JAK-STAT Research Articles

Epstein-Barr virus and immunity.

The Epstein-Barr virus has infected the vast majority of the world's population. EBV is the most common herpes virus in the human population. In childhood, viral infection is either asymptomatic or can lead to infectious mononucleosis. In a small proportion of latently infected people, especially in immunosuppressed patients, EBV causes lymphoid and epithelial malignancies and a number of autoimmune diseases, including one of the causes of the development of multiple sclerosis. Innate immunity is the first line of antiviral defense, which EBV evades using a number of strategies to achieve successful infection. EBV disrupts the innate immune signaling pathways activated by Toll-like, RIG-I-like, NOD-like and AIM2-like receptors, as well as cyclic GMP-AMP synthetase. EBV also antagonizes interferon production and signaling, including the TBK1-IRF3 and JAK-STAT pathways. Through differential modulation of proviral and antiviral mechanisms of action of caspases and other cellular life cycle regulators at different stages of infection, EBV actively interferes with apoptotic and inflammatory pathways to continue effective infection. By turning the activation of innate immunity to its advantage by triggering a pro-inflammatory reaction and proteolytic cleavage of caspases that exhibit proviral activity, the virus establishes latency or enters the lytic reactivation phase, which contributes to the development of serious life-threatening diseases, including oncogenesis. The outcome of EBV infection is regulated by a subtle interaction between innate, adaptive immunity, and viral replication. In the absence of licensed preventive vaccines, immunocorrection and antiviral therapy are the only possible ways to combat the virus and prevent diseases associated with it. Understanding the mechanisms of action of certain EBV genes involved in the life cycle at different stages of infection will help in the future to find the right approach to the development of preventive and therapeutic drugs against EBV.

Inhibition of peritendinous adhesion through targeting JAK2-STAT3 signaling pathway: The therapeutic potential of AG490

Peritendinous adhesion is a common complication following tendon injury repair, posing a significant clinical challenge that requires urgent attention. The primary cause of peritendinous adhesion is the excessive deposition of collagen matrix due to the abnormal proliferation of fibroblasts in an inflammatory state. Janus kinase2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) are key signaling molecules involved in cell proliferation and fibrosis development in various organs. However, the role of the JAK-2 and STAT3 signaling pathways in peritendinous adhesion fibrosis remains unclear. In our study, we first observed upregulation of p-JAK2 and p-STAT3 proteins in human peritendinous adhesion specimens and rat peritendinous adhesion models. In vitro, the JAK2/STAT3 pathway inhibitor AG490 effectively inhibited TGF-β1-induced fibroblast proliferation. Wound healing and transwell assays demonstrated that AG490 suppressed TGF-β1-induced fibroblast migration. Furthermore, we found that AG490 decreased the expression of pro-inflammatory factors, including IL-1β and TNF-α, as well as extracellular matrix (ECM) proteins in fibroblasts under TGF-β1 stimulation. In vivo, histological staining showed that AG490 prevented fibrous tissue formation in a rat model of tendon injury. Moreover, AG490 inhibited the overexpression of pro-inflammatory factors IL-1β and TNF-α, as well as ECM in the peritendinous adhesions. In conclusion, AG490 inhibited fibrosis and inflammation in injured tendons by targeting the JAK2-STAT3 signaling pathway, presenting a promising strategy for the prophylaxis of peritendinous adhesions.

041 Deciphering the contribution of JAK-STAT signaling pathway in inflammatory skin diseases

041 Deciphering the contribution of JAK-STAT signaling pathway in inflammatory skin diseases

Evidence from Machine Learning, Diagnostic Hub Genes in Sepsis and Diagnostic Models based on Xgboost Models, Novel Molecular Models for the Diagnosis of Sepsis.

