Strategies for non-viral vectors targeting organs beyond the liver – Nature Nanotechnology

Zhang, Y.-N., Poon, W., Tavares, A. J., McGilvray, I. D. & Chan, W. C. W. Nanoparticle–liver interactions: cellular uptake and hepatobiliary elimination. J. Control. Release 240, 332–348 (2016).

CAS  Google Scholar 

Akinc, A. et al. The Onpattro story and the clinical translation of nanomedicines containing nucleic acid-based drugs. Nat. Nanotechnol. 14, 1084–1087 (2019).

Article  CAS  Google Scholar 

Gillmore, J. D. et al. CRISPR–Cas9 in vivo gene editing for transthyretin amyloidosis. N. Engl. J. Med. 385, 493–502 (2021).

Article  CAS  Google Scholar 

Rotolo, L. et al. Species-agnostic polymeric formulations for inhalable messenger RNA delivery to the lung. Nat. Mater. 22, 369–379 (2023).

Article  CAS  Google Scholar 

Zhong, R. et al. Hydrogels for RNA delivery. Nat. Mater. 22, 818–831 (2023).

Article  CAS  Google Scholar 

Van Haasteren, J. et al. The delivery challenge: fulfilling the promise of therapeutic genome editing. Nat. Biotechnol. 38, 845–855 (2020).

Article  Google Scholar 

Poon, W., Kingston, B. R., Ouyang, B., Ngo, W. & Chan, W. C. W. A framework for designing delivery systems. Nat. Nanotechnol. 15, 819–829 (2020). This Review thoroughly discusses the characteristics of NPs required for effective delivery within a biological context.

Article  CAS  Google Scholar 

Patel, S. et al. Brief update on endocytosis of nanomedicines. Adv. Drug Deliv. Rev. 144, 90–111 (2019).

Article  CAS  Google Scholar 

Alameh, M.-G. et al. Lipid nanoparticles enhance the efficacy of mRNA and protein subunit vaccines by inducing robust T follicular helper cell and humoral responses. Immunity 54, 2877–2892.e7 (2021).

Article  CAS  Google Scholar 

Han, X. et al. Adjuvant lipidoid-substituted lipid nanoparticles augment the immunogenicity of SARS-CoV-2 mRNA vaccines. Nat. Nanotechnol. 18, 1105–1114 (2023).

Article  CAS  Google Scholar 

Tsoi, K. M. et al. Mechanism of hard-nanomaterial clearance by the liver. Nat. Mater. 15, 1212–1221 (2016).

Article  CAS  Google Scholar 

Klibanov, A. L., Maruyama, K., Torchilin, V. P. & Huang, L. Amphipathic polyethyleneglycols effectively prolong the circulation time of liposomes. FEBS Lett. 268, 235–237 (1990).

Article  CAS  Google Scholar 

Witzigmann, D. et al. Lipid nanoparticle technology for therapeutic gene regulation in the liver. Adv. Drug Deliv. Rev. 159, 344–363 (2020).

Article  CAS  Google Scholar 

Akinc, A. et al. Targeted delivery of RNAi therapeutics with endogenous and exogenous ligand-based mechanisms. Mol. Ther. 18, 1357–1364 (2010). This study discovered that the ApoE–LDLR pathway facilitates hepatocyte transfection when LNPs contain ionizable cationic lipids but not when permanently cationic lipids are used.

Article  CAS  Google Scholar 

Nair, J. K. et al. Multivalent N-acetylgalactosamine-conjugated siRNA localizes in hepatocytes and elicits robust RNAi-mediated gene silencing. J. Am. Chem. Soc. 136, 16958–16961 (2014).

Article  CAS  Google Scholar 

Kasiewicz, L. N. et al. GalNAc–lipid nanoparticles enable non-LDLR dependent hepatic delivery of a CRISPR base editing therapy. Nat. Commun. 14, 2776 (2023).

Article  CAS  Google Scholar 

Ozelo, M. C. et al. Valoctocogene roxaparvovec gene therapy for hemophilia A. N. Engl. J. Med. 386, 1013–1025 (2022).

Article  CAS  Google Scholar 

Sato, Y. et al. Resolution of liver cirrhosis using vitamin A-coupled liposomes to deliver siRNA against a collagen-specific chaperone. Nat. Biotechnol. 26, 431–442 (2008).

Article  CAS  Google Scholar 

Lawitz, E. J. et al. BMS‐986263 in patients with advanced hepatic fibrosis: 36‐week results from a randomized, placebo‐controlled phase 2 trial. Hepatology 75, 912–923 (2022).

Article  CAS  Google Scholar 

Han, X. et al. Ligand-tethered lipid nanoparticles for targeted RNA delivery to treat liver fibrosis. Nat. Commun. 14, 75 (2023).

Article  CAS  Google Scholar 

Paunovska, K. et al. Nanoparticles containing oxidized cholesterol deliver mrna to the liver microenvironment at clinically relevant doses. Adv. Mater. 31, 1807748 (2019).

Article  Google Scholar 

Eygeris, Y., Gupta, M., Kim, J. & Sahay, G. Chemistry of lipid nanoparticles for RNA delivery. Acc. Chem. Res. 55, 2–12 (2022).

Article  CAS  Google Scholar 

Zhang, Y., Sun, C., Wang, C., Jankovic, K. E. & Dong, Y. Lipids and lipid derivatives for RNA delivery. Chem. Rev. 121, 12181–12277 (2021).

Article  CAS  Google Scholar 

Viger-Gravel, J. et al. Structure of lipid nanoparticles containing sirna or mrna by dynamic nuclear polarization-enhanced NMR spectroscopy. J. Phys. Chem. B 122, 2073–2081 (2018).

Article  CAS  Google Scholar 

Goula, D. et al. Polyethylenimine-based intravenous delivery of transgenes to mouse lung. Gene Ther. 5, 1291–1295 (1998).

CAS  Google Scholar 

Green, J. J., Langer, R. & Anderson, D. G. A combinatorial polymer library approach yields insight into nonviral gene delivery. Acc. Chem. Res. 41, 749–759 (2008).

Article  CAS  Google Scholar 

Joubert, F. et al. Precise and systematic end group chemistry modifications on PAMAM and poly(l-lysine) dendrimers to improve cytosolic delivery of mRNA. J. Control. Release 356, 580–594 (2023).

