FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W3165161784> ?p ?o ?g. }

• W3165161784 endingPage "152" @default.
• W3165161784 startingPage "151.2" @default.
• W3165161784 abstract "Background: Traditional preclinical approaches, such as two-dimensional cell culture and animal models, are often inadequate to mimic the pathophysiological features of complex diseases such as systemic sclerosis (SSc). Human specific targets, such as the recently described pro-fibrotic long non coding RNA (lncRNA) H19X 1 , are becoming increasingly relevant in preclinical research, creating the need of new strategies and tools in translational medicine. The employment of novel three-dimensional (3D) culture systems, where multiple cell types are included, is filling an important gap left by the traditional preclinical methods. Objectives: To develop an easy to produce 3D fibrotic skin microtissues model for translational proof of concept studies. Methods: Two thousand five hundred dermal fibroblasts isolated from skin of SSc patients were seeded in ultra-low attachment 96-well plates. Fibroblast were let to aggregate into spheres for 48h. Two thousand five hundred primary normal human keratinocytes were added to the culture and let to layer onto the fibroblast spheres for 72h. H19X silencing experiments were used as proof of concept studies. H19X silencing with antisense oligonucleotides or transfections with a scrambled control were performed in fibroblasts prior to the sphere formation for 24h. TGFβ (10 ng/ml) was added to microtissue to exacerbate the fibrotic phenotype. Haematoxylin eosin staining as well as immunohistochemistry staining for vimentin and cytokeratin 10 was performed. Skin microtissues were processed for RNA and protein isolation. Pro-collagen Iα1 and fibronectin were quantified in the supernatants with ELISA. Results: The microtissues presented a core of SSc fibroblast as revealed by vimentin staining and an external layer of keratinocytes as revealed by cytokeratin 10 staining, mimicking the human skin architecture. Gene expression analysis following TGFβ stimulation displayed induced expression of extracellular matrix gene COL1A1 (p=0.044) and the myofibroblast marker ACTA2 (p=0.018), indicating that the microtissues were able to develop a fibrotic response. Microtissues, where H19X was silenced, displayed reduced gene expression of COL1A1 and ACTA2 after TGFβ stimulation ( COL1A1 p=0.007, ACTA2 p=0.045). Additionally, H19X silencing led to lower levels of αSMA protein expression (p=0.009) and pro-collagen1α1 secretion (p=0.039) in the supernatant of the microtissue cultures as revealed by Western Blot and ELISA, respectively. FN1 expression and fibronectin protein levels were not significantly reduced in the microtissues after H19X silencing. Conclusion: We were able to produce a 3D microtissue resembling skin architecture that can respond to fibrotic stimuli. Knockdown experiments of pro-fibrotic lncRNA H19X confirmed the potential of the model as screening platform for novel pro-fibrotic effectors. A future aim will be to optimize the model for high-throughput automated screening platforms. References: [1]Pachera, E., et al. (2020). “Long noncoding RNA H19X is a key mediator of TGF-β–driven fibrosis.” The Journal of Clinical Investigation 130(9): 4888-4905. Disclosure of Interests: Elena Pachera: None declared, Gabriela Kania: None declared, Astrid Juengel: None declared, Maurizio Calcagni Speakers bureau: Arthrex, Consultant of: Medartis, Arthrex, SilkBiomaterials, Grant/research support from: Medartis, Oliver Distler Speakers bureau: Actelion, Bayer, Boehringer Ingelheim, Medscape, Novartis, Roche, Consultant of: Abbvie, Actelion, Acceleron Pharma, Amgen, AnaMar, Arxx Therapeutics, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, ChemomAb, Corpuspharma, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, -Kymera, Medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi, UCB, Grant/research support from: Abbvie, Actelion, Bayer, Boehringer Ingelheim, Kymera Therapeutics, Mitsubishi Tanabe" @default.
• W3165161784 created "2021-06-07" @default.
• W3165161784 creator A5018529897 @default.
• W3165161784 creator A5033246902 @default.
• W3165161784 creator A5035867293 @default.
• W3165161784 creator A5064299522 @default.
• W3165161784 creator A5076353197 @default.
