FASEB bioAdvances最新文献

{"title":"Intestinal barrier dysfunction in murine sickle cell disease is associated with small intestine neutrophilic inflammation, oxidative stress, and dysbiosis","authors":"Caitlin V. Lewis,&nbsp;Hassan Sellak,&nbsp;Mariem A. Sawan,&nbsp;Giji Joseph,&nbsp;Trevor M. Darby,&nbsp;David VanInsberghe,&nbsp;Crystal R. Naudin,&nbsp;David R. Archer,&nbsp;Rheinallt M. Jones,&nbsp;W. Robert Taylor","doi":"10.1096/fba.2022-00121","DOIUrl":"10.1096/fba.2022-00121","url":null,"abstract":"<p>The intestinal microbiome has emerged as a potential contributor to the severity of sickle cell disease (SCD). We sought to determine whether SCD mice exhibit intestinal barrier dysfunction, inflammation, and dysbiosis. Using the Townes humanized sickle cell mouse model, we found a 3-fold increase in intestinal permeability as assessed via FITC-dextran (4 kDa) assay in SS (SCD) mice compared to AA (wild type) mice (<i>n</i> = 4, <i>p</i> &lt; 0.05). This was associated with 25 to 50% decreases in claudin-1, 3, and 15 and zonula occludens-1 gene expression (<i>n</i> = 8–10, <i>p</i> &lt; 0.05) in the small intestine. Increased Ly6G staining demonstrated more neutrophils in the SS small intestine (3-fold, <i>n</i> = 5, <i>p</i> &lt; 0.05) associated with increased expression of TNFα, IL-17A, CXCL1, and CD68 (2.5 to 5-fold, <i>n</i> = 7–10, <i>p</i> &lt; 0.05). In addition, we observed 30 to 55% decreases in superoxide dismutase-1, glutathione peroxidase-1, and catalase antioxidant enzyme expression (<i>n</i> = 7–8, <i>p</i> &lt; 0.05) concomitant to an increase in superoxide (2-fold, <i>n</i> = 4, <i>p</i> &lt; 0.05). Importantly, all significant observations of a leaky gut phenotype and inflammation were limited to the small intestine and not observed in the colon. Finally, characterization of the composition of the microbiome within the small intestine revealed dysbiosis in SS mice compared to their AA littermates with 47 phyla to species-level significant alterations in amplicon sequence variants. We conclude that the intestinal barrier is compromised in SCD, associated with decreased gene expression of tight junction proteins, enhanced inflammation, oxidative stress, and gut microbiome dysbiosis, all specific to the small intestine.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 5","pages":"199-210"},"PeriodicalIF":2.7,"publicationDate":"2023-02-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/e8/e6/FBA2-5-199.PMC10158626.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9423030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oxidative stress induces MUC5AC expression through mitochondrial damage-dependent STING signaling in human bronchial epithelial cells","authors":"Yutaka Nishida,&nbsp;Hisako Yagi,&nbsp;Masaya Ota,&nbsp;Atsushi Tanaka,&nbsp;Koichiro Sato,&nbsp;Takaharu Inoue,&nbsp;Satoshi Yamada,&nbsp;Naoya Arakawa,&nbsp;Takashi Ishige,&nbsp;Yasuko Kobayashi,&nbsp;Hirokazu Arakawa,&nbsp;Takumi Takizawa","doi":"10.1096/fba.2022-00081","DOIUrl":"https://doi.org/10.1096/fba.2022-00081","url":null,"abstract":"<p>Oxidative stress increases the production of the predominant mucin MUC5AC in airway epithelial cells and is implicated in the pathogenesis of bronchial asthma and chronic obstructive pulmonary disease. Oxidative stress impairs mitochondria, releasing mitochondrial DNA into the cytoplasm and inducing inflammation through the intracytoplasmic DNA sensor STING (stimulator of interferon genes). However, the role of innate immunity in mucin production remains unknown. We aimed to elucidate the role of innate immunity in mucin production in airway epithelial cells under oxidative stress. Human airway epithelial cell line (NCI-H292) and normal human bronchial epithelial cells were used to confirm <i>MUC5AC</i> expression levels by real-time PCR when stimulated with hydrogen peroxide (H₂O₂). <i>MUC5AC</i> transcriptional activity was increased and mitochondrial DNA was released into the cytosol by H₂O₂. Mitochondrial antioxidants were used to confirm the effects of mitochondrial oxidative stress where antioxidants inhibited the increase in <i>MUC5AC</i> transcriptional activity. Cyclic GMP-AMP synthase (cGAS) or STING knockout (KO) cells were generated to investigate their involvement. H₂O₂-induced <i>MUC5AC</i> expression was suppressed in STING KO cells, but not in cGAS KO cells. The epidermal growth factor receptor was comparably expressed in STING KO and wild-type cells. Thus, mitochondria and STING play important roles in mucin production in response to oxidative stress in airway epithelial cells.