Fusogenic Lipid Nanovesicle for Biomacromolecular Delivery.

Although biomacromolecules are promising cytosolic drugs which have attracted tremendous attention, the major obstacles were the cellular membrane hindering the entrance and the endosome entrapment inducing biomacromolecule degradation. How to avoid those limitations to realize directly cytosolic delivery was still a challenge. Here, we prepared oligoarginine modified lipid to assemble a nanovesicle for biomacromolecules delivery, including mRNA (mRNA) and proteins which could be directly delivered into the cytoplasm of dendritic cells through subendocytosis-mediated membrane fusion. We named this membrane fusion lipid nanovesicle as MF-LNV. The mRNA loaded MF-LNV as nanovaccines showed efficient antigen expression to elicit robust immuno responses for cancer therapy. What's more, the antigen protein loaded MF-LNV as nanovaccines elicits much stronger CD8+ T cell specific responses than lipid nanoparticles through normal uptake pathways. This MF-LNV represented a refreshing strategy for intracellular delivery of the biomacromolecule.

Adenovirus-Derived Nano-Capsid Platforms for Targeted Delivery and Penetration of Macromolecules into Resistant and Metastatic Tumors.

Macromolecular therapeutics such as nucleic acids, peptides, and proteins have the potential to overcome treatment barriers for cancer. For example, nucleic acid or peptide biologics may offer an alternative strategy for attacking otherwise undruggable therapeutic targets such as transcription factors and similar oncologic drivers. Delivery of biological therapeutics into tumor cells requires a robust system of cell penetration to access therapeutic targets within the cell interior. A highly effective means of accomplishing this may be borrowed from cell-penetrating pathogens such as viruses. In particular, the cell entry function of the adenovirus penton base capsid protein has been effective at penetrating tumor cells for the intracellular deposition of macromolecular therapies and membrane-impermeable drugs. Here, we provide an overview describing the evolution of tumor-targeted penton-base-derived nano-capsids as a framework for discussing the requirements for overcoming key barriers to macromolecular delivery. The development and pre-clinical testing of these proteins for therapeutic delivery has begun to also uncover the elusive mechanism underlying the membrane-penetrating function of the penton base. An understanding of this mechanism may unlock the potential for macromolecular therapeutics to be effectively delivered into cancer cells and to provide a treatment option for tumors resisting current clinical therapies.

Orderly Curled Silica Nanosheets with a Small Size and Macromolecular Loading Pores: Synthesis and Delivery of Macromolecules To Eradicate Drug-Resistant Cancer.

Hierarchically organized silica nanomaterials have shown great promise for nanomedicine. However, the synthesis of silica nanomaterials with a small size and macromolecular loading pore is still a big challenge. Herein, orderly curled silica nanosheets (OCSNs) with a ∼42 nm diameter and orderly connected large channels (∼13.4 nm) were successfully prepared for the first time. The key to the formation of the unique structure (OCSNs) is using an oil/water reaction system with high concentrations of the surfactant and alkali. The prepared OCSNs exhibit a long blood circulation halftime (0.97 h) and low internalization in the reticuloendothelial system. Notably, the large superficial channels can concurrently house large guest molecules (siRNA) and chemotherapeutic drugs. Furthermore, drug-loaded OCSNs modified with polyglutamic acids can greatly increase the accumulation of incorporated siRNA and doxorubicin in solid tumors and restrain the growth of drug-resistant orthotopic breast cancer by inducing cell apoptosis. Overall, we report the preparation of hierarchically OCSNs; their small size and macromolecular loading pores are very promising for the delivery of large guest molecules and chemotherapeutic drugs for cancer therapy.

Cationic copolymer augments membrane permeabilizing activity of an amphiphilic peptide.

Membrane disruptive peptides (also called membrane fusogenic peptides) have been employed for cytosolic delivery of macromolecules such as nucleic acids and proteins. We reported previously that the cationic graft copolymer, poly(allylamine)-graft-dextran (PAA-g-Dex), augments membrane disruptive activity of the negatively charged E5 peptide. Strong membrane disruptive activity was observed in the presence of the copolymer at both acidic and neutral pH. In this paper, activities of E5/PAA-g-Dex mixture were further explored. Membrane permeabilization activity of E5/PAA-g-Dex was dependent on concentrations of both E5 and PAA-g-Dex, indicating that a complex between E5 and PAA-g-Dex produced the activity. Since the activity of peptide/PAA-g-Dex was peptide sequence-specific, we reasoned that PAA-g-Dex activated membrane-permeabilization activity by facilitating folding of E5 into its active conformation. The membrane permeabilization activity of E5/PAA-g-Dex resulted in transportation of bovine serum albumin into HL-60 cells with less cellular toxicity than digitonin, a naturally occurring surfactant used for delivery of macromolecules into cells.

High intensity focused ultrasound hyperthermia for enhanced macromolecular delivery.

Mild hyperthermia has been used in combination with polymer therapeutics to further increase delivery to solid tumors and enhance efficacy. An attractive method for generating heat is through non-invasive high intensity focused ultrasound (HIFU). HIFU is often used for ablative therapies and must be adapted to produce uniform mild hyperthermia in a solid tumor. In this work a magnetic resonance imaging guided HIFU (MRgHIFU) controlled feedback system was developed to produce a spatially uniform 43°C heating pattern in a subcutaneous mouse tumor. MRgHIFU was employed to create hyperthermic conditions that enhance macromolecular delivery. Using a mouse model with two subcutaneous tumors, it was demonstrated that MRgHIFU enhanced delivery of both Evans blue dye (EBD) and Gadolinium-chelated N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers. The EBD accumulation in the heated tumors increased by nearly 2-fold compared to unheated tumors. The Gadolinium-chelated HPMA copolymers also showed significant enhancement in accumulation over control as evaluated through MRI T1-mapping measurements. Results show the potential of HIFU-mediated hyperthermia for enhanced delivery of polymer therapeutics.