Our overall goal is to create synthetic vesicles that mimic natural exosomes and microvesicles for the delivery of miRNA, antimiRNA and locked nucleic acids (LNA) to endothelial cells based on analyses of natural vesicles and imaging-based assessments of delivery. Membrane- encapsulated miRNAs circulating in human blood are known to be a mixture of microparticles (>100 nm) and exosomes (40-100 nm) which have long been thought to play a role in intercellular signaling. These microparticles contain a variety of biological components, including proteins and RNA molecules, and can effectively transfer these components from one cell type to the next. Importantly, recent evidence suggests that selective packaging of miRNAs into microparticles and exosomes is crucial to the specificity of biological function of secreted miRNAs. Preliminary data and a newly published paper from our group indicate that the anti- miR712 family can have a significant impact in the prevention of atherosclerosis. However, due to the potential off target effects of miRNA therapeutics, the creation of targeted vesicles that can enhance delivery at the target site is highly desirable. Preliminary data further demonstrate that: 1) unique antimiRNA-containing targeted vesicles produce effective knockdown in vitro and in vivo, 2) targeted synthetic vesicles accumulate with a 10-18 fold greater efficiency in regions of disturbance than surrounding tissue, 3) their accumulation is proportional to VCAM-1 expression in regions of flow disturbance (whereas the naked antimiRNA accumulates much less outside of the surgically modified carotid) and 4) the incorporation of phosphatidylserine (PS) enhances uptake of native and synthetic vehicles. We have developed the MRI, positron emission tomography and optical imaging methods required to quantify the pharmacokinetics and uptake of vesicles and miRNA. Within the proposed work, we will characterize the lipid and protein content of native vesicles and correlate these constituents with vesicle uptake. Here, we will accomplish the following aims: 1) based on an analysis of native vesicles, engineer antimiRNA and miRNA-loaded synthetic vesicles for uptake into endothelial cells and create functional knockdown. 2) Determine the pharmacokinetics, trafficking, safety and efficacy of antimiRNA- and miRNA-loaded synthetic vesicles in mouse and rabbit models.
Our overall goal is to create synthetic vesicles that mimic natural exosomes and microvesicles for the delivery of miRNA, creating an effective treatment for atherosclerosis. We will compare the efficiency of native and synthetic particles in the delivery of antimiRNA and miRNA and also specifically assess the therapeutic efficacy of miRNA therapeutics based on the miR712 family.
|Heath, Jack M; Fernandez Esmerats, Joan; Khambouneheuang, Lucky et al. (2018) Mechanosensitive microRNA-181b Regulates Aortic Valve Endothelial Matrix Degradation by Targeting TIMP3. Cardiovasc Eng Technol 9:141-150|
|Kim, Chan Woo; Pokutta-Paskaleva, Anastassia; Kumar, Sandeep et al. (2017) Disturbed Flow Promotes Arterial Stiffening Through Thrombospondin-1. Circulation 136:1217-1232|
|Kumar, Sandeep; Kang, Dong-Won; Rezvan, Amir et al. (2017) Accelerated atherosclerosis development in C57Bl6 mice by overexpressing AAV-mediated PCSK9 and partial carotid ligation. Lab Invest 97:935-945|
|Rathan, Swetha; Ankeny, Casey J; Arjunon, Sivakkumar et al. (2016) Identification of side- and shear-dependent microRNAs regulating porcine aortic valve pathogenesis. Sci Rep 6:25397|
|Kumar, Sandeep; Jang, In-Hwan; Kim, Chan Woo et al. (2016) Functional screening of mammalian mechanosensitive genes using Drosophila RNAi library- Smarcd3/Bap60 is a mechanosensitive pro-inflammatory gene. Sci Rep 6:36461|
|Simmons, Rachel D; Kumar, Sandeep; Jo, Hanjoong (2016) The role of endothelial mechanosensitive genes in atherosclerosis and omics approaches. Arch Biochem Biophys 591:111-31|
|Simmons, Rachel D; Kumar, Sandeep; Thabet, Salim Raid et al. (2016) Omics-based approaches to understand mechanosensitive endothelial biology and atherosclerosis. Wiley Interdiscip Rev Syst Biol Med 8:378-401|
|Mitchell, Adam J; Gray, Warren D; Schroeder, Max et al. (2016) Pleomorphic Structures in Human Blood Are Red Blood Cell-Derived Microparticles, Not Bacteria. PLoS One 11:e0163582|
|Mitchell, Adam J; Gray, Warren D; Hayek, Salim S et al. (2016) Platelets confound the measurement of extracellular miRNA in archived plasma. Sci Rep 6:32651|
|Fernández Esmerats, Joan; Heath, Jack; Jo, Hanjoong (2016) Shear-Sensitive Genes in Aortic Valve Endothelium. Antioxid Redox Signal 25:401-14|
Showing the most recent 10 out of 19 publications