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 4","pages":"171-181"},"PeriodicalIF":2.7,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00081","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50116087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"B cell-dependent subtypes and treatment-based immune correlates to survival in stage 3 and 4 lung adenocarcinomas","authors":"Susan Raju Paul,&nbsp;Ivan Valiev,&nbsp;Skylar E. Korek,&nbsp;Vladimir Zyrin,&nbsp;Diana Shamsutdinova,&nbsp;Olga Gancharova,&nbsp;Alexander Zaitsev,&nbsp;Ekaterina Nuzhdina,&nbsp;Diane L. Davies,&nbsp;Ibiayi Dagogo-Jack,&nbsp;Felix Frenkel,&nbsp;Jessica H. Brown,&nbsp;Joshua M. Hess,&nbsp;Sarah Viet,&nbsp;Jason L. Petersen,&nbsp;Cameron D. Wright,&nbsp;Harald C. Ott,&nbsp;Hugh G. Auchincloss,&nbsp;Ashok Muniappan,&nbsp;Toshihiro Shioda,&nbsp;Michael Lanuti,&nbsp;Christel M. Davis,&nbsp;Erik A. Ehli,&nbsp;Yin P. Hung,&nbsp;Mari Mino-Kenudson,&nbsp;Maria Tsiper,&nbsp;Ann E. Sluder,&nbsp;Patrick M. Reeves,&nbsp;Nikita Kotlov,&nbsp;Alexander Bagaev,&nbsp;Ravshan Ataullakhanov,&nbsp;Mark C. Poznansky","doi":"10.1096/fba.2023-00009","DOIUrl":"10.1096/fba.2023-00009","url":null,"abstract":"<p>Lung cancer is the leading cause of cancer-related deaths worldwide. Surgery and chemoradiation are the standard of care in early stages of non-small cell lung cancer (NSCLC), while immunotherapy is the standard of care in late-stage NSCLC. The immune composition of the tumor microenvironment (TME) is recognized as an indicator for responsiveness to immunotherapy, although much remains unknown about its role in responsiveness to surgery or chemoradiation. In this pilot study, we characterized the NSCLC TME using mass cytometry (CyTOF) and bulk RNA sequencing (RNA-Seq) with deconvolution of RNA-Seq being performed by Kassandra, a recently published deconvolution tool. Stratification of patients based on the intratumoral abundance of B cells identified that the B-cell rich patient group had increased expression of CXCL13 and greater abundance of PD1⁺ CD8 T cells. The presence of B cells and PD1⁺ CD8 T cells correlated positively with the presence of intratumoral tertiary lymphoid structures (TLS). We then assessed the predictive and prognostic utility of these cell types and TLS within publicly available stage 3 and 4 lung adenocarcinoma (LUAD) RNA-Seq datasets. As previously described by others, pre-treatment expression of intratumoral 12-chemokine TLS gene signature is associated with progression free survival (PFS) in patients who receive treatment with immune checkpoint inhibitors (ICI). Notably and unexpectedly pre-treatment percentages of intratumoral B cells are associated with PFS in patients who receive surgery, chemotherapy, or radiation. Further studies to confirm these findings would allow for more effective patient selection for both ICI and non-ICI treatments.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 4","pages":"156-170"},"PeriodicalIF":2.7,"publicationDate":"2023-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ed/74/FBA2-5-156.PMC10068771.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9250895","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Research productivity and training support for doctoral students in the biological and biomedical sciences","authors":"Michael D. Schaller","doi":"10.1096/fba.2022-00117","DOIUrl":"10.1096/fba.2022-00117","url":null,"abstract":"<p>Training of doctoral students as part of the next generation of the biomedical workforce is essential for sustaining the scientific enterprise in the United States. Training primarily occurs at institutions of higher education, and these trainees comprise an important part of the workforce at these institutions. Federal investment in the support of doctoral students in the biological and biomedical sciences is distributed differently than the distribution of students across different types of institutions, for example, public vs private. Institutions in states that historically receive less federal support for research also receive less support for doctoral student training. Doctorates at different types of institution exhibit little difference in research productivity, with the exception of citations, and subsequent receipt of additional NIH awards. Thus, training outcomes, which are related to the quality of the student and training environment, are similar across different institutions. Research productivity of doctoral students does not correlate with the number of F31s awarded to an institution. Factors that correlate with F31 funding include R01 funding levels and program size. The findings suggest strategies for institutions to increase success at securing F31s and modification of policy to promote more equitable distribution of F31s across institutions.