CAS  Google Scholar 

Yang, W., Mixich, L., Boonstra, E. & Cabral, H. Polymer-based mRNA delivery strategies for advanced therapies. Adv. Healthc. Mater. 12, 2202688 (2023).

Article  CAS  Google Scholar 

Cabral, H., Miyata, K., Osada, K. & Kataoka, K. Block copolymer micelles in nanomedicine applications. Chem. Rev. 118, 6844–6892 (2018).

Article  CAS  Google Scholar 

He, D. & Wagner, E. Defined polymeric materials for gene delivery. Macromol. Biosci. 15, 600–612 (2015).

Article  CAS  Google Scholar 

Reinhard, S. & Wagner, E. How to tackle the challenge of siRNA delivery with sequence-defined oligoamino amides. Macromol. Biosci. 17, 1600152 (2017).

Article  Google Scholar 

DeSimone, J. M. Co-opting Moore’s law: therapeutics, vaccines and interfacially active particles manufactured via PRINT®. J. Control. Release 240, 541–543 (2016).

CAS  Google Scholar 

Patel, A. K. et al. Inhaled nanoformulated mRNA polyplexes for protein production in lung epithelium. Adv. Mater. 31, 1805116 (2019). This study explored the application of polymeric NPs for inhaled mRNA delivery, highlighting the potential advantage of polymers for nebulization through their self-assembly.

Article  Google Scholar 

Kalra, H. et al. Vesiclepedia: a compendium for extracellular vesicles with continuous community annotation. PLoS Biol. 10, e1001450 (2012).

Article  CAS  Google Scholar 

Wahlgren, J. et al. Plasma exosomes can deliver exogenous short interfering RNA to monocytes and lymphocytes. Nucleic Acids Res. 40, e130–e130 (2012).

Article  CAS  Google Scholar 

Alvarez-Erviti, L. et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 29, 341–345 (2011).

Article  CAS  Google Scholar 

Ståhl, A. et al. A novel mechanism of bacterial toxin transfer within host blood cell-derived microvesicles. PLoS Pathog. 11, e1004619 (2015).

Article  Google Scholar 

Melamed, J. R. et al. Ionizable lipid nanoparticles deliver mRNA to pancreatic β cells via macrophage-mediated gene transfer. Sci. Adv. 9, eade1444 (2023).

Article  CAS  Google Scholar 

Wang, Q. et al. ARMMs as a versatile platform for intracellular delivery of macromolecules. Nat. Commun. 9, 960 (2018).

Article  Google Scholar 

Segel, M. et al. Mammalian retrovirus-like protein PEG10 packages its own mRNA and can be pseudotyped for mRNA delivery. Science 373, 882–889 (2021).

Article  CAS  Google Scholar 

Elsharkasy, O. M. et al. Extracellular vesicles as drug delivery systems: why and how? Adv. Drug Deliv. Rev. 159, 332–343 (2020).

Article  CAS  Google Scholar 

Klein, D. et al. Centyrin ligands for extrahepatic delivery of siRNA. Mol. Ther. 29, 2053–2066 (2021).

Article  CAS  Google Scholar 

Brown, K. M. et al. Expanding RNAi therapeutics to extrahepatic tissues with lipophilic conjugates. Nat. Biotechnol. 40, 1500–1508 (2022).

Article  CAS  Google Scholar 

Wels, M., Roels, D., Raemdonck, K., De Smedt, S. C. & Sauvage, F. Challenges and strategies for the delivery of biologics to the cornea. J. Control. Release 333, 560–578 (2021).

CAS  Google Scholar 

Baran-Rachwalska, P. et al. Topical siRNA delivery to the cornea and anterior eye by hybrid silicon-lipid nanoparticles. J. Control. Release 326, 192–202 (2020).

CAS  Google Scholar 

Bogaert, B. et al. A lipid nanoparticle platform for mRNA delivery through repurposing of cationic amphiphilic drugs. J. Control. Release 350, 256–270 (2022).

CAS  Google Scholar 

Kim, H. M. & Woo, S. J. Ocular drug delivery to the retina: current innovations and future perspectives. Pharmaceutics 13, 108 (2021).

Article  CAS  Google Scholar 

Yiu, G. et al. Suprachoroidal and subretinal injections of AAV using transscleral microneedles for retinal gene delivery in nonhuman primates. Mol. Ther. Methods Clin. Dev. 16, 179–191 (2020).

Article  CAS  Google Scholar 

Weng, C. Y. Bilateral subretinal voretigene neparvovec-rzyl (Luxturna) gene therapy. Ophthalmol. Retin. 3, 450 (2019).

Article  Google Scholar 

Jaskolka, M. C. et al. Exploratory safety profile of EDIT-101, a first-in-human in vivo CRISPR gene editing therapy for CEP290-related retinal degeneration. Invest. Ophthalmol. Vis. Sci. 63, 2836–A0352 (2022).

Google Scholar 

Chirco, K. R., Martinez, C. & Lamba, D. A. Advancements in pre-clinical development of gene editing-based therapies to treat inherited retinal diseases. Vis. Res. 209, 108257 (2023).

Article  Google Scholar 

Leroy, B. P. et al. Efficacy and safety of sepofarsen, an intravitreal RNA antisense oligonucleotide, for the treatment of CEP290-associated Leber congenital amaurosis (LCA10): a randomized, double-masked, sham-controlled, phase 3 study (ILLUMINATE). Invest. Ophthalmol. Vis. Sci. 63, 4536-F0323 (2022).

Google Scholar 

Ammar, M. J., Hsu, J., Chiang, A., Ho, A. C. & Regillo, C. D. Age-related macular degeneration therapy: a review. Curr. Opin. Ophthalmol. 31, 215–221 (2020).

Article  Google Scholar 

Goldberg, R. et al. Efficacy of intravitreal pegcetacoplan in patients with geographic atrophy (GA): 12-month results from the phase 3 OAKS and DERBY studies. Invest. Ophthalmol. Vis. Sci. 63, 1500–1500 (2022).

Google Scholar 

Shen, J. et al. Suprachoroidal gene transfer with nonviral nanoparticles. Sci. Adv. 6, eaba1606 (2020).

