Proteomics of Children Born After Intracytoplasmic Sperm Injection Reveal Indices of an Adverse Cardiometabolic Profile

ISSN 2472-1972 Proteomics of Children Born After Intracytoplasmic Sperm Injection Reveal Indices of an Adverse Cardiometabolic Profile Ioanna Kosteria,1 George Th. Tsangaris,3 Alexandra Gkourogianni,1 Athanasios Anagnostopoulos,3 Aggeliki Papadopoulou,3 Ioannis Papassotiriou,4 Dimitrios Loutradis,2 George P. Chrousos,1 and Christina Kanaka-Gantenbein1 1 Division of Endocrinology, Metabolism, and Diabetes, First Department of Pediatrics, and 2Division of In Vitro Fertilization, First Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens Medical School, Athens 11528, Greece; 3 Proteomics Research Unit, Biomedical Research Foundation of Academy of Athens, Athens 11527, Greece; and 4 Department of Clinical Biochemistry, Aghia Sophia Children’s Hospital, Athens 11527, Greece Context: Assisted reproduction technologies (ART), classic in vitro fertilization (IVF), and intracytoplasmic sperm injection (ICSI) are increasingly used. Several studies have demonstrated an unfavorable cardiometabolic profile of the ART offspring. Proteomics is a state-of-the-art technology used for the identification of early biomarkers of disease. Objectives: To investigate the proteomic profile of children born after ICSI compared with naturally conceived (NC) controls in search of cardiometabolic risk markers. Design: Cross-sectional case-control study: qualitative, comparative proteomic plasma analysis. Setting: Pediatric Endocrinology and IVF Outpatient Clinics, University of Athens and the Biomedical Research Foundation of the Academy of Athens. Participants: Forty-two sex- and age-matched couples of ICSI and NC children were assessed. Ten pairs additionally matched for birth weight and twin/single pregnancies were submitted to proteomic analysis. Intervention: Medical history, clinical examination, and blood biochemical, hormonal, and proteomic analyses. Main Outcome Measures: (1) Differences in auxological and laboratory data between groups. (2) Differences in plasma proteomic profile in 10 individual pairs and pooled samples. Results: The ICSI group had shorter gestation, more cesarean sections, smaller birth weight/length, and advanced maternal age. No major differences were observed regarding biochemical markers. Proteomic analysis revealed 19 over- and three underexpressed proteins in ICSI. Most overexpressed proteins are implicated in acute-phase reaction, blood coagulation, complement pathway activation, and iron and lipid metabolism, suggesting a subclinical unfavorable cardiometabolic profile. Conclusions: This study applies proteomics in ICSI-conceived children, providing evidence for an early adverse cardiometabolic profile and supporting the necessity of their long-term monitoring. Abbreviations: 2-DE, two-dimensional gel electrophoresis; ART, assisted reproduction technologies; hsCRP, high sensitivity C-reactive protein; ICSI, intracytoplasmic sperm injection; IVF, in vitro fertilization; NC, naturally conceived; RBP, retinol-binding protein; SDS, standard deviation score; SGA, small for gestational age. Received 15 November 2016 Accepted 22 February 2017 First Published Online 27 February 2017 April 2017 | Vol. 1, Iss. 4 doi: 10.1210/js.2016-1052 | Journal of the Endocrine Society | 288–301 doi: 10.1210/js.2016-1052 | Journal of the Endocrine Society | 289 Copyright © 2017 Endocrine Society This article is published under the terms of the Creative Commons Attribution-Non Commercial License (CC BY-NC-ND; https://creativecommons.org/licenses/by-nc-nd/4.0/). Freeform/Key Words: proteomics, ICSI, cardiometabolic, metabolism, IVF Assisted reproductive technologies (ART), comprising classic in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI), have significantly expanded over the years, aiming to overcome the growing problem of subfertility. ICSI, a technique first introduced in 1992, was primarily indicated for male subfertility, as it comprises the introduction of a single preselected sperm into the oocyte; however, it has progressively gained significant ground, constituting nowadays ~70% of all ART procedures performed [1, 2]. ART is usually associated with increased parental stress. Although it is still debated whether stress influences ART outcome, it is well established that glucocorticoids, by stimulating hepatic gluconeogenesis, inhibiting insulin actions on skeletal muscle, and potentiating its actions on adipose tissue, are the main mediators of the poor long-term outcome of an adverse intrauterine environment, ultimately promoting sarcopenia and visceral adiposity of the offspring and the occurrence of the metabolic syndrome in later life [3]. Infants born after ART are at higher risk of preterm delivery and intrauterine growth restriction, born small for gestational age (SGA) [4]. SGA, in turn, predisposes to adult disease and is associated with an unfavorable cardiometabolic profile in later life [5]. This alone could raise concern about the long-term health issues of children born after ART and may explain the observed poor cardiometabolic outcome of IVF children [6]. However, the latter might also be the result of the in vitro manipulations of the blastocyst and/or an adverse intratubal and/or intrauterine environment. This is in agreement with Barker’s fetal origin of adult disease hypothesis, which postulates development of epigenetic alterations occurring in prenatally distressed individuals [7]. Finally, there is increasing evidence for an association between ART and imprinting disorders, such as the Beckwith–Wiedemann or Angelmann syndromes, mostly in children born after ICSI [8] and less frequently after classic IVF [9], raising concern for genomic imprinting disorders and other harmful epigenetic changes in ART [10]. More specifically, reservations have been raised about the overall health of children born after ICSI, as this method has a greater risk for the introduction of a genetic error by bypassing all intrinsic barriers for the fertilization of abnormal gametes, thus eliminating sperm natural selection. Furthermore, embryo culture media and hormonal ovarian hyperstimulation have been shown to influence the imprinting status of some imprinted genes [11]. Indeed, epigenetic changes during the preimplantation period could be a potential mechanism for alterations in growth, development, and metabolism of ART-conceived children. Proteomics is a technology-driven science, which, in a high-throughput fashion, studies biologic fluid or tissue proteins, their posttranslational modifications, their interactions, changes in their expression levels, and their modifications in response to environmental factors, treatments, or disease [12]. To date, proteomic analysis has provided the opportunity to elucidate complex biological procedures and conditions, including fertility, fertilization, embryo implantation, and pregnancy. The advances in the use of mass spectral profiling in various biological fluids (...truncated)