# Endothelial Cell Analytic Core

> **NIH NIH P01** · AUGUSTA UNIVERSITY · 2024 · $392,700

## Abstract

CORE C PROJECT SUMMARY
The main goal of the Cell and Analytical Core (Core C) is to facilitate completion of the program investigators’
specific aims. To accomplish this goal, Core C will perform three specific aims, two of which are service
components and one of which is an academic component. Aim 1 - Service component 1 - Cell isolation and
characterization: a) To uniformly isolate primary mouse endothelial cells from wild type and transgenic mice and
from human tissue, using a calibrated system to make cell suspensions from tissue in combination with a
negative (CD45-based) and positive selection (CD31-based) endothelial cell purification method. Wild type or
transgenic mice will be received from animal Core B or from the three projects. b) To characterize the endothelial
character of the isolated EC. c) To expand and distribute purified endothelial cells in early passages to the three
research projects. Aim 2 - Service component 2 - Gene Delivery and deletion: a) To construct, purify and deliver
recombinant adenovirus, lentivirus and adeno-associated virus (AAV). b) To deplete and manipulate
endogenous genes in mouse and human EC, using CRISPR Cas9 and siRNA technology. Aim 3 -Academic
component 1- Analysis: a) To measure cell metabolism. O2 consumption rate (OCR) and extracellular
acidification rate (ECAR) using Seahorse and performing metabolomics. b) To evaluate oxidative and nitrosative
stress. c) To assess inflammation by measuring cytokine/chemokine profiles in cell supernatants using magnetic
bead-based Multiplex analysis and adhesion molecule expression relevant for endothelial-leukocyte interactions
using an in-house developed cell-based ELISA. This technology will also be applied to measure
cytokine/chemokine profile in plasma samples from mice. d) To evaluate endothelial-mesenchymal transition-
related genes in transcriptomics, using bulk and single cell RNASeq. e) To assess endothelial barrier function
by combining measurements of macromolecular permeability to a macromolecular tracer and electrical
impedance of endothelial monolayers. In summary, synergy with the three scientific projects, administrative Core
A and core B will be organized as follows: upon receiving wild type or transgenic mice from animal Core B and
from the three projects, Core C will isolate, characterize, expand and distribute early passages of primary mouse
endothelial cells and of well-characterized in house isolated or purchased human endothelial cells to all four
projects. The Core will moreover construct recombinant adenovirus, lentivirus and adeno-associated virus or use
CRISPR ON and specific siRNA to manipulate gene expression in human and mouse endothelial cells and will
coordinate scRNASeq and proteomics. The Core will moreover organize in a standardized manner cell analysis
for all parameters listed above. Finally, Core C will collaborate with administrative Core A and animal Core B to
coordinate dissemination of data generated through the collabor...

## Key facts

- **NIH application ID:** 10847546
- **Project number:** 1P01HL160557-01A1
- **Recipient organization:** AUGUSTA UNIVERSITY
- **Principal Investigator:** Rudolf Lucas
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $392,700
- **Award type:** 1
- **Project period:** 2024-06-05 → 2029-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10847546

## Citation

> US National Institutes of Health, RePORTER application 10847546, Endothelial Cell Analytic Core (1P01HL160557-01A1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10847546. Licensed CC0.

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