Modular functional peptides for the intracellular delivery of nanoparticles
Inventors
Delehanty, James B. • Medintz, Igor L. • Mattoussi, Hedi M. • Deschamps, Jeffrey R. • Dawson, Glyn • Dawson, Philip E. • Blanco-Canosa, Juan Bautista • Boeneman, Kelly
Assignees
University of Chicago • US Department of Navy
Publication Number
US-8796435-B2
Publication Date
2014-08-05
Expiration Date
2030-10-20
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Abstract
Described are nucleic acids encoding a polypeptide for delivery of a nanoparticle to the cytosol, the peptide comprising: (a) a nanoparticle association domain, (b) a spacer domain, (c) an uptake domain, and (d) a vesicle escape domain, wherein the domains (a) through (d) appear in the same order as listed above, and wherein the peptide, upon addition of a non-hydrolyzable lipophilic moiety to the vesicle escape domain and binding to a nanoparticle, is effective to induce uptake of a nanoparticle by a cell and delivery of the nanoparticle to the cytosol of the cell. Also described are methods of delivery of a nanoparticle to the cytosol of a cell, the method comprising providing to a cell a nanoparticle attached to such a peptide. Exemplary nanoparticles include quantum dots.
Core Innovation
Described is a nucleic acid encoding a polypeptide designed to deliver nanoparticles to the cytosol of cells. The polypeptide comprises multiple modular domains: a nanoparticle association domain, a spacer domain, an uptake domain, and a vesicle escape domain arranged sequentially. Upon attachment of a non-hydrolyzable lipophilic moiety to the vesicle escape domain and binding to a nanoparticle, the peptide induces cellular uptake and effective delivery of the nanoparticle to the cytosol.
The problem addressed is the limitation of existing methods for intracellular delivery of nanoparticles, such as quantum dots (QDs). Passive and facilitated delivery typically result in nanoparticles being sequestered within endolysosomal vesicles, limiting access to the cytosol. Active delivery methods like electroporation and microinjection, while delivering directly to the cytosol, are highly invasive and cause cellular damage and toxicity. Current strategies, including use of endosomal disruptors like sucrose or chloroquine, often involve toxic effects or limited cytosolic release of nanoparticles. Thus, there is a need for improved, efficient, and minimally toxic intracellular delivery approaches that enable cytosolic access.
This invention solves these problems by designing multifunctional modular peptides capable of mediating nanoparticle uptake and promoting endosomal escape to release nanoparticles into the cytosol. The polypeptide's modular domains act cooperatively: the nanoparticle association domain allows binding to the nanoparticle; the spacer reduces steric hindrance; the uptake domain facilitates cell membrane interaction and internalization; and the vesicle escape domain, bearing a non-hydrolyzable lipophilic moiety like palmitate, induces vesicular escape. This design enables the delivery of nanoparticles, including quantum dots, efficiently and with minimal cytotoxicity, overcoming deficiencies of previous methods.
Claims Coverage
The patent includes two independent claims directed to nucleic acids encoding specific multifunctional peptides for nanoparticle delivery.
Encoding multifunctional peptides for nanoparticle delivery
Nucleic acids encoding peptides selected from the group consisting of SEQ ID Nos: 3 through 6 and SEQ ID Nos: 9 through 36, which include modular multifunctional domains for nanoparticle association, cellular uptake, spacer function, and vesicle escape via a non-hydrolyzable lipid moiety.
Encoding of a specific peptide sequence for intracellular delivery
Nucleic acid encoding the specific peptide SEQ ID No. 3, which comprises a nanoparticle association domain (hexa-histidine), a proline-rich spacer domain, a positively charged lysine-rich uptake domain, and a vesicle escape domain with a non-hydrolyzable palmitoyl lipid moiety.
The claims cover nucleic acids encoding multifunctional modular peptides with defined sequence features that enable nanoparticle binding, cellular uptake, and endosomal escape, providing an effective system for intracellular nanoparticle delivery.
Stated Advantages
Efficient delivery of nanoparticles to the cytosol by peptides that mediate uptake and endosomal escape.
Minimal cytotoxicity observed upon delivery using the multifunctional peptides compared to other existing methods.
Long-term intracellular stability of peptide-nanoparticle assemblies even within acidic endolysosomal compartments.
The modular peptide design allows flexibility for iterative development and for assembly with various nanoparticle types.
Documented Applications
Cellular labeling including general labeling and labeling of specific subcellular compartments.
Cellular tagging for cell sorting and cataloging purposes.
Drug delivery and monitoring using nanoparticles coupled to multifunctional peptides.
In vivo imaging using biofunctionalized nanoparticles delivered intracellularly.
Delivery of various payloads such as proteins (e.g., 240-kDa B-phycoerythrin), carbohydrates, drugs, contrast agents, nucleic acids, and gene therapy agents into cells.
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