Abstract
Vascular dysfunctions are a common feature of multiple age-related diseases. However, modeling healthy and pathological aging of the human vasculature represents an unresolved experimental challenge. Here, we generated induced vascular endothelial cells (iVECs) and smooth muscle cells (iSMCs) by direct reprogramming of healthy human fibroblasts from donors of different ages and Hutchinson-Gilford Progeria Syndrome (HGPS) patients. iVECs induced from old donors revealed upregulation of GSTM1 and PALD1, genes linked to oxidative stress, inflammation and endothelial junction stability, as vascular aging markers. A functional assay performed on PALD1 KD VECs demonstrated a recovery in vascular permeability. We found that iSMCs from HGPS donors overexpressed bone morphogenetic protein (BMP)-4, which plays a key role in both vascular calcification and endothelial barrier damage observed in HGPS. Strikingly, BMP4 concentrations are higher in serum from HGPS vs. age-matched mice. Furthermore, targeting BMP4 with blocking antibody recovered the functionality of the vascular barrier in vitro, hence representing a potential future therapeutic strategy to limit cardiovascular dysfunction in HGPS. These results show that iVECs and iSMCs retain disease-related signatures, allowing modeling of vascular aging and HGPS in vitro.
Highlights
Physiological and pathological aging represent a major risk factor for the onset of cardiovascular diseases, the leading cause of death worldwide (Benjamin et al, 2019)
Since SMCs and VECs play an important role in human aging and Hutchinson-Gilford Progeria Syndrome (HGPS) (Hamczyk et al, 2019; Li et al, 2018), we directly induced iSMCs and induced vascular endothelial cells (iVECs) by isolating skin fibroblasts from human donors and overexpressing either the master regulators MYOCD or ETV2, respectively. iSMCs expressed cell-identity markers including alpha smooth muscle actin (Figure 1A, expressed by about 70% cells) and calponin (Figure 1B, expressed by about 80% cells), both in monoculture and when co-cultured with VECs (Figure 1C)
ASMA-expressing iSMCs contributed to the formation of microvascular networks in 3D fibrin matrices (Figure 1D), mimicking the behavior of fibroblasts and differentiated mesenchymal stem cells observed in previous vascular models developed by our (Bersini et al, 2016; Jeon et al, 2014) and other groups (Zheng et al, 2012)
Summary
Physiological and pathological aging represent a major risk factor for the onset of cardiovascular diseases, the leading cause of death worldwide (Benjamin et al, 2019). Direct reprogramming of skin fibroblasts into functional cells was previously reported for blood progenitor cells (Szabo et al, 2010), SMCs (van Tuyn et al, 2005) and VECs (Han et al, 2014; Morita et al, 2015). IPS cells have significantly contributed to the discovery of novel disease mechanisms (Park et al, 2019) as well as to the pre-clinical screening of promising drug candidates (Zhang et al, 2019), the differentiation process of iPS cells requires a mandatory transit through an embryonic-like state which was reported to reset the aging profile of Bersini et al eLife 2020;9:e54383.
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