Silencing of UTX Mitigates Aging-Associated Cardiac Fibrosis Via Blocking Cardiac Fibroblasts-to-Myofibroblasts Trans-Differentiation

Overview

Journal Anatol J Cardiol

Publisher Kare Publishing

Specialty Cardiology & Vascular Diseases

Date 2023 Jun 8

PMID 37288854

Authors

Chao Li

Tiantian Lin

Delin Li

Daoyuan Si

Huan Sun

Sibao Yang

Zhongfan Zhang

Qian Zhang

Kaiyao Shi

Affiliations

Soon will be listed here.

Abstract

Background: Cardiac fibrosis increases with age. Fibroblast activation plays an essential role in cardiac fibrosis. Histone modifications are involved in various chromatin-dependent processes. Attenuation of the histone H3 trimethylation on lysine 27 demethylase UTX by RNA interference or heterozygous mutation extends lifespan in worm. The objective of this study was to explore whether epigenetic silencing of UTX mitigates aging-associated cardiac fibrosis.

Methods: Middle-aged mice (15 months old) were used and started to receive adeno-associated virus-scrambled-small hairpin RNA and adeno-associated virus-UTX-small hairpin RNA every 3 months from 15 months to 21 months, respectively. The mice were euthanized at 24 months of age (length of the study).

Results: Adeno-associated virus-UTX-small hairpin RNA delivery significantly attenu-ated aging-associated increase in blood pressure, especially in diastolic blood pressure, indicating silencing of UTX rescued aging-associated cardiac dysfunction. Aging-associated cardiac fibrosis is characterized by fibroblast activation and abundant extracellular matrix deposition, including collagen deposition and alpha smooth muscle actin activation. Silencing of UTX abolished collagen deposition and alpha smooth muscle actin activation, decreased serum transforming growth factor β, blocked cardiac fibro blast s-to- myofi brobl asts trans-differentiation by elevation of cardiac resident mature fibroblast markers, TCF21, and platelet-derived growth factor receptor alpha, which are important proteins for maintaining cardiac fibroblast physiological function. In the mechanistic study, adeno-associated virus-UTX-small hairpin RNA blocked transforming growth factor β-induced cardiac fibro blast s-to- myofi brobl asts trans-differentiation in isolated fibroblasts from 24-month-old mouse heart. The same results demonstrated as the in vivo study.

Conclusions: Silencing of UTX attenuates aging-associated cardiac fibrosis via blocking cardiac fibroblasts-to-myofibroblasts transdifferentiation and consequently attenuates aging-associated cardiac dysfunction and cardiac fibrosis.

References

Rog-Zielinska E, Norris R, Kohl P, Markwald R . The Living Scar--Cardiac Fibroblasts and the Injured Heart. Trends Mol Med. 2016; 22(2):99-114. PMC: 4737999. DOI: 10.1016/j.molmed.2015.12.006. View

Agger K, Cloos P, Christensen J, Pasini D, Rose S, Rappsilber J . UTX and JMJD3 are histone H3K27 demethylases involved in HOX gene regulation and development. Nature. 2007; 449(7163):731-4. DOI: 10.1038/nature06145. View

Colella P, Ronzitti G, Mingozzi F . Emerging Issues in AAV-Mediated Gene Therapy. Mol Ther Methods Clin Dev. 2018; 8:87-104. PMC: 5758940. DOI: 10.1016/j.omtm.2017.11.007. View

Rohrer D, Hartong R, Dillmann W . Influence of thyroid hormone and retinoic acid on slow sarcoplasmic reticulum Ca2+ ATPase and myosin heavy chain alpha gene expression in cardiac myocytes. Delineation of cis-active DNA elements that confer responsiveness to thyroid hormone but not.... J Biol Chem. 1991; 266(13):8638-46. View

