PD-L1 on invasive fibroblasts drives fibrosis in a humanized model of idiopathic pulmonary fibrosis

Geng, Yan; Liu, Xue; Liang, Jiurong; Habiel, David M.; Kulur, Vrishika; Coelho, Ana Lúcia; Deng, Nan; Xie, Ting; Wang, Yizhou; Liu, Ningshan; Huang, Guanling; Kurkciyan, Adrianne; Liu, Zhenqiu; Tang, Jie; Hogaboam, Cory M.; Jiang, Dianhua; Noble, Paul W.

doi:10.1172/jci.insight.125326

Cited by 82 publications

(130 citation statements)

References 59 publications

(68 reference statements)

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“…The immune checkpoint ligand PD‐L1 has been reported to be upregulated and promote invasion of pulmonary fibroblasts . In that TGFβ is a regulatory cytokine that both suppresses immune function and is a primary mediator of organ fibrosis, we investigated whether these actions might be linked .…”

Section: Resultsmentioning

confidence: 99%

“…The immune checkpoint ligand PD-L1 has been reported to be upregulated and promote invasion of pulmonary fibroblasts. 21 In that TGFβ is a regulatory cytokine that both suppresses immune function and is a primary mediator of organ fibrosis, we investigated whether these actions might be linked. [28][29][30] In support of that hypothesis, TGFβ upregulated PD-L1 expression in both human lung (eg, MRC5, TIG-1) and murine (eg, AKR2B, Swiss3T3) fibroblast cell lines ( Figure 1A).…”

Section: Upregulation Of Pd-l1 By Tgfβmentioning

confidence: 99%

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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%

Section: Upregulation Of Pd-l1 By Tgfβmentioning

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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“…Integral to the discussion of the lung phenotype in IPF are the notions of fibroblast activation and myofibroblast accumulation. Surprisingly, despite significant progression in description of the molecular and metabolic changes required to drive a profibrotic profile of 205 fibroblasts in vitro or in animal models of disease (37)(38)(39)(40), little is known about the diversity of cellular populations in the IPF lung. This is because of the lack of specific markers for the distinct populations, as well as the phenotypic changes fibroblasts undergo during the common culturing methods (41).…”

Section: Discussionmentioning

confidence: 99%

Single Cell RNA-seq reveals ectopic and aberrant lung resident cell populations in Idiopathic Pulmonary Fibrosis

Adams

Schupp

Poli

et al. 2019

Preprint

247

499

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We provide a single cell atlas of Idiopathic Pulmonary Fibrosis (IPF), a fatal interstitial lung disease, focusing on resident lung cell populations. By profiling 312,928 cells from 32 IPF, 29 healthy control and 18 chronic obstructive pulmonary disease (COPD) lungs, we demonstrate that IPF is characterized by changes in discrete subpopulations of cells in the three major parenchymal compartments: the epithelium, endothelium and stroma. Among epithelial cells, we identify a novel population of IPF enriched aberrant basaloid cells that co-express basal epithelial markers, mesenchymal markers, senescence markers, developmental transcription factors and are located at the edge of myofibroblast foci in the IPF lung. Among vascular endothelial cells in the in IPF lung parenchyma we identify an expanded cell population transcriptomically identical to vascular endothelial cells normally restricted to the bronchial circulation. We confirm the presence of both populations by immunohistochemistry and independent datasets. Among stromal cells we identify fibroblasts and myofibroblasts in both control and IPF lungs and leverage manifold-based algorithms diffusion maps and diffusion pseudotime to infer the origins of the activated IPF myofibroblast. Our work provides a comprehensive catalogue of the aberrant cellular transcriptional programs in IPF, demonstrates a new framework for analyzing complex disease with scRNAseq, and provides the largest lung disease single-cell atlas to date.

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“…Such fibroblasts are reminiscent of fibrocytes, which express hematopoietic markers such as CD45, CD34, and MHCII (major histocompatibility complex class II).170 Fibroblasts can be reprogrammed into induced dendritic cells (iDCs) capable of cross-presenting antigen to CD8+ T cells.171 Interestingly, the immune checkpoint ligand PD-L1/CD274 was recently reported on fibroblasts in lung fibrosis, and this possibly plays a role in fibroblast invasion and progression of idiopathic pulmonary fibrosis. 172 Collectively, these results highlight fibroblast modulation of immune responses that extends beyond secretome influences on adaptive immune cells by engaging antigen-presentation and immune checkpoint signaling.…”

Section: Regulatory Functions Of Fibroblastsmentioning

confidence: 94%

Origin and functional heterogeneity of fibroblasts

2020

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The inherent plasticity and resiliency of fibroblasts make this cell type a conventional tool for basic research. But where do they come from, are all fibroblasts the same, and how do they function in disease? The first fibroblast lineages in mammalian development emerge from the ooze of primary mesenchyme during gastrulation. They are cells that efficiently create and negotiate the extracellular matrix of the mesoderm in order to migrate and meet their developmental fate. Mature fibroblasts in epithelial tissues live in the interstitial spaces between basement membranes that spatially delimit complex organ structures. While the function of resident fibroblasts in healthy tissues is largely conjecture, the accumulation of fibroblasts in pathologic lesions offers insight into biologic mechanisms that control their function; fibroblasts are poised to coordinate fibrogenesis in tissue injury, neoplasia, and aging. Here, we examine the developmental origin and plasticity of fibroblasts, their molecular and functional definitions, the epigenetic control underlying their identity and activation, and the evolution of their immune regulatory functions. These topics are reviewed through the lens of fate mapping using genetically engineered mouse models and from the perspective of single‐cell RNA sequencing. Recent observations suggest dynamic and heterogeneous functions for fibroblasts that underscore their complex molecular signatures and utility in injured tissues.

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PD-L1 on invasive fibroblasts drives fibrosis in a humanized model of idiopathic pulmonary fibrosis

Cited by 82 publications

References 59 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

Single Cell RNA-seq reveals ectopic and aberrant lung resident cell populations in Idiopathic Pulmonary Fibrosis

Origin and functional heterogeneity of fibroblasts

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