Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1

Abstract

NPs encapsulated with CHIR + FGF1 exerted substantial myocardial protective effects and represents a potentially novel strategy for improving postischemic myocardial protection. Results Identification of chemicals that promote cell cycle activity of human induced pluripotent stem cell-derived cardiomyocytes. Using the BrdU incorporation assay, we screened several chemicals for their capacity to enhance the cell cycle activity of cultured human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). We picked hiPSC-CMs because they are easy to handle and represent a practical system for drug testing and screening (5). These chemicals included Ly294002 (PI3K inhibitor) (6), FGF1 (6, 7), SB203580 and VX702 (p38 MAPK inhibitors) (6), KN93 (Ca 2+ /calmodulin-dependent protein kinase II inhibitor) (8), Su1498 (Flk-1 inhibitor) (8), and CHIR99021 (Wnt activator and GSK3α and 3β inhibitor) (8, 9). We found a combination of 5 μM CHIR99021 and 100 ng/mL FGF1 was the most potent treatment to induce cell cycle in hiPSC-derived cardiomyocytes (Supplemental Figure 1; supplemental material available online with this article; https://doi.org/10.1172/jci.insight.132796DS1). Characterization of CHIR-FGF1-NPs. We have previously shown that PLGA NPs can be used as a platform for slow release (up to 4 weeks) of chemicals to injured animal hearts and that they provide cardioprotection (2). To characterize the long-term cardioprotective function of the FGF/CHIR combination in vivo, we formulated the PLGA NPs with these 2 factors. The size of PLGA NPs was measured using scanning electron microscopy, for both CHIR99021-and FGF1-loaded NPs (Supplemental Figure 2, A and B). Quantification of particle diameter for CHIR-NPs (Supplemental Figure 2C) and FGF1-NPs (Supplemental Figure 2D) yielded values of 123.63 ± 44.48 nm and 129.57 ± 45.94 nm, respectively. The size and shape of CHIR-NPs and FGF1-NPs were uniform. The encapsulation efficiency of CHIR-NPs and FGF1-NPs, i.e., (the amount encapsulated/total amount available) × 100%, was 50.41% and 62.8 ± 1.6%, respectively. The concentration of encapsulated CHIR and FGF1 was 8.07 μg/mg and 1.26 ± 0.03 μg/mg, respectively. Determination of release kinetics of CHIR-and FGF1-loaded NPs as a function of time, using either NanoDrop via UV-Vis spectrophotometer (for CHIR) or ELISA (for FGF1), and the cumulative percentage of CHIR and FGF1 released from NPs, are shown in Supplemental Figure 2, E and F. When 1000 μg of CHIR-and FGF1-loaded NPs were incubated in 1000 μL of Dulbecco's phosphate-buffered saline (DPBS), pH 7.4 at 37°C, 55% of the encapsulated CHIR was released during the first day and 85% by day 15 (Supplemental Figure 2E). In contrast, 55% of the encapsulated FGF1 was released during the initial 3 days, and 63% was released by day 10. Notably, between day 1 and day 30, the release kinetics strictly followed the Korsmeyer-Peppas model for FGF1-NPs (Supplemental Figure 2F). Fitting this model, C t /C 0 = kt n , where C t = concentration at time t; C 0 = equilibrium concentration; ...

Nano-Medicine in the Cardiovascular System

“…High biocompatibility and low cytotoxicity are attributed to the degradation byproducts—lactate and glycolate—which can easily be incorporated into cellular metabolic pathways ( Figure 1 ) ( Chereddy et al, 2018 ). Due to these highly desirable properties, PLGA has been widely studied as both a therapeutic and diagnostic agent in the cardiac field as well ( Oduk et al, 2018 ; Zhang et al, 2019 ; Fan et al, 2020a ; Fan et al, 2020b ). With improvements in processing and production techniques, PLGA has also enjoyed a lot of attention due to the relative ease with which comparatively large batches of nano-medicines can be produced via emulsion polymerization.…”

“…Recent studies showing the synergistic effects of using a combination of extended delivery of CHIR99021 (a Wnt1 agonist/GSK-3β antagonist) and fibroblast growth factor 1 (FGF1) to protect ischemia-threatened cardiomyocytes from apoptosis, while accelerating angiogenesis through the promotion of endothelial and vascular SMC proliferation, and consequently enhance myocardial protection have been published ( Fan et al, 2020a ; Fan et al, 2020b ). These studies found that PLGA NPs loaded with CHIR99021 or FGF1 allowed for effective delayed release for up to 4 weeks.…”

Large Animal Models of Cell-Free Cardiac Regeneration

“…Fan et al identified acidic fibroblast growth factor (FGF-1) and CHIR99021 as synergistic enhancers of cardiomyocyte cell cycle activity [ 62 ]. CHIR99021 is an aminopyrimidine derivative that functions as a Wnt signaling activator, which has also been shown to be able to reprogram fibroblasts to a cardiomyocyte phenotype [ 63 ].…”

“…In their study, both molecules were loaded onto PLGA nanoparticles and injected intramyocardially 15 min after reperfusion of AMI in a pig model ( n = 12). This resulted in significantly reduced scarring as well as increased LVEF and decreased left ventricular end-diastolic volume (LVEDV) compared to control animals after 28 days of follow-up [ 62 ].…”

TMSB4 Overexpression Enhances the Potency of Marrow Mesenchymal Stromal Cells for Myocardial Repair

“…As the functionality of cardiomyocytes proliferation is limited in an adult mammal heart, promotion of angiogenesis remains the most crucial strategy in salvaging myocytes at the infarcted border zone. Thus, discovering practical or appropriate clinical interventions, sparing less severely damaged myocytes at border zones, could effectively reduce infarct size and save lives ( Fan et al, 2020a ). Mesenchymal stromal cells (MSCs) represent a promising tool for cell therapy, particularly for heart-related diseases.…”