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 3","pages":"131-148"},"PeriodicalIF":2.7,"publicationDate":"2023-01-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/12/11/FBA2-5-131.PMC9983073.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10849731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Erratum to “The roles of HMGB1-produced DNA gaps in DNA protection and aging biomarker reversal”","authors":"","doi":"10.1096/fba.1365","DOIUrl":"10.1096/fba.1365","url":null,"abstract":"<p>This erratum corrects the following:</p><p>Yasom, S., Watcharanurak, P., Bhummaphan, N., Thongsroy, J., Puttipanyalears, C., Settayanon, S., Chalertpet, K., Khumsri, W., Kongkaew, A., Patchsung, M., Siriwattanakankul, C., Pongpanich, M., Pin-on, P., Jindatip, D., Wanotayan, R., Odton, M., Supasai, S., Oo, T. T., Arunsak, B., Pratchayasakul, W., Chattipakorn, N., Chattipakorn, S., Mutirangura, A. (First published March 9, 2022) The roles of HMGB1-produced DNA gaps in DNA protection and aging biomarker reversal. <i>FASEB BioAdvances</i>. 2022;4:408–434. doi: 10.1096/fba.2021-00131</p><p>The authors report that inadvertent errors were made in assembling two of the figures submitted for publication. In both Figures 8 and 9, the left-hand row labels “Liver 1” to “Liver 4” are misleading and should not have been inserted. In Figure 8, the images in the 7m PC column should be the same as those shown in the 7m PC column in Figure 9, as the same group of rats is represented in both figures. In Figure 9, a different view of the image in row two of the 30m PC column was inadvertently placed in row four of the 30m PC column. The authors apologize for these oversights and for any confusion caused by the errors. These errors do not affect the results and conclusions reported in the article.</p><p>The correct versions of Figures 8 and 9 are as follows:</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 2","pages":"85-87"},"PeriodicalIF":2.7,"publicationDate":"2023-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://faseb.onlinelibrary.wiley.com/doi/epdf/10.1096/fba.1365","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10767158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of various buffers and weak bases on lysosomal and intracellular pH: Implications for infectivity of SARS-CoV-2","authors":"Jeffrey A. Kraut,&nbsp;Izaak J. Cheetham-Wilkinson,&nbsp;Laura E. Swan,&nbsp;Massimiliano Stagi,&nbsp;Ira Kurtz","doi":"10.1096/fba.2022-00062","DOIUrl":"10.1096/fba.2022-00062","url":null,"abstract":"<p>Acidification of the cellular lysosome is an important factor in infection of mammalian cells by SARS-CoV-2. Therefore, raising the pH of the lysosome would theoretically be beneficial in prevention or treatment of SARS-CoV-2 infection. Sodium bicarbonate, carbicarb, and THAM are buffers that can be used clinically to provide base to patients. To examine whether these bases could raise lysosomal pH and therefore be a primary or adjunctive treatment of SARS-CoV-2 infection, we measured lysosomal and intracellular pH of mammalian cells after exposure to each of these bases. Mammalian HEK293 cells expressing RpH-LAMP1-3xFLAG, a ratiometric sensor of lysosomal luminal pH, were first exposed to Hepes which was then switched to sodium bicarbonate, carbicarb, or THAM and lysosomal pH measured. In bicarbonate buffer the mean lysosomal pH was 4.3 ± 0.1 (<i>n =</i> 20); <i>p =</i> NS versus Hepes (<i>n =</i> 20). The mean lysosomal pH in bicarbonate/carbonate was 4.3 ± 0.1 (<i>n =</i> 21) versus Hepes (<i>n =</i> 21), <i>p =</i> NS. In THAM buffer the mean lysosomal pH was 4.7 ± 0.07 (<i>n =</i> 20) versus Hepes (4.6 ± 0.1, <i>n =</i> 20), <i>p =</i> NS. In addition, there was no statistical difference between pH_i in bicarbonate, carbicarb or THAM solutions. Using the membrane permeable base NH₄Cl (5 mM), lysosomal pH increased significantly to 5.9 ± 0.1 (<i>n =</i> 21) compared to Hepes (4.5 ± 0.07, <i>n =</i> 21); <i>p</i> &lt; 0.0001. Similarly, exposure to 1 mM hydroxychloroquine significantly increased the lysosomal pH to (5.9 ± 0.06, <i>n =</i> 20) versus Hepes (4.3 ± 0.1, <i>n =</i> 20), <i>p</i> &lt; 0.0001. Separately steady-state pHi was measured in HEK293 cells bathed in various buffers. In bicarbonate pH_i was 7.29 ± 0.02 (<i>n =</i> 12) versus Hepes (7.45 ± 0.03, [<i>n =</i> 12]), <i>p</i> &lt; 0.001. In cells bathed in carbicarb pH_i was 7.27 ± 0.02 (<i>n =</i> 5) versus Hepes (7.43 ± 0.04, [<i>n =</i> 5]), <i>p</i> &lt; 0.01. Cells bathed in THAM had a pH_i of 7.25 ± 0.03 (<i>n =</i> 12) versus Hepes (7.44 ± 0.03 [<i>n =</i> 12]), <i>p</i> &lt; 0.001. In addition, there was no statistical difference in pH_i in bicarbonate, carbicarb or THAM solutions. The results of these studies indicate that none of the buffers designed to provide base to patients alters lysosomal pH at the concentrations used in this study and therefore would be predicted to be of no value in the treatment of SARS-CoV-2 infection. If the goal is to raise lysosomal pH to decrease the infectivity of SARS-CoV-2, utilizing lysosomal permeable buffers at the appropriate dose that is non-toxic appears to be a useful approach to explore.