Fatty acid synthase is required for profibrotic TGF‐β signaling

Jung, Myungjin; Kang, Jeong Han; Hernandez, Danielle M.; Yin, Xueqian; Andrianifahanana, Mahefatiana; Wang, Youli; Gonzalez-Guerrico, Anatilde; Limper, Andrew H.; Lupu, Ruth; Leof, Edward B.

doi:10.1096/fj.201701187r

Cited by 59 publications

(47 citation statements)

References 79 publications

(104 reference statements)

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“…These alterations result in the activation of TGFβ and predispose to the development of pulmonary fibrosis . As metabolic reprograming can have a critical role in the modulation of immune checkpoint proteins and profibrotic TGFβ signaling utilizes metabolism relevant proteins including fatty acid synthase (FASN), hexokinase II (HKII), and glutaminase 1 (GLS1), we investigated the interrelation of PD‐L1 and metabolism using several metabolic enzyme inhibitors. As shown in Figure A, while PD‐L1 induction by TGFβ was independent of HKII, FASN, or GLS1 inhibition was as effective as previously reported indoleamine 2,3‐dioxygenase (IDO‐1) inhibition by INCB in preventing PD‐L1 expression in both human and murine fibroblasts.…”

Section: Resultsmentioning

confidence: 99%

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

et al. 2019

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Transforming growth factor-beta (TGFβ) is an enigmatic protein with various roles in healthy tissue homeostasis/development as well as the development or progression of cancer, wound healing, fibrotic disorders, and immune modulation, to name a few. As TGFβ is causal to various fibroproliferative disorders featuring localized or systemic tissue/organ fibrosis as well as the activated stroma observed in various malignancies, characterizing the pathways and players mediating its action is fundamental. In the current study, we found that TGFβ induces the expression of the immunoinhibitory molecule Programed death-ligand 1 (PD-L1) in human and murine fibroblasts in a Smad2/3-and YAP/TAZ-dependent manner. Furthermore, PD-L1 knockdown decreased the TGFβ-dependent induction of extracellular matrix proteins, including collagen Iα1 (colIα1) and alpha-smooth muscle actin (α-SMA), and cell migration/wound healing. In addition to an endogenous role for PD-L1 in profibrotic TGFβ signaling, TGFβ stimulated-human lung fibroblast-derived PD-L1 into extracellular vesicles (EVs) capable of inhibiting T cell proliferation in response to T cell receptor stimulation and mediating fibroblast cell migration. These findings provide new insights and potential targets for a variety of fibrotic and malignant diseases. K E Y W O R D Sfibroblast, checkpoint inhibitor, extracellular vesicles, signaling 2214 | KANG et Al. | 2223 KANG et Al. F I G U R E 6EVs from TGFβ treated fibroblasts inhibit T cell proliferation and cell migration. A, MRC5 cultures were treated in the absence(−) or presence (+) of TGFβ (10 ng/mL) for 3 days. Following the collection of the supernatant for EV isolation, the cells were lysed, and both were assessed for expression of PD-L1 or the EV-associated protein CD63 by Western blotting (representative of 3 independent experiments). B, EV particle size distribution was identified through nanoparticle tracking analysis and processed with NTA software (NanoSight). C, EVs were purified from MRC5 cells cultured in the absence (−) or presence (+) of TGFβ as in (A). Peripheral blood mononuclear cells (n = 5 volunteers) were then treated with anti-CD3 (+αCD3; 500 ng/mL) to activate T cell proliferation and incubated for 6 days with no EVs, EVs (10 μg/mL) isolated from cells ± TGFβ, or EVs isolated from cells ± TGFβ ± blocking antibody to PD-L1 (10 μg/mL Avelumab). The percent change in proliferation, measured by carboxyfluorescein diacetate succinimidyl ester (CFSE) content, compared to anti-CD3 treatment alone is shown. D, (Top 2 panels) Transwell invasion assays were performed in MRC5 cells treated in 0.1% FBS/DME alone (Con) or supplemented with 5 ng/mL TGFβ for 18 hours as described in Materials and Methods. (Bottom 3 panels) MRC5 cells were assessed as in the top panels except that the 0.1% FBS/DME contained EVs (10 μg/mL) isolated from MRC5 cells ± TGFβ or MRC5 cells which had been treated with siRNA to PD-L1 and then stimulated with TGFβ. E, Quantitation of transwell migration of cells treated in (D). Data reflect mean ±...

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Section: Resultsmentioning

confidence: 99%

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

et al. 2019

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“…[38, 39] Whereas, endogenous nitrated FAs exert anti-fibrotic effects by up-regulating peroxisome proliferator-activated receptor γ (PPARγ) in human lung fibroblasts another study showed that FASN is required for pro-fibrotic TGF-β signalling. [40]…”

Section: Discussionmentioning

confidence: 99%

Transcriptome analysis of IPF fibroblastic foci identifies key pathways involved in fibrogenesis

