Enabling customization of non-viral gene delivery systems for individual cell types by surface-induced mineralization

Abstract

Delivering genes to mediate functions of cells is a crucial technology for both basic science and clinical applications. Though numerous non-viral gene delivery systems have been developed, the diversity of mammalian cells poses a great challenge to the material design. Here, we demonstrate that surface-induced mineralization represents a promising approach to systematically customize DNA delivery with respect to the characteristics of cells. We initially examined gene transfer in nine cell types derived from different tissues and organisms by surface-induced DNA/calcium carbonate nanocomposites derived from a library of mineral solutions. Subsequently, we correlated gene transfer efficiency with cellular uptake, pH responsiveness of nanocomposites, and phagosomal pH of individual cell types. Based on the correlation, we were able to optimize the DNA delivery to the cell types of interest. Surface-induced mineralization possesses great potential for customizing gene transfer in realizing gene-and cell-based therapy and probing functions of genes.

Fabrication of a DNA-lipid-apatite composite layer for efficient and area-specific gene transfer

“…1), [CH 3 (CH 2 ) 17 ] 2 NCH 2 CH 2 CONHCH 2 CH 2 N(CH 2 CH 2 CONHCH 2 CH 2 NH 2 ) 2 (PD; modified from DL-G1 in [23], supplied by Katayama Chemical Industries, Japan), (2) 2,3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-n,n-dimethyl-1-propanaminium trifluoroacetate (DOSPA) and dioleoylphosphatidylethanolamine (DOPE) mixed at the mass ratio of 3: 1 (LF; Lipofectamine Ò , Life Technologies, USA), and (3) N-[1-(2,3-dioloyloxy) propyl]-N,N,N-trimethylammonium methylsulfate (DT; DOTAP, Roche, Switzerland). PD is a newly developed Lp [23][24][25], whereas LF and DT are conventional ones used in previous studies [19,20]. As a DNA source, pGL3 control vector (Promega, USA), which was propagated and purified in 0.7-1.2 mg/mL plasmid, was used.…”

“…We have also shown that area-specific gene transfer is feasible with the improved system using the DNA-fibronectin-apatite composite layer (DF-Ap layer) [18]. More recently, Sun et al and Luong et al [19,20] have shown that conjugation of DNA with a lipid transfection reagent (Lp) is also effective in improving the gene transfer efficiency of the D-Ap layer. Although Lps employed in these studies are commercially available and have long been used worldwide because of their excellent efficiency for a non-viral system, they have the drawback of reduced efficiency in the presence of serum.…”

“…Since the late 1990s, surface-mediated gene transfer systems using nonparticulate and planate substrates immobilizing DNA have been developed [11][12][13][14][15][16][17][18][19][20]. Among these substrate materials, apatite-based composite layers [15][16][17][18][19][20] consisting of a matrix layer of apatite nanocrystals have great potential in tissue engineering, especially for the bone tissue.…”

“…Among these substrate materials, apatite-based composite layers [15][16][17][18][19][20] consisting of a matrix layer of apatite nanocrystals have great potential in tissue engineering, especially for the bone tissue. This potential is because apatite is a main component of human hard tissues and hence exhibits good biocompatibility and bone-bonding ability [21,22].…”

Fabrication of DNA-antibody–apatite composite layers for cell-targeted gene transfer

“…However, the gene transfer efficiency of the calcium phosphate system is relatively low. It was improved by developing a non-particulate and surface-mediated gene transfer system using a DNA-apatite composite layer (D-Ap layer), in which DNA molecules are immobilized within a matrix of apatite nanocrystals [13,14]. Recently, we have further improved the efficiency of gene transfer by co-immobilizing a cell adhesion protein, in this case either laminin [15,16] or fibronectin [17,18], within the D-Ap layer.…”