# Integration of single-cell imaging and multi-omics sequencing to study EC mechano-pathophysiology

> **NIH NIH R01** · UNIVERSITY OF SOUTHERN CALIFORNIA · 2023 · $623,789

## Abstract

Summary
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 tra...

## Key facts

- **NIH application ID:** 10825307
- **Project number:** 7R01HL121365-10
- **Recipient organization:** UNIVERSITY OF SOUTHERN CALIFORNIA
- **Principal Investigator:** SHU CHIEN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $623,789
- **Award type:** 7
- **Project period:** 2023-05-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10825307

## Citation

> US National Institutes of Health, RePORTER application 10825307, Integration of single-cell imaging and multi-omics sequencing to study EC mechano-pathophysiology (7R01HL121365-10). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10825307. Licensed CC0.

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