Systemic multi-organ dysfunction resulting from dysregulated immune responses in the host triggered by microbial infection or other factors is a major cause of death in sepsis, and secretory pathways play an important role in it. GSE57065, GSE65682, GSE145227, and GSE54514 from Gene Expression Omnibus (GEO) were derived for this study. Secretory pathways single sample gene set enrichment analysis (ssGSEA) scores in sepsis and normal samples were exposed. Gene modules associated with secretory pathways were selected by weighted gene coexpression network analysis (WGCNA) for Protein-Protein Interaction Networks (PPI) assessment, and crossover genes in both were evaluated by eXtreme Gradient Boosting (XGBoost) model in feature selection to identify hub genes in sepsis. In addition, we explored the immune cells and signaling pathways regulated by hub genes. Remarkable dysregulation of secretory pathways was demonstrated in sepsis. The secretory pathways-associated gene modules were intimately involved in cytokine and immune responses in infection. Four crossover genes (CD163, FCER1G, C3AR1, ARG1) were present in WGCNA and PPI, and training in the XGBoost model revealed the best diagnostic performance of these 4 genes, meaning that these genes were the hub genes for sepsis. The 4-hub genes showed a significant negative correlation with T cell activity and a significant positive correlation with inflammatory immune cells. In addition, we found that the 4-hub genes markedly positively regulated INFLAMMATORY RESPONSE, IL6 JAK STAT3 SIGNALING. Based on WGCNA, PPI, and XGBoost models, we identified hub genes that play an important regulatory role in sepsis. We also developed novel molecular models for the diagnosis of sepsis.

Establishment of human minor salivary gland organoids in laminin-GelMA hydrogel from healthy individuals and Sjögren’s disease patients

Biopsies of human minor salivary glands (hMSGs), located beneath the oral mucosa, are crucial for a definitive clinical diagnosis of Sjögren’s Disease (SjD)。Although organoids from various exocrine glands have been cultured, organoids comprising the diverse cell types of hMSGs have not yet been successful. In this study, we established long-term cultures of hMSGs organoids derived from both healthy individuals and patients with SjD. The organoids, cultured in artificial laminin-GelMA hydrogel, retain distinct phenotypic and functional characteristics of the native glands, both when stimulated with neurotransmitters in vitro and upon transplantation in vivo. Single-cell RNA sequencing revealed that organoids from healthy individuals and from SjD patients exhibit unique phenotypic differences. Furthermore, treatment with tofacitinib enhanced the proliferation of patient organoids by inhibiting the JAK-STAT pathway, suggesting a potential therapeutic strategy for further investigation. Here, we derived organoids from healthy individuals as potential transplant material in regeneration medicine, and developed SjD hMSG organoids for use as a disease model for exploring SjD pathogenesis and developing therapeutic strategies.

The Role of Panax Notoginseng Saponins on Neuronal Oxidative Stress and Autophagy Response in Craniocerebral Injury Model Rats

This study assessed how IL-6 affects the body’s autophagy status by regulating JAK-STAT3 and explored the mechanism by which IL-6 inhibits JAK-STAT3 signaling pathway-mediated autophagy, and how Panax notoginseng total saponins promote neuronal cell regeneration in rats. A model of neuronal oxidative stress in craniocerebral injury model rats was established, and IL-6 levels were detected by ELISA. Rat neuronal cells were isolated and cultured, while dual-luciferase gene reporter experiments analyzed the targeting relationship between IL-6 and JAK. qRT-PCR detected expression of neuronal autophagy-related genes (JAK, STAT3, ULK1, OsATG7, FAM176A, and Beclin 1). The level of IL-6 in the craniocerebral injury model was significantly higher than that in the control group. IL-6 binds to 3′-UTR of JAK, and transfection with IL-6 inhibitor increases the relative luciferase activity. pMIR-JAK-mut group has no significant effect. There is a targeted regulatory relationship between IL-6 and JAK. Adding total saponins of Panax notoginseng combined with nursing intervention can promote reduction genetic expression related to neuronal autophagy damage. IL-6 can target and regulate JAK gene. IL-6 can promote neuronal development by regulating JAK-STAT3. Autophagy repairs damage and inhibits the autophagy state of cells. Panax notoginseng total saponins are effective medicinal components extracted from high quality panax notoginseng according to extraction and separation technology. The results from this study provide a better understanding of pathogenesis of neuronal oxidative stress in craniocerebral injury model rats and find potential intervention targets, which will provide more treatment methods for neuronal oxidative stress damage in craniocerebral injury model rats.

