RACK1 Contributes to the Upregulation of Embryonic Genes in a Model of Cardiac Hypertrophy

Overview

Journal Sci Rep

Specialty Science

Date 2024 Oct 28

PMID 39465301

Authors

Marcello Ceci

Davide Bonvissuto

Flavia Papetti

Federica Silvestri

Claudio Sette

Elisabetta Catalani

Davide Cervia

Rosalba Gornati

Nicla Romano

Affiliations

Soon will be listed here.

Abstract

Receptors for activated C kinases (RACKs) have been shown to coordinate PKC-mediated hypertrophic signalling in mice. However, little information is available on its participation in embryonic gene expression. This study investigated the involvement of RACK1 in the expression of embryonic genes in a zebrafish (ZF) ex vivo heart culture model by using phenylephrine (PE) or a growth factors cocktail (GFs) as a prohypertrophic/regeneration stimulus. Blebbistatin (BL) inhibition has also been studied for its ability to block the signal transduction actions of some PEs. qRT‒PCR and immunoblot analyses confirmed the upregulation of RACK1 in the PE- and GFs-treated groups. BL administration counteracted PE-induced hypertrophy and downregulated RACK1 expression. Immunohistochemical analyses of the heart revealed the colocalization of RACK1 and embryonic genes, namely, Gata4, Wt1, and Nfat2, under stimulation, whereas these genes were expressed at lower levels in the BL treatment group. Culturing ZF heart cells activated via GFs treatment increased the expression of RACK1. The overexpression of RACK1 induced by the transfection of recombinant RACK1 cDNA in ZF heart cells increased the expression of embryonic genes, especially after one week of GFs treatment. In summary, these results support the involvement of RACK1 in the induction of embryonic genes during cardiac hypertrophy/GFs stimulation in a fish heart model, which can be used as an alternative study model for mammals.

References

Buoso E, Galasso M, Serafini M, Ronfani M, Lanni C, Corsini E . Transcriptional regulation of RACK1 and modulation of its expression: Role of steroid hormones and significance in health and aging. Cell Signal. 2017; 35:264-271. DOI: 10.1016/j.cellsig.2017.02.010. View

Kadrmas J, Smith M, Pronovost S, Beckerle M . Characterization of RACK1 function in Drosophila development. Dev Dyn. 2007; 236(8):2207-15. DOI: 10.1002/dvdy.21217. View

Adams D, Ron D, Kiely P . RACK1, A multifaceted scaffolding protein: Structure and function. Cell Commun Signal. 2011; 9:22. PMC: 3195729. DOI: 10.1186/1478-811X-9-22. View

Bass-Stringer S, Tai C, McMullen J . IGF1-PI3K-induced physiological cardiac hypertrophy: Implications for new heart failure therapies, biomarkers, and predicting cardiotoxicity. J Sport Health Sci. 2020; 10(6):637-647. PMC: 8724616. DOI: 10.1016/j.jshs.2020.11.009. View

Romano N, Di Giacomo B, Nobile V, Borreca A, Willems D, Tilesi F . Ribosomal RACK1 Regulates the Dendritic Arborization by Repressing FMRP Activity. Int J Mol Sci. 2022; 23(19). PMC: 9569565. DOI: 10.3390/ijms231911857. View

Iwasaki S, Kawamata T, Tomari Y . Drosophila argonaute1 and argonaute2 employ distinct mechanisms for translational repression. Mol Cell. 2009; 34(1):58-67. DOI: 10.1016/j.molcel.2009.02.010. View

Jannot G, Bajan S, Giguere N, Bouasker S, Banville I, Piquet S . The ribosomal protein RACK1 is required for microRNA function in both C. elegans and humans. EMBO Rep. 2011; 12(6):581-6. PMC: 3128278. DOI: 10.1038/embor.2011.66. View

Suzuki H, Katanasaka Y, Sunagawa Y, Miyazaki Y, Funamoto M, Wada H . Tyrosine phosphorylation of RACK1 triggers cardiomyocyte hypertrophy by regulating the interaction between p300 and GATA4. Biochim Biophys Acta. 2016; 1862(9):1544-57. DOI: 10.1016/j.bbadis.2016.05.006. View

Volta V, Beugnet A, Gallo S, Magri L, Brina D, Pesce E . RACK1 depletion in a mouse model causes lethality, pigmentation deficits and reduction in protein synthesis efficiency. Cell Mol Life Sci. 2012; 70(8):1439-50. PMC: 11113757. DOI: 10.1007/s00018-012-1215-y. View

10.

Pass J, Gao J, Jones W, Wead W, Wu X, Zhang J . Enhanced PKC beta II translocation and PKC beta II-RACK1 interactions in PKC epsilon-induced heart failure: a role for RACK1. Am J Physiol Heart Circ Physiol. 2001; 281(6):H2500-10. DOI: 10.1152/ajpheart.2001.281.6.H2500. View

11.

Kong P, Christia P, Frangogiannis N . The pathogenesis of cardiac fibrosis. Cell Mol Life Sci. 2013; 71(4):549-74. PMC: 3769482. DOI: 10.1007/s00018-013-1349-6. View

12.

Frangogiannis N . Cardiac fibrosis: Cell biological mechanisms, molecular pathways and therapeutic opportunities. Mol Aspects Med. 2018; 65:70-99. DOI: 10.1016/j.mam.2018.07.001. View

13.

Care A, Catalucci D, Felicetti F, Bonci D, Addario A, Gallo P . MicroRNA-133 controls cardiac hypertrophy. Nat Med. 2007; 13(5):613-8. DOI: 10.1038/nm1582. View

14.

Yang W, Zhuang Y, Wu H, Su S, Li Y, Wang C . Substrate-dependent interaction of SPOP and RACK1 aggravates cardiac fibrosis following myocardial infarction. Cell Chem Biol. 2023; 30(10):1248-1260.e4. DOI: 10.1016/j.chembiol.2023.06.015. View

15.

Ceci M, Carlantoni C, Missinato M, Bonvissuto D, Di Giacomo B, Contu R . Micro RNAs are involved in activation of epicardium during zebrafish heart regeneration. Cell Death Discov. 2018; 4:41. PMC: 5849881. DOI: 10.1038/s41420-018-0041-x. View

16.

Hu B, Lelek S, Spanjaard B, El-Sammak H, Simoes M, Mintcheva J . Origin and function of activated fibroblast states during zebrafish heart regeneration. Nat Genet. 2022; 54(8):1227-1237. PMC: 7613248. DOI: 10.1038/s41588-022-01129-5. View

17.

Romano N, Ceci M . The face of epicardial and endocardial derived cells in zebrafish. Exp Cell Res. 2018; 369(1):166-175. DOI: 10.1016/j.yexcr.2018.05.022. View

18.

Dalby B, Cates S, Harris A, Ohki E, Tilkins M, Price P . Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications. Methods. 2004; 33(2):95-103. DOI: 10.1016/j.ymeth.2003.11.023. View

19.

Kowalewski J, Paris T, Gonzalez C, Lelievre E, Castano Valencia L, Boutrois M . Characterization of a member of the CEACAM protein family as a novel marker of proton pump-rich ionocytes on the zebrafish epidermis. PLoS One. 2021; 16(7):e0254533. PMC: 8274849. DOI: 10.1371/journal.pone.0254533. View

20.

Kalen M, Wallgard E, Asker N, Nasevicius A, Athley E, Billgren E . Combination of reverse and chemical genetic screens reveals angiogenesis inhibitors and targets. Chem Biol. 2009; 16(4):432-41. PMC: 3984492. DOI: 10.1016/j.chembiol.2009.02.010. View