCS-210-010 adipocytes at nontoxic doses (1.5–6 µM). At 3–6 µM, NMA downregulated adipogenic regulators, including PPARγ, C/EBPα, and SREBP1, along with adipogenic effectors, such as FABP4, adiponectin, LPL, PLIN1, and FAS. Mechanistic studies indicated that NMA treatment was associated with reduced phosphorylation of AKT, ERK, and p38, accompanied by cell cycle arrest and inhibition of mitotic clonal expansion. Meanwhile, activation of AMPK-ACC signaling, which may contribute to suppression of adipogenesis and reduced glucose uptake, was observed in differentiated 3T3-L1 cells after treatment with 6 µM NMA for 48 h. Additionally, molecular docking and molecular dynamics simulations suggested potential interaction between NMA and ERK1, supported by hydrogen bonding and hydrophobic contacts. Overall, these findings suggest that NMA exerts anti-adipogenic effects in vitro by modulating adipocyte proliferation, differentiation, and lipid metabolism. These findings highlight NMA as a promising acridone alkaloid scaffold for anti-adiposity applications, warranting further in vivo validation.

Fish offer an excellent source of high-quality protein with balanced nutrients and low fat content. However, the increasing global demand for food and the impacts of climate change have led to a significant decline in wild fish stocks. Cultivated fish meat has therefore emerged as a promising sustainable food alternative. In this review, we summarize the structural and physiological characteristics of fish muscle and highlight the methods used to establish primary muscle cell cultures, including explant outgrowth and enzymatic dissociation, alongside the optimization of environmental conditions and growth media composition. Particular attention is given to the isolation, development, and characterization of Cyprinidae muscle-derived cell lines via morphological assessments, gene expression profiling, and karyotyping. In addition, we discuss recent advances in scaffold-based and three-dimensional culture systems as well as the application of bioreactors for large-scale production. Current challenges include the limited availability of standardized muscle cell lines, dependence on serum-containing media, and the high cost of growth factors. Future progress will depend on innovations in serum-free formulations, cost-effective media, and reproducible culture protocols. Ultimately, fish muscle cell culture supports not only the development of alternative protein sources but also aquatic health research, disease modeling, and sustainable bioproduction.

Hepatitis B virus (HBV) persists in infected hepatocytes through covalently closed circular DNA (cccDNA), a stable episomal form that serves as the transcriptional template for viral replication. Accurate and sensitive quantification of intrahepatic cccDNA is crucial for evaluating antiviral therapies, particularly those targeting a functional cure. Here, we report the development of a novel, cccDNA-specific detection system combining recombinase polymerase amplification (RPA) with CRISPR/Cas12a-based fluorescence detection. We designed and validated CRISPR RNAs (crRNAs) targeting HBV cccDNA-specific regions conserved across genotypes A–D. Reaction conditions for both RPA and Cas12a detection were optimized to enhance sensitivity, specificity, and accuracy. The system reliably detected as few as 10 copies of cccDNA-containing plasmid per reaction and showed no cross-reactivity with non-cccDNA forms in serum or plasma, indicating assay specificity. When applied to liver tissue samples from 10 HBV-infected and 6 non-HBV patients, the RPA-CRISPR/Cas12a assay exhibited a high sensitivity (90%) and a strong correlation with qPCR results (R² = 0.9155), confirming its accuracy. In the conclusion, the RPA-CRISPR/Cas12a system provides a robust, cost-effective, and scalable platform for sensitive and specific quantification of intrahepatic HBV cccDNA. This method holds promises for research and high-throughput therapeutic screening applications targeting cccDNA clearance.