Sustained antiviral insulin signaling during West Nile virus infection results in viral mutations

Arthropod-borne viruses or arboviruses, including West Nile virus (WNV), dengue virus (DENV), and Zika virus (ZIKV) pose significant threats to public health. It is imperative to develop novel methods to control these mosquito-borne viral infections. We previously showed that insulin/insulin-like growth factor-1 signaling (IIS)-dependent activation of ERK and JAK-STAT signaling has significant antiviral activity in insects and human cells. Continuous immune pressure can lead to adaptive mutations of viruses during infection. We aim to elucidate how IIS-signaling in mosquitoes selects for West Nile virus escape variants, to help formulate future transmission blocking strategies. We hypothesize that passage of WNV under activation of IIS will induce adaptive mutations or escape variants in the infecting virus. To test our hypothesis, WNV was serially passaged through Culex quinquefasciatus Hsu cells in the presence or absence of bovine insulin to activate IIS antiviral pressure. We sequenced WNV genes encoding for E, NS2B, NS3, and NS5 and identified variants in E and NS5 arising from IIS antiviral pressure. In parallel to the genetic analyses, we also report differences in the levels of virus replication and Akt activation in human cells and mosquitoes using virus passaged in the presence or absence of insulin. Finally, using adult Culex quinquefasciatus, we demonstrated the enhancement of immune response gene expression in virus-infected mosquitoes fed on insulin, compared to control. Notably, virus collected from insulin-fed mosquitoes contained a non-synonymous mutation in NS3. These results contribute towards achieving our long-term goal of manipulating mosquito IIS-dependent antiviral immunity to reduce WNV or other flavivirus transmission to mammalian hosts.

Targeting the JAK-STAT pathway in colorectal cancer: mechanisms, clinical implications, and therapeutic potential

Colorectal cancer (CRC) remains one of the most prevalent and fatal malignancies worldwide, consistently ranking among the top three in terms of incidence and mortality. Despite notable advancements in early detection and therapeutic interventions, survival outcomes for advanced-stage CRC are still dismal, largely due to issues such as drug resistance and metastasis. Recent research has increasingly implicated the JAK-STAT signaling pathway as a pivotal contributor to CRC pathogenesis. This evolutionarily conserved pathway plays a key role in transmitting extracellular signals to the nucleus, thereby modulating gene expression involved in numerous fundamental biological processes. In CRC, dysregulation of the JAK-STAT pathway is frequently observed and is strongly associated with tumor progression, including processes such as cellular proliferation, apoptosis, metastasis, immune evasion, and the sustenance of cancer stem cells. Given its integral role in CRC advancement, the JAK-STAT pathway has gained recognition as a viable therapeutic target. Extensive evidence from preclinical and clinical models supports the efficacy and safety of targeting components of the JAK-STAT pathway, presenting new therapeutic possibilities for patients with CRC, particularly in addressing drug resistance and enhancing treatment outcomes. This review offers a detailed exploration of the JAK-STAT pathway, focusing on its regulatory mechanisms in CRC-related malignancies. Moreover, it examines the association between JAK-STAT protein expression, clinical features, prognosis, and its therapeutic potential in CRC management.

Full-length transcriptome sequencing of the longissimus dorsi muscle of yak and cattle-yak using nanopore technology

Short-read RNA sequencing has been used to sequence the transcriptome of the skeletal muscle of yak and cattle-yak; however, full-length transcripts cannot be obtained and alternative splicing (AS) events cannot be inferred using this sequencing approach. Here, we used Oxford Nanopore Technologies (ONT) full-length sequencing to sequence the transcriptome of the longissimus dorsi of yak and cattle-yak. A total of 20,323 novel genes and 172,870 novel transcripts were identified, and 159,700 novel transcripts were successfully annotated. A total of 157,812 AS events, 58,073 simple sequence repeats, 57,468 complete open reading frames, 2296 transcription factors, and 20,404 lncRNAs were detected. Differentially expressed transcripts (DETs) in the longissimus dorsi muscle of yak and cattle-yak were involved in the MAPK and JAK-STAT signaling pathways related to muscle development and growth. Protein–protein interaction analysis of DETs suggested that TNNI2 might make a major contribution to differences in muscle growth and meat quality traits between yak and cattle-yak. The results have enriched the transcriptome data of dorsal muscles, providing new ideas for the study of transcriptional regulation processes, and also providing useful information for the production of higher yields of yak meat.

Spatial dissection of tumour microenvironments in gastric cancers reveals the immunosuppressive crosstalk between CCL2+ fibroblasts and STAT3-activated macrophages

BackgroundA spatially resolved, niche-level analysis of tumour microenvironments (TME) can provide insights into cellular interactions and their functional impacts in gastric cancers (GC).ObjectiveOur goal was to translate the spatial organisation...

