Differential Secretion of Angiopoietic Factors and Expression of MicroRNA in Umbilical Cord Blood from Healthy Appropriate-For-Gestational-Age Preterm and Term Newborns—in Search of Biomarkers of Angiogenesis-Related Processes in Preterm Birth

Abstract

Objectives: Premature birth, defined as less than 37 weeks gestation, affects approximately 12% of all live births around the world. Advances in neonatal care have resulted in the increased survival of infants born prematurely. Although prematurity is a known risk factor for different cardiovascular diseases, little is known about the pathophysiology of vasculature during premature gestation and angiopoietic factors network during premature birth. Aims: The objective of this study was to determine whether the profile of several pro-angiogenic and anti-angiogenic factors in umbilical cord blood (UCB) is different in healthy appropriate-for-gestational-age preterm newborns and normal term babies. The second aim of this study was to investigate the microRNA (miRNAs) expression profile in UCB from preterm labor and to detect miRNAs potentially taking part in control of angogenesis-related processes (Angio-MiRs). Methods: Using an immunobead Luminex assay, we simultaneously measured the concentration of Angiogenin, Angiopoietin-1, FGF-acidic, FGF-basic, PDGF-aa, PlGF, VEGF, VEGF-D, Endostatin, Thrombospondin-2, NGF, BDNF, GDNF, and NT-4 in UCB samples collected from the preterm (n = 27) and term (n = 52) delivery. In addition, the global microRNA expression in peripheral blood mononuclear cells (PBMCs) circulating in such UCB samples was examined in this study using microarray MiRNA technique. Results: The concentrations of five from eight measured pro-angiogenic factors (VEGF, Angiopoietin-1, PDGF-AA, FGF-a, and FGF-b) were significantly lower in UCB from preterm newborns. On the contrary, two angiostatic factors (Endostatin and Thrombospondin-2) were significantly up-regulated in preterm UCB. Among analyzed neurotrophins in preterm newborns, the elevated UCB concentration was found only in the case of GDNF, whereas BDNF was significantly reduced. Moreover, two angiopoietic factors, VEGF-D and PlGF, and two neurotrophins, NT4 and NGF, did not differ in concentration in preterm and term babies. We also discovered that among the significantly down-regulated miRNAs, there were several classical Angio-MiRs (inter alia MiR-125, MiR-126, MiR-145, MiR-150, or MiR155), which are involved in angiogenesis regulation in newborn after preterm delivery. Conclusions: This is the first report of simultaneous measurements of several angiopoietic factors in UCB collected from infants during preterm and term labor. Here, we observed that several pro-angiogenic factors were at lower concentration in UCB collected from preterm newborns than term babies. In contrast, the two measured angiostatic factors, Endostatin and Thrombospondin-2, were significantly higher in UCB from preterm babies. This can suggest that distinct pathophysiological contributions from differentially expressed various angiopoietic factors may determine the clinical outcomes after preterm birth. Especially, our angiogenesis-related molecules analysis indicates that preterm birth of healthy, appropriate-for-gestational-age newborns is an “anti-angiogenic state” that may provide an increased risk for improper development and function of cardiovascular system in the adulthood. This work also contributes to a better understanding of the role of miRNAs potentially involved in angiogenesis control in preterm newborns.

Comparative Analysis of Global Gene Expression and Complement Components Levels in Umbilical Cord Blood from Preterm and Term Neonates: Implications for Significant Downregulation of Immune Response Pathways related to Prematurity

“…Total RNA isolated from UCB samples was pooled to generate one sample per group for microarray experiment. The procedure was performed according to the manufacturer's protocol, as previously described 21 . Generation of sense strand cDNA from the total RNA and subsequent fragmentation and labelling steps were made using GeneChip™ WT PLUS Reagent Kit (Thermo Fisher Scientific, Waltham, MA, USA).…”

“…In our previous studies, we have investigated umbilical cord blood (UCB) expression levels of small non-coding RNAs (miRNAs) potentially involved in angiogenesis control in preterm newborns and term neonates 21 . In this study, we aimed to investigate global gene expression in UCB samples from preterm neonates and compare it to term newborns.…”

Biomarkers of cardiovascular disease risk in the neonatal population

“…59 Prematurity was also associated with hypoproteinemia mediated impaired cardiovascular function and an "anti-angiogenic" status established by the downregulation of proangiogenic factors, angio-MiRs (inter alia MiR-125, MiR-126, MiR-145, MiR-150, or MiR155) and the upregulation of angiostatic factors endostatin and thrombospondin-2. 60,61 Serum proteomics have shown that the state of IUGR alters the expression of proteins such as lysophospholipid acyltransferase MBOAT7, serotransferrins, apolipoprotein E, apolipoprotein A-I, SERPIN1, with a known contribution to CVD pathways such as apoptosis, diabetes, dyslipidemia, inflammation, and arterial hypertension. 62 Concerning IUGR, renin, and angiotensin I levels were significantly elevated as well as systolic blood pressure and prolonged mean left ventricular isovolumic relaxation time.…”

MiR-125 Family in Cardiovascular and Cerebrovascular Diseases

“…Many angiogenesis related miRNAs, including miR-125, decreased significantly in cord blood miRNAs of preterm infants. This may increase the risk of abnormal development and function of the cardiovascular system after these premature infants reached adulthood ( Gródecka-Szwajkiewicz et al, 2020 ).…”