The hippo pathway is activated and is a causal mechanism for adipogenesis in arrhythmogenic cardiomyopathy.

Abstract

RATIONALE:

Mutations in the intercalated disc proteins, such as plakophilin 2 (PKP2), cause arrhythmogenic cardiomyopathy (AC). AC is characterized by the replacement of cardiac myocytes by fibro-adipocytes, cardiac dysfunction, arrhythmias, and sudden death.

OBJECTIVE:

To delineate the molecular pathogenesis of AC.

METHODS AND RESULTS:

Localization and levels of selected intercalated disc proteins, including signaling molecules, were markedly reduced in human hearts with AC. Altered protein constituents of intercalated discs were associated with activation of the upstream Hippo molecules in the human hearts, in Nkx2.5-Cre:Dsp(W/F) and Myh6:Jup mouse models of AC, and in the PKP2 knockdown HL-1 myocytes (HL-1(PKP2:shRNA)). Level of active protein kinase C-α isoform, which requires PKP2 for activity, was reduced. In contrast, neurofibromin 2 (or Merlin), a molecule upstream of the Hippo pathway and that is inactivated by protein kinase C-α isoform, was activated. Consequently, the downstream Hippo molecules mammalian STE20-like protein kinases 1/2 (MST1/2), large tumor suppressor kinases 1/2 (LATS1/2), and Yes-associated protein (YAP) (the latter is the effector of the pathway) were phosphorylated. Coimmunoprecipitation detected binding of phosphorylated YAP, phosphorylated β-catenin, and junction protein plakoglobin (the latter translocated from the junction). RNA sequencing, transcript quantitative polymerase chain reaction, and reporter assays showed suppressed activity of SV40 transcriptional enhancer factor domain (TEAD) and transcription factor 7-like 2 (TCF7L2), which are transcription factors of the Hippo and the canonical Wnt signaling, respectively. In contrast, adipogenesis was enhanced. Simultaneous knockdown of Lats1/2, molecules upstream to YAP, rescued inactivation of YAP and β-catenin and adipogenesis in the HL-1(PKP2:shRNA) myocytes.

CONCLUSIONS:

Molecular remodeling of the intercalated discs leads to pathogenic activation of the Hippo pathway, suppression of the canonical Wnt signaling, and enhanced adipogenesis in AC. The findings offer novel mechanisms for the pathogenesis of AC.

• Authors: Suet Nee Chen, Priyatansh Gurha, Raffaella Lombardi, Alessandra Ruggiero, James T Willerson, A J Marian
• Journal: Circulation research (Circ Res) Vol. 114 Issue 3 Pg. 454-68 (Jan 31 2014) ISSN: 1524-4571 [Electronic] United States
• PMID: 24276085 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Protein Serine-Threonine Kinases

Topics

• Adipogenesis (genetics)
• Animals
• Arrhythmogenic Right Ventricular Dysplasia (genetics, metabolism, pathology)
• Cell Line
• Hippo Signaling Pathway
• Humans
• Mice
• Mice, Knockout
• Protein Serine-Threonine Kinases (genetics, metabolism)
• Signal Transduction