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Open Access 01-12-2019 | Idiopathic Pulmonary Fibrosis | Research article

DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis

Authors: David M. Habiel, Miriam S. Hohmann, Milena S. Espindola, Ana Lucia Coelho, Isabelle Jones, Heather Jones, Richard Carnibella, Isaac Pinar, Freda Werdiger, Cory M. Hogaboam

Published in: BMC Pulmonary Medicine | Issue 1/2019

Abstract

Background

Recent studies have highlighted the contribution of senescent mesenchymal and epithelial cells in Idiopathic Pulmonary Fibrosis (IPF), but little is known regarding the molecular mechanisms that regulate the accumulation of senescent cells in this disease. Therefore, we addressed the hypothesis that the loss of DNA repair mechanisms mediated by DNA protein kinase catalytic subunit (DNA-PKcs) in IPF, promoted the accumulation of mesenchymal progenitors and progeny, and the expression of senescent markers by these cell types.

Methods

Surgical lung biopsy samples and lung fibroblasts were obtained from patients exhibiting slowly, rapidly or unknown progressing IPF and lung samples lacking any evidence of fibrotic disease (i.e. normal; NL). The expression of DNA-Pkcs in lung tissue was assessed by quantitative immunohistochemical analysis. Chronic inhibition of DNA-PKcs kinase activity was mimicked using a highly specific small molecule inhibitor, Nu7441. Proteins involved in DNA repair (stage-specific embryonic antigen (SSEA)-4⁺ cells) were determined by quantitative Ingenuity Pathway Analysis of transcriptomic datasets (GSE103488). Lastly, the loss of DNA-PKc was modeled in a humanized model of pulmonary fibrosis in NSG SCID mice genetically deficient in PRKDC (the transcript for DNA-PKcs) and treated with Nu7441.

Results

DNA-PKcs expression was significantly reduced in IPF lung tissues. Chronic inhibition of DNA-PKcs by Nu7441 promoted the proliferation of SSEA4⁺ mesenchymal progenitor cells and a significant increase in the expression of senescence-associated markers in cultured lung fibroblasts. Importantly, mesenchymal progenitor cells and their fibroblast progeny derived from IPF patients showed a loss of transcripts encoding for DNA damage response and DNA repair components. Further, there was a significant reduction in transcripts encoding for PRKDC (the transcript for DNA-PKcs) in SSEA4⁺ mesenchymal progenitor cells from IPF patients compared with normal lung donors. In SCID mice lacking DNA-PKcs activity receiving IPF lung explant cells, treatment with Nu7441 promoted the expansion of progenitor cells, which was observed as a mass of SSEA4⁺ CgA⁺ expressing cells.

Conclusions

Together, our results show that the loss of DNA-PKcs promotes the expansion of SSEA4⁺ mesenchymal progenitors, and the senescence of their mesenchymal progeny.

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Title: DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis
Authors: David M. Habiel
Miriam S. Hohmann
Milena S. Espindola
Ana Lucia Coelho
Isabelle Jones
Heather Jones
Richard Carnibella
Isaac Pinar
Freda Werdiger
Cory M. Hogaboam
Publication date: 01-12-2019
Publisher: BioMed Central
Keyword: Idiopathic Pulmonary Fibrosis
Published in: BMC Pulmonary Medicine / Issue 1/2019
Electronic ISSN: 1471-2466
DOI: https://doi.org/10.1186/s12890-019-0922-7

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DNA-PKcs modulates progenitor cell proliferation and fibroblast senescence in idiopathic pulmonary fibrosis

Abstract

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Methods

Results

Conclusions

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