HIPK2 attenuates bleomycin-induced pulmonary fibrosis by suppressing the Wnt/β-catenin signaling pathway
Abstract
Introduction. The present study aimed to investigate the effect of homeodomain interacting protein kinase 2 (HIPK2) on pulmonary fibrosis and the probable mechanisms.
Material and methods. We constructed a mouse model of bleomycin-induced pulmonary fibrosis and up-regulated the expression of HIPK2 in the lung by in vivo transfection. Lung tissues were collected for the detection of mesenchymal markers (α-SMA, collagen I, collagen III) and the expression of β-catenin as assessed by RT-PCR, western blot, and immunohistochemistry. Mouse lung fibroblasts (MLFs) with upregulation or downregulation of HIPK2 were successfully constructed and XAV939 was used to downregulate β-catenin expression. Then, we evaluated the activation of MLFs and the Wnt/β-catenin pathway under various conditions.
Results. The results showed that in the bleomycin-induced mouse model group, the lung alveolar structure was severely damaged, the amount of collagen fibers was increased in alveolar speta, and the expression of HIPK2 in the fibrotic area was found to be reduced. After upregulating HIPK2 in the lungs of the mouse fibrosis model we found that pulmonary fibrosis was attenuated and the expression of β-catenin and mesenchymal markers was reduced. The upregulation of HIPK2 inhibited the proliferation and migration of MLFs induced by TGF-β1, promoted apoptosis of MLFs, and reduced the expression of mesenchymal markers and β-catenin. Meanwhile, downregulation of HIPK2 promoted the proliferation and migration of MLFs, inhibited apoptosis, and promoted mesenchymal markers and β-catenin expression. XAV939 treatment of MLFs silencing HIPK2 inhibited their proliferation and activation via silencing HIPK2, promoted apoptosis, and reduced interstitial markers and β-catenin expression.
Conclusions. HIPK2 can attenuate bleomycin-induced pulmonary fibrosis by inhibiting the Wnt/β-catenin pathway in mouse lung fibroblasts.
Keywords: mousepulmonary fibrosismouse lung fibroblastHIPK2Wnt/β-cateninXAV939
References
- Deng KM, Yang XS, Luo Q, et al. Deleterious role of Th9 cells in pulmonary fibrosis. Cells. 2021; 10(11).
- Lee HY, Cho J, Kwak N, et al. Prognostic impact of malignant diseases in idiopathic pulmonary fibrosis. Sci Rep. 2020; 10(1): 18260.
- Sgalla G, Iovene B, Calvello M, et al. Idiopathic pulmonary fibrosis: pathogenesis and management. Respir Res. 2018; 19(1): 32.
- Raghu G, Richeldi L, Thomson CC, et al. authors of the guideline, Idiopathic Pulmonary Fibrosis (an Update) and Progressive Pulmonary Fibrosis in Adults: An Official ATS/ERS/JRS/ALAT Clinical Practice Guideline, American Thoracic Society, European Respiratory Society, Japanese Respiratory Society, and Latin American Thoracic Society. Diagnosis of idiopathic pulmonary fibrosis. An official ATS/ERS/JRS/ALAT clinical practice guideline. Am J Respir Crit Care Med. 2018; 198(5): e44–e68.
- Richeldi L, du Bois RM, Raghu G, et al. INPULSIS Trial Investigators. Efficacy of a tyrosine kinase inhibitor in idiopathic pulmonary fibrosis. N Engl J Med. 2014; 370(22): 2071–2082.
- King TE, Bradford WZ, Castro-Bernardini S, et al. ASCEND Study Group. A phase 3 trial of pirfenidone in patients with idiopathic pulmonary fibrosis. N Engl J Med. 2014; 370(22): 2083–2092.
- Raghu G, Rochwerg B, Zhang Y, et al. American Thoracic Society, European Respiratory society, Japanese Respiratory Society, Latin American Thoracic Association. An official ATS/ERS/JRS/ALAT clinical practice guideline: treatment of idiopathic pulmonary fibrosis. An update of the 2011 clinical practice guideline. Am J Respir Crit Care Med. 2015; 192(2): e3–19.
- Kim YH, Choi CY, Lee SJ, et al. Homeodomain-interacting protein kinases, a novel family of co-repressors for homeodomain transcription factors. J Biol Chem. 1998; 273(40): 25875–25879.
- Müller JP, Klempnauer KH. The CDC37-HSP90 chaperone complex co-translationally degrades the nascent kinase-dead mutant of HIPK2. FEBS Lett. 2021; 595(11): 1559–1568.
- Jin X, Qing S, Li Q, et al. Prostate cancer-associated SPOP mutations lead to genomic instability through disruption of the SPOP-HIPK2 axis. Nucleic Acids Res. 2021; 49(12): 6788–6803.
- Ricci A, Cherubini E, Ulivieri A, et al. Homeodomain-interacting protein kinase2 in human idiopathic pulmonary fibrosis. J Cell Physiol. 2013; 228(1): 235–241.
- Agnew C, Liu L, Liu S, et al. The crystal structure of the protein kinase HIPK2 reveals a unique architecture of its CMGC-insert region. J Biol Chem. 2019; 294(37): 13545–13559.
