-
Translational Research : the Journal of... Mar 2023Ubiquitination-mediated protein degradation is associated with the development of pulmonary fibrosis. We and others have shown that Nedd4L plays anti-inflammatory and...
Ubiquitination-mediated protein degradation is associated with the development of pulmonary fibrosis. We and others have shown that Nedd4L plays anti-inflammatory and anti-fibrotic roles by targeting lysophosphatidic acid receptor 1 (LPAR1), p-Smad2/3, and β-catenin, and other molecules for their degradation in lung epithelial cells and fibroblasts. However, the molecular regulation of Nedd4L expression in lung fibroblasts has not been studied. In this study, we find that Nedd4L levels are significantly suppressed in lung myofibroblasts in IPF patients and in experimental pulmonary fibrosis, and in TGF-β1-treated lung fibroblasts. Nedd4L knockdown promotes TGF-β1-mediated phosphorylation of Smad2/3 and lung myofibroblast differentiation. Mechanistically, Nedd4L targets TGF-β receptor II (TβRII), the first key enzyme of TGF-β1-mediated signaling, for its ubiquitination and degradation. Further, we show that inhibition of transcriptional factor E2F rescues Nedd4L levels and mitigates experimental pulmonary fibrosis. Together, our data reveal insight into mechanisms by which E2F-mediated Nedd4L suppression contributes to the pathogenesis of lung fibrosis. This study provides evidence showing that upregulation of Nedd4L is a potential therapeutic strategy to treat fibrotic disorders including lung fibrosis.
Topics: Humans; Pulmonary Fibrosis; Transforming Growth Factor beta1; Lung; Fibroblasts; Cell Differentiation; Myofibroblasts; Fibrosis; Bleomycin
PubMed: 36257596
DOI: 10.1016/j.trsl.2022.10.002 -
International Journal of Molecular... Mar 2021Systemic sclerosis (SSc) is an autoimmune disorder characterized by fibrosis of the skin and internal organs. Despite several studies on SSc treatments, effective...
Systemic sclerosis (SSc) is an autoimmune disorder characterized by fibrosis of the skin and internal organs. Despite several studies on SSc treatments, effective treatments for SSc are still lacking. Since evidence suggests an association between intestinal microbiota and SSc, we focused on butyrate, which has beneficial effects in autoimmune diseases as a bacterial metabolite. Here, we investigated the therapeutic potential of sodium butyrate (SB) using a bleomycin-induced fibrosis mouse model of SSc and human dermal fibroblasts (HDFs). SB attenuated bleomycin-induced dermal and lung fibrosis in mice. SB influenced fecal microbiota composition (phyla Actinobacteria and Bacteroidetes, genera and ). SB controlled macrophage differentiation in mesenteric lymph nodes, spleen, and bronchoalveolar lavage cells of mice with bleomycin-induced skin fibrosis. Profibrotic and proinflammatory gene expression was suppressed by SB administration in skin. Furthermore, SB inhibited transforming growth factor β1-responsive proinflammatory expression with increased acetylation of histone 3 in HDFs. Subcutaneous SB application had antifibrogenic effects on the skin. Butyrate ameliorated skin and lung fibrosis by improving anti-inflammatory activity in a mouse model of SSc. Butyrate may exhibit indirect and direct anti-fibrogenic action on fibroblasts by regulating macrophage differentiation and inhibition of histone deacetylase 3. These findings suggest butyrate as an SSc treatment.
Topics: Animals; Bleomycin; Butyrates; Disease Models, Animal; Dysbiosis; Male; Mice; Pulmonary Fibrosis; Skin Diseases
PubMed: 33803282
DOI: 10.3390/ijms22052765 -
Respiratory Research Apr 2024Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fatal, and aging-associated interstitial lung disease with a poor prognosis and limited treatment options,...
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fatal, and aging-associated interstitial lung disease with a poor prognosis and limited treatment options, while the pathogenesis remains elusive. In this study, we found that the expression of nuclear receptor subfamily 2 group F member 2 (NR2F2), a member of the steroid thyroid hormone superfamily of nuclear receptors, was reduced in both IPF and bleomycin-induced fibrotic lungs, markedly in bleomycin-induced senescent epithelial cells. Inhibition of NR2F2 expression increased the expression of senescence markers such as p21 and p16 in lung epithelial cells, and activated fibroblasts through epithelial-mesenchymal crosstalk, inversely overexpression of NR2F2 alleviated bleomycin-induced epithelial cell senescence and inhibited fibroblast activation. Subsequent mechanistic studies revealed that overexpression of NR2F2 alleviated DNA damage in lung epithelial cells and inhibited cell senescence. Adenovirus-mediated Nr2f2 overexpression attenuated bleomycin-induced lung fibrosis and cell senescence in mice. In summary, these data demonstrate that NR2F2 is involved in lung epithelial cell senescence, and targeting NR2F2 may be a promising therapeutic approach against lung cell senescence and fibrosis.
