Restricted accessResearch articleFirst published online 2025-07
Exosomal Prolactin-Induced Protein Inhibits the Activation of cGMP/PKG Pathway Mediated by ATP2B2 to Promote Myocardial Fibrosis in Atrial Fibrillation
Myocardial fibrosis is an important medium for atrial fibrillation (AF). Exosomes have been demonstrated to affect the development of AF. This study explored the molecular mechanism of exosomes from patients with AF (AF-exo) mediating myocardial fibrosis and thus affecting the development of AF.
Results:
Prolactin-induced protein (PIP) is highly expressed in AF-exo. AF-exo promoted the proliferation and activation of cardiac fibroblasts (CFs) as well as the migration and endothelial-to-mesenchymal transition (EndMT) of human umbilical vein endothelial cells (HUVECs). However, the effect of AF-exo on CFs and HUVECs was mitigated by PIP-specific short hairpin RNA (shPIP). Adeno-associated virus (AAV)-shPIP reduced the incidence and duration of AF in rats, and improved myocardial fibrosis and collagen deposition. ATPase plasma membrane Ca2+ transporting 2 (ATP2B2) overexpression or inhibition reverses the role of PIP or shPIP in CFs, HUVECs, and AF rats. Activation of the cyclic guanosine monophosphate/protein kinase G (cGMP/PKG) pathway is beneficial to alleviate myocardial fibrosis, but this effect is mitigated by shATP2B2.
Innovation:
Our investigation substantiates the pivotal role of the PIP/ATP2B2 axis in both HUVEC myocardial fibrosis and EndMT progression. Our findings suggest that AF-exo can suppress the activation of the cGMP/PKG pathway mediated by ATP2B2 through exosomal PIP, thus promoting myocardial fibrosis, indicating potential targets for novel antifibrotic drug development targeting either PIP or ATP2B2.
Conclusion:
Exosomal PIP can inhibit the activation of cGMP/PKG pathway mediated by ATP2B2, thus promoting the development of AF. Antioxid. Redox Signal. 43, 14–36.
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References
1.
AghaG, MendelsonMM, Ward-CavinessCK, et al.Blood leukocyte DNA methylation predicts risk of future myocardial infarction and coronary heart disease. Circulation, 2019; 140(8):645–657; doi: 10.1161/circulationaha.118.039357
DebilyMA, MarhomySE, BoulangerV, et al.A functional and regulatory network associated with PIP expression in human breast cancer. PLoS One, 2009; 4(3):e4696; doi: 10.1371/journal.pone.0004696
4.
GarcíaC, ArandaJ, ArnoldE, et al.Vasoinhibins prevent retinal vasopermeability associated with diabetic retinopathy in rats via protein phosphatase 2A-dependent eNOS inactivation. J Clin Invest, 2008; 118(6):2291–2300; doi: 10.1172/jci34508
5.
GonzalezC, Rosas-HernandezH, Jurado-ManzanoB, et al.The prolactin family hormones regulate vascular tone through NO and prostacyclin production in isolated rat aortic rings. Acta Pharmacol Sin, 2015; 36(5):572–586; doi: 10.1038/aps.2014.159
6.
GoversR, RabelinkTJ. Cellular regulation of endothelial nitric oxide synthase. American Journal of Physiology Renal Physiology, 2001; 280(2):F193–F206; doi: 10.1152/ajprenal.2001.280.2.F193
7.
HaagensenDEJr, GallSA, BrazyJE, et al.Analysis of amniotic fluid, maternal plasma, and cord blood for a human breast gross cystic disease fluid protein. Am J Obstet Gynecol, 1980; 138(1):25–32; doi: 10.1016/0002-9378(80)90007-1
8.
HaagensenDEJr, MazoujianG, DilleyWG, et al.Breast gross cystic disease fluid analysis. I. Isolation and radioimmunoassay for a major component protein. Journal of the National Cancer Institute, 1979; 62(2):239–247.
9.
HaoH, YanS, ZhaoX, et al.Atrial myocyte-derived exosomal microRNA contributes to atrial fibrosis in atrial fibrillation. J Transl Med, 2022; 20(1):407; doi: 10.1186/s12967-022-03617-y
10.
HuangQ, MaJ, WuH, et al.PDE-5-inhibited BMSCs alleviate high glucose-induced myocardial fibrosis and cardiomyocyte apoptosis by activating the cGMP/PKG pathway. Front Biosci (Landmark Ed), 2023; 28(7):155; doi: 10.3108/3/j.fbl2807155
11.
