gsk-j4 and Disease-Models--Animal

Chronic postsurgical pain (CPSP) is a serious problem. We developed a mouse model of CPSP induced by electrocautery and examined the mechanism of CPSP. In this mouse model, while both incision and electrocautery each produced acute allodynia, persistent allodynia was only observed after electrocautery. Under these conditions, we found that the mRNA levels of Small proline rich protein 1A (Sprr1a) and Annexin A10 (Anxa10), which are the key modulators of neuropathic pain, in the spinal cord were more potently and persistently increased by electrocautery than by incision. Furthermore, these genes were overexpressed almost exclusively in chronic postsurgical pain-activated neurons. This event was associated with decreased levels of tri-methylated histone H3 at Lys27 and increased levels of acetylated histone H3 at Lys27 at their promoter regions. On the other hand, persistent allodynia and overexpression of Sprr1a and Anxa10 after electrocautery were dramatically suppressed by systemic administration of GSK-J4, which is a selective H3K27 demethylase inhibitor. These results suggest that the effects of electrocautery contribute to CPSP along with synaptic plasticity and epigenetic modification.

Topics: Animals; Annexins; Benzazepines; Cornified Envelope Proline-Rich Proteins; Disease Models, Animal; Electrocoagulation; Female; Foot Injuries; Gene Expression Regulation; Gene Knock-In Techniques; Genes, fos; Genes, Reporter; Histone Code; Histones; Hyperalgesia; Jumonji Domain-Containing Histone Demethylases; Lysine; Male; Methylation; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins; Neuralgia; Neurons; Pain, Postoperative; Pyrimidines; RNA, Messenger; Spinal Cord; Tamoxifen

Streptococcus pneumoniae is a natural colonizer of the human respiratory tract and an opportunistic pathogen. Although epithelial cells are among the first to encounter pneumococci, the cellular processes and contribution of epithelial cells to the host response are poorly understood. Here, we show that a S. pneumoniae serotype 6B ST90 strain, which does not cause disease in a murine infection model, induces a unique NF-κB signature response distinct from an invasive-disease-causing isolate of serotype 4 (TIGR4). This signature is characterized by activation of p65 and requires a histone demethylase KDM6B. We show, molecularly, that the interaction of the 6B strain with epithelial cells leads to chromatin remodelling within the IL-11 promoter in a KDM6B-dependent manner, where KDM6B specifically demethylates histone H3 lysine 27 dimethyl. Remodelling of the IL-11 locus facilitates p65 access to three NF-κB sites that are otherwise inaccessible when stimulated by IL-1β or TIGR4. Finally, we demonstrate through chemical inhibition of KDM6B with GSK-J4 inhibitor and through exogenous addition of IL-11 that the host responses to the 6B ST90 and TIGR4 strains can be interchanged both in vitro and in a murine model of infection in vivo. Our studies therefore reveal how a chromatin modifier governs cellular responses during infection.

Topics: A549 Cells; Alveolar Epithelial Cells; Animals; Benzazepines; Chromatin Assembly and Disassembly; Disease Models, Animal; Enzyme Inhibitors; Epithelial Cells; Gene Expression Regulation; Host-Pathogen Interactions; Humans; Interleukin-11; Jumonji Domain-Containing Histone Demethylases; Mice; Mice, Inbred C57BL; NF-kappa B; Pneumococcal Infections; Promoter Regions, Genetic; Pyrimidines; Streptococcus pneumoniae

Topics: Adenosine; Animals; Apoptosis; Benzazepines; Bile Ducts; Carbon Tetrachloride; Cell Cycle; Cell Cycle Proteins; Cell Line; Cell Proliferation; Cellular Senescence; Cyclin-Dependent Kinase Inhibitor p21; Disease Models, Animal; Enhancer of Zeste Homolog 2 Protein; Enzyme Inhibitors; Hepatic Stellate Cells; Humans; Indoles; Interleukin-10; Jumonji Domain-Containing Histone Demethylases; Ligation; Liver; Liver Cirrhosis; Mice; Pyridones; Pyrimidines; Rats; RNA Interference

