A Tissue Engineered Blood Vessel Model of Hutchinson-Gilford Progeria Syndrome Using Human iPSC-derived Smooth Muscle Cells

Abstract

Hutchison-Gilford Progeria Syndrome (HGPS) is a rare, accelerated aging disorder caused by nuclear accumulation of progerin, an altered form of the Lamin A gene. The primary cause of death is cardiovascular disease at about 14 years. Loss and dysfunction of smooth muscle cells (SMCs) in the vasculature may cause defects associated with HGPS. Due to limitations of 2D cell culture and mouse models, there is a need to develop improved models to discover novel therapeutics. To address this need, we produced a functional three-dimensional model of HGPS that replicates an arteriole-scale tissue engineered blood vessel (TEBV) using induced pluripotent stem cell (iPSC)-derived SMCs from an HGPS patient. To isolate the effect of the HGPS iSMCs, the endothelial layer consisted of human cord blood-derived endothelial progenitor cells (hCB-EPCs) from a separate, healthy donor. TEBVs fabricated from HGPS iSMCs and hCB-EPCs show reduced vasoactivity, increased medial wall thickness, increased calcification and apoptosis relative to TEBVs fabricated from normal iSMCs or primary MSCs. Additionally, treatment of HGPS TEBVs with the proposed therapeutic Everolimus, increases HGPS TEBV vasoactivity and increases iSMC differentiation in the TEBVs. These results show the ability of this iPSC-derived TEBV to reproduce key features of HGPS and respond to drugs.

iPSC-Derived Endothelial Cells Affect Vascular Function in a Tissue-Engineered Blood Vessel Model of Hutchinson-Gilford Progeria Syndrome

“…These results are consistent with the HGPS cardiovascular disease phenotype as well as results from our previous HGPS iSMC TEBV model (Atchison et al, 2017).…”

“…After 4 weeks of perfusion, viSMCs in viTEBVs expressed not only the contractile proteins a-SMA and calponin as we had observed previously with the iSMC TEBVs (Atchison et al, 2017), but also MHC11, which was not present in iSMC TEBVs, indicating a more accurate vascular phenotype ( Figures 5C and S7C). HGPS viTEBVs showed reduced expression of all contractile proteins compared with healthy viTEBV controls ( Figure 5D) as well as increased expression of progerin and extracellular matrix proteins, collagen IV and fibronectin, compared with normal viTEBVs ( Figures 5C and S7D).…”

“…To study the effects of viSMCs and viECs on the cardiovascular phenotype of HGPS, we tested the structural and functional effects of these cells in TEBVs. After 1 week of perfusion, TEBVs fabricated from viSMCs from either normal or HGPS donors showed increased contractile response after exposure to 1 mM phenylephrine compared with TEBVs fabricated from normal or HGPS iSMCs used in our previous TEBV model (Atchison et al, 2017) (Figure 5A). In addition, vasoactivity of TEBVs with normal viSMCs and viECs responded at comparable levels to primary MSC-derived TEBVs, indicating that the Patsch et al differentiation protocol provides a more functional model.…”

“…Previously, we developed a 3D tissue-engineered blood vessel (TEBV) model of HGPS using iPS-derived SMCs (iSMCs) from HGPS patients and blood-derived endothelium from healthy individuals (Atchison et al, 2017). This model was capable of replicating the structure and function of small-diameter arterioles using healthy patient cells as well as exhibit known disease characteristics previously cited in HGPS (Fernandez et al, 2016).…”

Diminished Canonical β-Catenin Signaling During Osteoblast Differentiation Contributes to Osteopenia in Progeria

“…Normal and two HGPS(I) and HGPS(II) fibroblast cell lines (HGADFN168, HGADFN164, HGADFN167) were obtained from the Progeria Research Foundation cell bank (Peabody, MA, USA). These cell lines have been fully characterized, including the karyotypes . Induced pluripotent stem cells (iPSCs) were generated by reprogramming with KLF4 , SOX2 , OCT4 , and C‐MYC Yamanaka factors, as we have described previously .…”