Topics: Animals; Mice; Bleomycin; Cellular Senescence; Epithelial Cells; Idiopathic Pulmonary Fibrosis; Lung
PubMed: 38566093
DOI: 10.1186/s12931-024-02777-3 -
Scientific Reports Aug 2023Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive respiratory disease. Arguably, the complex interplay between immune cell subsets, coupled with an...
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive respiratory disease. Arguably, the complex interplay between immune cell subsets, coupled with an incomplete understanding of disease pathophysiology, has hindered the development of successful therapies. Despite efforts to understand its pathophysiology and develop effective treatments, IPF remains a fatal disease, necessitating the exploration of new treatment options. Mesenchymal stromal/stem cell (MSC) therapy has shown promise in experimental models of IPF, but further investigation is needed to understand its therapeutic effect. This study aimed to assess the therapeutic effect of adipose-derived mesenchymal stem cells in a bleomycin-induced pulmonary fibrosis model. First, MSC cells were obtained from mice and characterized using flow cytometry and cell differentiation culture methods. Then adult C57BL/6 mice were exposed to endotracheal instillation of bleomycin and concurrently treated with MSCs for reversal models on day 14. Experimental groups were evaluated on days 14, 21, or 28. Additionally, lung fibroblasts challenged with TGF-β1 were treated with MSCs supernatant or MSCs to explore the mechanisms underlying of pulmonary fibrosis reversal. Mesenchymal stem cells were successfully isolated from mouse adipose tissue and characterized based on their differentiation ability and cell phenotype. The presence of MSCs or their supernatant stimulated the proliferation and migration of lung fibrotic cells. MSCs supernatant reduced lung collagen deposition, improved the Ashcroft score and reduced the gene and protein expression of lung fibrosis-related substances. Bleomycin-challenged mice exhibited severe septal thickening and prominent fibrosis, which was effectively reversed by MSCs treatment. MSC supernatant could suppress the TGF-β1/Smad signaling pathway and supernatant promotes fibroblast autophagy. In summary, this study demonstrates that MSCs supernatant treatment is as effective as MSCs in revert the core features of bleomycin-induced pulmonary fibrosis. The current study has demonstrated that MSCs supernatant alleviates the BLM-induced pulmonary fibrosis in vivo. In vitro experiments further reveal that MSC supernatant could suppress the TGF-β1/Smad signaling pathway to inhibit the TGF-β1-induced fibroblast activation, and promotes fibroblast autophagy by Regulating p62 expression. These findings contribute to the growing body of evidence supporting the therapeutic application of MSCs in cell therapy medicine for IPF.
Topics: Adipocytes; Mesenchymal Stem Cells; Bleomycin; Idiopathic Pulmonary Fibrosis; Male; Female; Animals; Mice; Mice, Inbred C57BL; Cells, Cultured
PubMed: 37580529
DOI: 10.1038/s41598-023-40531-9 -
Radiology and Oncology Jun 2023Biomedical applications of electroporation are expanding out of the field of oncology into vaccination, treatment of arrhythmias and now in the treatment of vascular... (Review)
Review
Bleomycin electrosclerotherapy (BEST) for the treatment of vascular malformations. An International Network for Sharing Practices on Electrochemotherapy (InspECT) study group report.
BACKGROUND
Biomedical applications of electroporation are expanding out of the field of oncology into vaccination, treatment of arrhythmias and now in the treatment of vascular malformations. Bleomycin is a widely used sclerosing agent in the treatment of various vascular malformations. The application of electric pulses in addition to bleomycin enhances the effectiveness of the drug, as demonstrated by electrochemotherapy, which utilizes bleomycin in the treatment of tumors. The same principle is used in bleomycin electrosclerotherapy (BEST). The approach seems to be effective in the treatment of low-flow (venous and lymphatic) and, potentially, even high-flow (arteriovenous) malformations. Although there are only a few published reports to date, the surgical community is interested, and an increasing number of centers are applying BEST in the treatment of vascular malformations. Within the International Network for Sharing Practices on Electrochemotherapy (InspECT) consortium, a dedicated working group has been constituted to develop standard operating procedures for BEST and foster clinical trials.
