Disruption of Planar Cell Polarity Pathway Attributable to Valproic Acid-Induced Congenital Heart Disease through Hdac3 Participation in Mice.

Hong-yu Duan; Kai-yu Zhou; Tao Wang; Yi Zhang; Yi-fei LI; Yi-min Hua; Chuan Wang

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Disruption of Planar Cell Polarity Pathway Attributable to Valproic Acid-Induced Congenital Heart Disease through Hdac3 Participation in Mice.

Author: Hong-Yu DUAN ^{1
,

2} ; Kai-Yu ZHOU ^{1
,

3
,

4
,

5} ; Tao WANG ^{1
,

2} ; Yi ZHANG ^{4
,

5
,

6} ; Yi-Fei LI ^{1
,

2} ; Yi-Min HUA ^{1
,

3
,

4
,

5} ; Chuan WANG ^{1
,

2}
Author Information

1. Department of Pediatric Cardiology, West China Second University Hospital, Sichuan University
2. Cardiac Development and Early Intervention Unit, West China Institute of Women and Children's Health, West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
3. Cardiac Development and Early Intervention Unit, West China Institute of Women and Children's Health, West China Second University Hospital, Sichuan University
4. Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education
5. Key Laboratory of Development and Diseases of Women and Children of Sichuan Province, West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
6. Cardiac Development and Early Intervention Unit, West China Institute of Women and Children's Health, West China Second University Hospital, Sichuan University
Publication Type:Journal Article
Keywords: Congenital Heart Disease; Hdacs; Planar Cell Polarity; Valproic Acid
MeSH: Animals; Cell Polarity; Enzyme Inhibitors; adverse effects; Female; Fetal Heart; embryology; Heart Defects, Congenital; chemically induced; physiopathology; Histone Deacetylase Inhibitors; Histone Deacetylases; drug effects; physiology; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins; Pregnancy; Rats; Transfection; Valproic Acid; adverse effects
From: Chinese Medical Journal 2018;131(17):2080-2088
CountryChina
Language:English
Abstract: Background:Valproic acid (VPA) exposure during pregnancy has been proven to contribute to congenital heart disease (CHD). Our previous findings implied that disruption of planar cell polarity (PCP) signaling pathway in cardiomyocytes might be a factor for the cardiac teratogenesis of VPA. In addition, the teratogenic ability of VPA is positively correlated to its histone deacetylase (HDAC) inhibition activity. This study aimed to investigate the effect of the VPA on cardiac morphogenesis, HDAC1/2/3, and PCP key genes (Vangl2/Scrib/Rac1), subsequently screening out the specific HDACs regulating PCP pathway.
Methods:VPA was administered to pregnant C57BL mice at 700 mg/kg intraperitoneally on embryonic day 10.5. Dams were sacrificed on E15.5, and death/absorption rates of embryos were evaluated. Embryonic hearts were observed by hematoxylin-eosin staining to identify cardiac abnormalities. H9C2 cells (undifferentiated rat cardiomyoblasts) were transfected with Hdac1/2/3 specific small interfering RNA (siRNA). Based on the results of siRNA transfection, cells were transfected with Hdac3 expression plasmid and subsequently mock-treated or treated with 8.0 mmol/L VPA. Hdac1/2/3 as well as Vangl2/Scrib/Rac1 mRNA and protein levels were determined by real-time quantitative polymerase chain reaction and Western blotting, respectively. Total HDAC activity was detected by colorimetric assay.
Results:VPA could induce CHD (P < 0.001) and inhibit mRNA or protein expression of Hdac1/2/3 as well as Vangl2/Scrib in fetal hearts, in association with total Hdac activity repression (all P < 0.05). In vitro, Hdac3 inhibition could significantly decrease Vangl2/Scrib expression (P < 0.01), while knockdown of Hdac1/2 had no influence (P > 0.05); VPA exposure dramatically decreased the expression of Vanlg2/Scrib together with Hdac activity (P < 0.01), while overexpression of Hdac3 could rescue the VPA-induced inhibition (P > 0.05).
Conclusion:VPA could inhibit Hdac1/2/3, Vangl2/Scrib, or total Hdac activity both in vitro and in vivo and Hdac3 might participate in the process of VPA-induced cardiac developmental anomalies.