Mink1 Regulate Cardiac Left-Right Asymmetry by Modulating Cilium Assembly in Zebrafish

Proper establishment of cardiac left-right (LR) asymmetry is essential for vertebrate organ development, and its disruption frequently results in congenital heart disease (CHD). Although the central role of cilia in this process has been well established, the upstream regulatory factors governing ciliary function remain incompletely understood.In this study, we investigated the role of Misshapen-like kinase 1 (Mink1) in cardiac left-right asymmetric development in zebrafish. Using CRISPR/Cas9 technology, we generated a loss-of-function mink1 mutant line (mink1-/-). Phenotypic analysis revealed that approximately 40.7% of mutant embryos exhibited abnormal heart looping. In addition, mutant embryos exhibited cardiac chamber dilation, marked bradycardia, and caudal vein thrombosis. Transcriptome (RNA-seq) analysis showed significant downregulation of genes associated with ciliogenesis and LR asymmetry, including dzip1, snx10a, and nkx2.5, whereas genes involved in blood coagulation, such as itga2b, gp1bb, and f7l, were markedly upregulated. Gene set enrichment analysis (GSEA) further confirmed suppression of the "non-motile cilium assembly" pathway and activation of the "blood coagulation" pathway in mink1 mutants.Collectively, these results demonstrate that Mink1 contributes to the proper establishment of cardiac LR asymmetry in zebrafish by regulating the expression of cilia-related genes, and that its loss leads to cardiac dysfunction and a prothrombotic state. Our findings identify Mink1 as an important regulatory link between ciliary development, cardiac morphogenesis, and circulatory homeostasis, providing new molecular insight into the mechanisms underlying congenital heart disease.