Delivery of nanoparticles to neurons
Inventors
Delehanty, James B. • Medintz, Igor L. • Mattoussi, Hedi M. • Deschamps, Jeffrey R. • Dawson, Glyn • Dawson, Philip E. • Blanco-Canosa, Juan Bautista • Boeneman, Kelly • Susumu, Kimihiro • Stewart, Michael H. • Walters, Ryan
Assignees
University of Chicago • US Department of Navy
Publication Number
US-8859284-B2
Publication Date
2014-10-14
Expiration Date
2030-10-20
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Abstract
A peptide attached to a nanoparticles (such as quantum dots) selectively directs the nanoparticles to neurons in a tissue or organism.
Core Innovation
The invention relates to peptides attached to nanoparticles, such as quantum dots, which selectively direct the nanoparticles to neurons within a tissue or organism. A modular multifunctional peptide is described that includes distinct functional domains: a nanoparticle association domain, a spacer domain, an uptake domain, and a vesicle escape domain incorporating a non-hydrolyzable lipid moiety. When attached to extracellular nanoparticles, such peptides mediate cellular uptake and subsequent delivery of the nanoparticles to the cytosol of cells.
The problem addressed is the challenge of delivering nanoparticles, specifically quantum dots or other nanomaterials, effectively and selectively to the cytosol of neurons. Prior methods for nanoparticle delivery to cells suffer from deficiencies: passive and facilitated delivery typically result in entrapment of nanoparticles within endolysosomal vesicles, requiring toxic strategies to release payloads; active delivery techniques like electroporation and microinjection are invasive and harmful to cellular viability. Additionally, previous approaches lacked specificity to neurons and often encountered toxicity issues or aggregated nanoparticles. Thus, an improved method for selective, efficient, and non-toxic delivery of nanoparticles to neuronal cytosol is needed.
The disclosed approach employs a rationally-designed, modular peptide with multiple functional domains that collectively mediate nanoparticle binding, cellular uptake, endosomal escape, and ultimate cytosolic delivery, specifically demonstrating selective targeting and intracellular distribution in neuron-rich hippocampal slice cultures. The approach leverages a non-hydrolyzable palmitic acid attached to the peptide backbone to facilitate vesicle escape, a polyproline-rich spacer to extend functional domains beyond nanoparticle coatings, and a lysine-rich uptake sequence to enhance cellular interaction and internalization. This delivery system overcomes previous limitations by providing selective neuronal uptake, cytosolic release of nanoparticles, minimal cytotoxicity, and applicability to various nanoparticles and biological contexts.
Claims Coverage
The patent contains multiple inventive features focused on methods and peptides for delivery and labeling of nanoparticles to neurons, defined principally in two independent claims encompassing the method of delivery and the components of the multifunctional peptide.
Method of delivering nanoparticles to neurons using a compound of specified formula
The method involves providing to a tissue a nanoparticle bound to a compound of a particular chemical formula that facilitates selective delivery to neurons.
Use of a multifunctional peptide with defined domains for nanoparticle delivery
The nanoparticle is also bound to a peptide comprising (a) a nanoparticle association domain, (b) a spacer domain, (c) an uptake domain, and (d) a vesicle escape domain that includes a non-hydrolyzable lipid moiety, with the spacer domain positioned between the association domain and the uptake and vesicle escape domains.
Specific incorporation of Palm-1 peptide as delivery agent
The peptide used for delivery can comprise the Palm-1 sequence, a known multifunctional modular peptide with domains for nanoparticle association, spacer, uptake, and vesicle escape anchored by a non-hydrolyzable palmitic acid moiety.
Nanoparticle selection to include quantum dots
The nanoparticle to be delivered can be a quantum dot, a semiconductor nanocrystal specifically mentioned as an exemplary nanoparticle in the delivery methods.
Neuro-specific induction of nanoparticle uptake
The compound bound to the nanoparticle is effective to induce preferential uptake by neuronal cells, conferring cell type specificity within neural tissue.
Capability of delivering nanoparticle-bound drugs
The nanoparticle can be additionally bound to a drug, enabling therapeutic delivery to neurons alongside imaging or labeling capabilities.
Applicability of the delivery method to living organisms
The tissue targeted in the delivery method can be within a living organism, indicating applicability beyond in vitro or ex vivo systems.
Method of labeling neurons using nanoparticle conjugates
A method for labeling neurons includes providing to the tissue a nanoparticle bound to (1) the compound of the specified formula and (2) a multifunctional peptide with defined domains connecting via a non-hydrolyzable lipid moiety facilitating neuronal uptake and labeling.
Defined peptide spacer and sequences for enhanced delivery
The peptide includes a spacer domain comprising nine proline residues and can include sequences corresponding to SEQ ID No: 3 or SEQ ID No: 4, which contribute to the modular design for efficient cellular uptake and endosomal escape.
The claims collectively cover methods of delivering and labeling neurons with nanoparticles bound to specific multifunctional peptides and chemical compounds that allow selective neuronal targeting, cytosolic delivery, and potential drug loading, emphasizing a modular, multifunctional peptide design featuring a non-hydrolyzable lipid moiety, a proline-rich spacer, and lysine-rich uptake domain, with quantum dots as exemplar nanoparticles.
Stated Advantages
Selective targeting of nanoparticles to neurons in complex tissues, minimizing delivery to non-neuronal cells.
Efficient nanoparticle endosomal escape leading to cytosolic delivery, overcoming entrapment limitations of prior methods.
Minimal cytotoxicity resulting from the delivery system, preserving cell viability.
Versatile and modular peptide design allowing control over nanoparticle surface decoration, facilitating optimization for specific applications.
Applicability for in vivo use in living organisms.
Capability to deliver therapeutic agents or imaging labels specifically to neuronal cytosol.
Documented Applications
Selective delivery and labeling of neurons in tissue cultures and living organisms using nanoparticles conjugated with multifunctional peptides and chemical compounds.
Intracellular delivery of quantum dots for visualization, imaging, and biological labeling of neuronal cells.
Potential therapeutic delivery to neurons by conjugating drugs to nanoparticles decorated with the multifunctional peptides.
Use of quantum dots and similar nanoparticles for targeted neurotherapeutic applications in the central nervous system.
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