Nucleic acid delivery to retinal cells using lipopeptides as a potential tool towards ocular gene therapies

Lindomar J.C. Albuquerque; Fernando A. de Oliveira; Marcelo A. Christoffolete; Michelle Nascimento-Sales; Simone Berger; Ernst Wagner; Ulrich Lächelt; Fernando C. Giacomelli

doi:10.1016/j.jcis.2023.11.003

Nucleic acid delivery to retinal cells using lipopeptides as a potential tool towards ocular gene therapies

Lindomar J.C. Albuquerque
, Fernando A. de Oliveira
, Marcelo A. Christoffolete
, Michelle Nascimento-Sales
, Simone Berger
, Ernst Wagner
, Ulrich Lächelt
, Fernando C. Giacomelli

Department of Pharmaceutical Sciences

Publications: Contribution to journal › Article › Peer Reviewed

Abstract

We evaluated the use of lipopeptides capable to bind to nucleic acids towards plasmid DNA (pDNA) delivery. The investigations were particularly focused on arising retinal pigment epithelial cells (ARPE-19) as motivated by the considerable number of ocular disorders linked to gene aberrations. The lipopeptides comprised the artificial oligoamino acid succinyl-tetraethylene pentamine (Stp) as well as incorporated lysines, histidines, cysteines, fatty acids, and tyrosine trimers. Regardless of the structural differences, the lipopeptides demonstrated to efficiently condense pDNA at nitrogen-to-phosphate molar ratio (N/P) ≥ 6. Spheric nanoparticles were observed by cryo-TEM and dynamic light scattering determined hydrodynamic sizes ranging from 50 to 130 nm. The biological assays evidenced highly efficient pDNA delivery with a lower degree of cytotoxicity compared to the well-known transfecting agent linear polyethylenimine (LPEI). Although more efficient than LPEI, cysteine-containing carriers were demonstrated to be less efficient than the other counterparts possibly due to exceeding polyplex stabilization via disulfide cross links, which could hamper pDNA unpacking at the target site. Therefore, clearly a balance between complex stability and cargo release should be taken into account to optimize the transfection efficiency of the non-viral vectors. The gene transfer activity in ARPE-19 cells suggests the applicability of this kind of carrier for ocular treatments based on retinal gene delivery.

Original language	English
Pages (from-to)	346-356
Number of pages	11
Journal	Journal of Colloid and Interface Science
Volume	655
DOIs	https://doi.org/10.1016/j.jcis.2023.11.003
Publication status	Published - Feb 2024

Funding

This investigation was sponsored by FAPESP Grant 2019/20470-8 (linked to the European Union’s Horizon 2020 Research and Innovation Staff Exchange programme under the Marie Skłodowska-Curie Actions through grant no. 823883) and Grant 2021/12071-6. F.C.G acknowledges the productivity research fellowship granted by CNPq (Grant 303268/2020-4), F.A.O thanks the scholarship granted by FAPESP (Grant 2019/12944-0) and L.J.C.A. acknowledges the fellowships granted by FAPESP (Grant 2016/23844-8) and CAPES (Grant 88887.367963/2019-00). The LNNano (CNPEM, Brazil) is acknowledged for granting access to cryo-TEM facilities (proposal TEM-20210592). The CEM at UFABC is acknowledged for providing accessibility to the Malvern light scattering equipment. This investigation was sponsored by FAPESP Grant 2019/20470-8 (linked to the European Union's Horizon 2020 Research and Innovation Staff Exchange programme under the Marie Skłodowska-Curie Actions through grant no. 823883) and Grant 2021/12071-6. F.C.G acknowledges the productivity research fellowship granted by CNPq (Grant 303268/2020-4), F.A.O thanks the scholarship granted by FAPESP (Grant 2019/12944-0) and L.J.C.A. acknowledges the fellowships granted by FAPESP (Grant 2016/23844-8) and CAPES (Grant 88887.367963/2019-00). The LNNano (CNPEM, Brazil) is acknowledged for granting access to cryo-TEM facilities (proposal TEM-20210592). The CEM at UFABC is acknowledged for providing accessibility to the Malvern light scattering equipment.

