Epigenetic regulation of vascular functions has been found to play crucial roles in cardiovascular diseases. Vascular endothelial cells (ECs), which are exposed to different flow patterns, regulate vascular homeostasis. Differential epigenetic changes, e.g. histone modifications, caused by different flow patterns regulate EC gene expression profile and hence functional consequences. The coupling of histone phosphorylation, methylation, and acetylation have recently been identified to regulate gene expressions through the distinct chromatin remodeling complexes, which would alter the consequential phenotypic outcome. However, there is a paucity of study in the flow-regulation of histone modifications in vascular cells. We hypothesize that the coupling among epigenetic histone phosphorylation, methylation, and acetylation may serve as a transducing mechanism to regulate EC gene expressions under different patterns of flows. We will develop a directed evolution strategy for the systematic optimization and tuning of FRET biosensors with distinct colors to simultaneously monitor different histone modifications with high sensitivity and specificity. These biosensors will be used to track multiple histone modifications simultaneously in the same live cell and unravel the evolving multiplex landscape of histone modifications under different flows. We will further employ the endonuclease-deficient Cas9 (dCas9), small guide RNAs (sgRNAs) and split FPs to track the dynamics of histone modifications at the specific loci of EC phenotype marker genes. Our epigenetic manipulation system will then be employed to modulate epigenetics at these specific loci and determine their effects on gene expressions and consequent cellular functions in single live cells under different flows. The identified epigenetic profiles will then be modulated in vivo, and the consequent gene expression and phenotypic outcome examined.
Four specific aims are proposed: 1) Develop and optimize FRET biosensors to visualize the dynamic histone modifications in single cells, 2) Unravel the spatiotemporal coupling of histone phosphorylation-methylation-acetylation in regulating EC functions under different flows, 3) Establish the roles of locus-specific histone modifications in regulating EC gene expression under flows, 4) Elucidate the effect of histone modifications on gene expression and lesion formation in vivo. The simultaneous tracking of the spatiotemporal dynamics of histone modifications in the nucleus in conjunction with cell proliferation and inflammation in a single live cell will allow the elucidation of the spatiotemporal transducing mechanism in regulating epigenetic modulations and pathophysiological consequences upon the exposure of ECs to hemodynamic cues. The mechanistic insights obtained should allow us to identify the potential molecular targets and facilitate the design of pharmaceutical interventions for pathologic processes. As such, the project should have transformative impact in the field of vascular mechanobiology, particularly related to the molecular regulations of cell cycle and inflammation in mediating the development of atherosclerosis.
We will use fluorescence-based biosensors to monitor the dynamics of loci-specific epigenetic changes in order to elucidate their roles in the hemodynamic regulation of inflammatory gene expressions in endothelial cells. An epigenetic manipulation system will be developed to modulate endothelial histone modifications for the control of epigenetics and gene expressions. The results obtained will provide information to identify the potential molecular targets and facilitate the design of pharmaceutical interventions for epigenetic-mediated cardiovascular disease progression.
|Yeh, Yi-Ting; Serrano, Ricardo; François, Joshua et al. (2018) Three-dimensional forces exerted by leukocytes and vascular endothelial cells dynamically facilitate diapedesis. Proc Natl Acad Sci U S A 115:133-138|
|Pan, Yijia; Yoon, Sangpil; Zhu, Linshan et al. (2018) Acoustic mechanogenetics. Curr Opin Biomed Eng 7:64-70|
|Limsakul, Praopim; Peng, Qin; Wu, Yiqian et al. (2018) Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface. Cell Chem Biol 25:370-379.e4|
|Pan, Yijia; Yoon, Sangpil; Sun, Jie et al. (2018) Mechanogenetics for the remote and noninvasive control of cancer immunotherapy. Proc Natl Acad Sci U S A 115:992-997|
|Peng, Qin; Lu, Shaoying; Shi, Yuxin et al. (2018) Coordinated histone modifications and chromatin reorganization in a single cell revealed by FRET biosensors. Proc Natl Acad Sci U S A 115:E11681-E11690|
|Yoon, Sangpil; Wang, Pengzhi; Peng, Qin et al. (2017) Acoustic-transfection for genomic manipulation of single-cells using high frequency ultrasound. Sci Rep 7:5275|
|Vennin, Claire; Chin, Venessa T; Warren, Sean C et al. (2017) Transient tissue priming via ROCK inhibition uncouples pancreatic cancer progression, sensitivity to chemotherapy, and metastasis. Sci Transl Med 9:|
|Wan, Qiaoqiao; TruongVo, ThucNhi; Steele, Hannah E et al. (2017) Subcellular domain-dependent molecular hierarchy of SFK and FAK in mechanotransduction and cytokine signaling. Sci Rep 7:9033|
|Sun, Jie; Lei, Lei; Tsai, Chih-Ming et al. (2017) Engineered proteins with sensing and activating modules for automated reprogramming of cellular functions. Nat Commun 8:477|
|Seong, Jihye; Huang, Min; Sim, Kyoung Mi et al. (2017) FRET-based Visualization of PDGF Receptor Activation at Membrane Microdomains. Sci Rep 7:1593|
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