Fig. 7: Administration of MFSD7C mRNA improves lung damage in 7C−/− mice.
From: MFSD7C protects hemolysis-induced lung impairments by inhibiting ferroptosis
a Schematic illustration of lung targetability of hybrid 113-N16B lipid nanoparticles. hLNP was formulated by 113-N16B: cholesterol: phospholipids: DMG-PEG2000 (50:38.5:10:1.5). b Representative whole-body and organs bioluminescence images of mice by IVIS imaging system. Mice were intravenously administered with Control mRNA-loaded hLNPs or Luc+7C mRNA-loaded hLNPs at a single dose of 0.5 mg/kg. Images were taken at 6 h post injection. c Western blot analysis of MFSD7C expression in lung of 7Cflox/flox mice treated with 7C mRNA-loaded hLNPs at 6 h post injection. Three biological replicates were performed for each sample. d Treatment design for hemolytic mice and SCD mice. 12-month old HbAA- and HbSS-Townes mice and 8-week old 7Cflox/flox and 7C−/− mice (n = 6) received MFSD7C or control mRNA-loaded nanoparticles weekly for four weeks and were sacrificed for analysis at one week after last treatment. 7C−/− and 7Cflox/flox mice were additionally injected with 50 mg/kg of PHZ on day 0 and day 7 to induce hemolysis. e H&E staining of lung tissues (x4, x10, x40). f Cytokine levels, including TNFα, IL-1β, and IL-17 in murine BALF were measured by ELISA. n = 4. g Masson’s trichrome staining of lung tissues (x4, x10, x40). h Changes in the mRNA expression of Fibronectin, Collagen, α-SMA, and TGF-β were evaluated by RT-qPCR. n = 3. i Lipid ROS production in AMs were determined by flow cytometry using BODIPY C11. j Mitochondrial ROS measured by MitoSOX green in AMs. k Representative western blot of MFSD7C, HMOX1, ACSL4, GPX4, CD36 and FABP4 expression in AMs and endothelial cells isolated from HbSS-Townes mice and PHZ-challenged 7C−/− mice treated with MFSD7C mRNA or control mRNA. Data are presented as mean + standard deviation (SD). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