Article  CAS  Google Scholar 

Tan, G. et al. A core-shell nanoplatform as a nonviral vector for targeted delivery of genes to the retina. Acta Biomater. 134, 605–620 (2021).

Article  CAS  Google Scholar 

Jin, J. et al. Anti-inflammatory and antiangiogenic effects of nanoparticle-mediated delivery of a natural angiogenic inhibitor. Investig. Opthalmol. Vis. Sci. 52, 6230 (2011).

CAS  Google Scholar 

Keenan, T. D. L., Cukras, C. A. & Chew, E. Y. Age-related macular degeneration: epidemiology and clinical aspects. Adv. Exp. Med. Biol. 1256, 1–31 (2021).

Article  CAS  Google Scholar 

Chen, G. et al. A biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing. Nat. Nanotechnol. 14, 974–980 (2019).

Article  CAS  Google Scholar 

Mirjalili Mohanna, S. Z. et al. LNP-mediated delivery of CRISPR RNP for wide-spread in vivo genome editing in mouse cornea. J. Control. Release 350, 401–413 (2022).

CAS  Google Scholar 

Patel, S., Ryals, R. C., Weller, K. K., Pennesi, M. E. & Sahay, G. Lipid nanoparticles for delivery of messenger RNA to the back of the eye. J. Control. Release 303, 91–100 (2019).

CAS  Google Scholar 

Sun, D. et al. Non-viral gene therapy for stargardt disease with ECO/pRHO-ABCA4 self-assembled nanoparticles. Mol. Ther. 28, 293–303 (2020).

Article  CAS  Google Scholar 

Herrera-Barrera, M. et al. Peptide-guided lipid nanoparticles deliver mRNA to the neural retina of rodents and nonhuman primates. Sci. Adv. 9, eadd4623 (2023).

Article  Google Scholar 

Huertas, A. et al. Pulmonary vascular endothelium: the orchestra conductor in respiratory diseases: highlights from basic research to therapy. Eur. Respir. J. 51, 1700745 (2018).

Article  Google Scholar 

Hong, K.-H. et al. Genetic ablation of the Bmpr2 gene in pulmonary endothelium is sufficient to predispose to pulmonary arterial hypertension. Circulation 118, 722–730 (2008).

Article  CAS  Google Scholar 

Dahlman, J. E. et al. In vivo endothelial siRNA delivery using polymeric nanoparticles with low molecular weight. Nat. Nanotechnol. 9, 648–655 (2014).

Article  CAS  Google Scholar 

Cheng, Q. et al. Selective organ targeting (SORT) nanoparticles for tissue-specific mRNA delivery and CRISPR–Cas gene editing. Nat. Nanotechnol. 15, 313–320 (2020). This groundbreaking study found that incorporating differently charged (SORT) lipids into the conventional four-component LNPs shifts the location of mRNA transfection among the liver, spleen and lungs.

Article  CAS  Google Scholar 

Dilliard, S. A., Cheng, Q. & Siegwart, D. J. On the mechanism of tissue-specific mRNA delivery by selective organ targeting nanoparticles. Proc. Natl Acad. Sci. USA 118, e2109256118 (2021). This work thoroughly investigated the impact of SORT lipids added to LNPs on the formation of the biomolecular corona on the NP surface and its role in achieving organ-specific transfection.

Article  CAS  Google Scholar 

Kimura, S. & Harashima, H. On the mechanism of tissue-selective gene delivery by lipid nanoparticles. J. Control. Release https://doi.org/10.1016/j.jconrel.2023.03.052 (2023).

Qiu, M. et al. Lung-selective mRNA delivery of synthetic lipid nanoparticles for the treatment of pulmonary lymphangioleiomyomatosis. Proc. Natl Acad. Sci. USA 119, e2116271119 (2022).

Article  CAS  Google Scholar 

Kaczmarek, J. C. et al. Polymer–lipid nanoparticles for systemic delivery of mRNA to the lungs. Angew. Chem. Int. Ed. 55, 13808–13812 (2016).

Article  CAS  Google Scholar 

Shen, A. M. & Minko, T. Pharmacokinetics of inhaled nanotherapeutics for pulmonary delivery. J. Control. Release 326, 222–244 (2020).

CAS  Google Scholar 

Alton, E. W. F. W. et al. Repeated nebulisation of non-viral CFTR gene therapy in patients with cystic fibrosis: a randomised, double-blind, placebo-controlled, phase 2b trial. Lancet Respir. Med. 3, 684–691 (2015).

Article  CAS  Google Scholar 

Kim, J. et al. Engineering lipid nanoparticles for enhanced intracellular delivery of mRNA through inhalation. ACS Nano 16, 14792–14806 (2022).

CAS  Google Scholar 

Lokugamage, M. P. et al. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Nat. Biomed. Eng. 5, 1059–1068 (2021).

Article  CAS  Google Scholar 

Qiu, Y. et al. Effective mRNA pulmonary delivery by dry powder formulation of PEGylated synthetic KL4 peptide. J. Control. Release 314, 102–115 (2019).

CAS  Google Scholar 

Popowski, K. D. et al. Inhalable dry powder mRNA vaccines based on extracellular vesicles. Matter 5, 2960–2974 (2022).

Article  CAS  Google Scholar 

Telko, M. J. & Hickey, A. J. Dry powder inhaler formulation. Respir. Care 50, 1209 (2005).

Google Scholar 

Li, B. et al. Combinatorial design of nanoparticles for pulmonary mRNA delivery and genome editing. Nat. Biotechnol. https://doi.org/10.1038/s41587-023-01679-x (2023).

Fahy, J. V. & Dickey, B. F. Airway mucus function and dysfunction. N. Engl. J. Med. 363, 2233–2247 (2010).

Article  CAS  Google Scholar 

Schneider, C. S. et al. Nanoparticles that do not adhere to mucus provide uniform and long-lasting drug delivery to airways following inhalation. Sci. Adv. 3, e1601556 (2017).

Article  Google Scholar 

Wang, J. et al. Pulmonary surfactant–biomimetic nanoparticles potentiate heterosubtypic influenza immunity. Science 367, eaau0810 (2020).

