Multi-omics integration identifies key upstream regulators of pathomechanisms in hypertrophic cardiomyopathy due to truncating MYBPC3 mutations.

BACKGROUND: Hypertrophic cardiomyopathy (HCM) is the most common genetic disease of the cardiac muscle, frequently caused by mutations in MYBPC3. However, little is known about the upstream pathways and key regulators causing the disease. Therefore, we employed a multi-omics approach to study the pathomechanisms underlying HCM comparing patient hearts harboring MYBPC3 mutations to control hearts. RESULTS: Using H3K27ac ChIP-seq and RNA-seq we obtained 9310 differentially acetylated regions and 2033 differentially expressed genes, respectively, between 13 HCM and 10 control hearts. We obtained 441 differentially expressed proteins between 11 HCM and 8 control hearts using proteomics. By integrating multi-omics datasets, we identified a set of DNA regions and genes that differentiate HCM from control hearts and 53 protein-coding genes as the major contributors. This comprehensive analysis consistently points toward altered extracellular matrix formation, muscle contraction, and metabolism. Therefore, we studied enriched transcription factor (TF) binding motifs and identified 9 motif-encoded TFs, including KLF15, ETV4, AR, CLOCK, ETS2, GATA5, MEIS1, RXRA, and ZFX. Selected candidates were examined in stem cell-derived cardiomyocytes with and without mutated MYBPC3. Furthermore, we observed an abundance of acetylation signals and transcripts derived from cardiomyocytes compared to non-myocyte populations. CONCLUSIONS: By integrating histone acetylome, transcriptome, and proteome profiles, we identified major effector genes and protein networks that drive the pathological changes in HCM with mutated MYBPC3. Our work identifies 38 highly affected protein-coding genes as potential plasma HCM biomarkers and 9 TFs as potential upstream regulators of these pathomechanisms that may serve as possible therapeutic targets.

Schistosome infection is negatively associated with mite atopy, but not wheeze and asthma in Ghanaian schoolchildren.

BACKGROUND: Epidemiological evidence suggests that helminth infection and rural living are inversely associated with allergic disorders. OBJECTIVE: The aim of the study was to investigate the effect of helminth infections and urban versus rural residence on allergy in schoolchildren from Ghana. METHODS: In a cross-sectional study of 1385 children from urban-high socio-economic status (SES), urban-low SES and rural schools, associations between body mass index (BMI), allergen-specific IgE (sIgE), parasitic infections and allergy outcomes were analysed. Allergy outcomes were skin prick test (SPT) reactivity, reported current wheeze and asthma. RESULTS: Helminth infections were found predominantly among rural subjects, and the most common were hookworm (9.9%) and Schistosoma spp (9.5%). Being overweight was highest among urban-high SES (14.6%) compared to urban-low SES (5.5%) and rural children (8.6%). The prevalence of SPT reactivity to any allergen was 18.3%, and this was highest among rural children (21.4%) followed by urban-high SES (20.2%) and urban-low SES (10.5%) children. Overall, SPT reactivity to mite (12%) was most common. Wheeze and asthma were reported by 7.9% and 8.3%, respectively. In multivariate analyses, factors associated with mite SPT were BMI (aOR 2.43, 95% CI 1.28-4.60, P = 0.007), schistosome infection (aOR 0.15, 95% CI 0.05-0.41) and mite sIgE (aOR 7.40, 95% CI 5.62-9.73, P < 0.001) but not area. However, the association between mite IgE and SPT differed by area and was strongest among urban-high SES children (aOR = 15.58, 95% CI 7.05-34.43, P < 0.001). Compared to rural, urban-low SES area was negatively associated with current wheeze (aOR 0.41, 95% CI 0.20-0.83, P = 0.013). Both mite sIgE and mite SPT were significantly associated with current wheeze and asthma. CONCLUSION AND CLINICAL RELEVANCE: Infection with schistosomes appeared to protect against mite SPT reactivity. This needs to be confirmed in future studies, preferably in a longitudinal design where schistosome infections are treated and allergic reactions reassessed.

OS092. qPCR-based analysis of podocyturia is a feasible diagnostic tool in preeclampsia.

INTRODUCTION: Preeclampsia affects 5% of all pregnancies and is a significant cause of maternal and fetal morbidity and mortalityworldwide. A clinically useful screening test that can predict the development of preeclampsia at an early stage is urgently needed. The detection of podocyturia by immunohistochemistry following cell culture has been noted as a sensitive and specific marker for preeclampsia. However, this method is laborious and carries the risk of cell-culture contamination. OBJECTIVES: The aim of this study was to investigate the diagnostic value of qPCR as a rapid and sensitive method to detect podocyturia in women with preeclampsia. METHODS: Mid-stream urine samples were collected from preeclamptic [1] (n=35), healthy pregnant matched for gestational age (n=34), and healthy non-pregnant (n=12) women. mRNA was isolated using the Trizol method. qPCR analysis was performed for nephrin, VEGF, podocin, GAPDH and megalin transcripts. A ROC-curve analysis was performed. RESULTS: Significantly elevated mRNA expression levels of nephrin, podocin and VEGF were detected in preeclamptic women compared to healthy pregnant and healthy non-pregnant controls. A positive correlation (ρ=0.82, p<0.0001) was observed between nephrin and VEGF mRNA expression in preeclamptic women. ROC curve analyses demonstrated a strong ability of this method to discriminate between the different study groups. CONCLUSION: qPCR analysis of podocyte-specific molecules in urine samples is a rapid and reliable method to quantify podocyturia. We demonstrate that this method distinguishes preeclamptic patients from healthy controls at disease onset. This method may be a tool for the detection of preeclampsia at an earlier stage, thereby preventing maternal and fetal morbidity and mortality.

Cellular and humoral responses to tetanus vaccination in Gabonese children.

Protection to tetanus is often not optimal in developing countries due to incomplete vaccination schemes, or decreased efficacy of vaccination. In this study we investigated the immunological response to tetanus booster vaccination in school children living in a semi-urban or in a rural area of Gabon. Tetanus-specific total IgG as well as antibody subclasses of the IgG1, IgG2, IgG3 and IgG4 isotype and the avidity of the dominating IgG1 subclass were determined both before and 1 month after the booster vaccination. In addition, tetanus-specific cytokine responses were determined. We found a polarization towards a T helper 1 (Th1) profile in the semi-urban children, whereas the cytokine responses of the rural children showed a T helper 2 (Th2) skewed response. Furthermore, tetanus-specific antibodies of the different IgG subclasses were all increased upon a tetanus booster vaccination and levels of IgG1 and IgG3 were higher in the rural children. In conclusion, a tetanus booster vaccination induced a stronger Th2 over Th1 cytokine profile to tetanus toxoid (TT) in rural children who showed the highest levels of IgG1 and IgG3 anti-TT antibody responses.