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 4","pages":"149-155"},"PeriodicalIF":2.7,"publicationDate":"2023-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://faseb.onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00062","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9257153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exosomes from tubular epithelial cells undergoing epithelial-to-mesenchymal transition promote renal fibrosis by M1 macrophage activation","authors":"Yuqing Lu,&nbsp;Rui Zhang,&nbsp;Xiameng Gu,&nbsp;Xuerong Wang,&nbsp;Peipei Xi,&nbsp;Xiaolan Chen","doi":"10.1096/fba.2022-00080","DOIUrl":"10.1096/fba.2022-00080","url":null,"abstract":"<p>Kidney fibrosis is the common final pathway of chronic kidney disease (CKD), and it is distinguished by inflammation, mesenchymal transition with myofibroblast formation, and epithelial-to-mesenchymal transition (EMT). Macrophages are protuberant inflammatory cells in the kidney, and their roles are dependent on their phenotypes. However, it remains unclear whether tubular epithelial cells (TECs) undergoing EMT can influence the phenotypes of macrophages and the underlying mechanisms during the development of kidney fibrosis. Here, we investigated the characteristics of TECs and macrophages during kidney fibrosis with a focus on EMT and inflammation. We found that the coculture of exosomes from transforming growth factor-beta (TGF-β)-induced TECs with macrophages induced macrophage M1 polarization, while exosomes from TECs without TGF-β stimulation or stimulation with TGF-β alone did not induce an increase in M1 macrophage-related markers. Notably, TECs induced to undergo EMT by TGF-β treatment released more exosomes than the other groups. Furthermore, it is noteworthy that when we injected exosomes from TECs undergoing EMT into mice, in addition to the high level of inflammatory response and the activation of M1 macrophages, the indicators of EMT and renal fibrosis in mouse kidney tissue were correspondingly elevated. In summary, exosomes from TECs undergoing EMT by TGF-β treatment induced M1 polarization and led to a positive feedback effect for further EMT and the development of renal fibrosis. Therefore, the obstacle to the release of such exosomes may be a novel therapeutic strategy for CKD.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 3","pages":"101-113"},"PeriodicalIF":2.7,"publicationDate":"2023-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://faseb.onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00080","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10849728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analyzing the interactome of human CK2β in prostate carcinoma cells reveals HSP70-1 and Rho guanin nucleotide exchange factor 12 as novel interaction partners","authors":"Anna Nickelsen,&nbsp;Claudia Götz,&nbsp;Florian Lenz,&nbsp;Karsten Niefind,&nbsp;Simone König,&nbsp;Joachim Jose","doi":"10.1096/fba.2022-00098","DOIUrl":"10.1096/fba.2022-00098","url":null,"abstract":"<p>CK2β is the non-catalytic modulating part of the S/T-protein kinase CK2. However, the overall function of CK2β is poorly understood. Here, we report on the identification of 38 new interaction partners of the human CK2β from lysates of DU145 prostate cancer cells using photo-crosslinking and mass spectrometry, whereby HSP70-1 was identified with high abundance. The K_D value of its interaction with CK2β was determined as 0.57 μM by microscale thermophoresis, this being the first time, to our knowledge, that a K_D value of CK2β with another protein than CK2α or CK2α′ was quantified. Phosphorylation studies excluded HSP70-1 as a substrate or activity modulator of CK2, suggesting a CK2 activity independent interaction of HSP70-1 with CK2β. Co-immunoprecipitation experiments in three different cancer cell lines confirmed the interaction of HSP70-1 with CK2β in vivo. A second identified CK2β interaction partner was Rho guanin nucleotide exchange factor 12, indicating an involvement of CK2β in the Rho-GTPase signal pathway, described here for the first time to our knowledge. This points to a role of CK2β in the interaction network affecting the organization of the cytoskeleton.