Downstream targets of the homeobox gene DLX3 are differentially expressed in the placentae of pregnancies affected by human idiopathic fetal growth restriction

Impaired fetal growth is one of the most important causes of prematurity, stillbirth and infant mortality. The pathogenesis of idiopathic fetal growth restriction (FGR) is poorly understood but is thought to be multifactorial and comprise a range of genetic causes. This research aimed to investigate non-coding RNAs (lncRNAs) in the placentas of male and female fetuses affected by FGR. RNA-Seq data were analyzed to detect lncRNAs, their potential target genes and circular RNAs (circRNAs); a differential analysis was also performed. The multilevel bioinformatic analysis enabled the detection of 23,137 placental lncRNAs and 4263 of them were classified as novel. In FGR-affected female fetuses’ placentas (ff-FGR), among 19 transcriptionally active regions (TARs), five differentially expressed lncRNAs (DELs) and 12 differentially expressed protein-coding genes (DEGs) were identified. Within 232 differentially expressed TARs identified in male fetuses (mf-FGR), 33 encompassed novel and 176 known lncRNAs, and 52 DEGs were upregulated, while 180 revealed decreased expression. In ff-FGR ACTA2-AS1, lncRNA expression was significantly correlated with five DEGs, and in mf-FGR, 25 TARs were associated with DELs correlated with 157 unique DEGs. Backsplicing circRNA processes were detected in the range of H19 lncRNA, in both ff- and mf-FGR placentas. The performed global lncRNAs characteristics in terms of fetal sex showed dysregulation of DELs, DEGs and circRNAs that may affect fetus growth and pregnancy outcomes. In female placentas, DELs and DEGs were associated mainly with the vasculature, while in male placentas, disturbed expression predominantly affected immune processes.

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Unique microRNA Signals in Plasma Exosomes from Pregnancies Complicated by Preeclampsia

Hypertension ◽

10.1161/hypertensionaha.119.14081 ◽

2020 ◽

Vol 75 (3) ◽

pp. 762-771 ◽

Cited By ~ 11

Author(s):

Hui Li ◽

Yingshi Ouyang ◽

Elena Sadovsky ◽

W. Tony Parks ◽

Tianjiao Chu ◽

...

Keyword(s):

Fetal Growth ◽

Fetal Growth Restriction ◽

Quantitative Polymerase Chain Reaction ◽

Growth Restriction ◽

Differentially Expressed ◽

Outcome Variable ◽

Significant Difference ◽

Exosomal Mirna ◽

Plasma Mirna ◽

Insight Into

Although preeclampsia is a common and serious complication of pregnancy, insight into its pathobiology and diagnosis is lacking. Circulating plasma exosomes, which contain RNA and other molecules and have recently become accessible for diagnostics, may be informative in this regard. We tested the hypothesis that preeclampsia may affect the miRNA cargo within circulating maternal blood exosomes. We collected plasma from 60 pregnant women at term, including 20 women with pregnancy complicated by preeclampsia, and 20 women with fetal growth restriction and 20 with healthy pregnancy, serving as controls. We isolated exosomes from the maternal plasma by continuous density gradient ultracentrifugation. Our main outcome variable was exosomal miRNA cargo, analyzed by quantitative polymerase chain reaction-based TaqMan advanced miRNA assay in a card format and the expression of differentially expressed exosomal miRNA in whole plasma from the same participants. We found that 7 miRNA species were differentially expressed in exosomes from women with preeclampsia and those from controls. In contrast, there was no significant difference in exosomal miRNA expression between women with fetal growth restriction and controls. The results were not affected by fetal sex. Only one of the preeclampsia-related, differentially expressed exosomal miRNAs was significantly different in whole plasma miRNA analysis. We concluded that unlike whole plasma miRNA, exosomes extracted from the plasma of women with preeclampsia exhibit a unique miRNA profile, suggesting that plasma exosomal miRNA could provide insight into the pathophysiology of preeclampsia, and may play a role in disease diagnostics.

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Homeobox gene ESX1L expression is decreased in human pre-term idiopathic fetal growth restriction

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gal037 ◽

2006 ◽

Vol 12 (5) ◽

pp. 335-340 ◽

Cited By ~ 19

Author(s):

P. Murthi

Keyword(s):

Fetal Growth ◽

Fetal Growth Restriction ◽

Homeobox Gene ◽

Growth Restriction

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Homeobox Gene HLX1Expression Is Decreased in Idiopathic Human Fetal Growth Restriction

American Journal Of Pathology ◽

10.2353/ajpath.2006.050637 ◽

2006 ◽

Vol 168 (2) ◽

pp. 511-518 ◽

Cited By ~ 40

Author(s):

Padma Murthi ◽

Vicki Doherty ◽

Joanne Said ◽

Susan Donath ◽

Shaun P. Brennecke ◽

...

