Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads

Jayesh A. Kulkarni (Corresponding author), Dominik Witzigmann, Jerry Leung, Roy van der Meel, Josh Zaifman, Maria M. Darjuan, Hiu Man Grisch-Chan, Beat Thöny, Yuen Yi C. Tam, Pieter R. Cullis (Corresponding author)

Onderzoeksoutput: Bijdrage aan tijdschriftTijdschriftartikelAcademicpeer review

Uittreksel

The success of Onpattro™ (patisiran) clearly demonstrates the utility of lipid nanoparticle (LNP) systems for enabling gene therapies. These systems are composed of ionizable cationic lipids, phospholipid, cholesterol, and polyethylene glycol (PEG)-lipids, and are produced through rapid-mixing of an ethanolic-lipid solution with an acidic aqueous solution followed by dialysis into neutralizing buffer. A detailed understanding of the mechanism of LNP formation is crucial to improving LNP design. Here we use cryogenic transmission electron microscopy and fluorescence techniques to further demonstrate that LNP are formed through the fusion of precursor, pH-sensitive liposomes into large electron-dense core structures as the pH is neutralized. Next, we show that the fusion process is limited by the accumulation of PEG-lipid on the emerging particle. Finally, we show that the fusion-dependent mechanism of formation also applies to LNP containing macromolecular payloads including mRNA, DNA vectors, and gold nanoparticles.
TaalEngels
Pagina's9023-9031
Aantal pagina's9
TijdschriftNanoscale
Volume11
Nummer van het tijdschrift18
Vroegere onlinedatum22 apr 2019
DOI's
StatusGepubliceerd - 14 mei 2019

Vingerafdruk

Lipids
Fusion reactions
Nanoparticles
Polyethylene glycols
Gene therapy
Dialysis
Liposomes
Cholesterol
Dialysis Solutions
Phospholipids
Gold
Cryogenics
Buffers
DNA
Fluorescence
Transmission electron microscopy
Messenger RNA
Electrons

Citeer dit

Kulkarni, J. A., Witzigmann, D., Leung, J., van der Meel, R., Zaifman, J., Darjuan, M. M., ... Cullis, P. R. (2019). Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads. Nanoscale, 11(18), 9023-9031. DOI: 10.1039/C9NR02004G
Kulkarni, Jayesh A. ; Witzigmann, Dominik ; Leung, Jerry ; van der Meel, Roy ; Zaifman, Josh ; Darjuan, Maria M. ; Grisch-Chan, Hiu Man ; Thöny, Beat ; Tam, Yuen Yi C. ; Cullis, Pieter R./ Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads. In: Nanoscale. 2019 ; Vol. 11, Nr. 18. blz. 9023-9031
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abstract = "The success of Onpattro™ (patisiran) clearly demonstrates the utility of lipid nanoparticle (LNP) systems for enabling gene therapies. These systems are composed of ionizable cationic lipids, phospholipid, cholesterol, and polyethylene glycol (PEG)-lipids, and are produced through rapid-mixing of an ethanolic-lipid solution with an acidic aqueous solution followed by dialysis into neutralizing buffer. A detailed understanding of the mechanism of LNP formation is crucial to improving LNP design. Here we use cryogenic transmission electron microscopy and fluorescence techniques to further demonstrate that LNP are formed through the fusion of precursor, pH-sensitive liposomes into large electron-dense core structures as the pH is neutralized. Next, we show that the fusion process is limited by the accumulation of PEG-lipid on the emerging particle. Finally, we show that the fusion-dependent mechanism of formation also applies to LNP containing macromolecular payloads including mRNA, DNA vectors, and gold nanoparticles.",
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Kulkarni, JA, Witzigmann, D, Leung, J, van der Meel, R, Zaifman, J, Darjuan, MM, Grisch-Chan, HM, Thöny, B, Tam, YYC & Cullis, PR 2019, 'Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads' Nanoscale, vol. 11, nr. 18, blz. 9023-9031. DOI: 10.1039/C9NR02004G

Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads. / Kulkarni, Jayesh A. (Corresponding author); Witzigmann, Dominik; Leung, Jerry; van der Meel, Roy; Zaifman, Josh; Darjuan, Maria M.; Grisch-Chan, Hiu Man; Thöny, Beat; Tam, Yuen Yi C.; Cullis, Pieter R. (Corresponding author).

In: Nanoscale, Vol. 11, Nr. 18, 14.05.2019, blz. 9023-9031.

Onderzoeksoutput: Bijdrage aan tijdschriftTijdschriftartikelAcademicpeer review

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Kulkarni JA, Witzigmann D, Leung J, van der Meel R, Zaifman J, Darjuan MM et al. Fusion-dependent formation of lipid nanoparticles containing macromolecular payloads. Nanoscale. 2019 mei 14;11(18):9023-9031. Beschikbaar vanaf, DOI: 10.1039/C9NR02004G