FASEB bioAdvances最新文献

{"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}

{"title":"Excitation and contraction of cardiac muscle and coronary arteries of brain-dead pigs","authors":"Per Arlock,&nbsp;Mei Li,&nbsp;Benjamin Davis,&nbsp;Cecilia Lövdahl,&nbsp;Qiuming Liao,&nbsp;Trygve Sjöberg,&nbsp;Awahan Rahman,&nbsp;Björn Wohlfart,&nbsp;Stig Steen,&nbsp;Anders Arner","doi":"10.1096/fba.2022-00104","DOIUrl":"10.1096/fba.2022-00104","url":null,"abstract":"<p>Excitability and contraction of cardiac muscle from brain-dead donors critically influence the success of heart transplantation. Membrane physiology, Ca²⁺-handling, and force production of cardiac muscle and the contractile properties of coronary arteries were studied in hearts of brain-dead pigs. Cardiac muscle and vascular function after 12 h brain death (decapitation between C2 and C3) were compared with properties of fresh tissue. In both isolated cardiomyocytes (whole-cell patch clamp) and trabecular muscle (conventional microelectrodes), action potential duration was shorter in brain dead, compared to controls. Cellular shortening and Ca²⁺ transients were attenuated in the brain dead, and linked to lower mRNA expression of L-type calcium channels and a slightly lower I_Ca,_L, current, as well as to a lower expression of phospholamban. The current–voltage relationship and the current above the equilibrium potential of the inward K⁺ (I_K1) channel were altered in the brain-dead group, associated with lower mRNA expression of the Kir2.2 channel. Delayed K⁺ currents were detected (I_Kr, I_Ks) and were not different between groups. The transient outward K⁺ current (I_to) was not observed in the pig heart. Coronary arteries exhibited increased contractility and sensitivity to the thromboxane analogue (U46619), and unaltered endothelial relaxation. In conclusion, brain death involves changes in cardiac cellular excitation which might lower contractility after transplantation. Changes in the inward rectifier K⁺ channel can be associated with an increased risk for arrhythmia. Increased reactivity of coronary arteries may lead to increased risk of vascular spasm, although endothelial relaxant function was well preserved.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 2","pages":"71-84"},"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.2022-00104","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9328834","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":"Cell migration is impaired in XPA-deficient cells","authors":"Seiji Takeuchi,&nbsp;Takeshi Fukumoto,&nbsp;Chihiro Takemori,&nbsp;Naoaki Saito,&nbsp;Chikako Nishigori,&nbsp;Makoto Sato","doi":"10.1096/fba.2022-00084","DOIUrl":"10.1096/fba.2022-00084","url":null,"abstract":"<p>Xeroderma pigmentosum (XP) is a hereditary disorder characterized by photosensitivity, predisposition to skin cancers, and neurological abnormalities including microcephaly and progressive neurodegeneration. A lack of nucleotide excision repair (NER) in patients with XP can cause hypersensitivity to the sun, leading to skin cancer, whereas the etiology of the neuronal symptoms of XP remains ambiguous. There are various neurological disorders that perturb neuronal migration, causing mislocalization and disorganization of the cortical lamination. Here, we investigated the role of the XP group-A (<i>Xpa</i>) gene in directed cell migration. First, we adopted an in utero electroporation method to transduce shRNA vectors into the murine embryonic cerebral cortex for the in vivo knockdown of <i>Xpa</i>. <i>Xpa-</i>knockdown neurons in the embryonic cerebral cortex showed abnormal cell migration, cell cycle exit, and differentiation. The genotype–phenotype relationship between the lack of XPA and cell migration abnormalities was confirmed using both a scratch assay and time-lapse microscopy in XP-A patient-derived fibroblasts. Unlike healthy cells, these cells showed impairment in overall mobility and the direction of motility. Therefore, abnormal cell migration may explain neural tissue abnormalities in patients with XP-A.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 2","pages":"53-61"},"PeriodicalIF":2.7,"publicationDate":"2022-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/61/1b/FBA2-5-53.PMC9927838.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9328838","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":"Regulation of 5-fluorodeoxyuridine monophosphate-thymidylate synthase ternary complex levels by autophagy confers resistance to 5-fluorouracil","authors":"Nana Nishizawa,&nbsp;Chinatsu Kurasaka,&nbsp;Yoko Ogino,&nbsp;Akira Sato","doi":"10.1096/fba.2022-00099","DOIUrl":"10.1096/fba.2022-00099","url":null,"abstract":"<p>5-Fluorouracil (5-FU) is a cornerstone drug used to treat colorectal cancer (CRC). However, the prolonged exposure of CRC cells to 5-FU results in acquired resistance. We have previously demonstrated that levels of the 5-fluorodeoxyuridylate (FdUMP) covalent complex with thymidylate synthase (FdUMP-TS) and free-TS (native enzyme) are higher in 5-FU-resistant CRC cells than in the parental cell line (HCT116). Accordingly, resistant cells may have an efficient system for trapping and removing FdUMP-TS, thus imparting resistance. In this study, using a model of 5-FU-resistant CRC cells generated by repeated exposure, the role of autophagy in the elimination of FdUMP-TS in resistant cells was investigated. The resistant cells showed greater sensitivity to autophagy inhibitors than that of parental cells. Autophagy inhibition increased 5-FU cytotoxicity more substantially in resistant cells than in parental cells. Furthermore, autophagy inhibition increased FdUMP-TS protein accumulation in resistant cells. Our findings suggest that resistance to 5-FU is mediated by autophagy as a system to eliminate FdUMP-TS and may guide the use and optimization of combination therapies involving autophagy inhibitors.