Atherosclerosis assay development for drug efficacy and cytotoxicity screening

Project Summary
Cardiovascular disease, primarily driven by atherosclerosis, is the leading cause of death in the U.S. The recent
FDA Modernization Act 2.0 allows the use of alternatives to animal models in drug testing, spurring the
development of advanced in vitro models like cell-based assays and artificial intelligence-based systems to
streamline drug development processes. However, current atherosclerosis assays, mostly based on simple 2D cell
cultures, don't adequately replicate the disease's complexity, potentially leading to inaccurate results.
To address this issue, Endomimetics has developed innovative nanomatrix cell sheet technology and “cell-as-
glue” approach as well as formulated novel atherosclerotic inducing mediums, which can produce an in vitro 3D
nanomatrix vascular sheet (VS) and a novel 3D in vitro atherosclerosis model - nanomatrix vascular sheet with
atherosclerosis (VSA).
Therefore, our aim is to revolutionize the field by developing an automated, highly precise approach for fabricating
VS and VSA. This advancement will enable the use of VS and VSA in creating functional assays for efficient, high-
throughput drug testing in a 3D human atherosclerosis environment and allow for the evaluation of both the efficacy
and cytotoxicity of drugs across extensive sample sizes, while minimizing biological variability. Such an initiative,
unprecedented in its scope and precision, has not yet been achieved in any existing in vitro atherosclerosis assays.
Specifically, we will develop two innovative assays integrating BODIPY staining and ELISA methods, aimed at
determining the efficacy of atherosclerosis treatments by their capacity to suppress foam cell formation and curtail
inflammatory cytokine production.
Established anti-atherosclerosis and anti-inflammatory compounds will serve as positive controls, validating the
accuracy of the assays in detecting reductions in both foam cell formation and cytokine secretion. Simultaneously,
negative controls will be employed to evaluate assay specificity, ensuring the precision of our results, and
eliminating the potential for false positives.
To demonstrate the feasibility of our assays in evaluating new atherosclerosis therapeutics, we will assess the
efficacy of two histone deacetylases inhibitors at various doses using our assays. Additionally, we will develop
and execute cytotoxicity-based assays to investigate the safety of these inhibitors.

Grant Number: 1R43HL176310-01
NIH Institute/Center: NIH
Principal Investigator: Jun Chen