HuangS, DengY, XuJ, et al.The role of exosomes and their cargos in the mechanism, diagnosis, and treatment of atrial fibrillation. Front Cardiovasc Med, 2021; 8:712828; doi: 10.3389/fcvm.2021.712828
12.
HuangY, LiuY, HuangQ, et al.TMT-based quantitative proteomics analysis of synovial fluid-derived exosomes in inflammatory arthritis. Front Immunol, 2022; 13:800902; doi: 10.3389/fimmu.2022.800902
13.
InserteJ, Garcia-DoradoD. The cGMP/PKG pathway as a common mediator of cardioprotection: Translatability and mechanism. Br J Pharmacol, 2015; 172(8):1996–2009; doi: 10.1111/bph.12959
14.
JanuaryCT, WannLS, AlpertJS, et al;American College of Cardiology/American Heart Association Task Force on Practice Guidelines. 2014 AHA/ACC/HRS guideline for the management of patients with atrial fibrillation: A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines and the Heart Rhythm Society. Journal of the American College of Cardiology, 2014; 64(21):e1–e76; doi: 10.1016/j.jacc.2014.03.022
15.
JordanNP, TingleSJ, ShuttleworthVG, et al.MiR-126-3p is dynamically regulated in endothelial-to-mesenchymal transition during fibrosis. Int J Mol Sci, 2021; 22(16); doi: 10.3390/ijms22168629
16.
KatoT, SekiguchiA, SagaraK, et al.Endothelial-mesenchymal transition in human atrial fibrillation. J Cardiol, 2017; 69(5):706–711; doi: 10.1016/j.jjcc.2016.10.014
17.
LajinessJD, ConwaySJ. The dynamic role of cardiac fibroblasts in development and disease. J Cardiovasc Transl Res, 2012; 5(6):739–748; doi: 10.1007/s12265-012-9394-3
18.
LiCY, ZhangJR, HuWN, et al.Atrial fibrosis underlying atrial fibrillation (Review). Int J Mol Med, 2021; 47(3); doi: 10.3892/ijmm.2020.4842
19.
LiG, YangJ, ZhangD, et al.Research progress of myocardial fibrosis and atrial fibrillation. Front Cardiovasc Med, 2022; 9:889706; doi: 10.3389/fcvm.2022.889706
20.
LiangW, MaJX, VanL, et al.Prolactin-induced protein facilitates corneal wound healing. Exp Eye Res, 2022; 225:109300; doi: 10.1016/j.exer.2022.109300
21.
LiuL, LuoF, LeiK. Exosomes containing LINC00636 inhibit MAPK1 through the miR-450a-2-3p overexpression in human pericardial fluid and improve cardiac fibrosis in patients with atrial fibrillation. Mediators Inflamm, 2021; 2021:9960241; doi: 10.1155/2021/9960241
22.
LopreiatoR, GiacomelloM, CarafoliE. The plasma membrane calcium pump: New ways to look at an old enzyme. J Biol Chem, 2014; 289(15):10261–10268; doi: 10.1074/jbc.O114.555565
23.
LvX, LiJ, HuY, et al.Overexpression of miR-27b-3p targeting Wnt3a regulates the signaling pathway of Wnt/β-Catenin and attenuates atrial fibrosis in rats with atrial fibrillation. Oxid Med Cell Longev, 2019; 2019:5703764; doi: 10.1155/2019/5703764
24.
MengL, LuY, WangX, et al.NPRC deletion attenuates cardiac fibrosis in diabetic mice by activating PKA/PKG and inhibiting TGF-β1/Smad pathways. Sci Adv, 2023; 9(31):eadd4222; doi: 10.1126/sciadv.add4222
25.
NaderiA, MeyerM. Prolactin-induced protein mediates cell invasion and regulates integrin signaling in estrogen receptor-negative breast cancer. Breast Cancer Res, 2012; 14(4):R111; doi: 10.1186/bcr3232
26.
NattelS. Molecular and cellular mechanisms of atrial fibrosis in atrial fibrillation. JACC Clinical Electrophysiology, 2017; 3(5):425–435; doi: 10.1016/j.jacep.2017.03.002
27.