Article  CAS  Google Scholar 

Rock, J. R., Randell, S. H. & Hogan, B. L. M. Airway basal stem cells: a perspective on their roles in epithelial homeostasis and remodeling. Dis. Model. Mech. 3, 545–556 (2010).

Article  CAS  Google Scholar 

Getts, D. R. et al. Microparticles bearing encephalitogenic peptides induce T-cell tolerance and ameliorate experimental autoimmune encephalomyelitis. Nat. Biotechnol. 30, 1217–1224 (2012).

Article  CAS  Google Scholar 

Leuschner, F. et al. Therapeutic siRNA silencing in inflammatory monocytes in mice. Nat. Biotechnol. 29, 1005–1010 (2011).

Article  CAS  Google Scholar 

Rojas, L. A. et al. Personalized RNA neoantigen vaccines stimulate T cells in pancreatic cancer. Nature 618, 144–150 (2023).

Article  CAS  Google Scholar 

Bevers, S. et al. mRNA–LNP vaccines tuned for systemic immunization induce strong antitumor immunity by engaging splenic immune cells. Mol. Ther. 30, 3078–3094 (2022).

Article  CAS  Google Scholar 

Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941–951 (2015).

Article  CAS  Google Scholar 

Kranz, L. M. et al. Systemic RNA delivery to dendritic cells exploits antiviral defence for cancer immunotherapy. Nature 534, 396–401 (2016).

Article  Google Scholar 

Liu, S. et al. Membrane-destabilizing ionizable phospholipids for organ-selective mRNA delivery and CRISPR–Cas gene editing. Nat. Mater. 20, 701–710 (2021).

Article  CAS  Google Scholar 

Fenton, O. S. et al. Synthesis and biological evaluation of ionizable lipid materials for the in vivo delivery of messenger RNA to B lymphocytes. Adv. Mater. 29, 1606944 (2017).

Article  Google Scholar 

Zhao, X. et al. Imidazole‐based synthetic lipidoids for in vivo mRNA delivery into primary T lymphocytes. Angew. Chem. Int. Ed. 59, 20083–20089 (2020).

Article  CAS  Google Scholar 

LoPresti, S. T., Arral, M. L., Chaudhary, N. & Whitehead, K. A. The replacement of helper lipids with charged alternatives in lipid nanoparticles facilitates targeted mRNA delivery to the spleen and lungs. J. Control. Release 345, 819–831 (2022).

CAS  Google Scholar 

McKinlay, C. J., Benner, N. L., Haabeth, O. A., Waymouth, R. M. & Wender, P. A. Enhanced mRNA delivery into lymphocytes enabled by lipid-varied libraries of charge-altering releasable transporters. Proc. Natl Acad. Sci. USA 115, E5859–E5866 (2018).

Article  Google Scholar 

McKinlay, C. J. et al. Charge-altering releasable transporters (CARTs) for the delivery and release of mRNA in living animals. Proc. Natl Acad. Sci. USA 114, E448–E456 (2017).

Article  CAS  Google Scholar 

Ben-Akiva, E. et al. Biodegradable lipophilic polymeric mRNA nanoparticles for ligand-free targeting of splenic dendritic cells for cancer vaccination. Proc. Natl Acad. Sci. USA 120, e2301606120 (2023).

Article  Google Scholar 

Tombácz, I. et al. Highly efficient CD4⁺ T cell targeting and genetic recombination using engineered CD4+ cell-homing mRNA–LNPs. Mol. Ther. 29, 3293–3304 (2021).

Article  Google Scholar 

Rurik, J. G. et al. CAR T cells produced in vivo to treat cardiac injury. Science 375, 91–96 (2022).

Article  CAS  Google Scholar 

Kim, J., Eygeris, Y., Gupta, M. & Sahay, G. Self-assembled mRNA vaccines. Adv. Drug Deliv. Rev. 170, 83–112 (2021).

Article  CAS  Google Scholar 

Lindsay, K. E. et al. Visualization of early events in mRNA vaccine delivery in non-human primates via PET–CT and near-infrared imaging. Nat. Biomed. Eng. 3, 371–380 (2019). This pioneering study delved into the biodistribution of lipid-based mRNA vaccines after their intramuscular injection into non-human primates using a dual radionuclide–near-infrared probe.

Article  CAS  Google Scholar 

Alberer, M. et al. Safety and immunogenicity of a mRNA rabies vaccine in healthy adults: an open-label, non-randomised, prospective, first-in-human phase 1 clinical trial. Lancet 390, 1511–1520 (2017).

Article  CAS  Google Scholar 

Assessment Report: Comirnaty EMA/707383/2020 (European Medicines Agency, 2021); https://www.ema.europa.eu/en/documents/assessment-report/comirnaty-epar-public-assessment-report_en.pdf

Assessment Report: COVID-19 Vaccine Moderna EMA/15689/2021 (European Medicines Agency, 2021); https://www.ema.europa.eu/en/documents/assessment-report/spikevax-previously-covid-19-vaccine-moderna-epar-public-assessment-report_en.pdf

Ke, X. et al. Physical and chemical profiles of nanoparticles for lymphatic targeting. Adv. Drug Deliv. Rev. 151–152, 72–93 (2019).

Article  Google Scholar 

Hansen, K. C., D’Alessandro, A., Clement, C. C. & Santambrogio, L. Lymph formation, composition and circulation: a proteomics perspective. Int. Immunol. 27, 219–227 (2015).

Article  CAS  Google Scholar 

Chen, J. et al. Lipid nanoparticle-mediated lymph node-targeting delivery of mRNA cancer vaccine elicits robust CD8⁺ T cell response. Proc. Natl Acad. Sci. USA 119, e2207841119 (2022).

Article  CAS  Google Scholar 

Liu, S. et al. Zwitterionic phospholipidation of cationic polymers facilitates systemic mRNA delivery to spleen and lymph nodes. J. Am. Chem. Soc. 143, 21321–21330 (2021).

Article  CAS  Google Scholar 

Sahin, U. et al. Personalized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer. Nature 547, 222–226 (2017).

Article  CAS  Google Scholar 

Kreiter, S. et al. Intranodal vaccination with naked antigen-encoding rna elicits potent prophylactic and therapeutic antitumoral immunity. Cancer Res. 70, 9031–9040 (2010).

