Prolonged Scar-in-a-Jar: an in vitro screening tool for anti-fibrotic therapies using biomarkers of extracellular matrix synthesis

Rønnow, Sarah; Dabbagh, R; Genovese, Federica; Nanthakumar, Carmel B.; Barrett, Vikki; Good, Robert B.; Brockbank, Sarah; Cruwys, Simon; Jessen, Henrik; Sørensen, Grith Lykke; Karsdal, Morten A.; Leeming, Diana Julie; Sand, J.M.B.

doi:10.1186/s12931-020-01369-1

Cited by 26 publications

(37 citation statements)

References 57 publications

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“…The mouse serum samples were assessed for collagen degraded peptides (PRO-C3, C3M, C4M, C4G, PRO-C6, C6M, VICM, reC1M, PRO-C23) by immunoassays by Nordic Bioscience as previously described (Jensen, Madsen et al 2018, Ronnow, Dabbagh et al 2020.…”

Section: Collagen Degradation Peptide Analysismentioning

confidence: 99%

Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

Ramos

Tian²,

Ruiter

et al. 2021

eLife

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Collagens are a primary component of the extracellular matrix and are functional ligands for the inhibitory immune receptor leukocyte associated immunoglobulin-like receptor (LAIR)-1. LAIR-2 is a secreted protein that can act as a decoy receptor by binding collagen with higher affinity than LAIR-1. We propose that collagens promote immune evasion by interacting with LAIR-1 expressed on immune cells, and that LAIR-2 releases LAIR-1 mediated immune suppression. Analysis of public human datasets show that collagens, LAIR-1 and LAIR-2 have unique and overlapping associations with survival in certain tumors. We designed a dimeric LAIR-2 with a functional IgG1 Fc tail, NC410, and showed that NC410 increases human T cell expansion and effector function in vivo in a mouse xenogeneic-graft versus-host disease model. In humanized mouse tumor models NC410 reduces tumor growth that is dependent on T cells. Immunohistochemical analysis of human tumors shows that NC410 binds to collagen-rich areas where LAIR-1+ immune cells are localized. Our findings show that NC410 might be a novel strategy for cancer immunotherapy for immune-excluded tumors.

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Section: Collagen Degradation Peptide Analysismentioning

confidence: 99%

Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

Ramos

Tian²,

Ruiter

et al. 2021

eLife

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“…The function of fibroblasts can be studied in vitro by the scar-in-a-jar model (SiaJ). SiaJ was originally developed by Chen et al [40] and subsequently refined by us for pulmonary and skin fibrosis modelling by prolonged culture and the addition of ECM and collagen biomarkers for applicability as a pre-clinical drug screening tool to evaluate the fibrotic component [41,42]. Inclusion of ECM and collagen biomarkers is key for translational purposes and may ultimately aid in the selection of the best-in-class anti-CAF/anti-fibrosis compounds as well as selecting which patients to treat with these compounds as a prerequisite for the optimal clinical benefits for patients.…”

Section: Introductionmentioning

confidence: 99%

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Nissen

Johansen

Chen

et al. 2022

Cancers

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The use of novel tools to understand tumour-fibrosis in pancreatic ductal adenocarcinoma (PDAC) and novel anti-fibrotic treatments are highly needed. We established a pseudo-3D in vitro model including humane pancreatic fibroblasts (PFs) and pancreatic cancer-associated fibroblasts (CAFs) in combination with clinical collagen biomarkers, as a translational anti-fibrotic drug screening tool. Furthermore, we investigated the prognostic potential of serum collagen biomarkers in 810 patients with PDAC. PFs and CAFs were cultured in Ficoll-media. Cells were treated w/wo TGF-ß1 and the anti-fibrotic compound ALK5i. Biomarkers measuring the formation of type III (PRO-C3) and VI (PRO-C6) collagens were measured by ELISA in supernatant at days 3, 6, 9, and 12. PRO-C3 and PRO-C6, and their association with overall survival (OS), were evaluated in serum with PDAC (n = 810). PRO-C3 and PRO-C6 were upregulated in CAFs compared to PFs (p < 0.0001.). TGF-ß1 increased PRO-C3 in both PFs and CAFs (p < 0.0001). The anti-fibrotic compound ALK5i inhibited both PRO-C3 and PRO-C6 (p < 0.0001). High serum levels of PRO-C3 and PRO-C6 in patients with PDAC were associated with short OS (PRO-C3:HR= 1.48, 95%CI:1.29–1.71, p < 0.0001 and PRO-C6:HR = 1.31, 95%CI:1.14–1.50, p = 0.0002). PRO-C3 and PRO-C6 have the potential to be used both pre-clinically and clinically as a measure of tumor fibrosis and CAF activity.

