Spontaneous Extracellular Matrix Accumulation in a Human in vitro Model of Renal Fibrosis Is Mediated by αV Integrins

Bon, Hélène; Hales, Paul; Lumb, Simon; Holdsworth, Gill; Johnson, Tim; Qureshi, Omar; Twomey, Breda M.

doi:10.1159/000499506

“…The following drugs were used: Galunisertib (=LY2157299) (0.5, 1, 2.5 μM; 200-17, PeproTech); Omipalisib (=GSK2126458, GSK458, S2658) (0.01, 0.1, 1 μM; Selleck Chemicals); Imatinib (0.1, 0.5, 1 μM; I-5577, LC Laboratories, Woburn, MA); and Nintenadib (0.1, 0.5, 1 μM; Boehringer Ingelheim, Biberach, Germany). The used concentrations are in line with those of previous studies ( 45 – 49 ). Stock solutions of the drugs were prepared in dimethyl sulfoxide (DMSO; 100%) and stored at −20°C; the working solutions were diluted in culture medium with a final solvent concentration of ≤ 1%.…”

Section: Methodssupporting

confidence: 81%

Macromolecular Crowding as a Tool to Screen Anti-fibrotic Drugs: The Scar-in-a-Jar System Revisited

Cavanzo

¹

,

Bigaeva

²

,

Boersema

³

et al. 2021

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An unsolved therapeutic problem in fibrosis is the overproduction of collagen. In order to screen the effect of anti-fibrotic drugs on collagen deposition, the Scar-in-a-Jar approach has been introduced about a decade ago. With macromolecular crowding a rapid deposition of collagen is seen, resulting in a substantial decrease in culture time, but the system has never been tested in an adequate way. We therefore have compared six different macromolecular crowders [Ficoll PM 70 (Fc70), Ficoll PM 400 (Fc400), a mixture of Ficoll 70 and 400 (Fc70/400), polyvinylpyrrolidone 40 (PVP40), polyvinylpyrrolidone 360 (PVP360), neutral dextran 670 (ND670), dextran sulfate 500 (DxS500), and carrageenan (CR)] under profibrotic conditions (addition of TGFβ1) with primary human adult dermal fibroblasts in the presence of 0.5 and 10% FBS. We found that (1) collagen deposition and myofibroblast formation was superior with 0.5% FBS, (2) DxS500 and CR results in an aberrant collagen deposition pattern, (3) ND670 does not increase collagen deposition, and (4) CR, DxS500, and Fc40/700 affected important phenotypical properties of the cells when cultured under pro-fibrotic conditions, whereas PVP40 and PVP360 did less or not. Because of viscosity problems with PVP360, we conclude that PVP40 is the most optimal crowder for the screening of anti-fibrotic drugs. Finally, the effect of various concentrations of Imatinib, Galunisertib, Omipalisib or Nintedanib on collagen deposition and myofibroblast formation was tested with PVP40 as the crowder.

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“…This response was ALK5-dependent and significantly increased in Pkd1 −/− PTEC cells, suggesting a role for exaggerated ALK5-SMAD2/3 signalling in cystogenesis. ALK5 inhibition has been found to reduce fibrosis and extracellular matrix protein deposition in vitro 73 . These observations lend support to our findings that ALK5 inhibition can ameliorate the cystic phenotype.…”

Section: Discussionmentioning

confidence: 99%

A high throughput zebrafish chemical screen reveals ALK5 and non-canonical androgen signalling as modulators of the pkd2−/− phenotype

