Cardiac pericyte reprogramming by MEK inhibition promotes arteriologenesis and angiogenesis of the ischemic heart
Elisa Avolio, Rajesh Katare, Anita C. Thomas, Andrea Caporali, Daryl Schwenke, Michele Carrabba, Marco Meloni, Massimo Caputo, Paolo Madeddu
Elisa Avolio, Rajesh Katare, Anita C. Thomas, Andrea Caporali, Daryl Schwenke, Michele Carrabba, Marco Meloni, Massimo Caputo, Paolo Madeddu
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Research Article Angiogenesis Vascular biology

Cardiac pericyte reprogramming by MEK inhibition promotes arteriologenesis and angiogenesis of the ischemic heart

Abstract

Pericytes (PCs) are abundant yet remain the most enigmatic and ill-defined cell population in the heart. Here, we investigated whether PCs can be reprogrammed to aid neovascularization. Primary PCs from human and mouse hearts acquired cytoskeletal proteins typical of vascular smooth muscle cells (VSMCs) upon exclusion of EGF/bFGF, which signal through ERK1/2, or upon exposure to the MEK inhibitor PD0325901. Differentiated PCs became more proangiogenic, more responsive to vasoactive agents, and insensitive to chemoattractants. RNA sequencing revealed transcripts marking the PD0325901-induced transition into proangiogenic, stationary VSMC-like cells, including the unique expression of 2 angiogenesis-related markers, aquaporin 1 (AQP1) and cellular retinoic acid–binding protein 2 (CRABP2), which were further verified at the protein level. This enabled us to trace PCs during in vivo studies. In mice, implantation of Matrigel plugs containing human PCs plus PD0325901 promoted the formation of αSMA+ neovessels compared with PC only. Two-week oral administration of PD0325901 to mice increased the heart arteriolar density, total vascular area, arteriole coverage by PDGFRβ+AQP1+CRABP2+ PCs, and myocardial perfusion. Short-duration PD0325901 treatment of mice after myocardial infarction enhanced the peri-infarct vascularization, reduced the scar, and improved systolic function. In conclusion, myocardial PCs have intrinsic plasticity that can be pharmacologically modulated to promote reparative vascularization of the ischemic heart.

Authors

Elisa Avolio, Rajesh Katare, Anita C. Thomas, Andrea Caporali, Daryl Schwenke, Michele Carrabba, Marco Meloni, Massimo Caputo, Paolo Madeddu

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Figure 11

A 2-week treatment with PD0325901 improves left ventricular function and vascularization in a mouse MI model.

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A 2-week treatment with PD0325901 improves left ventricular function and...
(A) Cartoon summarizing the experimental design. Mice were given the MEKi (10 mg/kg/d) or DMSO vehicle orally for 14 days after MI induction. The drug was embedded in flavored jelly and eaten spontaneously by animals. (B–D) Graphs showing basal and final echocardiography indices. For vehicle, n = 12 mice basal and n = 8 final. For PD, n = 12 mice basal and n = 11 final. Individual values and mean ± SD. SV, stroke volume; CO, cardiac output. (E) Graph showing mouse survival. (F) Representative images showing the Azan-Mallory staining of the LV and bar graphs indicating the infarct size expressed as a percentage of the LV area. n = 8 mice for Veh, n = 10 mice for PD. (G) Representative immunofluorescence images showing arterioles (αSMA, red) and capillaries (isolectin B4, green) in the peri-infarct myocardium. The dashed line defines the infarct zone (IZ). Scale bars: 100 μm. (H–J) Graphs showing the quantification of arteriole (H) and capillary (I) densities and cardiomyocyte cross-sectional area (CSA) (J) in the LV. n = 7 mice. In F–J, individual values and mean ± SEM are shown. Veh, vehicle; PD, PD0325901. *P < 0.05, **P < 0.01, ***P < 0.001; #P < 0.05, ##P < 0.01 in the comparison between changes (Δ). In B–D, 2-way ANOVA (mixed effects model with Sidak’s multiple comparison test) was performed considering that there were missing data in the 2 treatment groups due to premature death after MI. In addition, we compared the changes (Δ) from basal to final times in the 2 groups using an unpaired Student’s t test. In F and H–J, an unpaired Mann-Whitney U test was used.