Endothelial targeting of nanocarriers loaded with antioxidant enzymes for protection against vascular oxidative stress and inflammation

Abstract

Endothelial-targeted delivery of antioxidant enzymes, catalase and superoxide dismutase (SOD), is promising strategy for protecting organs and tissues from inflammation and oxidative stress. Here we describe Protective Antioxidant Carriers for Endothelial Targeting (PACkET), the first carriers capable of targeted endothelial delivery of both catalase and SOD. PACkET formed through controlled precipitation loaded ~30% enzyme and protected it from proteolytic degradation, whereas attachment of PECAM monoclonal antibodies to surface of the enzyme-loaded carriers, achieved without adversely affecting their stability and functionality, provided targeting. Isotope tracing and microscopy showed that PACkET exhibited specific endothelial binding and internalization in vitro. Endothelial targeting of PACkET was validated in vivo by specific (vs IgG-control) accumulation in the pulmonary vasculature after intravenous injection achieving 33% of injected dose at 30 min. Catalase loaded PACkET protects endothelial cells from killing by H2O2 and alleviated the pulmonary edema and leukocyte infiltration in mouse model of endotoxin-induced lung injury, whereas SOD-loaded PACkET mitigated cytokine-induced endothelial pro-inflammatory activation and endotoxin-induced lung inflammation. These studies indicate that PACkET offers a modular approach for vascular targeting of therapeutic enzymes.

Translational research into vascular wall function: regulatory effects of systemic and specific factors

“…It should be noted that in an animal model of endotoxic shock SOD-CHS-CAT significantly increases viability after preventive and therapeutic administration [56]. These results are interesting in comparison with modeled therapeutic effects of modular approaches [51] and for a better understanding of the mechanism underlying the conjugate activity involving NO-dependent and NO-independent pathways for its therapeutic effect. The effectiveness of SOD-CHS-CAT conjugate urges further development of genetic and bioengineering technologies for production of three-functional enzymes with activities of SOD and CAT [61] and peroxidase and SOD [62] capable of entering the cell.…”

“…Superoxide dismutase and catalase (up to 30% total content) were precipitated on a protective antioxidant nanocarrier for endothelial targeting and bound to antibodies against endothelial adhesion molecules to provide the targeting [51]. In a mouse model of endotoxin-induced pulmonary damage CAT derivative protected endothelium, decreased lung edema and leukocyte infiltration.…”

“…SOD derivative reduced cytokine-induced proinflammatory activation of the endothelium and endotoxin-induced pneumonia. A modular approach has been suggested for targeting therapeutic enzymes at the vascular wall [51]. Among three concentration intervals of reactive forms of oxygen and nitrogen species (physiological, enhanced and toxic), there is the favourable effect on toxic interval with such approaches as center-specific, nanocarrier-based therapy [52], 3D supramolecular ensembles with antioxidants or enzymes [53] and nanotherapeutics with an antioxidant nucleus for blocking atherogenic oxidative stress and an amphiphilic polysaccharide covering targeted at scavenger receptor to inhibit athero-inflammatory cycle [54].…”

Nanomedicine for the molecular diagnosis of cardiovascular pathologies

“…The mechanisms of plaque development being understood, different molecular markers have been identified, among which: endothelial targets (VCAM-1, ICAM-1, E-and P-selectins) [7], macrophage targets (scavenger receptor class A (SR-A) and integrin MAC-1), fibrous cap and extracellular matrix (collagen types I, III, IV), apoptosis (expression of phosphatidylserine) and angiogenesis [8]. For preclinical studies, animal models have been developed to mimic the atherosclerotic lesions, such as Apolipoprotein E deficient mice (ApoEÀ/À mice) [9].…”

Angiopep-2-Mediated Delivery of Human Manganese Superoxide Dismutase in Brain Endothelial Cells and its Protective Effect Against Oxidative Stress

“…Several studies have focused on the utilization of various antioxidant enzymes as protective agents (Fennell et al 2002;He et al 2006;Hood et al 2014;Uyama et al 1992). Superoxide dismutase (SOD) is an endogenous enzyme that is a part of the first line of defense through elimination of O 2 -by converting it into H 2 O 2 and O 2 .…”