Inadequate response to tumor necrosis factor inhibitors (TNFi) in ankylosing spondylitis (AS) remains common. As the immunoregulatory and immunometabolic alterations across TNFi inadequate responder (TNFi-IR) subsets remain unexplored, we characterized molecular profiles across distinct immune subsets in TNFi-IRs and identified signatures linked to TNFi-IR. We compared subset-specific expression in 16 TNFi-IRs and 13 TNFi responders (TNFi-Rs). Gene expression demonstrated fair-good discriminative power (area under the curve: 0.70-0.87). TNFi-IRs displayed distinct transcriptomic profiles and aberrant transcription factor activity amplifying inflammatory responses across immune subsets. Pathway analysis revealed synergistic association between immune subsets and inadequate response, with recurrent alterations in mitochondrial respiration accompanied by metabolic reprogramming exacerbating immune activation. Altered cell-cell communication hinted at impaired IL-10 and IL-19 regulatory signaling. We identified candidates with repurposing potential for TNFi-IRs. These findings reveal crucial therapeutic targets spanning genes, transcription factors, pathways, metabolites, and cell-cell communication signaling for bench-to-bedside investigation.

Background: The gut virome is increasingly recognized as a crucial regulator of host immunity, yet its specific contribution to systemic autoimmune diseases remains obscure. We aimed to delineate the trans-kingdom crosstalk linking gut viral dysbiosis to the pathogenesis of autoimmune uveitis in Behçet’s disease (BD). Methods: We performed metagenomic virome sequencing and parallel 16S rRNA gene sequencing on 59 patients with Behçet’s disease and 51 healthy controls. To investigate the downstream immunometabolic mechanisms, we utilized Cd4Cre Ldhafl/fl mice alongside in vitro multi-omics and epigenetic approaches to trace the impact of virome-derived metabolic signals on CD4+ T cell polarization. Findings: Patients with Behçet’s uveitis exhibited a specific depletion of Escherichia-targeting phages, notably Straboviridae and Drexlerviridae. This viral dysbiosis triggered a systemic accumulation of lactate. We discovered that this systemic lactate acts as a metabolic signal inducing p300-mediated Creb1 lactylation at lysine 325 in CD4+ T cells. Intrinsic LDHA activity served as an essential metabolic checkpoint for this modification, while SIRT1 was identified as the specific delactylase. Epigenetically, Creb1 lactylation remodeled chromatin accessibility to directly activate Rorα transcription, thereby enforcing a pathogenic Th17 program. Interpretation: Our findings establish that gut viral perturbations orchestrate T cell fate via a systemic metabolic-epigenetic relay. Targeting this pathway to restore viral-metabolic homeostasis represents a promising therapeutic strategy for Behçet’s disease and potentially other systemic autoimmune conditions.

Plasmid dosages used for HCC murine models

Histology analysis of liver tissues from Akt/NRas and sgGSK3α/β/NRas mice

Gene expression pattern in Akt/NRas and Akt/NRas/pT3-FoxoAAA liver tumors

Pathways commonly upregulated and co-downregulated in Akt/NRas, Tsc2-/-/NRas, and sgGSK3α/β/NRas liver tumors