Austrian Fields of Science 2012

301208 Pharmaceutical technology

Keywords

Cytotoxicity
Lipopeptides
Nanomedicine
Non-viral gene delivery
Ocular diseases

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10.1016/j.jcis.2023.11.003

Cite this

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title = "Nucleic acid delivery to retinal cells using lipopeptides as a potential tool towards ocular gene therapies",

abstract = "We evaluated the use of lipopeptides capable to bind to nucleic acids towards plasmid DNA (pDNA) delivery. The investigations were particularly focused on arising retinal pigment epithelial cells (ARPE-19) as motivated by the considerable number of ocular disorders linked to gene aberrations. The lipopeptides comprised the artificial oligoamino acid succinyl-tetraethylene pentamine (Stp) as well as incorporated lysines, histidines, cysteines, fatty acids, and tyrosine trimers. Regardless of the structural differences, the lipopeptides demonstrated to efficiently condense pDNA at nitrogen-to-phosphate molar ratio (N/P) ≥ 6. Spheric nanoparticles were observed by cryo-TEM and dynamic light scattering determined hydrodynamic sizes ranging from 50 to 130 nm. The biological assays evidenced highly efficient pDNA delivery with a lower degree of cytotoxicity compared to the well-known transfecting agent linear polyethylenimine (LPEI). Although more efficient than LPEI, cysteine-containing carriers were demonstrated to be less efficient than the other counterparts possibly due to exceeding polyplex stabilization via disulfide cross links, which could hamper pDNA unpacking at the target site. Therefore, clearly a balance between complex stability and cargo release should be taken into account to optimize the transfection efficiency of the non-viral vectors. The gene transfer activity in ARPE-19 cells suggests the applicability of this kind of carrier for ocular treatments based on retinal gene delivery.",

keywords = "Cytotoxicity, Lipopeptides, Nanomedicine, Non-viral gene delivery, Ocular diseases",

author = "Albuquerque, \{Lindomar J.C.\} and \{de Oliveira\}, \{Fernando A.\} and Christoffolete, \{Marcelo A.\} and Michelle Nascimento-Sales and Simone Berger and Ernst Wagner and Ulrich L{\"a}chelt and Giacomelli, \{Fernando C.\}",

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AU - Albuquerque, Lindomar J.C.

AU - de Oliveira, Fernando A.

AU - Christoffolete, Marcelo A.

AU - Nascimento-Sales, Michelle

AU - Berger, Simone

AU - Wagner, Ernst

AU - Lächelt, Ulrich

AU - Giacomelli, Fernando C.

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AB - We evaluated the use of lipopeptides capable to bind to nucleic acids towards plasmid DNA (pDNA) delivery. The investigations were particularly focused on arising retinal pigment epithelial cells (ARPE-19) as motivated by the considerable number of ocular disorders linked to gene aberrations. The lipopeptides comprised the artificial oligoamino acid succinyl-tetraethylene pentamine (Stp) as well as incorporated lysines, histidines, cysteines, fatty acids, and tyrosine trimers. Regardless of the structural differences, the lipopeptides demonstrated to efficiently condense pDNA at nitrogen-to-phosphate molar ratio (N/P) ≥ 6. Spheric nanoparticles were observed by cryo-TEM and dynamic light scattering determined hydrodynamic sizes ranging from 50 to 130 nm. The biological assays evidenced highly efficient pDNA delivery with a lower degree of cytotoxicity compared to the well-known transfecting agent linear polyethylenimine (LPEI). Although more efficient than LPEI, cysteine-containing carriers were demonstrated to be less efficient than the other counterparts possibly due to exceeding polyplex stabilization via disulfide cross links, which could hamper pDNA unpacking at the target site. Therefore, clearly a balance between complex stability and cargo release should be taken into account to optimize the transfection efficiency of the non-viral vectors. The gene transfer activity in ARPE-19 cells suggests the applicability of this kind of carrier for ocular treatments based on retinal gene delivery.

KW - Cytotoxicity

KW - Lipopeptides

KW - Nanomedicine

KW - Non-viral gene delivery

KW - Ocular diseases

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JO - Journal of Colloid and Interface Science

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ER -

Nucleic acid delivery to retinal cells using lipopeptides as a potential tool towards ocular gene therapies

Abstract

Funding

Austrian Fields of Science 2012

Keywords

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