Figure 5. Miro1-overexpressing MSC are better mitochondrial donors to asthmatic airways.
A Scheme for development of acute mouse model of asthma and delivery of mGFP labeled MSC into the mice lungs. Mice were Ova-sensitized with weekly 3 successive intraperitoneal (i.p.) injections of ovalbumin (50 μg) allergen and then intranasally challenged with 3% aerosolized ovalbumin for a week on alternate days to develop acute airway allergy. MSC were injected after two challenges on 23rd day post ovalbumin challenge and further after subsequent challenges, the mice were sacrificed.
B Representative images of TFAM expression in mice lungs. Increased mitochondrial biogenesis indicated by higher expression of TFAM in Ova mice compared to Sham as indicated by immunohistochemistry for TFAM. Scale bars, 50 μm.
C Percentage of mGFP in different cell populations of the mice lung under normal and Ova condition treated with MSC, quantified by FACS. BE, bronchial epithelial cells stained with CCSP; ECs, epithelial cells stained with EpCAM; AE, alveolar epithelial cells stained with SPC; Ma, macrophages stained with F4/80; ME, mesenchymal cells stained with alpha smooth muscle actin.
D Percentage of mGFP signal in bronchial epithelial cells as quantified by FACS. Treatment with Miro1-overexpressing MSC (MSCmiroHi) significantly increased percentage of mGFP positive cells, while Miro1 knockdown MSC (MSCmiroLo) was associated with decreased mGFP positive cells compared to control-cDNA or control shRNA-treated cells.
E Airway hyperresponsiveness (AHR) was significantly increased in Ova and shows no improvement with administration of MSCmiroLo. MSCmiroHi treatment led to a significant decrease in AHR compared to Ova mice or Ova mice treated with control MSC (MSCmiroCc, MSCmiroSc).
F, G IL-5 and IL-13 were significantly upregulated in Ova mice compared to Sham mice. MSCmiroHi treatment significantly decreased the levels of both inflammatory cytokines, while there was no effect with administration of MSCmiroLo.
H ATP levels were significantly decreased in Ova mice compared to Sham mice. Miro1 overexpressing MSC (MSCmiroHi) significantly restored ATP levels, while there was no effect with Miro1-downregulated MSC (MSCmiroLo).
Data information: In (C-H): data are of two independent experiments, *P < 0.05 versus Sham, $P < 0.05 versus Ova and #P < 0.05 versus Ova, Ova + MSCmiroCc or Ova + MSCmiroSc.