scholarly journals Identification of Hepatic Fibrosis Inhibitors Through Morphometry Analysis of Hepatic Multicellular Spheroids Models

2020 ◽  
Author(s):  
Haengran Seo ◽  
Yeonhwa Song ◽  
Jinyeong Heo ◽  
David Shum ◽  
Su-Yeon Lee ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Hepatic Fibrosis ◽  
High Throughput Screening ◽  
Epithelial To Mesenchymal Transition ◽  
New Drugs ◽  
Multicellular Spheroids ◽  
Mesenchymal Transition ◽  
Morphometry Analysis ◽  
Endothelial To Mesenchymal Transition ◽  
Approved Drugs

Abstract Background: A chronic, local inflammatory milieu can cause tissue fibrosis that results in epithelial-to-mesenchymal transition (EMT), endothelial-to-mesenchymal transition (EndMT), increased abundance of fibroblasts, and further acceleration of fibrosis. Methods: In this study, we aimed to identify potential mechanisms and inhibitors of fibrosis using 3D model-based phenotypic screening. We established liver fibrosis models using multicellular tumor spheroids (MCTSs) composed of hepatocellular carcinoma (HCC) and stromal cells such as fibroblasts (WI38), hepatic stellate cells (LX2), and endothelial cells (HUVEC) seeded at constant ratios. Results: Through high-throughput screening of FDA-approved drugs, we identified retinoic acid and forskolin as candidates to attenuate the compactness of MCTSs as well as inhibit the expression of ECM-related proteins. Additionally, retinoic acid and forskolin induced reprogramming of fibroblast and cancer stem cells in the HCC microenvironment. Of interest, retinoic acid and forskolin had anti-fibrosis effects by decreasing expression of a-SMA and F-actin in LX2 cells and HUVEC cells. Moreover, when sorafenib was added along with retinoic acid and forskolin, apoptosis was increased, suggesting that anti-fibrosis drugs may improve tissue penetration to support the efficacy of anti-cancer drugs. Conclusion: Collectively, these findings support the potential utility of morphometric analyses of hepatic multicellular spheroid models in the development of new drugs with novel mechanisms for the treatment of hepatic fibrosis and HCCs.

scholarly journals Identification of hepatic fibrosis inhibitors through morphometry analysis of a hepatic multicellular spheroids model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yeonhwa Song ◽  
Sanghwa Kim ◽  
Jinyeong Heo ◽  
David Shum ◽  
Su-Yeon Lee ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Hepatic Fibrosis ◽  
High Throughput Screening ◽  
Epithelial To Mesenchymal Transition ◽  
New Drugs ◽  
Multicellular Spheroids ◽  
Mesenchymal Transition ◽  
Morphometry Analysis ◽  
Endothelial To Mesenchymal Transition ◽  
Approved Drugs

AbstractA chronic, local inflammatory milieu can cause tissue fibrosis that results in epithelial-to-mesenchymal transition (EMT), endothelial-to-mesenchymal transition (EndMT), increased abundance of fibroblasts, and further acceleration of fibrosis. In this study, we aimed to identify potential mechanisms and inhibitors of fibrosis using 3D model-based phenotypic screening. We established liver fibrosis models using multicellular tumor spheroids (MCTSs) composed of hepatocellular carcinoma (HCC) and stromal cells such as fibroblasts (WI38), hepatic stellate cells (LX2), and endothelial cells (HUVEC) seeded at constant ratios. Through high-throughput screening of FDA-approved drugs, we identified retinoic acid and forskolin as candidates to attenuate the compactness of MCTSs as well as inhibit the expression of ECM-related proteins. Additionally, retinoic acid and forskolin induced reprogramming of fibroblast and cancer stem cells in the HCC microenvironment. Of interest, retinoic acid and forskolin had anti-fibrosis effects by decreasing expression of α-SMA and F-actin in LX2 cells and HUVEC cells. Moreover, when sorafenib was added along with retinoic acid and forskolin, apoptosis was increased, suggesting that anti-fibrosis drugs may improve tissue penetration to support the efficacy of anti-cancer drugs. Collectively, these findings support the potential utility of morphometric analyses of hepatic multicellular spheroid models in the development of new drugs with novel mechanisms for the treatment of hepatic fibrosis and HCCs.

scholarly journals Noncoding RNAs as Promising Diagnostic Biomarkers and Therapeutic Targets in Intestinal Fibrosis of Crohn’s Disease: The Path From Bench to Bedside

2020 ◽  
Author(s):  
Long-Yuan Zhou ◽  
Si-Nan Lin ◽  
Florian Rieder ◽  
Min-Hu Chen ◽  
Sheng-Hong Zhang ◽  
...  
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
Transforming Growth Factor ◽  
Noncoding Rnas ◽  
Transforming Growth Factor Β ◽  
Circular Rnas ◽  
Diagnostic Biomarkers ◽  
Mesenchymal Transition ◽  
Intestinal Fibrosis ◽  
Fibrotic Diseases ◽  
Endothelial To Mesenchymal Transition

