Fig. 6: High-density lipoprotein functioned as a potent LNP efficacy modulator.

a The spike-in of HDL, but not VLDL modulated LNP-mediated eGFP expression at 50, 100, and 200 ng/well mRNA dose with LNPs formulated with MC3 (lipid:mRNA = 1:10 or 1:20), cKK-E12 or c12-200 CILs in Huh7 hepatocytes. The error bars represent standard deviation of the mean values derived from raw images (n = 3 experimental replicates). Source data are provided as a Source Data file. b The intensity of corona apolipoprotein E, B, AII and M with various amounts of spike-in HDL(H) or VLDL (V). Source data are provided as a Source Data file. c The expression of the HDL receptors LDLr and SRB1 in Huh7 cells were inhibited separately or simultaneously with siRNA. LNPs were dosed to cells supplemented with LPDS. In the absence of HDL, inhibition of LDLr, but not SRB1, significantly reduced LNP uptake. With spiked HDL, the inhibition of both LDLr and SRB1 lessened LNP uptake. The type of siRNA treatments are indicated as +, treated and −, untreated. A scrambled sequence with no cellular target was used as the siRNA negative control (ctrl). The error bars represent standard deviation of the mean values derived from raw images (n = 3 experimental replicates). Significance P values are determined by unpaired two-tailed t-test. Source data are provided as a Source Data file. d Lean Zucker rats were dosed with eGFP mRNA containing LNPs or PolyA mRNA containing LNPs (Neg Ctrl, n = 5 independent rat) at a dose of 0.1 mg mRNA/kg body weight. The eGFP LNPs were preincubated in 1% LPDS with additional HDL (HDL group, n = 5 independent rat), or without HDL (Non-HDL, n = 5 independent rat) for 4 h to allow corona formation. The ratio of additional HDL to LNPs used was equivalent to the 8.0 × 10⁷ HDL particles/well condition used in (a, b). 6 h post administration, the rats were sacrificed and the expression of eGFP in liver, spleen and kidney was quantified using ELISA. With the addition of HDL, LNPs were more exclusively delivered to liver, with approximately four-fold higher GFP expression in liver and decreased expression in spleen and kidney. The eGFP signal in control group was below the detection limit. The error bars represent standard deviation of the mean values derived individual rats. Significance P values are determined by unpaired two-tailed t-test. Source data are provided as a Source Data file.