The overall goal of this proposal is to develop exosome-like nanoparticles as a delivery vehicle for targetable delivery of extracellular RNA (exRNA) primarily focusing on miRNAs which modulate gene expression and induce immune responses upon delivery in mouse tumor models. Development of exRNA-based therapeutics must overcome challenges including issues of delivery, potential off-target effects, safety, toxicity, the cost of large scale production, elimination of potential biohazards to the environmen and tissue specificity. Unlike the situation with artificially synthesized nanoparticles or liposomes, naturally derived nanoparticle-size exosomes are released from many different types of cells. miRNAs and RNAs can be encapsulated in naturally derived exosomes and the RNAs are stable, and a number of miRNAs can be naturally encapsulated simultaneously in these exosomes. Recently, data published by ours and other groups have demonstrated that mammalian exosomes can deliver poorly soluble agents, chemotherapeutic drugs, and siRNA in vivo. A limitation of this mode is the need for large-scale production of exosomes, and potential biosafety. Our preliminary data show that exosome-like nanoparticles can be isolated in large quantities from the tissue of edible plants, including grapes. We have demonstrated that exosome-like nanoparticles from grapes are composed of small RNAs, proteins, and lipids. An edible plant-derived nano-vector (EPNV) assembled from grape exosome-like derived lipids is capable of encapsulating siRNAs. Co-delivery of folic acid, with the chemotherapy drug paclitaxel by EPNVs results in an increased efficiency in tumor tissue targeting. Reducing the size of EPNVs by passing them through an appropriate pore size filter significantly enhances EPNV translocation to the brain and oral administration of the EPNVs results in their migration to the liver. Based on these preliminary data and the current literature, we propose to: (1) Determine whether an EPNV can deliver therapeutic miR17 via oral administration to target metastatic colon tumor in the liver, subsequently inducing NK cell activation to kill metastatic tumor cells;(2) Determine whether miR155 encapsulated in EPNV promotes differentiation of myeloid derived suppressor cells (MDSCs) into mature dendritic cells in mouse breast/brain tumor models;(3) Determine whether a therapeutic dose of miRNA17 or miRNA155 delivered by EPNV induces no side-effects in mouse tumor models;and (4) Determine whether EPNV can be produced in large scale amounts economically for use in clinical settings. Demonstration of using a EPNV-based delivery system loaded with a desired therapeutic extracellular RNA (exRNA) including microRNA to target to inflammatory cells and tumor cells in vivo would not only be a significant step forward in the treatment of cancer, but also provide a powerful tool for identifying the role and effects of each small RNA encapsulated in resident or circulating microvesicles on the recipient cells. We will accomplish this in our study by knock-out and knock-in strategies.

Public Health Relevance

The concept that extracellular RNA (exRNA) may alter target cell phenotypes by way of intercellular signaling is becoming universally accepted in science. Whether exRNA can be used as a therapeutic agent is not known. The overall goal of this study is to develop grape lipid derived nano vectors for the delivery of exRNAs as therapeutic agents to target tumor cells as well as inflammatory cells in animal tumor models. Additionally we will assess the therapeutic potential of exRNA used in a clinical setting.

Agency
National Institute of Health (NIH)
Institute
National Center for Advancing Translational Sciences (NCATS)
Type
Exploratory/Developmental Cooperative Agreement Phase I (UH2)
Project #
3UH2TR000875-02S1
Application #
8840378
Study Section
Special Emphasis Panel (ZRG1 (50))
Program Officer
Tagle, Danilo A
Project Start
2013-08-01
Project End
2015-07-31
Budget Start
2014-08-28
Budget End
2015-07-31
Support Year
2
Fiscal Year
2014
Total Cost
$74,993
Indirect Cost
$24,998
Name
University of Louisville
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
057588857
City
Louisville
State
KY
Country
United States
Zip Code
40202
Guo, Peixuan; Noji, Hiroyuki; Yengo, Christopher M et al. (2016) Biological Nanomotors with a Revolution, Linear, or Rotation Motion Mechanism. Microbiol Mol Biol Rev 80:161-86
Zhuang, Xiaoying; Teng, Yun; Samykutty, Abhilash et al. (2016) Grapefruit-derived Nanovectors Delivering Therapeutic miR17 Through an Intranasal Route Inhibit Brain Tumor Progression. Mol Ther 24:96-105
Deng, Z; Rong, Y; Teng, Y et al. (2016) Exosomes miR-126a released from MDSC induced by DOX treatment promotes lung metastasis. Oncogene :
Wang, Qilong; Ren, Yi; Mu, Jingyao et al. (2015) Grapefruit-Derived Nanovectors Use an Activated Leukocyte Trafficking Pathway to Deliver Therapeutic Agents to Inflammatory Tumor Sites. Cancer Res 75:2520-9
Quesenberry, Peter J; Aliotta, Jason; Camussi, Giovanni et al. (2015) Potential functional applications of extracellular vesicles: a report by the NIH Common Fund Extracellular RNA Communication Consortium. J Extracell Vesicles 4:27575
Deng, Zhongbin; Mu, Jingyao; Tseng, Michael et al. (2015) Enterobacteria-secreted particles induce production of exosome-like S1P-containing particles by intestinal epithelium to drive Th17-mediated tumorigenesis. Nat Commun 6:6956
Jiang, Hong; Wang, Ping; Li, Xiaohua et al. (2014) Restoration of miR17/20a in solid tumor cells enhances the natural killer cell antitumor activity by targeting Mekk2. Cancer Immunol Res 2:789-99
Jiang, Hong; Wang, Ping; Wang, Qilong et al. (2014) Quantitatively controlling expression of miR-17~92 determines colon tumor progression in a mouse tumor model. Am J Pathol 184:1355-68
Mu, Jingyao; Zhuang, Xiaoying; Wang, Qilong et al. (2014) Interspecies communication between plant and mouse gut host cells through edible plant derived exosome-like nanoparticles. Mol Nutr Food Res 58:1561-73
Deng, Zhong-Bin; Zhuang, Xiaoying; Ju, Songwen et al. (2013) Exosome-like nanoparticles from intestinal mucosal cells carry prostaglandin E2 and suppress activation of liver NKT cells. J Immunol 190:3579-89

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