Blood vessels present a great barrier for delivering therapeutic nanoparticles; however the existing approaches are not sufficient to resolve this problem. Neutrophils are the most abundant white blood cells circulating in bloodstream, and are capable of transmigrating into tissues to rapidly respond tissue injury and infection. Therefore, neutrophils could be an excellent carrier to mediate nanoparticle transport across vessel barrier. Using intravital microscopy and an acute lung inflammation mouse model we have demonstrated that nanoparticles made from albumin protein can specifically bind activated neutrophils leading to the transport of these nanoparticles across endothelial barrier via neutrophil-mediated transmigration. Thus, we hypothesize that neutrophils can transport drugs loaded in albumin nanoparticles across a blood-vessel barrier and deposit the drugs in diseased tissues. Furthermore, using this novel approach we could prevent acute lung injury, a devastating disease currently without effective pharmacological therapies. We have proposed three aims:
Aim 1 : Define the properties of albumin nanoparticle platforms required for targeting of activated neutrophils in vivo. We will develop the methods to generate bright fluorescently-labeled albumin nanoparticles for tracking nanoparticles in cells and live mice. We will address the roles of particle size, density, surface charge and albumin properties in affecting nanoparticle uptake. We will determine drug loading efficiency in albumin nanoparticles and drug release profiles in vitro and in vivo using HPLC/mass spectroscopy.
Aim 2 : Dissect the mechanism of neutrophil-mediated nanoparticle transport across the blood- vasculature barrier. Using intravital microscopy of mouse cremaster tissues, we will real-time visualize the uptake of albumin nanoparticles by neutrophils and neutrophils carrying the nanoparticles transmigrate across endothelial vessels and move to inflammatory tissues. In a tracheal-LPS induced acute lung inflammation model, we will investigate whether nanoparticle uptake by neutrophils affects neutrophil biological functions (transport and activity) using confocal microscopy, biochemical approaches and knockout mouse models.
Aim 3 : Determine the efficacy of our nanoparticle platforms in treatment of acute lung injury. By measuring cytokines, neutrophil infiltration and plasma protein accumulation in mouse lungs, and histochemistry, we will evaluate whether albumin nanoparticles loaded with NF-?B inhibitors can dramatically mitigate lung inflammation/injury compared to the free drugs. At the end of these studies, we will develop the novel strategies of delivering therapeutic into targets based on neutrophil transmigration to diseased locations, and create a way to deliver nanoparticles across endothelial barrier. Our studies will change current strategies to treat inflammatory disorders underlying acute and chronic diseases including cancer. Specifically, our approach will significantly impact on the development of new pharmacological therapies to treat acute lung injury, and a severe form, acute respiratory distress syndrome (ARDS) because there are no effective therapies to treat these devastating diseases with 40% mortality.

Public Health Relevance

Therapeutic nanoparticles should cross the blood vessel barrier for medical benefits, but little is known about how to overcome this barrier. We propose a novel strategy to deliver therapeutic nanoparticles across this barrier into diseased locations based on neutrophil-mediated transmigration which is a natural immune defense pathway. We will employ multidisciplinary approaches (intravital microscopy, nanotechnology and molecular biology) to demonstrate that therapeutic nanoparticles are able to transport across the blood vessel barrier in animal models. The results from our proposed studies will be transformative and have considerable impact on nanomedicine and pharmaceutical industries to improve treatments of a range of inflammatory disorders, such as acute lung injury and cancer.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Gene and Drug Delivery Systems Study Section (GDD)
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Garcia, Martha
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Washington State University
Schools of Pharmacy
United States
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Wang, Sihan; Gao, Jin; Wang, Zhenjia (2018) Outer membrane vesicles for vaccination and targeted drug delivery. Wiley Interdiscip Rev Nanomed Nanobiotechnol :e1523
Chu, Dafeng; Dong, Xinyue; Shi, Xutong et al. (2018) Neutrophil-Based Drug Delivery Systems. Adv Mater 30:e1706245
Wang, Sihan; Gao, Jin; Li, Mo et al. (2018) A facile approach for development of a vaccine made of bacterial double-layered membrane vesicles (DMVs). Biomaterials 187:28-38
Wang, Sihan; Dong, Xinyue; Gao, Jin et al. (2018) Targeting Inflammatory Vasculature by Extracellular Vesicles. AAPS J 20:37
Dong, Xinyue; Chu, Dafeng; Wang, Zhenjia (2018) Neutrophil-mediated delivery of nanotherapeutics across blood vessel barrier. Ther Deliv 9:29-35
Zhang, Can Yang; Gao, Jin; Wang, Zhenjia (2018) Bioresponsive Nanoparticles Targeted to Infectious Microenvironments for Sepsis Management. Adv Mater 30:e1803618
Gao, Jin; Wang, Sihan; Wang, Zhenjia (2017) High yield, scalable and remotely drug-loaded neutrophil-derived extracellular vesicles (EVs) for anti-inflammation therapy. Biomaterials 135:62-73
Dong, Xinyue; Chu, Dafeng; Wang, Zhenjia (2017) Leukocyte-mediated Delivery of Nanotherapeutics in Inflammatory and Tumor Sites. Theranostics 7:751-763
Chu, Dafeng; Dong, Xinyue; Zhao, Qi et al. (2017) Photosensitization Priming of Tumor Microenvironments Improves Delivery of Nanotherapeutics via Neutrophil Infiltration. Adv Mater 29:
Gao, Jin; Chu, Dafeng; Wang, Zhenjia (2016) Cell membrane-formed nanovesicles for disease-targeted delivery. J Control Release 224:208-216

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