- Zhu W, Li J, Zhang Y, et al. Inhibition of HMGB1 Suppresses Hepatocellular Carcinoma Progression HIPK2-Mediated Autophagic Degradation of ZEB1. Front Oncol. 2021; 11: 599124.
- Kuwano Y, Nishida K, Akaike Y, et al. Homeodomain-interacting protein kinase-2: a critical regulator of the DNA damage response and the epigenome. Int J Mol Sci. 2016; 17(10).
- Liu R, Das B, Xiao W, et al. A novel inhibitor of homeodomain interacting protein kinase 2 mitigates kidney fibrosis through inhibition of the tgf-1/smad3 pathway. J Am Soc Nephrol. 2017; 28(7): 2133–2143.
- Jin Y, Ratnam K, Chuang PY, et al. A systems approach identifies HIPK2 as a key regulator of kidney fibrosis. Nat Med. 2012; 18(4): 580–588.
- He P, Yu ZJ, Sun CY, et al. Knockdown of HIPK2 attenuates the pro-fibrogenic response of hepatic stellate cells induced by TGF-β1. Biomed Pharmacother. 2017; 85: 575–581.
- Zheng H, Sun Y, Shu X, et al. Overexpression of microRNA-100-5p attenuates the endothelial cell dysfunction by targeting HIPK2 under hypoxia and reoxygenation treatment. J Mol Histol. 2021; 52(5): 1115–1125.
- Zhang F, Qi L, Feng Q, et al. HIPK2 phosphorylates HDAC3 for NF-κB acetylation to ameliorate colitis-associated colorectal carcinoma and sepsis. Proc Natl Acad Sci U S A. 2021; 118(28).
- Fan Y, Wang N, Chuang P, et al. Role of HIPK2 in kidney fibrosis. Kidney Int Suppl (2011). 2014; 4(1): 97–101.
- Kim EA, Kim JiE, Sung KiSa, et al. Homeodomain-interacting protein kinase 2 (HIPK2) targets beta-catenin for phosphorylation and proteasomal degradation. Biochem Biophys Res Commun. 2010; 394(4): 966–971.
- Cheng W, Wang F, Feng A, et al. CXXC5 attenuates pulmonary fibrosis in a bleomycin-induced mouse model and MLFs by suppression of the CD40/CD40L pathway. Biomed Res Int. 2020; 2020: 7840652.
- Yu W, Song X, Liu Y. TRB3 regulates pulmonary interstitial fibrosis through the MAPK signaling pathway. Int J Clin Exp Pathol. 2019; 12(9): 3247–3257.
- Huang SMA, Mishina YM, Liu S, et al. Tankyrase inhibition stabilizes axin and antagonizes Wnt signalling. Nature. 2009; 461(7264): 614–620.
- Spagnolo P, Sverzellati N, Rossi G, et al. Idiopathic pulmonary fibrosis: an update. Ann Med. 2015; 47(1): 15–27.
- Chanda D, Otoupalova E, Smith SR, et al. Developmental pathways in the pathogenesis of lung fibrosis. Mol Aspects Med. 2019; 65: 56–69.
- Blaquiere JA, Verheyen EM. Homeodomain-interacting protein kinases: diverse and complex roles in development and disease. Curr Top Dev Biol. 2017; 123: 73–103.
- Cao H, Wang C, Chen X, et al. Inhibition of Wnt/β-catenin signaling suppresses myofibroblast differentiation of lung resident mesenchymal stem cells and pulmonary fibrosis. Sci Rep. 2018; 8(1): 13644.
- Lam AP, Flozak AS, Russell S, et al. Nuclear β-catenin is increased in systemic sclerosis pulmonary fibrosis and promotes lung fibroblast migration and proliferation. Am J Respir Cell Mol Biol. 2011; 45(5): 915–922.
- Swann JB, Happe C, Boehm T. Elevated levels of Wnt signaling disrupt thymus morphogenesis and function. Sci Rep. 2017; 7(1): 785.
- Li C, Zheng Xu, Han Y, et al. XAV939 inhibits the proliferation and migration of lung adenocarcinoma A549 cells through the WNT pathway. Oncol Lett. 2018; 15(6): 8973–8982.
- Wei G, Ku S, Ma GK, et al. HIPK2 represses beta-catenin-mediated transcription, epidermal stem cell expansion, and skin tumorigenesis. Proc Natl Acad Sci U S A. 2007; 104(32): 13040–13045.
- Hikasa H, Sokol S. Phosphorylation of TCF proteins by homeodomain-interacting protein kinase 2. J Biol Chem. 2011; 286(14): 12093–12100.
- Kulak O, Chen H, Holohan B, et al. Disruption of wnt/β-catenin signaling and telomeric shortening are inextricable consequences of tankyrase inhibition in human cells. Mol Cell Biol. 2015; 35(14): 2425–2435.
- Wang C, Zhu H, Sun Z, et al. Inhibition of Wnt/β-catenin signaling promotes epithelial differentiation of mesenchymal stem cells and repairs bleomycin-induced lung injury. Am J Physiol Cell Physiol. 2014; 307(3): C234–C244.
