Balkan Journal of Medical Genetics

DIFFERENTIALLY EXPRESSED CIRCULATING LONG-NONCODING RNAS IN PREMATURE INFANTS WITH RESPIRATORY DISTRESS SYNDROME
Bao ZD, Wan J, Zhu W, Shen JX, Yang Y, Zhou XY
*Corresponding Author: Dr. Yang Yang and Dr. Zhou Xiao‑Yu, E‑mail: yy860507@126.com (YY) and xyzhou161@163.com (XYZ), Tel:+ 86-25-83117362, Department of Neonatology, Children’s Hospital of Nanjing Medical University, Nanjing, Jiangsu 210008, P.R. China
page: 11

RESULTS

Clinical characteristics of the premature infants (G1, G2 and G3) This present study was comprised of 15 premature infants in total, 5 cases without RDS for control (named as Group 1, G1), 5 cases with mild RDS (named as Group 2, G2) and 5 with severe RDS (named as Group 3, G3). The recruitment procedures are shown in Figure 1, and clinical characteristics of the infants are summarized in table 2. There were no significant differences in gestational age, birth weight, 5 min Apgar score, gender, mode of delivery, twin pregnancy, mother of gestational diabetes, and prenatal glucocorticoid (P>0.05). Expression profile of lncRNAs and mRNAs in three groups (G1, G2, and G3) Affymetrix Human GeneChip was utilized to determine the expression spectrum of lncRNAs. As a result, the G1 vs. G2 comparison showed a total of 10112 differentially expressed lncRNAs, while the G2 vs. G3 comparison showed a total of 4663 differentially expressed lncRNAs. Of them, 135 lncRNAs were indicated to be differentially expressed among all three groups (G1, G2, and G3) after fold‑change filtering (adjusted P value<0.05 and |log2-fold change|>2). The information of the top 10 upregulated and downregulated lncRNAs are listed in table 3. A hierarchical clustering map is presented to distinguish lncRNA expression profiles among the three groups. (Figure 2) Construction of the lncRNA-mRNA co-expression network (G1, G2, and G3) Furthermore, differentially expressed mRNAs were compared for target prediction. Of them, the comparison between G1 and G2 showed a total of 2520 differentially expressed mRNAs, while the comparison between G2 and G3 showed a total of 530 mRNAs. The comparison of the three groups showed a total of 616 differentially expressed mRNAs. The lncRNA-mRNA co-expression network was constructed and showed a complex interaction between lncRNAs and mRNAs. Our analysis finally identified a total of 278 mRNAs closely related to 108 upregulated lncRNAs and 27 downregulated lncRNAs. These mRNAs with FC> 2 are shown in table 4. GO and KEGG analysis of lncRNAs and mRNAs GO and KEGG analysis were further performed to annotate the biological functions of differentially expressed mRNAs. The GO analysis indicated that the mRNAs co‑expressed with 108 upregulated lncRNAs were associated with 247 GO terms. The top 25 enriched terms are shown in Figures 3A and 3B. Additionally, a KEGG pathway analysis was performed to investigate the possible roles of the lncRNA‑associated mRNA genes. The most significant pathways enriched in the set of upregulated protein‑coding genes included PI3 kinase/Akt (PI3K-Akt), RAS, and mitogenactivated protein kinase (MAPK) signal pathways, while the most significant KEGG pathways of the downregulated protein‑coding genes were mainly related to metabolic pathways, etc. The bubble diagrams of the top KEGG pathways of mRNAs co‑expressed with upregulated and downregulated lncRNAs are shown in Figure 3C and 3D. Differentially expressed lncRNAs verified by qRT-PCR Following the screening, four lncRNAs including ENST00000470527.1, ENST00000504497.1, ENST00000417781.5, and ENST00000440408.5 were further confirmed by qRT-PCR. Compared with G2 and G1, the expression levels of these four lncRNAs were increased in G3, which is consistent with the results of RNA sequencing. The relative expression levels are shown in Figure 4. In-depth bioinformatics analysis of lncRNAs showed all the four lncRNAs were involved in the MAPK signaling pathway by down-regulating gene GRB2 and MECOM. Moreover, ENST00000417781.5 and ENST00000440408.5 may regulate the MAPK signaling pathway by down-regulating gene IGF2, while ENST00000440408.5 and ENST00000504497.1 may target the MAPK signaling pathway by up-regulating gene EFNA1 and PLA2G4F, respectively. This study also showed that the above four lncRNAs participate in PI3KAkt and RAS signaling pathway by down-regulating GRB2, while ENST00000417781.5 and ENST00000440408.5 could regulate PI3K-Akt pathway by down-regulating IGF2 and up-regulating ITGB8 and TCL1B. In addition, three of lncRNAs including ENST00000417781.5, ENST00000470527.1, and ENST00000504497.1 could target the RAS signaling pathway by up-regulating RASAL1, while ENST000004177, as well as ENST00000440408.5 could regulate RAS pathway by down-regulating IGF2. ENST00000440408.5 and ENST00000504497.1 may be involved in the RAS signaling pathway by up-regulating EFNA1 and PLA2G4F, respectively. LncRNAs including ENST00000417781.5, ENST00000470527.1, and ENST00000504497.1 could participate in the TGF-β signaling pathway by promoting gene expression of AMH and inhibiting TGIF2. In addition, ENST00000470527.1 and ENST00000504497.1 could be involved in the TGF-β pathway by up-regulating GDF7 and down-regulating GDF6, while ENST00000440408.5 may down-regulate FST to be involved in TGF-β pathway. The pathway regulatory network of four validated lncRNAs are shown in Figure 5.

