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Principal Investigator: Stephen Patrick Walton
Organization: MICHIGAN STATE UNIVERSITY
Fiscal Year: 2024
Award: $289,946
Funding agency: National Institute of General Medical Sciences
Project Summary
Intercellular communication based on the delivery of miRNAs from one cell to adjacent or remote cells is now
known to be a critical part of normal cellular/organismal function. Recent work has demonstrated that a new
class of extracellular nanoparticles, exomeres, may be more important for miRNA-mediated intercellular
communication than extracellular vesicles (EVs). Exomeres are small, non-membranous secreted particles
that are different from EVs. Compared to EVs, exomeres are smaller, have distinct protein and nucleic acid
content, and are not encapsulated by lipid membranes. Also, exomeres contain Argonaute-2 (Ago2), one of
the principal proteins of the RNA interference (RNAi) pathway. With respect to intercellular communication,
exomeres contain miRNAs, which are likely bound by Ago2, and the EGFR ligand, amphiregulin, which could
initiate receptor-mediated endocytosis. Thus, exomeres are a natural RNA-containing nanoparticle evolved to
have molecular and physical properties that facilitate efficient endocytosis and intracellular processing of the
miRNA cargo by recipient cells. In this work, we will begin to explore whether exomeres could be used for
delivery of exogenous miRNAs (or siRNAs) for therapeutic applications.
The mechanisms of exomere biogenesis, secretion, endocytosis, and intracellular trafficking remain poorly
understood. This is due, in part, to the challenges of isolating exomeres. Current exomere isolation approaches
have allowed for preliminary characterization of exomeres, but further study of exomeres, and their critical
roles in RNA-mediated intercellular communication, will require approaches for isolating exomeres that yield
high concentrations of reasonably homogeneous complexes. In this work, we will develop a novel, scalable
exomere isolation approach based on tangential flow filtration (TFF) that will enhance our ability to study
exomeres and the endocytosis and intracellular trafficking of exomeres by recipient cells. The overarching
goal of this work is to demonstrate the utility of TFF for exomere isolation and to demonstrate that exomeres
isolated by TFF are identical to those isolated by current approaches. Given that TFF is compatible with
possible industrial production methods for exomeres, our results will provide the foundation for future efforts to
use exomeres for RNA delivery in clinical applications.
Terms: <Address><Amphiregulin><Antisense Agent><Antisense Oligonucleotides><Binding><Biogenesis><Biotech><Biotechnology><Body Tissues><CRGF><CRISPR><CRISPR/Cas system><Categories><Cell Body><Cell Culture Techniques><Cell Membrane Lipids><Cell to Cell Communication and Signaling><Cell-Cell Signaling><Cells><Centrifugation><Centrifugation Fractionation><Charge><Clustered Regularly Interspaced Short Palindromic Repeats><Colorectum Cell-Derived Growth Factor><Complement><Complement Proteins><Complex><Development><Disease><Disorder><EGF Receptor><EGFR><ERBB Protein><Encapsulated><Endocytosis><Endosomes><Epidermal Growth Factor Receptor><Epidermal Growth Factor Receptor Kinase><Epidermal Growth Factor Receptor Protein-Tyrosine Kinase><Epidermal Growth Factor-Urogastrone Receptors><Field Flow Fractionation><Filtration><Filtration Fractionation><Formulation><Foundations><Future><Goals><HER1><Health><Human><Keratinocyte-Derived Autocrine Factor><Laboratories><Ligands><Location><Mediating><Membrane Lipids><Messenger RNA><Methods><Micro RNA><MicroRNAs><Mission><Modern Man><Molecular><Molecular Interaction><NIH><National Institutes of Health><Non-Polyadenylated RNA><Nucleic Acids><Origin of Life><Post-Transcriptional Gene Silencing><Posttranscriptional Gene Silencing><Process><Production><Property><Proteins><Protocol><Protocols documentation><Publishing><RNA><RNA Gene Products><RNA Interference><RNA Interference Pathway><RNA Silencing><RNA based therapeutics><RNA based therapy><RNA delivery><RNA therapy><RNAi><Receptosomes><Repression><Reproducibility><Ribonucleic Acid><Role><Schwannoma-Derived Growth Factor><Sequence-Specific Posttranscriptional Gene Silencing><Shapes><Short interfering RNA><Small Interfering RNA><TGF-alpha Receptor><Technology><Therapeutic><Tissues><Transforming Growth Factor alpha Receptor><United States National Institutes of Health><Urogastrone Receptor><Work><antisense oligo><bioprocess><c-erbB-1><c-erbB-1 Protein><cell culture><cell cultures><cell type><clinical applicability><clinical application><colorectal cell-derived growth factor><colorectal-associated growth factor><colorectum-associated growth factor><complementation><cost effective><deliver short interfering RNA><deliver siRNA><deliver small interfering RNA><delivery system for siRNA><delivery system for small interfering RNA><delivery vector><delivery vectors for siRNA><delivery vehicle><developmental><erbB-1><erbB-1 Proto-Oncogene Protein><erbBl><extracellular><extracellular vesicles><human disease><improved><industrial production><innovate><innovation><innovative><intercellular communication><keratinocyte autocrine factor><lipid based nanoparticle><lipid nanoparticle><mRNA><miRNA><miRNA delivery><miRNAs><microRNA delivery><nano particle><nano-sized particle><nanoparticle><nanosized particle><new approaches><novel><novel approaches><novel strategies><novel strategy><nucleic acid delivery><nucleic acid therapy><nucleic acid-based therapeutics><particle><physical property><prevent><preventing><proto-oncogene protein c-erbB-1><receptor mediated endocytosis><scale up><short interfering RNA delivery><siRNA><siRNA delivery><site targeted delivery><small interfering RNA delivery><social role><success><targeted delivery><therapeutic RNA><therapeutic nucleic acids><therapeutically effective><trafficking><uptake>