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 1","pages":"43-51"},"PeriodicalIF":2.7,"publicationDate":"2022-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://faseb.onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00099","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10589587","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":"FtsZ phosphorylation brings about growth arrest upon DNA damage in Deinococcus radiodurans","authors":"Reema Chaudhary,&nbsp;Shruti Mishra,&nbsp;Ganesh K. Maurya,&nbsp;Yogendra S. Rajpurohit,&nbsp;Hari S. Misra","doi":"10.1096/fba.2022-00082","DOIUrl":"https://doi.org/10.1096/fba.2022-00082","url":null,"abstract":"<p>The polymerization/depolymerization dynamics of FtsZ play a pivotal role in cell division in the majority of the bacteria. <i>Deinococcus radiodurans</i>, a radiation-resistant bacterium, shows an arrest of growth in response to DNA damage with no change in the level of FtsZ. This bacterium does not deploy LexA/RecA type of DNA damage response and cell cycle regulation, and its genome does not encode SulA homologues of <i>Escherichia coli</i>, which attenuate FtsZ functions in response to DNA damage in other bacteria. A radiation-responsive Ser/Thr quinoprotein kinase (RqkA), characterized for its role in radiation resistance in this bacterium, could phosphorylate several cognate proteins, including FtsZ (drFtsZ) at Serine 235 (S235) and Serine 335 (S335) residues. Here, we reported the detailed characterization of S235 and S335 phosphorylation effects in the regulation of drFtsZ functions and demonstrated that the phospho-mimetic replacements of these residues in drFtsZ had grossly affected its functions that could result in cell cycle arrest in response to DNA damage in <i>D. radiodurans</i>. Interestingly, the phospho-ablative replacements were found to be nearly similar to drFtsZ, whereas the phospho-mimetic mutant lost the wild-type protein's signature characteristics, including its dynamics under normal conditions. The kinetics of post-bleaching recovery for drFtsZ and phospho-mimetic mutant were nearly similar at 2 h post-irradiation recovery but were found to be different under normal conditions. These results highlighted the role of S/T phosphorylation in the regulation of drFtsZ functions and cell cycle arrest in response to DNA damage, which is demonstrated for the first time, in any bacteria.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 1","pages":"27-42"},"PeriodicalIF":2.7,"publicationDate":"2022-10-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fba.2022-00082","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50138241","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":"Early developmental phenotypes in the cystic fibrosis sheep model","authors":"Arnaud J. Van Wettere,&nbsp;Shih-Hsing Leir,&nbsp;Calvin U. Cotton,&nbsp;Misha Regouski,&nbsp;Iuri Viotti Perisse,&nbsp;Jenny L. Kerschner,&nbsp;Alekh Paranjapye,&nbsp;Zhiqiang Fan,&nbsp;Ying Liu,&nbsp;Makayla Schacht,&nbsp;Kenneth L. White,&nbsp;Irina A. Polejaeva,&nbsp;Ann Harris","doi":"10.1096/fba.2022-00085","DOIUrl":"10.1096/fba.2022-00085","url":null,"abstract":"<p>Highly effective modulator therapies for cystic fibrosis (CF) make it a treatable condition for many people. However, although CF respiratory illness occurs after birth, other organ systems particularly in the digestive tract are damaged before birth. We use an ovine model of CF to investigate the in utero origins of CF disease since the sheep closely mirrors critical aspects of human development. Wildtype (WT) and <i>CFTR</i> ^-/- sheep tissues were collected at 50, 65, 80, 100, and 120 days of gestation and term (147 days) and used for histological, electrophysiological, and molecular analysis. Histological abnormalities are evident in <i>CFTR-/-</i> ^-/-  animals by 80 days of gestation, equivalent to 21 weeks in humans. Acinar and ductal dilation, mucus obstruction, and fibrosis are observed in the pancreas; biliary fibrosis, cholestasis, and gallbladder hypoplasia in the liver; and intestinal meconium obstruction, as seen at birth in all large animal models of CF. Concurrently, cystic fibrosis transmembrane conductance regulator (CFTR)-dependent short circuit current is present in WT tracheal epithelium by 80 days gestation and is absent from <i>CFTR</i> ^-/- tissues. Transcriptomic profiles of tracheal tissues confirm the early expression of <i>CFTR</i> and suggest that its loss does not globally impair tracheal differentiation.</p>","PeriodicalId":12093,"journal":{"name":"FASEB bioAdvances","volume":"5 1","pages":"13-26"},"PeriodicalIF":2.7,"publicationDate":"2022-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/54/b9/FBA2-5-13.PMC9832529.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10535609","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}