Abstract Fibrosis is a major pathway to organ injury and failure, accounting for more than one-third of deaths worldwide. Intestinal fibrosis causes irreversible and serious clinical complications, such as strictures and obstruction, secondary to a complex pathogenesis. Under the stimulation of profibrotic soluble factors, excessive activation of mesenchymal cells causes extracellular matrix deposition via canonical transforming growth factor-β/Smads signaling or other pathways (eg, epithelial-to-mesenchymal transition and endothelial-to-mesenchymal transition) in intestinal fibrogenesis. In recent studies, the importance of noncoding RNAs (ncRNAs) stands out in fibrotic diseases in that ncRNAs exhibit a remarkable variety of biological functions in modulating the aforementioned fibrogenic responses. In this review, we summarize the role of ncRNAs, including the emerging long ncRNAs and circular RNAs, in intestinal fibrogenesis. Notably, the translational potential of ncRNAs as diagnostic biomarkers and therapeutic targets in the management of intestinal fibrosis is discussed based on clinical trials from fibrotic diseases in other organs. The main points of this review include the following: • Characteristics of ncRNAs and mechanisms of intestinal fibrogenesis • Wide participation of ncRNAs (especially the emerging long ncRNAs and circular RNAs) in intestinal fibrosis, including transforming growth factor-β signaling, epithelial-to-mesenchymal transition/endothelial-to-mesenchymal transition, and extracellular matrix remodeling • Translational potential of ncRNAs in the diagnosis and treatment of intestinal fibrosis based on clinical trials from fibrotic diseases in other organs

scholarly journals Vimentin as a target for the treatment of COVID-19

2020 ◽  
Vol 7 (1) ◽  
pp. e000623
Author(s):  
Zhenlin Li ◽  
Denise Paulin ◽  
Patrick Lacolley ◽  
Dario Coletti ◽  
Onnik Agbulut
Keyword(s):  
Viral Infection ◽  
Lung Inflammation ◽  
Epithelial To Mesenchymal Transition ◽  
Attachment Site ◽  
Vimentin Expression ◽  
Mesenchymal Transition ◽  
Cellular Target ◽  
Innovative Idea ◽  
Life Threatening ◽  
Approved Drugs

We and others propose vimentin as a possible cellular target for the treatment of COVID-19. This innovative idea is so recent that it requires further attention and debate. The significant role played by vimentin in virus-induced infection however is well established: (1) vimentin has been reported as a co-receptor and/or attachment site for SARS-CoV; (2) vimentin is involved in viral replication in cells; (3) vimentin plays a fundamental role in both the viral infection and the consequent explosive immune-inflammatory response and (4) a lower vimentin expression is associated with the inhibition of epithelial to mesenchymal transition and fibrosis. Moreover, the absence of vimentin in mice makes them resistant to lung injury. Since vimentin has a twofold role in the disease, not only being involved in the viral infection but also in the associated life-threatening lung inflammation, the use of vimentin-targeted drugs may offer a synergistic advantage as compared with other treatments not targeting vimentin. Consequently, we speculate here that drugs which decrease the expression of vimentin can be used for the treatment of patients with COVID-19 and advise that several Food and Drug Administration-approved drugs be immediately tested in clinical trials against SARS-CoV-2, thus broadening therapeutic options for this type of viral infection.

scholarly journals Tenascin C promotes valvular remodeling in two large animal models of ischemic mitral regurgitation

2020 ◽  
Vol 115 (6) ◽  
Author(s):  
Ouafa Hamza ◽  
Attila Kiss ◽  
Anne-Margarethe Kramer ◽  
Sandra Trojanek ◽  
Dietmar Abraham ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Mitral Regurgitation ◽  
Epithelial To Mesenchymal Transition ◽  
Ischemic Mitral Regurgitation ◽  
Large Animal ◽  
Mesenchymal Transition ◽  
Mitral Valves ◽  
Tenascin C ◽  
Large Animal Models ◽  
Endothelial To Mesenchymal Transition