Number 27 VOL. 27 (2), 2024	Number 27 VOL. 27 (1), 2024
Number 26 Number 26 VOL. 26(2), 2023 All in one	Number 26 VOL. 26(2), 2023
Number 26 VOL. 26, 2023 Supplement	Number 26 VOL. 26(1), 2023
Number 25 VOL. 25(2), 2022	Number 25 VOL. 25 (1), 2022
Number 24 VOL. 24(2), 2021	Number 24 VOL. 24(1), 2021
Number 23 VOL. 23(2), 2020	Number 22 VOL. 22(2), 2019
Number 22 VOL. 22(1), 2019	Number 22 VOL. 22, 2019 Supplement
Number 21 VOL. 21(2), 2018	Number 21 VOL. 21 (1), 2018
Number 21 VOL. 21, 2018 Supplement	Number 20 VOL. 20 (2), 2017
Number 20 VOL. 20 (1), 2017	Number 19 VOL. 19 (2), 2016
Number 19 VOL. 19 (1), 2016	Number 18 VOL. 18 (2), 2015
Number 18 VOL. 18 (1), 2015	Number 17 VOL. 17 (2), 2014
Number 17 VOL. 17 (1), 2014	Number 16 VOL. 16 (2), 2013
Number 16 VOL. 16 (1), 2013	Number 15 VOL. 15 (2), 2012
Number 15 VOL. 15, 2012 Supplement	Number 15 Vol. 15 (1), 2012
Number 14 14 - Vol. 14 (2), 2011	Number 14 The 9th Balkan Congress of Medical Genetics
Number 14 14 - Vol. 14 (1), 2011	Number 13 Vol. 13 (2), 2010
Number 13 Vol.13 (1), 2010	Number 12 Vol.12 (2), 2009
Number 12 Vol.12 (1), 2009	Number 11 Vol.11 (2),2008
Number 11 Vol.11 (1),2008	Number 10 Vol.10 (2), 2007
Number 10 10 (1),2007	Number 9 1&2, 2006
Number 9 3&4, 2006	Number 8 1&2, 2005
Number 8 3&4, 2004	Number 7 1&2, 2004
Number 6 3&4, 2003	Number 6 1&2, 2003
Number 5 3&4, 2002	Number 5 1&2, 2002
Number 4 Vol.3 (4), 2000	Number 4 Vol.2 (4), 1999
Number 4 Vol.1 (4), 1998	Number 4 3&4, 2001
Number 4 1&2, 2001	Number 3 Vol.3 (3), 2000
Number 3 Vol.2 (3), 1999	Number 3 Vol.1 (3), 1998
Number 2 Vol.3(2), 2000	Number 2 Vol.1 (2), 1998
Number 2 Vol.2 (2), 1999	Number 1 Vol.3 (1), 2000
Number 1 Vol.2 (1), 1999	Number 1 Vol.1 (1), 1998

About the journal ::: Editorial ::: Subscription ::: Information for authors ::: Contact