A Two‐Pronged Pulmonary Gene Delivery Strategy: A Surface‐Modified Fullerene Nanoparticle and a Hypotonic Vehicle

Abstract

Inhaled gene therapy poses a unique potential of curing chronic lung diseases, which are currently managed primarily by symptomatic treatments. However, it has been challenging to achieve therapeutically relevant gene transfer efficacy in the lung due to the presence of numerous biological delivery barriers. Here, we introduce a simple approach that overcomes both extracellular and cellular barriers to enhance gene transfer efficacy in the lung in vivo. We endowed tetra(piperazino)fullerene epoxide (TPFE)‐based nanoparticles with non‐adhesive surface polyethylene glycol (PEG) coatings, thereby enabling the nanoparticles to cross the airway mucus gel layer and avoid phagocytic uptake by alveolar macrophages. In parallel, we utilized a hypotonic vehicle to facilitate endocytic uptake of the PEGylated nanoparticles by lung parenchymal cells via the osmotically driven regulatory volume decrease (RVD) mechanism. We demonstrate that this two‐pronged delivery strategy provides safe, wide‐spread and high‐level transgene expression in the lungs of both healthy mice and mice with chronic lung diseases characterized by reinforced delivery barriers.

Development of nintedanib nanosuspension for inhaled treatment of experimental silicosis

“…Furthermore, nonadhesive surface F127 coating enhances lung retention of our formulation by minimizing the adhesive interactions with airway mucus and lung‐resident macrophages 16–20 that promote the clearance of inhaled foreign matters from the lung as natural host defense mechanisms 11 . Of note, we have demonstrated that F127‐coated model NS formulations efficiently traverse human mucus samples harvested from various mucosal surfaces 31 and that covalent surface shielding of nanoparticles with the hydrophilic segment of F127 (i.e., polyethylene glycol) markedly reduces particle phagocytosis by alveolar macrophages 46 …”

“…11 Of note, we have demonstrated that F127-coated model NS formulations efficiently traverse human mucus samples harvested from various mucosal surfaces 31 and that covalent surface shielding of nanoparticles with the hydrophilic segment of F127 (i.e., polyethylene glycol) markedly reduces particle phagocytosis by alveolar macrophages. 46 Albeit not primarily for inhaled use, there are more than 20 marketed NS products as of 2020. 45 On the other hand, aqueous drug suspensions are widely used in clinic for the inhaled treatment of patients with numerous lung diseases, including asthma, chronic obstructive pulmonary disease, and cystic fibrosis.…”

pH‐Responsive Polymer Nanoparticles for Efficient Delivery of Cas9 Ribonucleoprotein With or Without Donor DNA

“…To further enhance the gene editing efficiency in lung epithelium, NHEJ‐NP was prepared in a hypotonic aqueous solution, because previous studies demonstrated that a hypotonic shock could force cells to undergo intensive endocytosis and thus enhance the cellular uptake of nanoparticles via the osmotically‐driven regulatory volume decrease mechanism. [ 16 ] It was found that hypotonic NHEJ‐NP led to significant enhancement of the tdTomato fluorescence intensity in comparison with the PBS‐treated group (Figure 4C). Consistent with the IVIS results, more tdTomato expression was found in the hypotonic NHEJ‐NP‐treated lung epithelium than the isotonic NHEJ‐NP‐treated one (Figure 4D).…”

Molecule-to-Material-to-Bio Nanoarchitectonics with Biomedical Fullerene Nanoparticles

“…Shu, Suk, and coworkers investigate localized gene therapy via inhalation to treat refractory lung diseases ( Figure 17 ) [ 172 ]. This is advantageous in mitigating risks of systemic adverse effects caused by extra-pulmonary gene transfer.…”