Research Projects

Congenital Heart Disease (CHD) represents a major cause of infant morbidity and mortality, with its etiology involving both genetic and epigenetic components. Emerging evidence suggests that post-transcriptional RNA modifications, particularly mRNA methylation (m6A, m5C, m1A, and m7G), may play a critical regulatory role in heart development and CHD pathogenesis. However, the mechanistic contribution of these modifications remains poorly understood. This project aims to identify and functionally validate dysregulated mRNA methylation signatures in CHD. The central hypothesis is that specific mRNA methylation signatures are dysregulated in CHD and that these epitranscriptomic changes contribute to aberrant cardiac development. Using Nanopore Direct RNA Sequencing, transcriptome-wide methylation profiles will be generated from cardiac tissues of patients with ASD, VSD, TOF, and non-CHD controls. Integrating this with differentially expressed RNAs will help identify differentially methylated and expressed transcripts relevant to heart development. Candidate methylated mRNAs will be selected based on significance, expression pattern, and relevance to heart development. Functional validation will be performed through siRNA-mediated knockdown of selected transcripts in zebrafish embryos, a well-established model for cardiac morphogenesis. The phenotypic impact of knockdown will be assessed via morphological, histological, and molecular assays. Complementary in vitro validation in AC16 cardiac cell lines will assess effects on apoptosis, proliferation, and cardiac gene expression. This study will advance the understanding of the epitranscriptomic basis of CHD and may identify novel biomarkers for early diagnosis and therapeutic targeting.