Adverse immunological responses against non-viral nanoparticle (NP) delivery systems in the lung

Figures & data

Table 1. Current candidate gene therapies for COPD, asthma, and IPF.

Disease	Gene therapy approach	Outcome of intervention	References
COPD	Aerosol delivery of cationic liposome complexed to plasmid encoding recombinant human α1-anti-trypsin gene to New Zealand White rabbits	Transfection and expression of human α1-antitrypsin in alveolar epithelial cells for 7 d after single dose	(Canonico et al. 1994)
	Transfection of human bronchial epithelial cells with p65 siRNA and treatment with NF-κB small molecule inhibitor	Attenuation of NF-κB pathway Suppression of pro-inflammatory cytokine IL-1β- and IL-17A-induced expression of mucin (MUC)5AC gene	(Fujisawa et al. 2009)
	Treatment of human alveolar epithelial lung cells with cationic polymeric NP attached to miR-146a	Inhibited expression of IL-1 receptor-associated kinase (IRAK1) genes Reduced IL-1β-dependent activation of NF-κB	(Mohamed et al. 2019)
Asthma	Intratracheal instillation of dexamethasone attached to cationic polymeric NP loaded with Vitamin D binding protein (VDBP) to OVA-sensitized mice model of allergic asthma	Down-regulated alveolar macrophage VDBP expression Reduced airway inflammation Decreased mucus hypersecretion Expression of TH2 cytokines	(Choi et al. 2017)
	Intranasal delivery of recombinant vaccinia vector encoding TH1 cytokine IL-12 to OVA-sensitized mice	Inhibited local Type 2 cytokine production Suppressed allergy and airway hyper-reactivity mediated by boosting endogenous IFNγ expression Reversed suppression of local cell-mediated immunity	(Hogan et al. 1998)
	Intranasal delivery of cationic chitosan NP loaded with IFNγ pDNA to OVA-sensitized mice	Reduced airway hyper-responsiveness to methacholine Reduced IFNγ production by OVA-sensitized CD8^+ T-cells Decreased DC activity	(Kumar et al. 2003; Kong et al. 2008)
	Intravenous delivery of IFNγ pDNA to OVA-sensitized mice	Decrease in TH2 immune response Reduced IFNγ production by reduction in DC activity and OVA-sensitized CD8^+ T-cell activation	(Nakagome et al. 2009)
IPF	Inhaled delivery of cationic nanostructure lipid carriers loaded with prostaglandin E (PGE2) and ECM-degrading enzyme MMP3, CCL12 and hypoxia-inducible factor 1α (HIF1A) siRNA to mice model of IPF	Reduced expression and production of MMP3, CCL12 and HIF1A Decreased lung inflammation and fibrotic tissue damage Down-regulated expression of connective tissue growth factor (CTGF), TGFB, and TGFBR genes Reduced proliferation, activation and migration of fibroblasts Decreased collagen production and secretion by fibroblasts	(Garbuzenko et al. 2017)

Table 2. Gene therapy modalities used in disease models.

Gene therapy approach	Disease model	Outcome of intervention	References
mRNA	Congenital surfactant protein B (SP-B) deficiency	Decreased interaction with TLR3, TLR7, TLR8 and RIG-I Increased stability Low immunogenicity Prolonged gene expression	(Kormann et al. 2011)
mRNA	n/a	Increased systemic production of serum protein erythropoietin Increased circulating reticulocytes Increased % of circulating RBC No immunogenicity Does not interfere with potency of regulatory sequences	(Thess et al. 2015)
mRNA	n/a	Leukotriene inhibitors for lung cancer treatment increase transfection efficacy by 200%	(Patel et al. 2017)
siRNA	Hereditary transthyretin-mediated amyloidosis	Reduced production of misfolded transthyretin protein in the liver Slow progression of hereditary transthyretin-mediated amyloidosis	(Kristen et al. 2019)
siRNA	Multi-drug resistant lung, breast, colon, and ovarian cancer	Suppress genes of pump drug resistance (drug efflux) Suppress genes of non-pump resistance (anti-apoptosis) Apoptosis induction in cancer cells Increased toxicity of anti-cancer drug to cancer cells Near complete tumor regression	(Saad et al. 2008; Taratula et al. 2013)
DNA	Lung cancer	Increased anti-cancer drug toxicity Passive and active tumor-targeting ability Increased in vivo gene transfection efficiency Enhanced anti-tumor activity	(Shao et al. 2015)
ASO	Drug-sensitive and multi-drug resistance small-cell lung cancer	Suppress genes of pump drug resistance (drug efflux) Suppress genes of non-pump resistance (anti-apoptosis) Enhanced apoptosis induction in cancer cells Increased anti-cancer drug toxicity Significant tumor regression	(Garbuzenko et al. 2010; Pakunlu et al. 2004)

Table 3. Effect of physicochemical properties of NP on the immune system.

Physicochemical property	Effects	References
Size	Dictates endocytic uptake route and subcellular localization Smaller NP have higher surface-to-volume ratios and higher charge densities therefore induce stronger pro-inflammatory responses, potentially more cytotoxic Larger NP are retained in lung, likely to be internalized by macrophages Smaller NP can enter the systemic circulation through the lung, likely to be internalized by DC	(Dobrovolskaia et al. 2016; Hoshyar et al. 2016; Liu et al. 2017; Muhammad et al. 2020; Thorp et al. 2020)
Shape and stiffness	Spherical shapes are more rapidly and efficiently taken up by macrophages than non-spherical shapes (fibre, worm-like, rod, cylinder) Rigid NP cause damage to cell membrane during internalization	(Champion and Mitragotri 2009; Patel et al. 2015; Thorp et al. 2020)
Charge	Positive surface charges enhance internalization by macrophages and DC due to electrostatic interaction with negatively-charged cell membrane High surface charge densities, either too negatively- or positively-charged, enhance cellular uptake	(Patel et al. 2015; Dobrovolskaia et al. 2016; Liu et al. 2017; Muhammad et al. 2020; Thorp et al. 2020)
Hydrophobicity	Increased hydrophobicity enhances opsonization, cellular internalization, induces danger signal pathways, stimulates immune cells and innate and adaptive immune responses	(Patel et al. 2015; Liu et al. 2017; Muhammad et al. 2020)
Composition	Polymeric NP can carry a wide range of molecules and can control release of their load thus are potent, versatile therapeutics but are cytotoxic and often lack degradability Lipid NP are biocompatible and less cytotoxic but attract negatively-charged biomolecules forming corona, may be pro-inflammatory Peptides enhance transfection efficiency but are unstable in vivo due to susceptibility to protease degradation hence premature release of payload	(Chen et al. 2017; Thorp et al. 2020)

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