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“…In accordance with previous publications, several other organspecific fibrotic models induced by MMC supplementation have been developed (Chen et al, 2009;Graupp et al, 2015;Graupp et al, 2018;Fan et al, 2019;Good et al, 2019;Fan et al, 2020;Rønnow et al, 2020;De Pieri et al, 2021). It is worth noting that cocktails of pro-inflammatory cytokines have been used for more accurate recapitulation of fibrosis in vitro (Chawla and Ghosh, 2018) and such approach should be studied further in the future in combination with MMC.…”

Section: Discussionmentioning

confidence: 60%

“…In 2009, the first pathophysiologically relevant in vitro fibrosis model (termed Scar-in-the-Jar ) was published that utilised the principles of MMC (to enhance and accelerate ECM deposition) and TGF β 1 (to induce myofibroblast transformation of WI-38 lung fibroblasts) ( Chen et al, 2009 ). Since then, several fibrotic models based on MMC have been developed for screening anti-fibrotics in different fibrotic diseases (e.g., dermal ( Fan et al, 2019 ; Fan et al, 2020 ), lung ( Good et al, 2019 ; Rønnow et al, 2020 ), vocal fold ( Graupp et al, 2015 ; Graupp et al, 2018 ) scarring). Unfortunately, these dermal scar models might be incomplete as the optimal crowding molecule was not used ( Chen et al, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

Adapting the Scar-in-a-Jar to Skin Fibrosis and Screening Traditional and Contemporary Anti-Fibrotic Therapies

Coentro

May

Prince

et al. 2021

Front. Bioeng. Biotechnol.

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Skin fibrosis still constitutes an unmet clinical need. Although pharmacological strategies are at the forefront of scientific and technological research and innovation, their clinical translation is hindered by the poor predictive capacity of the currently available in vitro fibrosis models. Indeed, customarily utilised in vitro scarring models are conducted in a low extracellular matrix milieu, which constitutes an oxymoron for the in-hand pathophysiology. Herein, we coupled macromolecular crowding (enhances and accelerates extracellular matrix deposition) with transforming growth factor β1 (TGFβ1; induces trans-differentiation of fibroblasts to myofibroblasts) in human dermal fibroblast cultures to develop a skin fibrosis in vitro model and to screen a range of anti-fibrotic families (corticosteroids, inhibitors of histone deacetylases, inhibitors of collagen crosslinking, inhibitors of TGFβ1 and pleiotropic inhibitors of fibrotic activation). Data obtained demonstrated that macromolecular crowding combined with TGFβ1 significantly enhanced collagen deposition and myofibroblast transformation. Among the anti-fibrotic compounds assessed, trichostatin A (inhibitors of histone deacetylases); serelaxin and pirfenidone (pleiotropic inhibitors of fibrotic activation); and soluble TGFβ receptor trap (inhibitor of TGFβ signalling) resulted in the highest decrease of collagen type I deposition (even higher than triamcinolone acetonide, the gold standard in clinical practice). This study further advocates the potential of macromolecular crowding in the development of in vitro pathophysiology models.

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Prolonged Scar-in-a-Jar: an in vitro screening tool for anti-fibrotic therapies using biomarkers of extracellular matrix synthesis

Cited by 26 publications

References 57 publications

Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

Collagen Biomarkers Quantify Fibroblast Activity In Vitro and Predict Survival in Patients with Pancreatic Ductal Adenocarcinoma

Adapting the Scar-in-a-Jar to Skin Fibrosis and Screening Traditional and Contemporary Anti-Fibrotic Therapies

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