Phage-Display-Guided Nanocarrier Targeting to Atheroprone Vasculature

Abstract

In regions of the circulation where vessels are straight and unbranched, blood flow is laminar and unidirectional. In contrast, at sites of curvature, branch points and regions distal to stenoses blood flow becomes disturbed. Atherosclerosis preferentially develops in these regions of disturbed blood flow. Current therapies for atherosclerosis are systemic, and may not sufficiently target these atheroprone regions. In this study, we sought to leverage the alterations on the luminal surface of endothelial cells caused by this atheroprone flow for nanocarrier targeting. In vivo phage display was used to discover unique peptides that selectively bind to atheroprone regions in the mouse partial carotid artery ligation model. The peptide GSPREYTSYMPH (PREY) was found to bind 4.5-fold more avidly to the region of disturbed flow, and was used to form targeted liposomes. When administered intravenously, PREY-targeted liposomes preferentially accumulated in endothelial cells in the partially occluded carotid artery and other areas of disturbed flow. Proteomic analysis and immunoblotting indicated that fibronectin and Filamin A were preferentially bound by PREY-nanocarriers in vessels with disturbed flow. In additional experiments, PREY-nanocarriers were used therapeutically to deliver the nitric oxide synthase co-factor tetrahydrobiopterin (BH4), which we have previously shown to be deficient in regions of disturbed flow. This intervention increased vascular BH4 and reduced vascular superoxide in the partially ligated artery in wild-type mice, and reduced plaque burden in the partially ligated left carotid artery of fat fed atheroprone mice (ApoE−/−). Targeting atheroprone sites of the circulation with functionalized nanocarriers provides a new approach for prevention of early atherosclerotic lesion formation.

Oxidative Stress and Hypertensive Diseases

“…19 Moreover, administration of BH 4 also prevents the development of endothelium dysfunction, vascular inflammation and atherosclerosis induced by disturbed flow. 20, 21 Acute and short-term administration of BH 4 improves endothelial function in humans with rheumatoid arthritis. 22 There is also convincing evidence that eNOS uncoupling occurs in humans with obstructive sleep apnea, leading to increased vascular superoxide production and endothelial dysfunction.…”

Anti‐Atherogenic Effect of Stem Cell Nanovesicles Targeting Disturbed Flow Sites

“…[ 5 ] Hence, the anti‐inflammatory effects ( Figure A) of BMSC‐NVs, ASC‐NVs, and PMSC‐NVs were examined. In particular, since our previous study reported that monocyte/macrophage could uptake PREY‐liposome, [ 12 ] mouse bone marrow‐derived macrophages (BMDMs) obtained after primary culture or the mouse macrophage cell line (Raw 246.7) were treated with the MSC‐NVs to inhibit their inflammatory responses. First, the effective cellular uptake of the MSC‐NVs (ASC‐NVs, BMSC‐NVs, and PMSC‐NVs) by the activated BMDMs was confirmed by visualization with DiI and DiO, respectively (Figure 2B).…”

“…An in vivo imaging system (IVIS) was used to visualize their biodistribution at 24 h post‐injection ( Figure A,B; Figure S10, Supporting Information). Since we previously demonstrated that a scrambled peptide of the PREY sequence had no significant targeting effect, [ 12 ] no peptide control group was included in this study. The LCA‐targeting efficiency of PMSC‐NVs was better than that of MSC‐NVs or PMSCs, indicating the synergistic roles of PREY and NV to hinder the pulmonary capillary entrapment of cells and thereby to effectively target disturbed flow sites.…”

“…In the present study, we developed human MSC‐derived NVs that are functionalized with GSPREYTSYMPH (PREY), a disturbed flow‐targeting peptide that was previously identified from the in vivo phage display screening of a hundred million candidates (Figure S1, Supporting Information). [ 12 ] We used plasmid DNA that was designed to specifically functionalize the MSC membrane with PREY and produced NVs through physical cell crushing and subsequent self‐assembly. Here, we demonstrate the effective anti‐inflammatory and pro‐endothelial recovery effects of these PREY‐expressing MSC (PMSC)‐NVs targeting disturbed flow regions in mouse and porcine partial carotid artery ligation (PCL) models.…”

Bacteriophages and phage-inspired nanocarriers for targeted delivery of therapeutic cargos

“…Chen’s group[35] used the synthetic peptide ACSSSPSKHCG to improve the transdermal delivery of proteins such as human growth hormone and insulin. Hofmeister et al[36]found that the peptide GSPREYTSYMPH (PREY) could guide the nanocarriers to bind to atherosclerotic vasculature.…”