AbstractIschemic mitral regurgitation (MR) is a frequent complication of myocardial infarction (MI) characterized by adverse remodeling both at the myocardial and valvular levels. Persistent activation of valvular endothelial cells leads to leaflet fibrosis through endothelial-to-mesenchymal transition (EMT). Tenascin C (TNC), an extracellular matrix glycoprotein involved in cardiovascular remodeling and fibrosis, was also identified in inducing epithelial-to-mesenchymal transition. In this study, we hypothesized that TNC also plays a role in the valvular remodeling observed in ischemic MR by contributing to valvular excess EMT. Moderate ischemic MR was induced by creating a posterior papillary muscle infarct (7 pigs and 7 sheep). Additional animals (7 pigs and 4 sheep) served as controls. Pigs and sheep were sacrificed after 6 weeks and 6 months, respectively. TNC expression was upregulated in the pig and sheep experiments at 6 weeks and 6 months, respectively, and correlated well with leaflet thickness (R = 0.68; p < 0.001 at 6 weeks, R = 0.84; p < 0.001 at 6 months). To confirm the translational potential of our findings, we obtained mitral valves from patients with ischemic cardiomyopathy presenting MR (n = 5). Indeed, TNC was also expressed in the mitral leaflets of these. Furthermore, TNC induced EMT in isolated porcine mitral valve endothelial cells (MVEC). Interestingly, Toll-like receptor 4 (TLR4) inhibition prevented TNC-mediated EMT in MVEC. We identified here for the first time a new contributor to valvular remodeling in ischemic MR, namely TNC, which induced EMT through TLR4. Our findings might set the path for novel therapeutic targets for preventing or limiting ischemic MR.

scholarly journals Collagen-based three-dimensional culture microenvironment promotes epithelial to mesenchymal transition and drug resistance of human ovarian cancerin vitro

RSC Advances ◽  
2018 ◽  
Vol 8 (16) ◽  
pp. 8910-8919 ◽  
Author(s):  
Ming Liu ◽  
Xiuzhen Zhang ◽  
Canling Long ◽  
Hong Xu ◽  
Xu Cheng ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Epithelial To Mesenchymal Transition ◽  
Three Dimensional ◽  
Collagen I ◽  
Multicellular Spheroids ◽  
Mesenchymal Transition ◽  
Hydrogel Scaffolds ◽  
Three Dimensional Culture ◽  
Cells Cultured

OV-NC and OV-206 cells cultured in collagen I hydrogel scaffolds, could gradually generate multicellular spheroids.

scholarly journals Pathogenesis of Type 2 Epithelial to Mesenchymal Transition (EMT) in Renal and Hepatic Fibrosis

2015 ◽  
Vol 5 (1) ◽  
pp. 4 ◽  
Author(s):  
Anusha Tennakoon ◽  
Takeshi Izawa ◽  
Mitsuru Kuwamura ◽  
Jyoji Yamate
Keyword(s):  
Hepatic Fibrosis ◽  
Epithelial To Mesenchymal Transition ◽  
Mesenchymal Transition

High Throughput Screening Reveals Unique Sensitivity of CD34+CD38- AML Cells to Isotretinoin and Tretinoin

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1430-1430
Author(s):  
Wen Xie ◽  
Duane C Hassane ◽  
Craig T. Jordan ◽  
Gail J. Roboz ◽  
Monica L. Guzman
Keyword(s):  
Flow Cytometry ◽  
Retinoic Acid ◽  
High Throughput ◽  
High Throughput Screening ◽  
Conflicts Of Interest ◽  
Chemotherapy Resistance ◽  
Multiparameter Flow Cytometry ◽  
Cell Subpopulations ◽  
Us Fda ◽  
Approved Drugs

Abstract Abstract 1430 Acute myeloid leukemia (AML) is a heterogeneous disease, originating from leukemic stem cells (LSCs). This relatively rare sub-population shows mostly a CD34+CD38- immunophenotype and has been proposed to be responsible for disease relapse and chemotherapy resistance. In order to identify effective drug(s) for clinical use that can eliminate LSCs, we have screened a library of 2000 compounds from the SPECTRUM collection (MicroSource Discovery Systems, Inc.) using a multi-parameter flow cytometry high-throughput screening. The drugs compiled in the library are 40% US FDA approved drugs, 30% natural products, 10% approved drugs for use in Europe and Japan, and the remainder consistent of bioactive compounds. The initial screen aimed to identify potential compounds that can decrease viability or increase differentiation of AML cells at 24 and 72 hours of treatment. Compounds demonstrating autofluorescence were eliminated as false positive hits. We found 137 compounds that decreased viability in all populations. Furthermore, 37 drugs demonstrated ability to increase cell death (more than 50%) in phenotypically described LSCs, progenitor and blasts populations, but with viability over 50% for normal residual lymphocytes within the same AML specimen. Interestingly, only 4 drugs displayed higher specificity to LSCs when compared to total blast populations. Surprisingly, two of these drugs are different forms of retinoic acid (isotretinoin and tretinoin). The selectivity of isotretinoin and tretinoin was confirmed in different primary AML samples. We observed that phenotypically described stem cell subpopulations were affected when exposed to retinoic acid in 4 out of 5 AML samples. LSC populations display increased apoptosis and differentiation assessed by flow cytometry. Characterization of what confers sensitivity to these types of drugs is under investigation. Together, a multiparameter flow cytometry screen revealed specific anti-LSC activity for retinoid acid, which was not detectable when analyzing total populations and would have been missed using traditional screening. These studies underscore the utility of LSC-based assays for defining novel therapies and suggest that retinoid-based therapies merit further investigation in AML. Disclosures: No relevant conflicts of interest to declare.

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