Gene expression pattern in Akt/NRas and Tsc2-/-/NRas/sgGSK3α/β liver tumors

Background Drug resistance remains a major challenge in multiple myeloma (MM). This study explores the role of the oncogenic long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in MM pathogenesis and lenalidomide resistance. Methods MALAT1 expression and function were examined in MM cell lines and patient-derived samples using antisense oligonucleotide (ASO) knockdown. Single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing (RNA-seq) profiled transcriptional changes. Mitochondrial membrane potential and reactive oxygen species (ROS) were measured by fluorescence microscopy. Cellular adenosine triphosphate (ATP) and oxidative phosphorylation (OXPHOS) activity were quantified by colorimetric and luminescence assays. Cytokine profiling of patient bone marrow mononuclear cells (BMMCs) was performed with a 48-plex MILLIPLEX assay. Functional assays were performed in primary patient samples and lenalidomide-resistant MM cell lines, and combination therapy experiments were conducted in resistant MM cell lines. Results MALAT1 was significantly overexpressed in lenalidomide-resistant MM cells and positively correlated with CD38 expression. scRNA-seq analysis revealed elevated CD38 expression and increased OXPHOS activity in relapsed MM plasma cells. Analysis of the MMRF CoMMpass dataset confirmed a positive correlation between CD38 and MALAT1 expression. MALAT1 knockdown reduced CD38 via PRC2-mediated epigenetic repression and impaired mitochondrial function, leading to reduced OXPHOS activity, decreased ATP production, and increased ROS accumulation. These effects were confirmed in patient-derived BMMCs. MALAT1 inhibition reduced viability of drug-resistant MM cells and patient-derived BMMCs, and disrupted the tumor-supportive bone marrow microenvironment by lowering pro-tumorigenic and immunosuppressive cytokine secretion. Combining anti-MALAT1 ASO with lenalidomide, pomalidomide, or carfilzomib synergistically enhanced cytotoxicity in lenalidomide-resistant MM cells. Conclusions MALAT1 promotes drug resistance in MM through epigenetic regulation, metabolic reprogramming, and microenvironmental remodeling. Targeting MALAT1 with ASOs sensitizes MM cells to standard therapies and disrupts tumor-supportive signaling, highlighting its potential as a therapeutic target to help overcome drug resistance in MM.

Main characteristics of the plasmids used in the experiments

List of up-regulated genes in Akt/NRas and Tsc2-/-/NRas, but not in sgGSK3α/β/NRas liver tumors

List of gene expression patterns correlated with Foxm1 expression in human HCC samples

Diabetic foot ulcers (DFUs) are chronic complications of diabetes characterized by impaired healing due to poor angiogenesis and fibroblast dysfunction. This study aimed to evaluate a novel thermosensitive, conductive hybrid dressing composed of PEI-polypyrrole-Pluronic F127 loaded with recombinant human platelet-derived growth factor-BB (rhPDGF-BB) for enhancing diabetic wound healing. We developed a conductive PEI-polypyrrole-based hydrogel incorporating rhPDGF-BB. Physicochemical properties, rheology, and electrical conductivity were characterized. Biocompatibility and cell migration were evaluated using human dermal fibroblasts (CCD966SK) and epidermal keratinocytes (HaCaT). In vivo wound healing efficacy was evaluated in a diabetic mouse dorsal wound model. The hydrogel exhibited body-temperature-triggered gelation and promoted fibroblast proliferation. In vitro scratch assays showed the hybrid dressing significantly enhanced cell migration. In vivo diabetic wound model, the hybrid dressing significantly accelerated wound closure (p < 0.0001), achieving results comparable to Regranex gel while using only 1/200th of the rhPDGF-BB dose. Histological analysis revealed increased collagen deposition and angiogenesis in treated wounds. These findings highlight the innovative and efficient therapeutic potential of incorporating rhPDGF-BB into a polypyrrole-PF127-based hybrid conductive dressing. This approach not only accelerates diabetic wound healing, also addresses a critical gap in existing treatment strategies, offering a promising solution for chronic DFUs.

BACKGROUND:Vogt-Koyanagi-Harada (VKH) disease causes vitiligo, poliosis, and alopecia. OBJECTIVE:To investigate the effect of prednisone plus either adalimumab or cyclosporine-based immunosuppression on VKH dermatologic disorders using ancillary data from a VKH eye disease trial. METHODS:Patients with VKH disease treated with individualized prednisone tapering (maximum daily dose, 40 mg; maximum duration, 6 months) were randomized to adjunctive adalimumab (N = 54) or cyclosporine (N = 56). Outcomes included changes in vitiligo, poliosis, and alopecia at the sixth month. RESULTS:Overall, there was a decrease in the percentage of alopecia but no change in the presence of vitiligo or poliosis at the sixth month. The adalimumab group showed no nominally significant differences in the percentage changes of each dermatologic manifestation but a greater reduction in the number of affected dermatologic categories compared with the cyclosporine group. LIMITATIONS:This is a secondary analysis of a VKH eye disease trial. Six-month follow-up may not fully assess effects on vitiligo and poliosis. CONCLUSIONS:In conjunction with prednisone tapering, both adalimumab and cyclosporine similarily improved alopecia but had no obvious effect on the presence of vitiligo or poliosis at 6 months. Adalimumab was associated with a greater decrease in the number of affected dermatologic conditions compared with cyclosporine.