Chronic cystitis is characterized by the hyperplasia and fibrosis of the bladder wall as well as attenuated compliance of the bladder. To further unravel its underlying molecular mechanism, the role of NFκB-JMJD3 signaling pathway in cystitis induced bladder fibrosis was investigated. Jmjd3 and Col1/3 expression was detected in a cystitis mouse model that was developed by intraperitoneal injection of cyclophosphamide (CYP). Human bladder smooth muscle cells (hBSMCs) were stimulated in vitro with lipopolysaccharide (LPS), and the cell proliferation and collagen accumulation were detected using EdU, CCK8, flow cytometry, qPCR, western blotting and immunofluorescence assays. Furthermore, the effects of NFκB and JMJD3 on cell proliferation and collagen accumulation were investigated using its selective antagonists, JSH23 and GSK-J4, respectively. CYP induced cystitis significantly increased Jmjd3, Col1 and Col3 expression in the bladder muscle cells. Furthermore, LPS stimulation markedly activated NFκB signaling and elevated JMJD3 expression in hBSMCs, and the activation of NFκB-JMJD3 signaling significantly promoted cell proliferation and collagen accumulation by upregulating CCND1 and COL1/3 expression, respectively. Our study reveals the critical role of NFκB-JMJD3 signaling in cystitis induced bladder reconstruction by regulating hBSMC proliferation and extracellular matrix (ECM) deposition, and these findings provide an avenue for effective treatment of patients with cystitis.

Topics: Animals; Benzazepines; Cell Proliferation; Collagen; Cystitis; Disease Models, Animal; Female; Fibrosis; Humans; Jumonji Domain-Containing Histone Demethylases; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Myocytes, Smooth Muscle; NF-kappa B; Phenylenediamines; Pyrimidines; Signal Transduction; Up-Regulation; Urinary Bladder

Radiotherapy (RT) has long been and remains the only treatment option for diffuse intrinsic pontine glioma (DIPG). However, all patients show evidence of disease progression within months of completing RT. No further clinical benefit has been achieved using alternative radiation strategies. Here, we tested the hypothesis that histone demethylase inhibition by GSK-J4 enhances radiation-induced DNA damage, making it a potential radiosensitizer in the treatment of DIPG.. GSK-J4 significantly reduced the expression of DNA DSB repair genes and DNA accessibility in DIPG cells. GSK-J4 sustained high levels of γH2AX and 53BP1 in irradiated DIPG cells, thereby inhibiting DNA DSB repair through homologous recombination pathway. GSK-J4 reduced clonogenic survival and enhanced radiation effect in DIPG cells.. Together, these results highlight GSK-J4 as a potential radiosensitizer and provide a rationale for developing combination therapy with radiation in the treatment of DIPG.

Topics: Animals; Benzazepines; Cell Line, Tumor; Diffuse Intrinsic Pontine Glioma; Disease Models, Animal; DNA Damage; DNA Repair; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Female; Histone Demethylases; Homologous Recombination; Humans; Mice; Prognosis; Pyrimidines; Radiation Tolerance; Radiation-Sensitizing Agents; Xenograft Model Antitumor Assays

The rapid and dynamic transcriptional changes of Schwann cells in response to injury are critical to peripheral nerve repair, yet the epigenomic reprograming that leads to the induction of injury-activated genes has not been characterized. Polycomb Repressive Complex 2 (PRC2) catalyzes the trimethylation of lysine 27 of histone H3 (H3K27me3), which produces a transcriptionally repressive chromatin environment. We find that many promoters and/or gene bodies of injury-activated genes of mature rat nerves are occupied with H3K27me3. In contrast, the majority of distal enhancers that gain H3K27 acetylation after injury are not repressed by H3K27 methylation before injury, which is normally observed in developmentally poised enhancers. Injury induces demethylation of H3K27 in many genes, such as Sonic hedgehog (Shh), which is silenced throughout Schwann cell development before injury. In addition, experiments using a Schwann cell-specific mouse knock-out of the Eed subunit of PRC2 indicate that demethylation is a rate-limiting step in the activation of such genes. We also show that some transcription start sites of H3K27me3-repressed injury genes of uninjured nerves are bound with a mark of active promoters H3K4me3, for example, Shh and Gdnf, and the reduction of H3K27me3 results in increased trimethylation of H3K4. Our findings identify reversal of polycomb repression as a key step in gene activation after injury.. Peripheral nerve regeneration after injury is dependent upon implementation of a novel genetic program in Schwann cells that supports axonal survival and regeneration. Identifying means to enhance Schwann cell reprogramming after nerve injury could be used to foster effective remyelination in the treatment of demyelinating disorders and in identifying pathways involved in regenerative process of myelination. Although recent progress has identified transcriptional determinants of successful reprogramming of the Schwann cell transcriptome after nerve injury, our results have highlighted a novel epigenomic pathway in which reversal of the Polycomb pathway of repressive histone methylation is required for activation of a significant number of injury-induced genes.