Article  CAS  Google Scholar 

Fan, C.-H. et al. Folate-conjugated gene-carrying microbubbles with focused ultrasound for concurrent blood–brain barrier opening and local gene delivery. Biomaterials 106, 46–57 (2016).

Article  CAS  Google Scholar 

Yu, Y. J. et al. Boosting brain uptake of a therapeutic antibody by reducing its affinity for a transcytosis target. Sci. Transl. Med. 3, 84ra44 (2011).

Article  Google Scholar 

Yu, Y. J. et al. Therapeutic bispecific antibodies cross the blood–brain barrier in nonhuman primates. Sci. Transl. Med. 6, 261ra154 (2014).

Article  Google Scholar 

Kariolis, M. S. et al. Brain delivery of therapeutic proteins using an Fc fragment blood–brain barrier transport vehicle in mice and monkeys. Sci. Transl. Med. 12, eaay1359 (2020).

Article  CAS  Google Scholar 

Ullman, J. C. et al. Brain delivery and activity of a lysosomal enzyme using a blood–brain barrier transport vehicle in mice. Sci. Transl. Med. 12, eaay1163 (2020).

Article  CAS  Google Scholar 

Ma, F. et al. Neurotransmitter-derived lipidoids (NT-lipidoids) for enhanced brain delivery through intravenous injection. Sci. Adv. 6, eabb4429 (2020). This study suggests that designing lipids to mimic neurotransmitters and incorporating them into NPs can enhance the delivery of nucleic acids and proteins to the brain following IV injection.

Article  CAS  Google Scholar 

Zhou, Y. et al. Blood–brain barrier-penetrating siRNA nanomedicine for Alzheimer’s disease therapy. Sci. Adv. 6, eabc7031 (2020).

Article  CAS  Google Scholar 

Li, W. et al. BBB pathophysiology-independent delivery of siRNA in traumatic brain injury. Sci. Adv. 7, eabd6889 (2021).

Article  CAS  Google Scholar 

Nance, E. A. et al. A dense poly(ethylene glycol) coating improves penetration of large polymeric nanoparticles within brain tissue. Sci. Transl. Med. 4, 149ra119 (2012).

Article  Google Scholar 

Thorne, R. G. & Nicholson, C. In vivo diffusion analysis with quantum dots and dextrans predicts the width of brain extracellular space. Proc. Natl Acad. Sci. USA 103, 5567–5572 (2006).

Article  CAS  Google Scholar 

Kim, M. et al. Delivery of self-replicating messenger RNA into the brain for the treatment of ischemic stroke. J. Control. Release 350, 471–485 (2022).

CAS  Google Scholar 

Willerth, S. M. & Sakiyama-Elbert, S. E. Approaches to neural tissue engineering using scaffolds for drug delivery. Adv. Drug Deliv. Rev. 59, 325–338 (2007).

Article  CAS  Google Scholar 

Saucier-Sawyer, J. K. et al. Distribution of polymer nanoparticles by convection-enhanced delivery to brain tumors. J. Control. Release 232, 103–112 (2016).

Article  CAS  Google Scholar 

Dhaliwal, H. K., Fan, Y., Kim, J. & Amiji, M. M. Intranasal delivery and transfection of mRNA therapeutics in the brain using cationic liposomes. Mol. Pharm. 17, 1996–2005 (2020).

Article  CAS  Google Scholar 

Frangoul, H. et al. CRISPR–Cas9 gene editing for sickle cell disease and β-thalassemia. N. Engl. J. Med. 384, 252–260 (2021).

Article  CAS  Google Scholar 

Hirabayashi, H. & Fujisaki, J. Bone-specific drug delivery systems: approaches via chemical modification of bone-seeking agents. Clin. Pharmacokinet. 42, 1319–1330 (2003).

Article  CAS  Google Scholar 

Wang, G., Mostafa, N. Z., Incani, V., Kucharski, C. & Uludağ, H. Bisphosphonate-decorated lipid nanoparticles designed as drug carriers for bone diseases. J. Biomed. Mater. Res. A 100, 684–693 (2012).

Article  Google Scholar 

Giger, E. V. et al. Gene delivery with bisphosphonate-stabilized calcium phosphate nanoparticles. J. Control. Release 150, 87–93 (2011).

CAS  Google Scholar 

Xue, L. et al. Rational design of bisphosphonate lipid-like materials for mRNA delivery to the bone microenvironment. J. Am. Chem. Soc. 144, 9926–9937 (2022). This study proposes that improving lipid design to mimic bisphosphates can improve LNP-mediated mRNA delivery to the bone microenvironment after IV injection.

Article  CAS  Google Scholar 

Liang, C. et al. Aptamer-functionalized lipid nanoparticles targeting osteoblasts as a novel RNA interference-based bone anabolic strategy. Nat. Med. 21, 288–294 (2015).

Article  Google Scholar 

Zhang, Y., Wei, L., Miron, R. J., Shi, B. & Bian, Z. Anabolic bone formation via a site-specific bone-targeting delivery system by interfering with semaphorin 4D expression. J. Bone Miner. Res. 30, 286–296 (2015).

Article  CAS  Google Scholar 

Zhang, G. et al. A delivery system targeting bone formation surfaces to facilitate RNAi-based anabolic therapy. Nat. Med. 18, 307–314 (2012).

Article  Google Scholar 

Shi, D., Toyonaga, S. & Anderson, D. G. In vivo RNA delivery to hematopoietic stem and progenitor cells via targeted lipid nanoparticles. Nano Lett. 23, 2938–2944 (2023).

CAS  Google Scholar 

Sago, C. D. et al. Nanoparticles that deliver RNA to bone marrow identified by in vivo directed evolution. J. Am. Chem. Soc. 140, 17095–17105 (2018).

Article  CAS  Google Scholar 

Zhang, X., Li, Y., Chen, Y. E., Chen, J. & Ma, P. X. Cell-free 3D scaffold with two-stage delivery of miRNA-26a to regenerate critical-sized bone defects. Nat. Commun. 7, 10376 (2016).

Article  CAS  Google Scholar 

Wang, P. et al. In vivo bone tissue induction by freeze-dried collagen–nanohydroxyapatite matrix loaded with BMP2/NS1 mRNAs lipopolyplexes. J. Control. Release 334, 188–200 (2021).