Lan F, Bayliss P, Rinn J, Whetstine J, Wang J, Chen S . A histone H3 lysine 27 demethylase regulates animal posterior development. Nature. 2007; 449(7163):689-94. DOI: 10.1038/nature06192. View

Tallquist M, Molkentin J . Redefining the identity of cardiac fibroblasts. Nat Rev Cardiol. 2017; 14(8):484-491. PMC: 6329009. DOI: 10.1038/nrcardio.2017.57. View

Trial J, Heredia C, Taffet G, Entman M, Cieslik K . Dissecting the role of myeloid and mesenchymal fibroblasts in age-dependent cardiac fibrosis. Basic Res Cardiol. 2017; 112(4):34. PMC: 5591578. DOI: 10.1007/s00395-017-0623-4. View

Rohr U, Wulf M, Stahn S, Steidl U, Haas R, Kronenwett R . Fast and reliable titration of recombinant adeno-associated virus type-2 using quantitative real-time PCR. J Virol Methods. 2002; 106(1):81-8. DOI: 10.1016/s0166-0934(02)00138-6. View

Wipff P, Rifkin D, Meister J, Hinz B . Myofibroblast contraction activates latent TGF-beta1 from the extracellular matrix. J Cell Biol. 2007; 179(6):1311-23. PMC: 2140013. DOI: 10.1083/jcb.200704042. View

10.

MacKenna D, Summerour S, Villarreal F . Role of mechanical factors in modulating cardiac fibroblast function and extracellular matrix synthesis. Cardiovasc Res. 2000; 46(2):257-63. DOI: 10.1016/s0008-6363(00)00030-4. View

11.

Rinn J, Chang H . Genome regulation by long noncoding RNAs. Annu Rev Biochem. 2012; 81:145-66. PMC: 3858397. DOI: 10.1146/annurev-biochem-051410-092902. View

12.

Sun Y, Kiani M, Postlethwaite A, Weber K . Infarct scar as living tissue. Basic Res Cardiol. 2002; 97(5):343-7. DOI: 10.1007/s00395-002-0365-8. View

13.

Hermens W, ter Brake O, Dijkhuizen P, Sonnemans M, Grimm D, Kleinschmidt J . Purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. Hum Gene Ther. 1999; 10(11):1885-91. DOI: 10.1089/10430349950017563. View

14.

Balakrishnan B, Jayandharan G . Basic biology of adeno-associated virus (AAV) vectors used in gene therapy. Curr Gene Ther. 2014; 14(2):86-100. DOI: 10.2174/1566523214666140302193709. View

15.

Jin C, Li J, Green C, Yu X, Tang X, Han D . Histone demethylase UTX-1 regulates C. elegans life span by targeting the insulin/IGF-1 signaling pathway. Cell Metab. 2011; 14(2):161-72. DOI: 10.1016/j.cmet.2011.07.001. View

16.

Brummelkamp T, Bernards R, Agami R . A system for stable expression of short interfering RNAs in mammalian cells. Science. 2002; 296(5567):550-3. DOI: 10.1126/science.1068999. View

17.

Paddison P, Caudy A, Bernstein E, Hannon G, Conklin D . Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. Genes Dev. 2002; 16(8):948-58. PMC: 152352. DOI: 10.1101/gad.981002. View

18.

Olson L, Soriano P . Increased PDGFRalpha activation disrupts connective tissue development and drives systemic fibrosis. Dev Cell. 2009; 16(2):303-13. PMC: 2664622. DOI: 10.1016/j.devcel.2008.12.003. View

19.

Pieraggi M, BOUISSOU H, Angelier C, Uhart D, Magnol J, Kokolo J . [The fibroblast]. Ann Pathol. 1985; 5(2):65-76. View

20.

Barsha G, Denton K, Mirabito Colafella K . Sex- and age-related differences in arterial pressure and albuminuria in mice. Biol Sex Differ. 2016; 7:57. PMC: 5109725. DOI: 10.1186/s13293-016-0110-x. View