Comprehensive maternal serum proteomics identifies the cytoskeletal proteins as non-invasive biomarkers in prenatal diagnosis of congenital heart defects.

Lizhu Chen,Wei Ma,Hui Gu,Tianchu Huang,Xiaowei Wei,Jun Li,Xiao Sun,Ze-Yu Yang,Li Zhang,Zhengwei Yuan,Henan Zhang,Jianing Miao,Lina Wu,Yili Zhao,Songying Cao,Liping Shan

doi:10.1038/srep19248

Abstract

Congenital heart defects (CHDs) are the most common group of major birth defects. Presently there are no clinically used biomarkers for prenatally detecting CHDs. Here, we performed a comprehensive maternal serum proteomics assessment, combined with immunoassays, for the discovery of non-invasive biomarkers for prenatal diagnosis of CHDs. A total of 370 women were included in this study. An isobaric tagging for relative and absolute quantification (iTRAQ) proteomic approach was used first to compare protein profiles in pooled serum collected from women who had CHD-possessing or normal fetuses, and 47 proteins displayed significant differential expressions. Targeted verifications were performed on 11 proteins using multiple reaction monitoring mass spectrometry (MRM-MS), and the resultant candidate biomarkers were then further validated using ELISA analysis. Finally, we identified a biomarker panel composed of 4 cytoskeletal proteins capable of differentiating CHD-pregnancies from normal ones [with an area under the receiver operating characteristic curve (AUC) of 0.938, P < 0.0001]. The discovery of cytoskeletal protein changes in maternal serum not only could help us in prenatal diagnosis of CHDs, but also may shed new light on CHD embryogenesis studies.

Highlights

Maternal blood is accepted as the “holy grail” for prenatal diagnosis
Comparative proteomic analysis recently has been used in prenatal diagnosis of fetal abnormalities, and found a number of differentially expressed proteins for non-invasive diagnosis of preeclampsia[10,11] and chromosome disorders[12,13,14,15]
We identified a panel consisting of 4 proteins (LMNA, FLNA, Tropomyosin α -4 chain (TPM4), and ACTG1) with fair robustness in regard to specificity and sensitivity in differentiating Congenital heart defects (CHDs) from controls

Summary

Introduction

It offers the opportunity to eliminate risk-associated invasive procedures, such as amniocentesis and chorionic villus sampling[5] It could be performed even in the primary hospitals as a screening method, and may help to set indications for referral for fetal echocardiography. This method probably could provide insight into the pathophysiologic basis for CHDs6, which may provide additional information to guide clinical interventions or target therapies before the development of CHDs. Comparative proteomic analysis is a powerful diagnostic tool to determine the onset, progression, and prognosis of human diseases[7]. This is the first comprehensive proteomic study aimed at prenatally diagnosing CHDs, and we identified a biomarker panel capable of differentiating CHD-pregnancies from normal ones These candidates were all involved in the cytoskeleton pathway. CHDs were associated with a specific pattern of changes in nuclear and sarcomeric cytoskeletons, which could have important implications in understanding the mechanisms involved

Methods

Results

Conclusion