Topics: Animals; Benzazepines; Cellular Reprogramming; Chromatin Immunoprecipitation; Computational Biology; Disease Models, Animal; Enzyme Inhibitors; Epigenomics; Gene Expression Regulation; Histones; In Vitro Techniques; Jumonji Domain-Containing Histone Demethylases; Mice; Mice, Inbred C57BL; Polycomb Repressive Complex 2; Pyrimidines; Rats; Rats, Sprague-Dawley; Regeneration; RNA, Messenger; Schwann Cells; Sciatic Neuropathy; Signal Transduction

Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood cancer. We performed a chemical screen in patient-derived DIPG cultures along with RNA-seq analyses and integrated computational modeling to identify potentially effective therapeutic strategies. The multi-histone deacetylase inhibitor panobinostat demonstrated therapeutic efficacy both in vitro and in DIPG orthotopic xenograft models. Combination testing of panobinostat and the histone demethylase inhibitor GSK-J4 revealed that the two had synergistic effects. Together, these data suggest a promising therapeutic strategy for DIPG.

Topics: Animals; Benzazepines; Brain Stem Neoplasms; Disease Models, Animal; Drug Synergism; Glioma; Humans; Hydroxamic Acids; Indoles; Panobinostat; Pyrimidines; Sequence Analysis, RNA; Xenograft Model Antitumor Assays

Cardio-facio-cutaneous (CFC) syndrome is one of the 'RASopathies', a group of phenotypically overlapping syndromes caused by germline mutations that encode components of the RAS-MAPK pathway. Germline mutations in BRAF cause CFC syndrome, which is characterized by heart defects, distinctive facial features and ectodermal abnormalities. To define the pathogenesis and to develop a potential therapeutic approach in CFC syndrome, we here generated new knockin mice (here Braf(Q241R/+)) expressing the Braf Q241R mutation, which corresponds to the most frequent mutation in CFC syndrome, Q257R. Braf(Q241R/+) mice manifested embryonic/neonatal lethality, showing liver necrosis, edema and craniofacial abnormalities. Histological analysis revealed multiple heart defects, including cardiomegaly, enlarged cardiac valves, ventricular noncompaction and ventricular septal defects. Braf(Q241R/+) embryos also showed massively distended jugular lymphatic sacs and subcutaneous lymphatic vessels, demonstrating lymphatic defects in RASopathy knockin mice for the first time. Prenatal treatment with a MEK inhibitor, PD0325901, rescued the embryonic lethality with amelioration of craniofacial abnormalities and edema in Braf(Q241R/+) embryos. Unexpectedly, one surviving pup was obtained after treatment with a histone 3 demethylase inhibitor, GSK-J4, or NCDM-32b. Combination treatment with PD0325901 and GSK-J4 further increased the rescue from embryonic lethality, ameliorating enlarged cardiac valves. These results suggest that our new Braf knockin mice recapitulate major features of RASopathies and that epigenetic modulation as well as the inhibition of the ERK pathway will be a potential therapeutic strategy for the treatment of CFC syndrome.

Topics: Animals; Benzamides; Benzazepines; Diphenylamine; Disease Models, Animal; Drug Synergism; Ectodermal Dysplasia; Embryo, Mammalian; Facies; Failure to Thrive; Female; Gene Expression Regulation; Gene Knock-In Techniques; Genes, Lethal; Heart Defects, Congenital; Histone Deacetylase Inhibitors; Histone Demethylases; Humans; Liver; Male; MAP Kinase Kinase Kinases; Mice; Mice, Transgenic; Mutation; Myocardium; Protein Kinase Inhibitors; Proto-Oncogene Proteins B-raf; Pyrimidines; Signal Transduction; Skull