Exosomal LncRNA and CircRNA Regulate Peripheral Blood Mononuclear Cell Function through a Competitive Endogenous RNA Mechanism in Allergic Rhinitis.

Peripheral blood mononuclear cells (PBMCs) dysfunction is involved in the pathogenesis and progression of allergic rhinitis (AR). This study aims to investigate the competing endogenous RNA (ceRNA) networks in PBMCs influenced by differentially expressed long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) found in plasma exosomes induced by AR. All subjects were from the Affiliated Taizhou People's Hospital of Nanjing Medical University. Differential expression of mRNAs in PBMCs and lncRNAs/circRNAs in plasma exosomes was analyzed using high-throughput sequencing. Differentially expressed lncRNAs and circRNAs that target mRNAs were identified using bioinformatics methods. The predicted target mRNAs were intersected with the differentially expressed mRNAs in PBMCs to construct ceRNA networks. The subcellular localizations of lncRNAs and circRNAs within the ceRNA networks were determined using RNA fluorescence in situ hybridization or bioinformatics methods. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of differentially expressed mRNAs in PBMCs were conducted using the clusterProfiler R package. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to validate the expression levels of each molecule within the constructed ceRNA networks in clinical samples, along with receiver operating characteristic (ROC) curve analysis to assess diagnostic value. Further validation was performed using in vitro cultured PBMCs and dual-luciferase reporter assays. Five differentially expressed circRNAs and 31 differentially expressed lncRNAs were identified in exosomes. In PBMCs, 130 differentially expressed mRNAs were identified. Six ceRNA networks were constructed, affecting PBMCs chemorepellent activity, JAK-STAT signaling pathway, and other functions or pathways. The expression level of ENST00000650850 in plasma exosomes was significantly lower in AR patients, suggesting its potential diagnostic value. The expression level of ENST00000650850 in plasma exosomes was positively correlated with the expression levels of ENST00000650850 and IL6 mRNA in PBMCs. PBMCs from healthy individuals were stimulated with plasma exosomes isolated from AR patients, leading to a reduction in IL6R mRNA expression levels in the PBMCs. Differentially expressed lncRNA (ENST00000650850) in plasma-derived exosomes of AR patients may regulate IL6R mRNA expression in PBMCs via miR-6747-3p, thereby influencing PBMC function and contributing to the pathogenesis and progression of AR.

Structural Evaluation of Interleukin-19 Cytokine and Interleukin-19-Bound Receptor Complex Using Computational Immuno-Engineering Approach

Interleukin 19 (IL-19) is an anti-inflammatory cytokine that belongs to the IL-10 family, where IL-20 and IL-24 also exist. While IL-19 and IL-20 share some comparable structural folds, there are certain structural divergences in their N-terminal ends. To date, there are no reported IL-19 receptors; although, it has been suggested in the literature that IL-19 would bind to lL-20 receptor (IL-20R) and trigger the JAK-STAT signaling pathways. The present report examines the structure of the IL-19 cytokine and its receptor complex using a computational approach. Specifically, the postulated modes of interactions for IL-20R as an IL-19 receptor are examined on the basis of a set of computational findings. The author has used molecular docking and molecular dynamics simulation to generate a 3D model for the IL-19 complex with IL-20R. When a protein’s crystal structure is not available in the literature, predictive modeling is often employed to determine the protein’s 3D structure. The model assessment can be based on various factors, which include stability analysis using RMSD calculations, tracking changes in time-based secondary structures and the associated Gibbs energies, ΔG. Since one model complex (referred to as model A throughout this paper) can be used as a working hypothesis for future experiments, this structure has been explored here in detail to check its stability, subunit interfaces, and binding residues. The information gathered in this approach can potentially help to design specific experiments to test the validity of the model protein structure. Additionally, the results of this research should be relevant for understanding anti-inflammatory mechanisms and, eventually, could contribute to the efforts for therapeutic developments and targeted therapy.

Transdermal Delivery of Tofacitinib Citraste via Mannose-Decorated Transferosomes Loaded with Tofacitinib Citrate in Arthritic Joints.