Guillotin

Taylor

Platé

et al. 2020

Preprint

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Fibroblastic foci (FF) represent the cardinal pathogenic lesion in idiopathic pulmonary fibrosis (IPF) and comprise activated fibroblasts and myofibroblasts, the key effector cells responsible for dysregulated extracellular matrix deposition in multiple fibrotic conditions. The aim of this study was to define the major transcriptional programmes involved in fibrogenesis in IPF by profiling un-manipulated myo/fibroblasts within FF in situ by laser capture microdissection.The challenges associated with deriving gene calls from low amounts of RNA and the absence of a meaningful comparator cell type were overcome by adopting novel data mining strategies and by using weighted gene co-expression network analysis (WGCNA), as well as an eigengene-based approach to identify transcriptional signatures which correlate with fibrillar collagen gene expression. WGCNA identified prominent clusters of genes associated with cell cycle, inflammation/differentiation, translation and cytoskeleton/cell adhesion. Collagen eigengene analysis revealed that TGF-β1, RhoA kinase and the TSC2/RHEB axis formed major signalling clusters associated with collagen gene expression. Functional studies using CRISPR-Cas9 gene edited cells demonstrated a key role for the TSC2/RHEB axis in regulating TGF-β1-induced mTORC1 activation and collagen I deposition in mesenchymal cells reflecting IPF and other disease settings, including cancer-associated fibroblasts. These data provide strong support for the human tissue-based and bioinformatics approaches adopted to identify critical transcriptional nodes associated with the key pathogenic cell responsible for fibrogenesis in situ and further identifies the TSC2/RHEB axis as a potential novel target for interfering with excessive matrix deposition in IPF and other fibrotic conditions.What is the key question?Can we identify a transcriptional signature associated with collagen gene expression in the fibrotic focus, the cardinal fibrotic lesion in IPF?What is the bottom line?We herein define the major transcriptional programmes involved in fibrogenesis in IPF by profiling myo/fibroblasts within FF in situ by laser capture microdissection.Why read on?The data provide strong support for a human tissue-based approach to identify critical transcriptional nodes associated with fibrogenesis in situ and further identifies the TSC2/RHEB axis as a potential novel target for interfering with excessive matrix deposition in IPF and other fibrotic conditions.

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“…Further studies are required to support the role of these factors in the activation of COX-2 during MDV infection. COX-2-inducing factors such as TGF-β (32), which also activates fatty acid synthesis (31, 35), might be involved in activation of COX-2 in MDV-infected cells. In fact, we have recently shown that MDV infection increases the induction of TGF-β in vivo (36); however, further studies are required to examine the role of MDV-induced TGF-β in the induction of COX-2 during MDV infection.…”

Section: Discussionmentioning

confidence: 99%

Replication of Marek's Disease Virus Is Dependent on Synthesis of De Novo Fatty Acid and Prostaglandin E ₂

Boodhoo

Kamble

Kaufer

et al. 2019

J Virol

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Marek’s disease virus (MDV) causes deadly lymphoma and induces an imbalance of the lipid metabolism in infected chickens. Here, we discovered that MDV activates the fatty acid synthesis (FAS) pathway in primary chicken embryo fibroblasts (CEFs). In addition, MDV-infected cells contained high levels of fatty acids and showed increased numbers of lipid droplets (LDs). Chemical inhibitors of the FAS pathway (TOFA and C75) reduced MDV titers by approximately 30-fold. Addition of the downstream metabolites, including malonyl-coenzyme A and palmitic acid, completely restored the inhibitory effects of the FAS inhibitors. Furthermore, we could demonstrate that MDV infection activates the COX-2/prostaglandin E2 (PGE2) pathway, as evident by increased levels of arachidonic acid, COX-2 expression, and PGE2 synthesis. Inhibition of the COX-2/PGE2 pathway by chemical inhibitors or knockdown of COX2 using short hairpin RNA reduced MDV titers, suggesting that COX-2 promotes virus replication. Exogenous PGE2 completely restored the inhibition of the COX-2/PGE2 pathway in MDV replication. Unexpectedly, exogenous PGE2 also partially rescued the inhibitory effects of FAS inhibitors on MDV replication, suggesting that there is a link between these two pathways in MDV infection. Taken together, our data demonstrate that the FAS and COX-2/PGE2 pathways play an important role in the replication of this deadly pathogen. IMPORTANCE Disturbances of the lipid metabolism in chickens infected with MDV contribute to the pathogenesis of disease. However, the role of lipid metabolism in MDV replication remained unknown. Here, we demonstrate that MDV infection activates FAS and induces LD formation. Moreover, our results demonstrate that MDV replication is highly dependent on the FAS pathway and the downstream metabolites. Finally, our results reveal that MDV also activates the COX-2/PGE2 pathway, which supports MDV replication by activating PGE2/EP2 and PGE2/EP4 signaling pathways.

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Fatty acid synthase is required for profibrotic TGF‐β signaling

Cited by 59 publications

References 79 publications

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

Transcriptome analysis of IPF fibroblastic foci identifies key pathways involved in fibrogenesis

Replication of Marek's Disease Virus Is Dependent on Synthesis of De Novo Fatty Acid and Prostaglandin E ₂

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Fatty acid synthase is required for profibrotic TGF‐β signaling

Cited by 59 publications

References 79 publications

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

Transforming growth factor beta induces fibroblasts to express and release the immunomodulatory protein PD‐L1 into extracellular vesicles

Transcriptome analysis of IPF fibroblastic foci identifies key pathways involved in fibrogenesis

Replication of Marek's Disease Virus Is Dependent on Synthesis of De Novo Fatty Acid and Prostaglandin E 2

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Replication of Marek's Disease Virus Is Dependent on Synthesis of De Novo Fatty Acid and Prostaglandin E ₂