Mechanical Force and Vascular Remodeling
Chih-Wen Ni, Ph.D.
Department of Bioengineering
UC Merced
In vertebrate animals, blood flow provides oxygen to cells, removes metabolic waste, and supports immune and endocrine systems. Hemodynamic forces generated by blood flow provide environmental cues guiding vascular development and remodeling, which also play unique roles to maintain physiological function of endothelial cells (ECs). Given the fact that many genes and pathways have been implied in the process of vascular morphogenesis and patterning, the detail mechanisms have not been fully elucidated. Despite many studies utilized cultured cells in flow apparatus or surgical procedures in rodents to shed light on this subject, limitations on current models still prevent the understanding of mechanobiology, especially in the early developmental stages. Recently, zebrafish has been used as a model system to identify flow-responsive genes in ECs during embryonic development. We developed a method to identify transcriptome profiles in ECs from zebrafish embryos in the presence or absence of blood flow. We performed pulled-down experiments using EGFP antibody to precipitate polysome-EGFP complex expressed in tissue-specific L10a-EGFP transgenic line. The acquired RNA samples were subjected to RNA-seq analysis by which the data reveals a group of genes showing significant expression alteration when the blood flow was abolished comparing to normal condition. In this presentation, several newly identified flow-responsive genes will be presented as examples to demonstrate the importance of their function in vascular remodeling. Importantly, establishing reverse genetic screening pipeline using the approach of genetic engineering such as CRISPR/Cas9 on selected genes in our gene list allows us to identify the regulatory networks in response to blood flow. Moreover, examples will also be given on the applications of genetic engineering to generate transgenic lines, which are used in studying the behavior of ECs in response to different flow patterns. Those include the indicator lines for endothelial calcium signaling and the presence of primary cilia in ECs. In summary, this presentation would provide an idea of using genetic tools and zebrafish model to study endothelial cells
in vivo, which should lead to the understanding of the effect of mechanical force on vascular remodeling.
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