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Deletion of PDK1 Caused Cardiac Malmorphogenesis and Heart Defects Due to Profound Protein Phosphorylation Changes Mediated by SHP2

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Abstract

Phosphoinositide-dependent protein kinase-1 (PDK1), a master kinase and involved in multiple signaling transduction, participates in regulating embryonic cardiac development and postnatal cardiac remodeling. Germline PDK1 knockout mice displayed no heart development; in this article, we deleted PDK1 in heart tissue with different cre to characterize the temporospatial features and find the relevance with congenital heart disease(CHD), furthermore to investigate the underlying mechanism. Knocking out PDK1 with Nkx2.5-cre, the heart showed prominent pulmonic stenosis. Ablated PDK1 with Mef2cSHF-cre, the second heart field (SHF) exhibited severe hypoplasia. And deleted PDK1 with αMHC-cre, the mice displayed dilated heart disease, protein analysis indicated PI3K and ERK were activated; meanwhile, PDK1-AKT-GSK3, and S6K-S6 were disrupted; phosphorylation level of Akt473, S6k421/424, and Gsk3α21 enhanced; however, Akt308, S6k389, and Gsk3β9 decreased. In mechanism investigation, we found SHP2 membrane localization and phosphorylation level of SHP2542 elevated, which suggested SHP2 likely mediated the disruption.

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Abbreviations

PDK1 :

Phosphoinositide-dependent protein kinase-1

CHD:

Congenital heart disease

FHF:

First heart field

SHF:

Second heart field

OFT:

Outflow tract

LV:

Left ventricle

RV:

Right ventricle

VSD:

Ventricular septum defect

ASD:

Atrium septum defect

PI3K:

Phosphoinositide-3 kinase

PIP3:

PtdIns(3,4,5)P3

SHP2 :

Src homology-2-containing protein tyrosine phosphatase 2

PS:

Pulmonic stenosis

PTA:

Persistent truncus arteriosus

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Funding

This study was supported by grants from National Natural Science Foundation of China (Nos. 31930029).

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Authors and Affiliations

  1. Guangdong Medical University, Guangdong Dongguan, 523808, China

    Hongmei Luo

  2. Model Animal Research Center, Nanjing University, Jiangsu Nanjing, 210028, China

    Hongmei Luo, Zhongzhou Yang, Jie Li, Hengwei Jin, Mingyang Jiang & Congjia Shan

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  1. Hongmei Luo
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Correspondence to Hongmei Luo.

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Associate Editor Nicola Smart oversaw the review of this article.

Clinical Relevance

Knocking out PDK1 with Nkx2.5-cre, Mef2cSHF-cre, and αMHC-cre, hearts manifested individual different phenotypes, which are similar to the clinical pulmonic stenosis (PS), persistent truncus arteriosus (PTA), and dilated cardiomyopathy (DCM) respectively; this means PDK1 plays an essential role in embryonic cardiac development and postnatal cardiac remodeling. PS is a common congenital heart disease and accounts for approximately 8% of all congenital heart defects. It includes pulmonary valvular stenosis, infundibular stenosis, and trunk stenosis, which can exist simultaneously or solely, and clinical symptoms may vary from critical stenosis in the new born to mild asymptomatic stenosis without need for therapy throughout life. Knocking out PDK1 with Nkx2.5-cre could induce typical pulmonic stenosis. PTA is a rare severe congenital cardiac disease occupying 0.7–1.4 % of all congenital cardiac malformations. It occurs as a result of the failure of the conotruncal separation during fetal development and is categorized into three subtypes. Half of the clinical cases belong to type I, in which the pulmonary artery arises from lateral aspect of the arterial trunk and then branches into the left and right pulmonary arteries. The truncal valve is usually larger than the normal semilunar valve, number may vary from single to rarely six, and ventricular septal defect is always presented. The phenotype of PDK1 removal with Mef2cSHF-cre mimics persistent truncus arteriosus type I highly. Dilated cardiomyopathy is one of the most common myocardial structural abnormalities associated with heart failure. Knocking out PDK1 with αMHC-cre led to DCM, downregulation of PDK1 and its downstream substrates such as Akt, S6k was the main cause. From this research work, it helps us to understand the pathogenesis of PS, PTA and DCM, and provides some clues for their prevention and diagnosis.

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Luo, H., Yang, Z., Li, J. et al. Deletion of PDK1 Caused Cardiac Malmorphogenesis and Heart Defects Due to Profound Protein Phosphorylation Changes Mediated by SHP2. J. of Cardiovasc. Trans. Res. 16, 1220–1231 (2023). https://doi.org/10.1007/s12265-023-10380-y

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