CONCLUSIONS
By treatment standardization and successful completion of clinical trials demonstrating the effectiveness and safety of the approach, higher quality data and better clinical outcomes may be achieved.
Topics: Humans; Electrochemotherapy; Vascular Malformations; Electroporation Therapies; Electroporation; Bleomycin
PubMed: 37341196
DOI: 10.2478/raon-2023-0029 -
Bioelectrochemistry (Amsterdam,... Aug 2021Nanosecond electric pulses have several potential advantages in electroporation-based procedures over the conventional micro- and millisecond pulses including low level...
Nanosecond electric pulses have several potential advantages in electroporation-based procedures over the conventional micro- and millisecond pulses including low level of heating, reduced electrochemical reactions and reduced muscle contractions making them alluring for use in biomedicine and food industry. The aim of this study was to evaluate if nanosecond pulses can enhance the cytotoxicity of chemotherapeutics bleomycin and cisplatin in vitro and to quantify metal release from electrodes in comparison to 100 μs pulses commonly used in electrochemotherapy. The effects of nanosecond pulse parameters (voltage, pulse duration, number of pulses) on cell membrane permeabilization, resealing and on cell survival after electroporation only and after electrochemotherapy with bleomycin and cisplatin were evaluated on Chinese hamster ovary cells. Application of permeabilizing nanosecond pulses in combination with chemotherapeutics resulted in successful cell kill. Higher extracellular concentrations of bleomycin - but not cisplatin - were needed to achieve the same decrease in cell survival with nanosecond pulses as with eight 100 μs pulses, however, the tested bleomycin concentrations were still considerably lower compared to doses used in clinical practice. Decreasing the pulse duration from microseconds to nanoseconds and concomitantly increasing the amplitude to achieve the same biological effect resulted in reduced release of aluminum ions from electroporation cuvettes.
Topics: Aluminum; Animals; Antineoplastic Agents; Apoptosis; Bleomycin; CHO Cells; Cell Survival; Cisplatin; Cricetulus; Electrochemotherapy; Electrodes; Neoplasms
PubMed: 33743336
DOI: 10.1016/j.bioelechem.2021.107798 -
Biochemistry Nov 2014The bleomycin (BLM) family of glycopeptide-derived antitumor antibiotics consists of BLMs, tallysomycins (TLMs), phleomycins (PLMs), and zorbamycin (ZBM). The...
The bleomycin (BLM) family of glycopeptide-derived antitumor antibiotics consists of BLMs, tallysomycins (TLMs), phleomycins (PLMs), and zorbamycin (ZBM). The self-resistant elements BlmB and TlmB, discovered from the BLM- and TLM-producing organisms Streptomyces verticillus ATCC15003 and Streptoalloteichus hindustanus E465-94 ATCC31158, respectively, are N-acetyltransferases that provide resistance to the producers by disrupting the metal-binding domain of the antibiotics required for activity. Although each member of the BLM family of antibiotics possesses a conserved metal-binding domain, the structural differences between each member, namely, the bithiazole moiety and C-terminal amine of BLMs, have been suggested to instill substrate specificity within BlmB. Here we report that BlmB and TlmB readily accept and acetylate BLMs, TLMs, PLMs, and ZBM in vitro but only in the metal-free forms. Kinetic analysis of BlmB and TlmB reveals there is no strong preference or rate enhancement for specific substrates, indicating that the structural differences between each member of the BLM family play a negligible role in substrate recognition, binding, or catalysis. Intriguingly, the zbm gene cluster from Streptomyces flavoviridis ATCC21892 does not contain an N-acetyltransferase, yet ZBM is readily acetylated by BlmB and TlmB. We subsequently established that S. flavoviridis lacks the homologue of BlmB and TlmB, and ZbmA, the ZBM-binding protein, alone is sufficient to provide ZBM resistance. We further confirmed that BlmB can indeed confer resistance to ZBM in vivo in S. flavoviridis, introduction of which into wild-type S. flavoviridis further increases the level of resistance.
Topics: Acetylation; Acetyltransferases; Anti-Bacterial Agents; Bacterial Proteins; Bleomycin; Drug Resistance, Bacterial; Kinetics; Microbial Sensitivity Tests; Streptomyces; Substrate Specificity
PubMed: 25299801
DOI: 10.1021/bi501121e -
JPMA. the Journal of the Pakistan... Oct 2023To evaluate the effectiveness of intralesional bleomycin in comparison to cryotherapy in the treatment of cutaneous warts. (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
To evaluate the effectiveness of intralesional bleomycin in comparison to cryotherapy in the treatment of cutaneous warts.