QinL, ZangM, XuY, et al.Chlorogenic acid alleviates hyperglycemia-induced cardiac fibrosis through activation of the NO/cGMP/PKG pathway in cardiac fibroblasts. Mol Nutr Food Res, 2021; 65(2):e2000810; doi: 10.1002/mnfr.202000810
28.
RainerPP, KassDA. Old dog, new tricks: Novel cardiac targets and stress regulation by protein kinase G. Cardiovasc Res, 2016; 111(2):154–162; doi: 10.1093/cvr/cvw107
29.
SagrisM, VardasEP, TheofilisP, et al.Atrial fibrillation: Pathogenesis, predisposing factors, and genetics. Int J Mol Sci, 2021; 23(1); doi: 10.3390/ijms23010006
30.
SuzukiK, WashioT, TsukamotoS, et al.Habitual cigarette smoking attenuates shear-mediated dilation in the brachial artery but not in the carotid artery in young adults. Physiol Rep, 2020; 8(3):e14369; doi: 10.1481/4/phy2.14369
31.
TaoY, LamasV, DuW, et al.Treatment of monogenic and digenic dominant genetic hearing loss by CRISPR-Cas9 ribonucleoprotein delivery in vivo. Nat Commun, 2023; 14(1):4928; doi: 10.1038/s41467-023-40476-7
32.
UrbaniakA, JablonskaK, Podhorska-OkolowM, et al.Prolactin-induced protein (PIP)-characterization and role in breast cancer progression. Am J Cancer Res, 2018; 8(11):2150–2164.
33.
VenugopalH, HannaA, HumeresC, et al.Properties and functions of fibroblasts and myofibroblasts in myocardial infarction. Cells, 2022; 11(9); doi: 10.3390/cells11091386
WangS, MinJ, YuY, et al.Differentially expressed miRNAs in circulating exosomes between atrial fibrillation and sinus rhythm. J Thorac Dis, 2019; 11(10):4337–4348; doi: 10.2103/7/jtd.2019.09.50
36.
WangW, LiX, NingZ. The study of endothelial-mesenchymal transition in an atrial fibrillation rat model. Alternative Therapies in Health and Medicine, 2024; 30(1):179–185.
37.
WuX, ShowiheenSAA, SunAR, et al.Exosomes extraction and identification. Methods Mol Biol, 2019; 2054:81–91; doi: 10.1007/978-1-4939-9769-5_4
38.
XiaW, WanY, LiYY, et al.PFOS prenatal exposure induce mitochondrial injury and gene expression change in hearts of weaned SD rats. Toxicology, 2011; 282(1–2):23–29; doi: 10.1016/j.tox.2011.01.011
39.
XuY, GuoX, NingS, et al.Inhibition of IP(3)R3 attenuates endothelial to mesenchymal transition induced by TGF-β1 through restoring mitochondrial function. Biochem Biophys Res Commun, 2022; 619:144–150; doi: 10.1016/j.bbrc.2022.06.033
40.
YuLM, DiWC, DongX, et al.Melatonin protects diabetic heart against ischemia-reperfusion injury, role of membrane receptor-dependent cGMP-PKG activation. Biochim Biophys Acta Mol Basis Dis, 2018; 1864(2):563–578; doi: 10.1016/j.bbadis.2017.11.023
41.
YuanJW, ZhangYN, LiuYR, et al.Diffusion behaviors of integrins in single cells altered by epithelial to mesenchymal transition. Small, 2022; 18(5):e2106498; doi: 10.1002/smll.202106498
42.
ZeisbergEM, TarnavskiO, ZeisbergM, et al.Endothelial-to-mesenchymal transition contributes to cardiac fibrosis. Nat Med, 2007; 13(8):952–961; doi: 10.1038/nm1613
43.
ZhangL, WangS, LiY, et al.Cardioprotective effect of icariin against myocardial fibrosis and its molecular mechanism in diabetic cardiomyopathy based on network pharmacology: Role of ICA in DCM. Phytomedicine, 2021; 91:153607; doi: 10.1016/j.phymed.2021.153607
44.
ZhuP, LiH, ZhangA, et al.MicroRNAs sequencing of plasma exosomes derived from patients with atrial fibrillation: MiR-124-3p promotes cardiac fibroblast activation and proliferation by regulating AXIN1. J Physiol Biochem, 2022; 78(1):85–98; doi: 10.1007/s13105-021-00842-9
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