CAS  Google Scholar 

Athirasala, A. et al. Matrix stiffness regulates lipid nanoparticle-mRNA delivery in cell-laden hydrogels. Nanomed. Nanotechnol. Biol. Med. 42, 102550 (2022).

Article  CAS  Google Scholar 

Nims, R. J., Pferdehirt, L. & Guilak, F. Mechanogenetics: harnessing mechanobiology for cellular engineering. Curr. Opin. Biotechnol. 73, 374–379 (2022).

Article  CAS  Google Scholar 

O’Driscoll, C. M., Bernkop-Schnürch, A., Friedl, J. D., Préat, V. & Jannin, V. Oral delivery of non-viral nucleic acid-based therapeutics—do we have the guts for this? Eur. J. Pharm. Sci. 133, 190–204 (2019).

Article  Google Scholar 

Ball, R. L., Bajaj, P. & Whitehead, K. A. Oral delivery of siRNA lipid nanoparticles: fate in the GI tract. Sci. Rep. 8, 2178 (2018).

Article  Google Scholar 

Attarwala, H., Han, M., Kim, J. & Amiji, M. Oral nucleic acid therapy using multi-compartmental delivery systems. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 10, e1478 (2018).

Article  Google Scholar 

Abramson, A. et al. An ingestible self-orienting system for oral delivery of macromolecules. Science 363, 611–615 (2019).

Article  CAS  Google Scholar 

Abramson, A. et al. Oral mRNA delivery using capsule-mediated gastrointestinal tissue injections. Matter 5, 975–987 (2022). This study shows the potential for delivery of mRNA-loaded PBAE NPs directly to the submucosa of the stomach using orally ingested robotic pills.

Article  CAS  Google Scholar 

Doll, S. et al. Region and cell-type resolved quantitative proteomic map of the human heart. Nat. Commun. 8, 1469 (2017).

Article  Google Scholar 

Xin, M., Olson, E. N. & Bassel-Duby, R. Mending broken hearts: cardiac development as a basis for adult heart regeneration and repair. Nat. Rev. Mol. Cell Biol. 14, 529–541 (2013).

Article  CAS  Google Scholar 

Zangi, L. et al. Modified mRNA directs the fate of heart progenitor cells and induces vascular regeneration after myocardial infarction. Nat. Biotechnol. 31, 898–907 (2013).

Article  CAS  Google Scholar 

Tang, R., Long, T., Lui, K. O., Chen, Y. & Huang, Z.-P. A roadmap for fixing the heart: RNA regulatory networks in cardiac disease. Mol. Ther. Nucleic Acids 20, 673–686 (2020).

Article  CAS  Google Scholar 

Han, P. et al. A long noncoding RNA protects the heart from pathological hypertrophy. Nature 514, 102–106 (2014).

Article  CAS  Google Scholar 

Anttila, V. et al. Direct intramyocardial injection of VEGF mRNA in patients undergoing coronary artery bypass grafting. Mol. Ther. 31, 866–874 (2023).

Article  CAS  Google Scholar 

Täubel, J. et al. Novel antisense therapy targeting microRNA-132 in patients with heart failure: results of a first-in-human phase 1b randomized, double-blind, placebo-controlled study. Eur. Heart J. 42, 178–188 (2021).

Article  Google Scholar 

Nishiyama, T. et al. Precise genomic editing of pathogenic mutations in RBM20 rescues dilated cardiomyopathy. Sci. Transl. Med. 14, eade1633 (2022).

Article  CAS  Google Scholar 

Reichart, D. et al. Efficient in vivo genome editing prevents hypertrophic cardiomyopathy in mice. Nat. Med. 29, 412–421 (2023).

Article  CAS  Google Scholar 

Chai, A. C. et al. Base editing correction of hypertrophic cardiomyopathy in human cardiomyocytes and humanized mice. Nat. Med. 29, 401–411 (2023).

Article  CAS  Google Scholar 

Rubin, J. D. & Barry, M. A. Improving molecular therapy in the kidney. Mol. Diagn. Ther. 24, 375–396 (2020).

Article  Google Scholar 

Oroojalian, F. et al. Recent advances in nanotechnology-based drug delivery systems for the kidney. J. Control. Release 321, 442–462 (2020).

CAS  Google Scholar 

Jiang, D. et al. DNA origami nanostructures can exhibit preferential renal uptake and alleviate acute kidney injury. Nat. Biomed. Eng. 2, 865–877 (2018).

Article  CAS  Google Scholar 

Xu, Y. et al. NIR-II photoacoustic-active DNA origami nanoantenna for early diagnosis and smart therapy of acute kidney injury. J. Am. Chem. Soc. 144, 23522–23533 (2022).

Article  CAS  Google Scholar 

Stribley, J. M., Rehman, K. S., Niu, H. & Christman, G. M. Gene therapy and reproductive medicine. Fertil. Steril. 77, 645–657 (2002).

Article  Google Scholar 

Boekelheide, K. & Sigman, M. Is gene therapy for the treatment of male infertility feasible? Nat. Clin. Pract. Urol. 5, 590–593 (2008).

Article  CAS  Google Scholar 

Rodríguez-Gascón, A., del Pozo-Rodríguez, A., Isla, A. & Solinís, M. A. Vaginal gene therapy. Adv. Drug Deliv. Rev. 92, 71–83 (2015).

Article  Google Scholar 

Lindsay, K. E. et al. Aerosol delivery of synthetic mRNA to vaginal mucosa leads to durable expression of broadly neutralizing antibodies against HIV. Mol. Ther. 28, 805–819 (2020).

Article  CAS  Google Scholar 

Poley, M. et al. Nanoparticles accumulate in the female reproductive system during ovulation affecting cancer treatment and fertility. ACS Nano 16, 5246–5257 (2022).

CAS  Google Scholar 

DeWeerdt, S. Prenatal gene therapy offers the earliest possible cure. Nature 564, S6–S8 (2018).

Article  CAS  Google Scholar 

Palanki, R., Peranteau, W. H. & Mitchell, M. J. Delivery technologies for in utero gene therapy. Adv. Drug Deliv. Rev. 169, 51–62 (2021).