Keyword(s):

Fetal Growth ◽

Fetal Growth Restriction ◽

Homeobox Gene ◽

Growth Restriction

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220. Homeobox gene HLX is a regulator of HGF/c-met mediated trophoblast migration

Reproduction Fertility and Development ◽

10.1071/srb08abs220 ◽

2008 ◽

Vol 20 (9) ◽

pp. 20

Author(s):

G. Rajaraman ◽

P. Murthi ◽

S. P. Brennecke ◽

B. Kalionis

Keyword(s):

Cell Migration ◽

Mrna Expression ◽

Fetal Growth ◽

Fetal Growth Restriction ◽

Homeobox Gene ◽

Trophoblast Cell ◽

Growth Restriction ◽

Trophoblast Cells ◽

Cytotrophoblast Cell ◽

Trophoblast Migration

Homeobox gene transcription factors play critical roles in normal placental development and are expressed in specialised trophoblast cells. Abnormal trophoblast cell function is associated with pregnancy disorders including fetal growth restriction. Our studies show homeobox gene HLX expression in trophoblast cells (1) and that HLX is significantly decreased in fetal growth restriction (2). HLX gene inactivation in cultured trophoblast cells shows that HLX is a regulator of cytokine-dependent trophoblast proliferation (3). Hepatocyte growth factor (HGF) activates trophoblast cell migration in a paracrine fashion and its receptor, c-met, is expressed in trophoblast cells. This study investigates the regulation of HGF/c-met mediated trophoblast migration by HLX, in two human trophoblast cell lines SGHPL-4 and HTR-8/SVNeo. HGF stimulation significantly increased HLX mRNA expression (e.g. 43.2 ± 2.5, HGF v. 18.4 ± 1.7 control, densitometric units, P < 0.001, n = 3). siRNA-mediated inactivation of HLX resulted in significantly decreased trophoblast migration (e.g. 32 ± 4, siRNA v. 127 ± 12 control, migrated cells, P < 0.05, n = 4). When HLX was inactivated in the presence of HGF stimulation, migration remained significantly decreased (e.g. 112 ± 15, siRNA + HGF v. 368 ± 44 HGF, migrated cells, P < 0.05, n = 4). In order to determine if HGF is acting via the c-met receptor, the Met tyrosine kinase inhibitor, SU11274, was employed to inhibit c-met activity. c-met inhibition resulted in significantly reduced HLX mRNA expression (e.g. 2.1 ± 0.32, SU11274 v. 12.3 ± 1.4 control, densitometric units, P < 0.05, n = 3). HLX expression remained significantly reduced with HGF stimulation and SU11274 mediated c-met inhibition (e.g. 8.02 ± 1.3, SU11274 v. 38.3 ± 5.4 HGF, densitometric units, P < 0.05, n = 3). This is the first study to show that homeobox gene HLX is a downstream effector gene of HGF, that HLX regulates trophoblast migration and that HGF, via its receptor c-met, acts through HLX to control cell migration. (1) Rajaraman G, Murthi P, Quinn L, Brennecke SP, Kalionis B. Homeodomain protein HLX is expressed primarily in cytotrophoblast cell types in the early human placenta. (2008) Reproduction, Fertility a (2) Murthi P, Doherty V, Said J, Donath S, Brennecke SP, Kalionis B. Homeobox gene HLX1 expression is decreased in idiopathic human fetal growth restriction. (2006) Am J Pathol. 2006 Feb;168(2):511–8. (3) Rajaraman G, Murthi P, Leo B, Brennecke SP, Kalionis B. Homeobox gene HLX1 is a regulator of colony stimulating factor-1 dependent cell proliferation. (2007) Placenta Volume 28, Issue 10, October

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205. Homeobox gene TGIF is increased in human idiopathic fetal growth restriction

Reproduction Fertility and Development ◽

10.1071/srb08abs205 ◽

2008 ◽

Vol 20 (9) ◽

pp. 5

Author(s):

P. Murthi ◽

N. Pathirage ◽

A. Borg ◽

S. Brennecke ◽

B. Kalionis

Keyword(s):

Fetal Growth ◽

Fetal Growth Restriction ◽

Homeobox Gene ◽

Growth Restriction ◽

Growth Potential ◽

Mrna Levels ◽

Developmental Abnormalities ◽

Clinical Criteria ◽

Targeted Mutation ◽

Clinically Significant

Fetal Growth Restriction (FGR) is a clinically significant pregnancy disorder in which the fetus fails to achieve its full growth potential in utero. Recently, we identified a novel homeobox gene TGIF, in the placenta using microarray expression profiling (1). Targeted mutation of tgif in mouse results in placental dysfunction (2). In this study, we have investigated TGIF expression levels in idiopathic FGR. FGR-affected placental samples were collected based on strict clinical criteria to ensure inclusion of cases at the severe end of the spectrum of the disease. TGIF mRNA expression was analysed in placentae obtained from pregnancies complicated by idiopathic FGR and gestation-matched control pregnancies (n = 25 each). Real-time PCR showed a significant increase in TGIF mRNA levels in FGR-affected placentae and gestation-matched controls [1.29 ± 0.06 FGR v. 0.78 ± 0.04 Control, P < 0.001]. western blotting using a TGIF polyclonal antibody revealed significantly increased levels of TGIF protein in term FGR-affected placentae compared with term controls [3970 ± 1101 (n = 10) v. 2323 ± 644 (n = 10), P < 0.05]. The spatial distribution of TGIF protein by immunohistochemistry revealed immunoreactive TGIF protein in residual cytotrophoblast cells, syncytiotrophoblast cells, microvascular endothelial cells and in stromal cells. We conclude that increased expression of homeobox gene TGIF may be a contributing factor to the developmental abnormalities seen in the FGR-affected placentae. (1) Murthi P, Hiden U, Rajaraman G, Kalionis B. Placenta May 29, [Epub ahead of print]. (2) Bartholin L, Melhuish TA et al. Dev Biol. 2008 May 2. [Epub ahead of print].

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