Metzner

¹

,

Griffiths

²

,

Streets

³

et al. 2020

Sci Rep

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Autosomal dominant polycystic kidney disease (ADPKD) is the most common monogenic cause of end-stage renal failure in humans and results from germline mutations in PKD1 or PKD2. Despite the recent approval of tolvaptan, safer and more effective alternative drugs are clearly needed to slow disease progression. As a first step in drug discovery, we conducted an unbiased chemical screen on zebrafish pkd2 mutant embryos using two publicly available compound libraries (Spectrum, PKIS) totalling 2,367 compounds to identify novel treatments for ADPKD. Using dorsal tail curvature as the assay readout, three major chemical classes (steroids, coumarins, flavonoids) were identified from the Spectrum library as the most promising candidates to be tested on human PKD1 cystic cells. Amongst these were an androgen, 5α−androstane 3,17-dione, detected as the strongest enhancer of the pkd2 phenotype but whose effect was found to be independent of the canonical androgen receptor pathway. From the PKIS library, we identified several ALK5 kinase inhibitors as strong suppressors of the pkd2 tail phenotype and in vitro cyst expansion. In summary, our results identify ALK5 and non-canonical androgen receptors as potential therapeutic targets for further evaluation in drug development for ADPKD.

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“…Activation of NF- κ B can also promote excessive secretion of adhesion molecules and chemokines, leading to ECM deposition and RIF. Overdeposition of ECM is one of the most important factors leading to RIF [ 26 ], which is mainly composed of collagen, FN, and other substances. Col- IV is the main collagen in the glomerular basement membrane, constructing its scaffold.…”

Section: Discussionmentioning

confidence: 99%

Effects of Qingshen Granules on Immune Function in Patients with Comorbid Chronic Renal Failure and Damp‐Heat Syndrome: A Multicenter, Randomized, Controlled Trial

Wang

¹

,

Wang

²

,

Li

³

et al. 2020

Evidence-Based Complementary and Alternative Medicine

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Objective. The current study sought to compare the effects of the addition of Qingshen granules to conventional Western medicine on immune function in patients with comorbid chronic renal failure and damp-heat syndrome and to explore the possible mechanisms responsible for any differences observed. Methods. Through a multicenter, randomized, controlled study, a total of 282 eligible patients were divided into experimental (n = 136) and control groups (n = 146). All of the patients were treated with conventional Western medical therapy. The experimental group also received Qingshen granules three times daily for 12 weeks. Clinical efficacy was observed in the two groups. Peripheral blood levels of CD4+ T cells, CD8+ T cells, Th17 cells, nuclear factor-κB p65 (NF-κB p65) activity, serum interleukin-17 (IL-17), serum interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), tumor necrosis factor receptor-associated factor 6 (TRAF6), fibronectin (FN), and type IV collagen (Col-IV) were detected in both groups. Results. The total clinical curative effective rate was significantly higher p < 0.05 in the experimental group (79.41%) than in the control group (67.12%). Before treatment, there were no significant differences in CD4+/CD8+ T cell ratio, Th17 cell level, NF-κB p65 activity, serum IL-17, IL-6, TNF-α, TRAF6, FN, and Col-IV between the experimental and control groups p > 0.05 ; however, all of the measures were significantly higher than those observed in a healthy comparison group ( p < 0.05 or p < 0.01 ). After treatment, the above indexes in the experimental group were significantly lower than those before treatment ( p < 0.05 or p < 0.01 ). Similarly, NF-κB p65 activity, serum IL-17, TNF-α, TRAF6, FN, and Col-IV in the control group were significantly lower than the levels observed prior to treatment ( p < 0.05 or p < 0.01 ); however, while all of the other indexes were lower than those observed before treatment, the differences were not statistically significant p > 0.05 . Conclusion. Qingshen granules adjust immune dysfunction, improve immunity mediated inflammatory response, and attenuate renal fibrosis in patients with comorbid chronic renal failure and damp-heat syndrome.

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Spontaneous Extracellular Matrix Accumulation in a Human in vitro Model of Renal Fibrosis Is Mediated by αV Integrins

Cited by 31 publications

References 42 publications

Macromolecular Crowding as a Tool to Screen Anti-fibrotic Drugs: The Scar-in-a-Jar System Revisited

Macromolecular Crowding as a Tool to Screen Anti-fibrotic Drugs: The Scar-in-a-Jar System Revisited

A high throughput zebrafish chemical screen reveals ALK5 and non-canonical androgen signalling as modulators of the pkd2−/− phenotype

Effects of Qingshen Granules on Immune Function in Patients with Comorbid Chronic Renal Failure and Damp‐Heat Syndrome: A Multicenter, Randomized, Controlled Trial

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