Article  CAS  Google Scholar 

Riley, R. S. et al. Ionizable lipid nanoparticles for in utero mRNA delivery. Sci. Adv. 7, 1028–1041 (2021).

Article  Google Scholar 

Swingle, K. L. et al. Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery. J. Control. Release 341, 616–633 (2022).

CAS  Google Scholar 

Ricciardi, A. S. et al. In utero nanoparticle delivery for site-specific genome editing. Nat. Commun. 9, 2481 (2018). This study presents in utero gene editing of a disease-causing β-thalassemia mutation in foetal mice.

Article  Google Scholar 

Chaudhary, N. et al. Lipid nanoparticle structure and delivery route during pregnancy dictates mRNA potency, immunogenicity, and health in the mother and offspring. Preprint at bioRxiv https://doi.org/10.1101/2023.02.15.528720 (2023).

Young, R. E. et al. Lipid nanoparticle composition drives mRNA delivery to the placenta. Preprint at bioRxiv https://doi.org/10.1101/2022.12.22.521490 (2022).

Swingle, K. L. et al. Ionizable lipid nanoparticles for in vivo mRNA delivery to the placenta during pregnancy. J. Am. Chem. Soc. 145, 4691–4706 (2023).

Article  CAS  Google Scholar 

Lan, Y. et al. Recent development of AAV-based gene therapies for inner ear disorders. Gene Ther. 27, 329–337 (2020).

CAS  Google Scholar 

Delmaghani, S. & El-Amraoui, A. Inner ear gene therapies take off: current promises and future challenges. J. Clin. Med. 9, 2309 (2020).

Article  CAS  Google Scholar 

Wang, L., Kempton, J. B. & Brigande, J. V. Gene therapy in mouse models of deafness and balance dysfunction. Front. Mol. Neurosci. 11, 300 (2018).

Article  CAS  Google Scholar 

Du, X. et al. Regeneration of cochlear hair cells and hearing recovery through Hes1 modulation with siRNA nanoparticles in adult guinea pigs. Mol. Ther. 26, 1313–1326 (2018).

Article  CAS  Google Scholar 

Gao, X. et al. Treatment of autosomal dominant hearing loss by in vivo delivery of genome editing agents. Nature 553, 217–221 (2018).

Article  CAS  Google Scholar 

Jero, J. et al. Cochlear gene delivery through an intact round window membrane in mouse. Hum. Gene Ther. 12, 539–548 (2001).

CAS  Google Scholar 

Egeblad, M., Nakasone, E. S. & Werb, Z. Tumors as organs: complex tissues that interface with the entire organism. Dev. Cell 18, 884–901 (2010).

Article  CAS  Google Scholar 

El-Sawy, H. S., Al-Abd, A. M., Ahmed, T. A., El-Say, K. M. & Torchilin, V. P. Stimuli-responsive nano-architecture drug-delivery systems to solid tumor micromilieu: past, present, and future perspectives. ACS Nano 12, 10636–10664 (2018).

CAS  Google Scholar 

Hansen, A. E. et al. Positron emission tomography based elucidation of the enhanced permeability and retention effect in dogs with cancer using copper-64 liposomes. ACS Nano 9, 6985–6995 (2015).

CAS  Google Scholar 

Zhou, Q. et al. Enzyme-activatable polymer–drug conjugate augments tumour penetration and treatment efficacy. Nat. Nanotechnol. 14, 799–809 (2019).

Article  CAS  Google Scholar 

Sindhwani, S. et al. The entry of nanoparticles into solid tumours. Nat. Mater. 19, 566–575 (2020).

Article  CAS  Google Scholar 

Wilhelm, S. et al. Analysis of nanoparticle delivery to tumours. Nat. Rev. Mater. 1, 16014 (2016). This Review deeply explores the possible factors behind the ineffective tumour-targeting of NPs, uncovering that only a small fraction of the administered NP dose reaches a solid tumour.

Article  CAS  Google Scholar 

Schroeder, A. et al. Treating metastatic cancer with nanotechnology. Nat. Rev. Cancer 12, 39–50 (2012).

Article  CAS  Google Scholar 

Chan, W. C. W. Principles of nanoparticle delivery to solid tumors. BME Front. 4, 0016 (2023). This Review delineates key principles for designing tumour-targeting NPs, considering both macro- and micro-level analysis of the environment surrounding NPs and their physicochemical attributes.

Article  CAS  Google Scholar 

Kingston, B. R. et al. Specific endothelial cells govern nanoparticle entry into solid tumors. ACS Nano 15, 14080–14094 (2021).

CAS  Google Scholar 

Boehnke, N. et al. Massively parallel pooled screening reveals genomic determinants of nanoparticle delivery. Science 377, eabm5551 (2022).

Article  CAS  Google Scholar 

Li, Y. et al. Multifunctional oncolytic nanoparticles deliver self-replicating IL-12 RNA to eliminate established tumors and prime systemic immunity. Nat. Cancer 1, 882–893 (2020).

Article  CAS  Google Scholar 

Hotz, C. et al. Local delivery of mRNA-encoded cytokines promotes antitumor immunity and tumor eradication across multiple preclinical tumor models. Sci. Transl. Med. 13, eabc7804 (2021).

Article  CAS  Google Scholar 

Li, W. et al. Biomimetic nanoparticles deliver mRNAs encoding costimulatory receptors and enhance T cell mediated cancer immunotherapy. Nat. Commun. 12, 7264 (2021).

Article  CAS  Google Scholar 

Van Lint, S. et al. Intratumoral delivery of TriMix mRNA results in T-cell activation by cross-presenting dendritic cells. Cancer Immunol. Res. 4, 146–156 (2016).

Article  Google Scholar 

Oberli, M. A. et al. Lipid nanoparticle assisted mRNA delivery for potent cancer immunotherapy. Nano Lett. 17, 1326–1335 (2017).

CAS  Google Scholar 

Huayamares, S. G. et al. High-throughput screens identify a lipid nanoparticle that preferentially delivers mRNA to human tumors in vivo. J. Control. Release 357, 394–403 (2023).

