Plasma Metabolomic Profiling Suggests Early Indications for Predisposition to Latent Insulin Resistance in Children Conceived by ICSI

et al. (2014) Plasma Metabolomic Profiling Suggests Early Indications for Predisposition to Latent Insulin Resistance in Children Conceived by ICSI. PLoS ONE 9(4): e94001. doi:10.1371/journal.pone.0094001 Plasma Metabolomic Profiling Suggests Early Indications for Predisposition to Latent Insulin Resistance in Children Conceived by ICSI Alexandra Gkourogianni 0 Ioanna Kosteria 0 Aristeidis G. Telonis 0 Alexandra Margeli 0 Emilia Mantzou 0 Maria Konsta 0 Dimitrios Loutradis 0 George Mastorakos 0 Ioannis Papassotiriou 0 Maria I. Klapa 0 Christina Kanaka-Gantenbein 0 George P. Chrousos 0 Ioannis P. Androulakis, Rutgers University, United States of America 0 1 Division of Endocrinology, Diabetes and Metabolism, First Department of Pediatrics, University of Athens Medical School , Athens , Greece , 2 Metabolic Engineering and Systems Biology Laboratory, Institute of Chemical Engineering Sciences, Foundation for Research and Technology-Hellas (FORTH/ICE-HT) , Patras , Greece , 3 Department of Clinical Biochemistry, ''Aghia Sophia'' Children's Hospital , Athens , Greece , 4 Endocrine Unit, Department of Endocrinology and Metabolism, Evgenidion Hospital , Athens , Greece , 5 Division of In Vitro Fertilization, First Department of Obstetrics and Gynecology, University of Athens Medical School , Athens , Greece , 6 Division of Endocrinology, Second Department of Obstetrics and Gynecology, University of Athens Medical School , Athens , Greece , 7 Graduate Program ''Biological Technology'', Department of Biology, University of Patras , Greece Background: There have been increasing indications about an epigenetically-based elevated predisposition of assisted reproductive technology (ART) offspring to insulin resistance, which can lead to an unfavorable cardio-metabolic profile in adult life. However, the relevant long-term systematic molecular studies are limited, especially for the IntraCytoplasmic Sperm Injection (ICSI) method, introduced in 1992. In this study, we carefully defined a group of 42 prepubertal ICSI and 42 naturally conceived (NC) children. We assessed differences in their metabolic profile based on biochemical measurements, while, for a subgroup, plasma metabolomic analysis was also performed, investigating any relevant insulin resistance indices. Methods & Results: Auxological and biochemical parameters of 42 6.862.1 yrs old ICSI-conceived and 42 age-matched controls were measured. Significant differences between the groups were determined using univariate and multivariate statistics, indicating low urea and low-grade inflammation markers (YKL-40, hsCRP) and high triiodothyronine (T3) in ICSIchildren compared to controls. Moreover, plasma metabolomic analysis carried out for a subgroup of 10 ICSI- and 10 NC girls using Gas Chromatography-Mass Spectrometry (GC-MS) indicated clear differences between the two groups, characterized by 36 metabolites linked to obesity, insulin resistance and metabolic syndrome. Notably, the distinction between the two girl subgroups was accentuated when both their biochemical and metabolomic measurements were employed. Conclusions: The present study contributes a large auxological and biochemical dataset of a well-defined group of prepubertal ICSI-conceived subjects to the research of the ART effect to the offspring's health. Moreover, it is the first time that the relevant usefulness of metabolomics was investigated. The acquired results are consistent with early insulin resistance in ICSI-offspring, paving the way for further systematic investigations. These data support that metabolomics may unravel metabolic differences before they become clinically or biochemically evident, underlining its utility in the ART research. - Funding: The study was funded by University of Athens, First Pediatric Clinic and FORTH/ICE-HT internal funds and a fellowship of Panhellenic Endocrinology Society to Dr. Alexandra Gkourogianni. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. . These authors contributed equally to this work. Assisted reproductive technology (ART) comprises the classic in vitro fertilization (IVF) and the intra-cytoplasmic sperm injection (ICSI), the latter primarily developed and indicated for the treatment of male infertility [13]. ICSI bypasses several natural selection barriers, as it involves the injection of a single spermatozoon or even spermatid directly into the oocyte through the zona pellucida. The spermatozoa and/or spermatids currently used in ICSI are usually obtained from oligozoospermic men [3]. The ovarian stimulation protocols and the in vitro manipulation of both gametes and the blastocyst raise serious concerns about the genetic, epigenetic and developmental risks of the ICSI method to the offspring [46]. Despite these concerns, the European Society of Human Reproduction and Embryology (ESHRE) reported a recent marked increase in the use of ICSI [1], even in cases of no apparent medical indication [7]. Presently, long-term detailed systematic prospective studies of the ART offspring regarding potential health risks are limited, especially for the ICSI method due to its recent (i.e. in 1992) introduction as ART method. The existing studies have mainly investigated auxological data, indicating a higher risk of preterm delivery and intrauterine growth restriction (IUGR) for the ART offspring, leading to children born prematurely and/or small for gestational age (SGA) [8]. Predisposition for adult disease is per se increased by being born SGA [911], further accentuated by the fact that ART pregnancies are characterized by older mothers, an increased anxiety level before and during pregnancy, and a higher rate of multiple gestations, all risk factors for numerous adverse effects [12]. However, our group as well as others have extended the investigation to cardiometabolic markers reporting that children born after classic IVF show early indices of cardiometabolic derangements, such as higher blood pressure (BP) or higher triglycerides, irrespectively of having been born SGA or not, indicating thus that ART per se is a predisposing factor for unfavorable cardiometabolic outcomes [13,14]. Glucocorticoids are considered the main mediators of the poor long-term outcome of an adverse intrauterine environment, by stimulating hepatic gluconeogenesis, inhibiting insulin actions on skeletal muscle and potentiating insulins actions on visceral adipose tissue, ultimately promoting visceral adiposity and sarcopenia leading to metabolic syndrome [15]. Recent evidence suggests that ART may be associated with epigenetic modifications in several genes that could have long-term impact on the health of the offspring [16] and that both obesity and metabolic syndrome may have their basis in epigenetic modifications [17]. Thus, systematic studies of the ART offspring are req (...truncated)