METHODS
The randomized controlled trial was conducted at the Department of Dermatology, Jinnah Postgraduate Medical Centre, Karachi, from January to July 2021, and comprised patients of either gender aged 18-60 years who had cutaneous warts for 1-48 weeks. The subjects were divided into group A treated with 0.1% intralesional bleomycin, and group B were treated with cryotherapy through non probability consecutive sampling. Follow-up examination was done at the 2, 4 and 6 weeks. Data was analysed using SPSS 23.
RESULTS
Of the 154 patients, 96(62.3%) were male and 58(37.7%) were females. The overall mean age was 33.253±6.726 years. There were 77(50%) patients in each of the two groups. The therapy after 6 weeks was found to be effective 73(94.8%). group A patients and 57(74%) group B patients (p=0.001).
CONCLUSIONS
Intralesional bleomycin was found to be more successful than cryotherapy in the treatment of cutaneous warts.
Topics: Female; Humans; Male; Adult; Bleomycin; Warts; Cryotherapy; Injections, Intralesional; Hospitals; Treatment Outcome
PubMed: 37876049
DOI: 10.47391/JPMA.6372 -
IEEE Transactions on Bio-medical... Nov 2022This paper presents a 40 GHz microwave biosensor used to monitor and characterize single cells (THP-1) subjected to electrochemotherapy and obtain an electronic...
This paper presents a 40 GHz microwave biosensor used to monitor and characterize single cells (THP-1) subjected to electrochemotherapy and obtain an electronic signature of the treatment efficiency. This biosensor proposes a non-destructive and label-free technique that first allows, with the rapid measurement of single untreated cells in their culture medium, the extraction of two frequency-dependent dielectric parameters, the capacitance (C (f)) and the conductance (G (f)). Second, this technique can powerfully reveal the effects of a chemical membrane permeabilizing treatment (Saponin). At last, it permits us to detect, and predict, the potentiation of a molecule classically used in chemotherapy (Bleomycin) when combined with the application of electric pulses (principle of electrochemotherapy). Treatment-affected cells show a decrease in the capacitive and conductive contrasts, indicating damages at the cellular levels. Along with these results, classical biological tests are conducted. Statistical analysis points out a high correlation rate (R>0.97), which clearly reveals the reliability and efficacy of our technique and makes it an attractive technique for biology related researches and personalized medicine.
Topics: Electrochemotherapy; Microwaves; Reproducibility of Results; Bleomycin; Biosensing Techniques; Saponins
PubMed: 35471894
DOI: 10.1109/TBME.2022.3170267 -
Translational Research : the Journal of... Jul 2023Dysregulation of type II alveolar epithelial cells (AECII) plays a vital role in the initiation and development of pulmonary fibrosis (PF). Dachshund homolog 1 (Dach1),...
Dysregulation of type II alveolar epithelial cells (AECII) plays a vital role in the initiation and development of pulmonary fibrosis (PF). Dachshund homolog 1 (Dach1), frequently expressed in epithelial cells with stem cell potential, controls cell proliferation, apoptosis, and cell cycle in tissue development and disease process. In this study, we demonstrated that the lungs collected from PF patients and mice of Bleomycin (BLM)-treated were characterized by low expression of Dachshund homolog 1 (Dach1), especially in AECII. Dach1 deficiency in the alveolar epithelium exacerbated PF in BLM-treated mice, as evidenced by reduced pulmonary function and increased expression of fibrosis markers. Rather, treatment with lung-specific overexpression of Dach1 alleviated histopathological damage, lung compliance, and fibrosis in BLM-treated mice. Moreover, overexpression of Dach1 could inhibit epithelial apoptosis in vitro. Conversely, primary AECII with Dach1 depletion were more susceptible to apoptosis in vivo. Mechanically, Dach1 combined with C-Jun protooncogene selectively bound to the promoter of B-cell lymphoma 2 interacting mediators of cell death (Bim), by which it repressed Bim expression and alleviated epithelial apoptosis. Taken together, our data support that Dach1 in AECII contributes to the progression of PF and may be a viable target for the prevention and treatment of PF.
Topics: Animals; Mice; Alveolar Epithelial Cells; Apoptosis; Bleomycin; Epithelium; Eye Proteins; Lung; Pulmonary Fibrosis
PubMed: 36754276
DOI: 10.1016/j.trsl.2023.01.006