CAR (CARSKNKDC) Peptide Modified ReNcell-Derived Extracellular Vesicles as a Novel Therapeutic Agent for Targeted Pulmonary Hypertension Therapy

Abstract

In recent years, mesenchymal stem cells (MSCs)–derived extracellular vesicles (EVs) are emerging as a potential therapeutic agent for pulmonary hypertension (PH). However, the full realization of MSCs–derived EVs therapy has been hampered by the absence of standardization in MSCs culture and the challenges of industrial scale–up. The study was to exploit an alternative replacement for MSCs using currently commercialized stem cell lines for effective targeted PH therapy. ReNcell VM—a human neural stem cell line—has been utilized here as a reliable and easily adoptable source of EVs. We first demonstrated that ReNcell-derived EVs (ReNcell-EVs) pretreatment effectively prevented Su/Hx (SU5416/hypoxia)–induced PH in mice. Then for targeted therapy, we conjugated ReNcell-EVs with CAR (CARSKNKDC) peptide (CAR-EVs)—a peptide identified to specifically target hypertensive pulmonary arteries, by bio-orthogonal chemistry. Intravenous administration of CAR-EVs selectively targeted hypertensive pulmonary artery lesions especially pulmonary artery smooth muscle cells. Moreover, compared with unmodified ReNcell-EVs, CAR-EVs treatment significantly improved therapeutic effect in reversing Su/Hx-induced PH in mice. Mechanistically, ReNcell-EVs inhibited hypoxia-induced proliferation, migration, and phenotype switch of pulmonary artery smooth muscle cells, at least in part, via the delivery of its endogenous highly expressed miRNAs, let-7b-5p, miR-92b-3p, and miR-100-5p. In addition, we also found that ReNcell-EVs inhibited hypoxia-induced cell apoptosis and endothelial-mesenchymal transition in human microvascular endothelial cells. Taken together, our results provide an alternative to MSCs-derived EVs–based PH therapy via using ReNcell as a reliable source of EVs. Particularly, our CAR-conjugated EVs may serve as a novel drug carrier that enhances the specificity and efficiency of drug delivery for effective PH-targeted therapy.

Extracellular Vesicles Derived From Human Umbilical Cord Mesenchymal Stem Cells Protect Against DOX-Induced Heart Failure Through the miR-100-5p/NOX4 Pathway

“…The expression of miR-100-5p is decreased by hypoxia. EVs derived a human neural stem cell line that inhibits hypoxia-induced proliferation and migration through EV miR-100-5-p in pulmonary artery smooth muscle cells ( Wang et al, 2020 ). Hromadnikova et al found that downregulation of miR-100-5p was associated with gestational hypertension and preeclampsia ( Hromadnikova et al, 2016 ).…”

MicroRNA‑100 inhibits breast cancer cell proliferation, invasion and migration by targeting FOXA1

“…In addition, the results of the CCK-8, wound healing and Transwell assays demonstrated that overexpression of miR-100 inhibited the proliferation, migration and invasion of breast cancer cells, respectively. miR-100 has been reported to play an important role in diseases, including atherosclerosis (16), acute lung injury (17), pulmonary hypertension (18), childhood acute lymphoblastic leukemia (19) and osteoarthritis of the temporomandibular joint (20). In addition, miR-100 has been suggested to serve as a diagnostic and therapeutic biomarker in epithelial ovarian cancer (8) and gastric and esophageal cancers (9).…”

Selective Targeting and Tissue Penetration to the Retina by a Systemically Administered Vascular Homing Peptide in Oxygen Induced Retinopathy (OIR)

“…We demonstrate homing of the CAR peptide to OIR neovasculature at the peak of angiogenesis. More recently, studies have demonstrated that the CAR peptide also homes to diseases with a clear inflammatory component such as PAH [29,34,[38][39][40][41][42][43][44]47,48], vascular (aorta) aneurysms [35], muscular dystrophies [37] and myocardial infarction [49]. Due to its homing and tissue penetrating capabilities, CAR peptide has been used as a targeting device for a large number of different therapeutics in different experimental pulmonary disease models [29,34,[38][39][40][41][42][43][44]47].…”