Modulation of ADAR mRNA expression in patients with congenital heart defects

Figures

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  Fig 1. ADAR2 expression is reduced in CHD patients. (A) ADAR1 p110 is strongly expressed in brain cerebral hemispheres as compared to left heart ventricle. The Y-axis represented the log10 of the transcript per million (TPM) value. The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. (B) The expression of ADAR1 p150 is strongest in the bladder, followed by the cervix and tibial artery. ADAR1p150 expression is slightly higher in the left heart ventricle as compared to cerebral hemisphere. The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. (C) ADAR2 expression is strongest in the tibial artery, followed by the bladder, stomach aorta and coronary artery. The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. (D) FOXP1 expression is strongest in the aorta, followed by the tibial artery and bladder. The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. (E) Bar graph showing the relative RNA levels of ADAR1, ADAR2 and FOXP1 in patients compared to controls. ADAR2 and FOXP1…

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  Fig 2. Heart defect-specific function. (A& B) ADAR1 p110 and ADAR1 p150 showed higher expression in the atrial appendage than in the left ventricle. The Y-axis represented the log10 of the transcript per million (TPM) value. The expression of ADAR1 p150 (log10TPM 0.5) is lower in heart atrium as compared to ADARp110 (log10TPM 1.0). The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. This indicated that ADAR1 isoforms express strongly in the heart atrium as compared to ventricles. (C&D) ADAR2 and FOXP1 showed higher expression in the atrial appendage than in the left ventricle (log10TPM 0.7). The expression of ADAR2 and FOXP1 is lower as compared to ADAR1. The whiskers represent the maximum and minimum values. The median is represented as a black line in the box. (E& F) Bar graph showing upregulation of ADAR1 p110 (ADAR1c) and p150 (ADAR1i) in different CHD cases. Both isoforms are significantly upregulated in ASD and VSD. This indicated that both ADAR1 isoforms have potential role in atrial heart tissue. However, a 3- to 4-fold increase in RNA levels was found in AVSD and TOF cases compared to controls. This implies that ADAR1…

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  Fig 3. Expression analysis of the miR-29b target genes COL1A2 and IGF1 from ADAR2-KO mouse hearts in three different biological replicates. COL1A2 and IGF1 are cardiac hypertrophy and fibrosis related miR-29b target genes. miR-29b has been found downregulated ~2.0 fold in the absence of ADAR2. Bar graph showing slight but not significant changes in the expression of these genes. The error bars indicate the mean +/-SD. *P < 0.05 (versus control), as determined by the Mann–Whitney U test.

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  Fig 4. FLNB editing and ADAR1 expression in ADAR2-KO mice. (A) Electropherogram showing a 24% decrease in editing in ADAR2-KO compared to WT mouse tissues. This indicated that FLNB is primarily edited at the Q/R site by ADAR2. (B) Expression analysis of FLNB. The impact of FLNB editing on its expression was evaluated by semi-quantitative real time PCR. The bar graph showed that there was no significant difference in the expression of FLNB in the hearts of ADAR2-/- mice compared to those of WT mice. This implies that editing of FLNB did not alter its expression. The error bars indicate the mean +/-SD. (C) ADAR1 isoform expression in ADAR2-/- mouse tissue. The upregulation of ADAR1 in CHD is either because of the absence of ADAR2 or CHD occurrence. Therefore expression of ADAR1 isoforms was determined in the ADAR2KO mouse heart tissues. The bar graph showed no significant difference in the expression of the ADAR1 isoforms (p110 and p150) in the hearts of ADAR2-KO mice compared to those of WT mice. This implies that the upregulation of ADAR1 is due to CHD. The error bars indicate the mean +/-SD.