Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare autosomal dominant disease of accelerated aging which leads to death between 7 and 20 years of age. The disease arises from point mutations that produce an alternately spliced form of the nuclear protein lamin A, known as progerin, that accumulates in the cell nucleus. Mouse models of HGPS exhibit many phenotypical similarities with the HGPS lamin gene mutation, but atherosclerosis does not develop, suggesting a limit to the suitability of animal models. Since cardiovascular disease represents the primary cause of death among those with HGPS, we propose to use a novel tissue engineered blood vessel microphysiological system to develop biomarkers for the disease and assess the effectiveness of treatment against relevant physiological measurements. We have developed arteriolar-scale endothelialized tissue-engineered blood vessels (TEBVs) using smooth muscle cells (SMCs) derived from induced pluripotent stem cells (iPSCs) using healthy and HGPS cells. The TEBVs can be produced and perfused at physiological flow conditions within a few hours of preparation and exhibit endothelial-mediated vasoactivity and respond to inflammatory mediators. We can perform standard functional tests and examine the effects of inflammatory signals, thus tracking the progression of the disease in the same vessel. The HGPS-TEBVs provide a more realistic in vitro environment than cells cultured on plastic and can help advance the process of discovering novel therapeutics and identification of biomarkers. In the overall project, we will test the hypotheses that tissue-engineered blood vessels made with cells derived from individuals with HGPS recapitulate in vitro the structure and activity found in vivo and can aid in assessing the effectiveness and mode of action suitable drug candidates for clinical studies. The goal of this diversity supplement is to support Ms. Crystal Kennedy to examine how the vascular endothelium is altered by HGPS and how the function of endothelial cells and smooth muscle cells is altered a gene editing mechanism to correct the disease.
In Aim 1 of the Supplement we will use RNASeq to examine the differential change in gene expression of HGPS endothelial cells and compare with the results from healthy endothelium.
This aim complements AIM 1.2 of the parent grant and will enable us to identify functional pathways altered by HGPS.
In Aim 2, we will correct HGPS using gene editing and assess the impact on the function of TEBVs and endothelium. This study is consistent with Specific Aim 3 of the parent grant, which aims to assess novel HGPS treatments.
This project involves using engineered human tissue created with induced pluripotent stem cells as a model for disease processes and development of new therapies. In this supplement, we will further explore how one of the cell types, the arterial endothelium, influences the progress of the rare disease progeria. These studies will provide new insight into disease mechanisms.
|Truskey, George A (2018) Human Microphysiological Systems and Organoids as in Vitro Models for Toxicological Studies. Front Public Health 6:185|