Transdermal drug delivery systems are a promising option for the treatment of rheumatoid arthritis (RA) because they can lower systemic adverse effects of immunosuppressants. Janus kinase (JAK) inhibitors were found to be effective for the treatment of RA by inhibiting the JAK-STAT pathway and preventing autoimmune joint destruction. The aim of this study is to deliver tofacitinib (a JAK 1 and 3 inhibitor) through mannose-decorated transferosomes (MDTs) directly to inflamed joints. Transferosomes are composed of phospholipids, Cremophor A25, PEG400, Labrafac lipophile, and oleic acid to enhance the permeation of tofacitinib and control nanovesicle size (∼70-200 nm). Permeation through rat skin was evaluated, where the skin permeation of MDTs (Q24: 38.8 ± 9.82 μg/cm2) and flux (0.5311 ± 0.072 μg/cm2/h) were significantly greater than those of the uncoated transferosomes (Q24 of T1: 1.522 ± 0.329 μg/cm2, Q24 of T2: 3.5002 ± 0.998 μg/cm2, and Q24 of T3: 18.226 ± 5.25 μg/cm2). In addition, MDTs seem to permeate the skin intact, as shown by the transmission electron microscopy (TEM) images of the recipient buffer removed from the Franz diffusion cell. A histopathology assay was performed during the in vivo evaluation of MDTs in an arthritic rat model, in which, significantly less inflammation was observed when MDTs were applied directly to the joint compared to when applied to the dorsal skin and untreated arthritic joints. Furthermore, significantly lower tumor necrosis factor-α (TNFα), IL-6, and IL-1β levels (P < 0.05) were detected by enzyme-linked immunosorbent assay (ELISA) in homogenates of the joints treated with MDTs than in untreated arthritic joints. In conclusion, this study proposed effective MDTs that could deliver tofacitinib directly to inflamed joints possibly by targeting the macrophages circulating in the proximity of the site of inflammation.

Exploring the Antiviral Potential of Artemisia annua Through JAK-STAT Pathway Targeting: A Network Pharmacology Approach.

Artemisia annua, a plant with antiviral potential, has shown promise against various viral infections, yet its mechanisms of action are not fully understood. This study explores A. annua's antiviral effects using network pharmacology and molecular docking, focusing on key active compounds and their interactions with viral protein targets, particularly within the JAK-STAT signaling pathway-a critical mediator of immune responses to viral infections. From the TCMSP database, we identified eight active compounds and 335 drug targets for A. annua, with 19 intersecting targets between A. annua compounds and viral proteins. A protein-protein interaction (PPI) network highlighted 10 key hub genes, analyzed further through Gene Ontology (GO) and KEGG pathways to understand their immune and antiviral roles. ADMET properties of the active compound Patuletin (MOL004112) were assessed, followed by 200 ns molecular dynamics simulations to examine its stability in complex with JAK2. PPI analysis identified JAK2, MAPK3, MAPK1, JAK1, PTPN1, HSPA8, TYK2, RAF1, MAPT, and HMOX1 as key hub genes, with JAK2 emerging as a critical regulator of immune and antiviral pathways. ADMET analysis confirmed Patuletin's favorable pharmacokinetic properties, and molecular dynamics simulations showed a stable Patuletin-JAK2 complex, with FEL analysis indicating minimal disruption to JAK2's intrinsic flexibility. These findings highlight JAK2 as a promising target in the antiviral activity of A. annua compounds, particularly Patuletin, supporting its potential as an antiviral agent and providing a foundation for further research on A. annua's therapeutic applications.

Integrative Multi-PTM Proteomics Reveals Dynamic Global, Redox, Phosphorylation, and Acetylation Regulation in Cytokine-Treated Pancreatic Beta Cells

Studying regulation of protein function at a systems level necessitates an understanding of the interplay among diverse post-translational modifications (PTMs). A variety of proteomics sample processing workflows are currently used to study specific PTMs but rarely characterize multiple types of PTMs from the same sample inputs. Method incompatibilities and laborious sample preparation steps complicate large-scale physiological investigations and can lead to variations in results. The single-pot, solid-phase-enhanced sample preparation (SP3) method for sample cleanup is compatible with different lysis buffers and amenable to automation, making it attractive for high-throughput multi-PTM profiling. Herein, we describe an integrative SP3 workflow for multiplexed quantification of protein abundance, cysteine thiol oxidation, phosphorylation, and acetylation. The broad applicability of this approach is demonstrated using cell and tissue samples, and its utility for studying interacting regulatory networks is highlighted in a time-course experiment of cytokine-treated β-cells. We observed a swift response in global regulation of protein abundances consistent with rapid activation of JAK-STAT and NF-κB signaling pathways. Regulators of these pathways as well as proteins involved in their target processes displayed multi-PTM dynamics indicative of a complex cellular response stages: acute, adaptation, and chronic (prolonged stress). PARP14, a negative regulator of JAK-STAT, had multiple co-localized PTMs that may be involved in intraprotein regulatory crosstalk. Our workflow provides a high-throughput platform that can profile multi-PTMomes from the same sample set, which is valuable in unraveling the functional roles of PTMs and their co-regulation.