BACKGROUND:Vogt-Koyanagi-Harada (VKH) is a multisystemic autoimmune disorder characterized by bilateral panuveitis frequently accompanied by neurologic manifestations. While metabolic dysregulation is increasingly recognized in the context of autoimmune diseases, the role of specific metabolites in VKH disease remains unexplored. METHODS:Non-targeted and targeted metabolomics analysis, phospho-antibody array, proteome microarray, surface plasmon resonance, and molecular simulation were used to identify molecular target of OA. RESULTS:We investigated metabolic profile of VKH disease and found that oleic acid (OA) was enriched in this disease. A series of functional assays showed that OA could exacerbate experimental autoimmune uveitis (EAU) in association with increased frequency of Th1 and Th17 cells and decreased proportion of Treg cells in vitro. However, the specific molecular target of OA remains elusive. Through proteome microarrays, molecular simulations and surface plasmon resonance assays, Ornithine decarboxylase 1 (ODC1) was identified as target protein of OA. OA could bind to ODC1, increase ODC1 protein expression in both a time- and concentration-dependent manner and promote subsequently putrescine production. Phospho-antibody array analysis revealed that OA inhibited phosphorylation of STAT5A (Y694) in CD4+T cells, leading to imbalance of Th1/Th17 and Treg cells and decreased transcription of IL-10. OA upregulated ODC1 protein and putrescine levels through binding to LYS-78, inhibited phosphorylation of STAT5A protein and subsequently decreased binding of STAT5A at IL-10 promoter. CONCLUSION:These results reveals that OA could be a crucial metabolite for modulation of CD4+T cell differentiation and that ODC1-mediated phosphorylation and transcriptional activity of STAT5A contributes to development of VKH disease progression, highlighting ODC1 as a novel therapeutic target in VKH disease.

Gene list up- and down-regulated by Foxo1 in Akt/NRas/Foxo1AAA liver tumors

Basic information of HCC murine models

Pathways involved in Akt/NRas liver tumors, but not engaged in Tsc2-/-/NRas/sgGSK3α/β liver tumors

Background & aims: Hepatoblastoma (HB) is the predominant primary malignant liver tumor in childhood. Concomitant YAP and β-Catenin activation occurs in most HB. However, the signaling pathways distinctively regulated by YAP and β-Catenin protooncogenes in HB remain unexplored. Approach and results: We engineered an inducible HB murine model using hydrodynamic injection to deliver transposon plasmids encoding constitutive YAP and doxycycline (Dox)-inducible ΔN90-β-Catenin(YAP/TRE-β-Catenin). Gene expression patterns in mouse HB lesions upon short-term Dox withdrawal, i.e., tumors still existed, but ΔN90-β-Catenin was turned OFF, were analyzed. Mice rapidly developed aggressive HB lesions when fed doxycycline. However, upon Dox withdrawal, HB regressed, although tumors did not completely disappear over a long time. At the molecular level, YAP and β-Catenin were found to regulate distinct gene expression programs in HB. Specifically, YAP controls the Hippo and metabolism-related pathways, whereas β-Catenin modulates immune-related pathways contributing to immune exclusion in the tumor microenvironment. Furthermore, we identified the transcription factor ONECUT1 as a tumor suppressor gene downregulated by activated β-Catenin in HB. Low ONECUT1 expression also characterizes human HB, and co-expression of ONECUT1 strongly suppressed YAP/β-Catenin-driven HB formation in mice. Mechanistically, ONECUT1 functions downstream of activated β-Catenin and negatively regulates tumor cell glycolysis. Conclusions: We show that suppressing activated β-Catenin could hamper HB progression in vivo by affecting pathways distinct from those regulated by YAP in HB. Inhibition of ONECUT1 expression by β-Catenin might represent a critical molecular event leading to HB formation.

Unique pathways downregulated in Tsc2-/-/NRas and sgGSK3α/β/NRas liver tumors

Correlation analysis between p-AKTSer473 and FOXM1 in human HCC samples from TCPA database