• W3165161784 date "2021-05-19" @default.
• W3165161784 modified "2023-09-27" @default.
• W3165161784 title "OP0248 DEVELOPMENT OF A 3D SKIN MICROTISSUE MODEL FOR FIBROTIC DISEASES" @default.
• W3165161784 doi "https://doi.org/10.1136/annrheumdis-2021-eular.3306" @default.
• W3165161784 hasPublicationYear "2021" @default.
• W3165161784 type Work @default.
• W3165161784 sameAs 3165161784 @default.
• W3165161784 citedByCount "0" @default.
• W3165161784 crossrefType "journal-article" @default.
• W3165161784 hasAuthorship W3165161784A5018529897 @default.
• W3165161784 hasAuthorship W3165161784A5033246902 @default.
• W3165161784 hasAuthorship W3165161784A5035867293 @default.
• W3165161784 hasAuthorship W3165161784A5064299522 @default.
• W3165161784 hasAuthorship W3165161784A5076353197 @default.
• W3165161784 hasBestOaLocation W31651617841 @default.
• W3165161784 hasConcept C142724271 @default.
• W3165161784 hasConcept C147447768 @default.
• W3165161784 hasConcept C189165786 @default.
• W3165161784 hasConcept C204232928 @default.
• W3165161784 hasConcept C2777459323 @default.
• W3165161784 hasConcept C2778005187 @default.
• W3165161784 hasConcept C2779751288 @default.
• W3165161784 hasConcept C2780381497 @default.
• W3165161784 hasConcept C2780559512 @default.
• W3165161784 hasConcept C54355233 @default.
• W3165161784 hasConcept C71924100 @default.
• W3165161784 hasConcept C74864618 @default.
• W3165161784 hasConcept C81885089 @default.
• W3165161784 hasConcept C86492073 @default.
• W3165161784 hasConcept C86803240 @default.
• W3165161784 hasConcept C95444343 @default.
• W3165161784 hasConceptScore W3165161784C142724271 @default.
• W3165161784 hasConceptScore W3165161784C147447768 @default.
• W3165161784 hasConceptScore W3165161784C189165786 @default.
• W3165161784 hasConceptScore W3165161784C204232928 @default.
• W3165161784 hasConceptScore W3165161784C2777459323 @default.
• W3165161784 hasConceptScore W3165161784C2778005187 @default.
• W3165161784 hasConceptScore W3165161784C2779751288 @default.
• W3165161784 hasConceptScore W3165161784C2780381497 @default.
• W3165161784 hasConceptScore W3165161784C2780559512 @default.
• W3165161784 hasConceptScore W3165161784C54355233 @default.
• W3165161784 hasConceptScore W3165161784C71924100 @default.
• W3165161784 hasConceptScore W3165161784C74864618 @default.
• W3165161784 hasConceptScore W3165161784C81885089 @default.
• W3165161784 hasConceptScore W3165161784C86492073 @default.
• W3165161784 hasConceptScore W3165161784C86803240 @default.
• W3165161784 hasConceptScore W3165161784C95444343 @default.
• W3165161784 hasIssue "Suppl 1" @default.
• W3165161784 hasLocation W31651617841 @default.
• W3165161784 hasOpenAccess W3165161784 @default.
• W3165161784 hasPrimaryLocation W31651617841 @default.
• W3165161784 hasRelatedWork W2012526429 @default.
• W3165161784 hasRelatedWork W2051244420 @default.
• W3165161784 hasRelatedWork W2121529430 @default.
• W3165161784 hasRelatedWork W2369020123 @default.
• W3165161784 hasRelatedWork W2466727286 @default.
• W3165161784 hasRelatedWork W2905016282 @default.
• W3165161784 hasRelatedWork W2949592293 @default.
• W3165161784 hasRelatedWork W3026506096 @default.
• W3165161784 hasRelatedWork W3165135049 @default.
• W3165161784 hasRelatedWork W3165161784 @default.
• W3165161784 hasVolume "80" @default.
• W3165161784 isParatext "false" @default.
• W3165161784 isRetracted "false" @default.
• W3165161784 magId "3165161784" @default.
• W3165161784 workType "article" @default.