Harnessing Tumor Cell-Derived Exosomes for Immune Rejection Management in Corneal Transplantation.

Transplantation remains the definitive treatment for end-stage organ failures, but its efficacy is frequently compromised by immune rejection. This study introduces a novel strategy by utilizing tumor-derived exosomes from B16-F10 melanoma cells (B16-Exo), diverging from the conventional use of immune cell-derived exosomes, to alleviate post-transplantation immune rejection. Utilizing murine corneal transplantation as a model, it is demonstrated that B16-Exo significantly reduces immune rejection, evidenced by decreased corneal opacity, neovascularization, and immune dysregulation, while enhancing postoperative survival. Proteomic analyses reveal differential expression of pivotal proteins in B16-Exo, notably the JAK2 protein within the JAK-STAT signaling pathway, which has been mechanistically demonstrated to amplify the activity of myeloid-derived suppressor cells (MDSCs) and inhibit T cell proliferation. These findings demonstrate the significant immunomodulatory effect of B16-Exo in transplant immunology, supporting the continued exploration of tumor-derived exosomes as a platform to uncover novel immunosuppressive mechanisms in transplantation.

Effect of the Intake of Solid Block Dairy Products Like Cheese on Serum Uric Acid in Children: A Preliminary Mechanistic Investigation.

This study aimed to investigate the relationship between the intake of solid block dairy products like cheese and serum uric acid levels, along with its potential physiological mechanisms. Data for our study were obtained from the Chinese Children and Lactating Women Nutrition and Health Surveillance. Generalized linear models and restricted cubic splines were employed to analyze the relationship between the intake of solid block dairy products like cheese and serum uric acid levels. Two-sample Mendelian randomization (TSMR) analysis was conducted to infer causality, based on a large sample size and robust methodology. Gene Ontology (GO) enrichment analysis was also performed to identify potential biological pathways. Among all types of dairy products, a significant negative association with serum uric acid levels was observed only for the intake of solid block dairy products like cheese, regardless of covariate adjustment (β = -0.182, p < 0.001). TSMR results supported a negative causal relationship between cheese intake and serum uric acid levels (β = -0.103, 95% CI: -0.149 to -0.057; p = 0.002). The JAK-STAT signaling pathway and autophagy regulation were identified as potential physiological mechanisms underlying this relationship. The intake of solid block dairy products like cheese was found to result in decreased levels of serum uric acid, with potential mechanisms involving the JAK-STAT signaling pathway and the regulation of autophagy.

Abstract 4120687: Multi-Omics Insights into Recovery from Acute Fulminant Myocarditis Treated with Ruxolitinib

Background: There are currently no approved medical therapies for acute fulminant myocarditis (AFM). Janus kinase (JAK) inhibitors target the JAK-STAT signaling pathway, which is crucial in immune activation. We report the first use of ruxolitinib, a JAK inhibitor, for treatment of AFM. Multi-omics single-cell technologies, including RNA-seq, T-/B-cell receptor seq, and CITE-seq, were employed to analyze immune profiles pre- and post-ruxolitinib treatment. Results: A 20-year-old female presented with AFM with cardiogenic shock and was supported by VA-ECMO and impella CP. Endomyocardial biopsy showed lymphocytic myocarditis. The patient received pulsed steroids and was listed for orthotopic heart transplant. Due to deteriorating conditions, ruxolitinib (10 mg BID) was added with immediate improvement in cardiac and inflammatory biomarkers and hemodynamics. ECMO was decannulated on day 6 and impella CP was removed on day 8. Repeat TTE on day 9 showed normalization of cardiac function (LVEF 58%, increased from 10%). She was discharged on ruxolitinib and is doing well in follow-up. Multi-omics single-cell technologies were employed on PBMCs collected at four time points: prior to ruxolitinib treatment (but after treatment with corticosteroids), and post-ruxolitinib treatment on day 5, day 8, and 2-months. At baseline, prior to treatment, scRNA-Seq and CITE-seq analysis revealed upregulated JAK-STAT signaling in pathogenic immune cells such as NK cells, CCR2 + /CCR5 + /HLA-DR + monocytes and activated T cells. Ruxolitinib treatment significantly decreased these pathogenic immune cells, with inhibition of STAT1/STAT3 signaling. Ruxolitinib also significantly decreased key cytotoxic genes PRF1, GZMB, and TBX21 in T and NK cells. TCR sequencing revealed clonal expansions of activated T cells with high levels of pro-inflammatory genes (IL2/IL6/IFNg) at baseline which were dramatically reduced post-treatment with ruxolitinib. Longitudinal data indicated normalization of naive T and B cell levels and clonal diversity, mirroring her clinical improvement. Conclusions: Ruxolitinib significantly modulates immune profiles and disrupts pathogenic signaling in AFM. This first reported use of ruxolitinib in AFM, coupled with our multi-omics analysis, highlights profound immune reprogramming and supports targeted immune modulation for rapid recovery, underscoring ruxolitinib’s therapeutic efficacy.