CAS  Google Scholar 

Vetter, V. C. & Wagner, E. Targeting nucleic acid-based therapeutics to tumors: challenges and strategies for polyplexes. J. Control. Release 346, 110–135 (2022).

CAS  Google Scholar 

Yong, S. et al. Dual‐targeted lipid nanotherapeutic boost for chemo‐immunotherapy of cancer. Adv. Mater. 34, 2106350 (2022).

Article  CAS  Google Scholar 

Kedmi, R. et al. A modular platform for targeted RNAi therapeutics. Nat. Nanotechnol. 13, 214–219 (2018). This study developed a modular, ligand-based RNA delivery platform that avoids the chemical conjugation of antibodies by using linkers that bind to the Fc region, ensuring precise antibody orientation on the NP surface.

Article  CAS  Google Scholar 

Mitchell, M. J. et al. Engineering precision nanoparticles for drug delivery. Nat. Rev. Drug Discov. 20, 101–124 (2021).

Article  CAS  Google Scholar 

Adachi, K., Enoki, T., Kawano, Y., Veraz, M. & Nakai, H. Drawing a high-resolution functional map of adeno-associated virus capsid by massively parallel sequencing. Nat. Commun. 5, 3075 (2014).

Article  Google Scholar 

Dahlman, J. E. et al. Barcoded nanoparticles for high throughput in vivo discovery of targeted therapeutics. Proc. Natl Acad. Sci. USA 114, 2060–2065 (2017). This work presents the remarkable capabilities of DNA barcoding and deep sequencing in conducting high-throughput screening of NPs, assessing their effectiveness in target-specific gene delivery in vivo.

Article  CAS  Google Scholar 

Da Silva Sanchez, A. J. et al. Universal barcoding predicts in vivo ApoE-independent lipid nanoparticle delivery. Nano Lett. 22, 4822–4830 (2022).

Google Scholar 

Guimaraes, P. P. G. et al. Ionizable lipid nanoparticles encapsulating barcoded mRNA for accelerated in vivo delivery screening. J. Control. Release 316, 404–417 (2019).

CAS  Google Scholar 

Dobrowolski, C. et al. Nanoparticle single-cell multiomic readouts reveal that cell heterogeneity influences lipid nanoparticle-mediated messenger RNA delivery. Nat. Nanotechnol. 17, 871–879 (2022).

Article  CAS  Google Scholar 

Rhym, L. H., Manan, R. S., Koller, A., Stephanie, G. & Anderson, D. G. Peptide-encoding mRNA barcodes for the high-throughput in vivo screening of libraries of lipid nanoparticles for mRNA delivery. Nat. Biomed. Eng. 7, 901–910 (2023).

Article  CAS  Google Scholar 

Stoeckius, M. et al. Simultaneous epitope and transcriptome measurement in single cells. Nat. Methods 14, 865–868 (2017).

Article  CAS  Google Scholar 

Keenum, M. C. et al. Single-cell epitope-transcriptomics reveal lung stromal and immune cell response kinetics to nanoparticle-delivered RIG-I and TLR4 agonists. Biomaterials 297, 122097 (2023).

Article  CAS  Google Scholar 

Grandi, F. C., Modi, H., Kampman, L. & Corces, M. R. Chromatin accessibility profiling by ATAC-seq. Nat. Protoc. 17, 1518–1552 (2022).

Article  CAS  Google Scholar 

Rao, N., Clark, S. & Habern, O. Bridging genomics and tissue pathology: 10x Genomics explores new frontiers with the Visium Spatial Gene Expression Solution. Genet. Eng. Biotechnol. News 40, 50–51 (2020).

Article  Google Scholar 

Francia, V., Schiffelers, R. M., Cullis, P. R. & Witzigmann, D. The biomolecular corona of lipid nanoparticles for gene therapy. Bioconjug. Chem. 31, 2046–2059 (2020).

Article  CAS  Google Scholar 

Shao, D. et al. HBFP: a new repository for human body fluid proteome. Database 2021, baab065 (2021).

Article  CAS  Google Scholar 

Greener, J. G., Kandathil, S. M., Moffat, L. & Jones, D. T. A guide to machine learning for biologists. Nat. Rev. Mol. Cell Biol. 23, 40–55 (2022).

Article  CAS  Google Scholar 

Zhang, H. et al. Algorithm for optimized mRNA design improves stability and immunogenicity. Nature 621, 396–403 (2023).

Article  CAS  Google Scholar 

Wang, W. et al. Prediction of lipid nanoparticles for mRNA vaccines by the machine learning algorithm. Acta Pharm. Sin. B 12, 2950–2962 (2022).

Article  CAS  Google Scholar 

Xu, Y. et al. AGILE platform: a deep learning-powered approach to accelerate LNP development for mRNA delivery. Preprint at bioRxiv https://doi.org/10.1101/2023.06.01.543345 (2023). This work implements artificial intelligence in ionizable lipid design for intramuscular mRNA delivery.

Gong, D. et al. Machine learning guided structure function predictions enable in silico nanoparticle screening for polymeric gene delivery. Acta Biomater. 154, 349–358 (2022).

Article  CAS  Google Scholar 

Reker, D. et al. Computationally guided high-throughput design of self-assembling drug nanoparticles. Nat. Nanotechnol. 16, 725–733 (2021).

Article  CAS  Google Scholar 

Yamankurt, G. et al. Exploration of the nanomedicine-design space with high-throughput screening and machine learning. Nat. Biomed. Eng. 3, 318–327 (2019).

Article  CAS  Google Scholar 

Lazarovits, J. et al. Supervised learning and mass spectrometry predicts the in vivo fate of nanomaterials. ACS Nano 13, 8023–8034 (2019).

CAS  Google Scholar 

Goodfellow, I. et al. Generative adversarial networks. Commun. ACM 63, 139–144 (2020).

Article  Google Scholar 

Repecka, D. et al. Expanding functional protein sequence spaces using generative adversarial networks. Nat. Mach. Intell. 3, 324–333 (2021).

Article  Google Scholar 

De Backer, L., Cerrada, A., Pérez-Gil, J., De Smedt, S. C. & Raemdonck, K. Bio-inspired materials in drug delivery: exploring the role of pulmonary surfactant in siRNA inhalation therapy. J. Control. Release 220, 642–650 (2015).

Google Scholar