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 3","pages":"114-130"},"PeriodicalIF":2.7,"publicationDate":"2023-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ca/dc/FBA2-5-114.PMC9983076.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10849733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nobiletin, a NF-κB signaling antagonist, promotes BMP-induced bone formation","authors":"Thira Rojasawasthien,&nbsp;Michihiko Usui,&nbsp;William N. Addison,&nbsp;Takuma Matsubara,&nbsp;Tomohiko Shirakawa,&nbsp;Toshiyuki Tsujisawa,&nbsp;Keisuke Nakashima,&nbsp;Shoichiro Kokabu","doi":"10.1096/fba.2022-00093","DOIUrl":"10.1096/fba.2022-00093","url":null,"abstract":"<p>The NF-κB family of transcription factors plays an important role in skeletal development and bone homeostasis. In osteoblast cells, NF-κB signaling has been shown to suppress survival, proliferation, and differentiation. Furthermore, pharmacological suppression of NF-κB enhances osteoblast differentiation and bone formation. Thus, NF-κB antagonists are promising candidates as anabolic agents for enhancing bone mass. In this study, we describe the mechanism by which nobiletin, an inhibitor of NF-κB activity, regulates osteoblast differentiation and mineralization. We found that in MC3T3-E1 osteoblast cells, nobiletin inhibited a TNF-α responsive NF-κB luciferase reporter and also decreased the induction of classical NF-κB target genes by TNF-α. Consistent with this, nobiletin prevented TNF-α -mediated suppression of osteogenesis and potently enhanced the differentiation and mineralization of MC3T3-E1 cells. Likewise, in an in vivo BMP2-induced ectopic bone formation assay, nobiletin markedly enhanced ossicle bone volume. Western blotting and SMAD-responsive luciferase assays also demonstrated that NF-κB suppression of BMP signaling could be inhibited by nobiletin. Thus, our data suggest that mechanistically, nobiletin prevents the endogenous repression of BMP signaling by TNF-α, thereby enhancing osteoblast activity. In conclusion, nobiletin is a novel NF-κB antagonist that may be a useful anabolic agent for bone formation.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 2","pages":"62-70"},"PeriodicalIF":2.7,"publicationDate":"2022-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ac/20/FBA2-5-62.PMC9927861.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9328835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the potential of human cultured nasal epithelial cell sheets to differentiate into airway epithelium","authors":"Yoshiyuki Kasai,&nbsp;Tsunetaro Morino,&nbsp;Tsuguhisa Nakayama,&nbsp;Kazuhisa Yamamoto,&nbsp;Hiromi Kojima","doi":"10.1096/fba.2022-00106","DOIUrl":"10.1096/fba.2022-00106","url":null,"abstract":"<p>Understanding the expected efficacy and safety of a new regenerative therapy requires analysis of the fate of the transplanted cell graft. We have shown that transplantation of autologous cultured nasal epithelial cell sheets onto the middle ear mucosa can improve middle ear aeration and hearing. However, it remains unknown whether cultured nasal epithelial cell sheets have the potential to gain mucociliary function in the environment of the middle ear because sampling cell sheets after transplantation is challenging. The present study re-cultured cultured nasal epithelial cell sheets in different culture media and evaluated whether the sheets have the potential to differentiate into airway epithelium. Before re-cultivation, cultured nasal epithelial cell sheets fabricated in keratinocyte culture medium (KCM) contained no FOXJ1-positive and acetyl-α-tubulin-positive multiciliated cells or MUC5AC-positive mucus cells. Interestingly, multiciliated cells and mucus cells were observed when the cultured nasal epithelial cell sheets were re-cultured in conditions that promote differentiation of airway epithelium. However, multiciliated cells, mucus cells and CK1-positive keratinized cells were not observed when cultured nasal epithelial cell sheets were re-cultured in conditions that promote epithelial keratinization. These findings support the suggestion that cultured nasal epithelial cell sheets have the ability to differentiate and gain mucociliary function in response to an appropriate environment (possibly including the environment found in the middle ear) but are unable to develop into an epithelial type that differs from its origins.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 3","pages":"89-100"},"PeriodicalIF":2.7,"publicationDate":"2022-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://faseb.onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00106","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10849734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}