Analyzing immune cell infiltrates in skeletal muscle of infantile-onset Pompe disease using bioinformatics and machine learning

Pompe disease, a severe lysosomal storage disorder, is marked by heart problems, muscle weakness, and respiratory difficulties. This study aimed to identify novel markers for infantile-onset Pompe disease by analyzing key genes and immune cells infiltrating affected skeletal muscles, building on existing research linking its progression to the immune cell infiltration. The datasets GSE38680 and GSE159062 were downloaded and differential expression genes (DEGs) were identified. Potential markers were screened using support vector machine recursive feature elimination (SVM-RFE) analysis and least absolute shrinkage and selection operator (LASSO) regression models. To analyze DEGs and immune processes, 22 immune cell types in affected tissues were examined using CIBERSORT. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed significant enrichment in the calcium signaling pathway, phosphatidylinositol signaling system, ubiquitin-mediated proteolysis and JAK-STAT signaling pathway. Machine learning identified GPNMB, CALML6 and TRIM7 as key genes. Immune cell infiltration analysis showed that the expression levels of the above three genes were closely related to immune cells in our target population under study, including T cells regulatory (Tregs), monocytes, macrophages M0, resting and activated dendritic cells, naive and memory B cells. Overall, our study results may provide new clues for exploring markers related to Pompe disease by highlighting key genes and their relationship with immune infiltration, offering insights into the disease’s development. However, previous studies have shown limited evidence of significant immune cell infiltration in muscle biopsies from Pompe patients, and this lack of direct evidence necessitates further investigation and experimental validation in laboratory settings to confirm these associations.

DDDR-01. TUMOR ASSOCIATED MACROPHAGES VIS-À-VIS TMZ RESISTANCE IN PTEN-DEFICIENT GLIOMA

Abstract Drug resistance is one of the most important obstacles during chemotherapy. Currently temozolomide (TMZ) is still first line chemotherapy agent for gliomas, while drug resistance is one of the main reasons for chemotherapy failure. It has been reported that tumor-associated macrophages (TAMs) could be related to drug resistance, and thus we have investigated the association between TAMs and TMZ response in gliomas. Firstly, our study have found that PTEN-deficient glioma can specifically induce a large number of M2 phenotype TAMs, which can attenuate the TMZ-mediated the killing effect on glioma cells. Further search for the possible mechanism, we have found TAMs may secrete cytokine IL-6 in PTEN-deficient gliomas, which in turn mediate TMZ resistance. We also revealed that IL-6 highly secreted by TAMs can enhance the stemness of glioma cells through activation of the JAK-STAT3 signaling pathway in PTEN-deficient glioma cells. Secondly, have found valproic acid (VPA) can promote the transformation of TAMs from M2 to M1, reduce its IL-6 secretion, and in turn enhancing sensitivity of TMZ in PTEN-deficient gliomas. Our study revealed that TMZ resistance may modulated through soluble factors released by TAMs. Conventional anti-epilepsy drug- VPA may enhance TMZ sensitivity in PTEN-deficient gliomas, and worth to further clinical investigation. Keywords: PTEN-deficient gliomas; tumor-associated macrophages; valproic acid; temozolomide resistance