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

Author(s)

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

• Document type: Journal article
• Source: Journal of Colloid and Interface Science; v. 655, p. 11-pg., 2023-11-08.

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, cysteinecontaining 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. (AU)

• FAPESP's process: 21/12071-6 - Tailoring colloids through supramolecular interactions: from fundamentals to applications
• Grantee: Watson Loh
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/23844-8 - Biodegradable and pH-responsive polymersomes for gene and protein delivery
• Grantee: Lindomar Jose Calumby Albuquerque
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 19/20470-8 - Environmentally-responsive supramolecular polymeric assemblies for the intracellular delivery of therapeutic DNA: towards a toolbox for novel generation of nanopharmaceuticals in ophthalmology
• Grantee: Fernando Carlos Giacomelli
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/12944-0 - Structural and biological evaluations of polyplexes obtained by the complexation of DNA and polyethylenimine derivatives containing alkyl chains and lactose towards the treatment of genetic eye disorders
• Grantee: Fernando Augusto de Oliveira
• Support Opportunities: Scholarships in Brazil - Doctorate