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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, <t>picrosirius</t> red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.
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An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, picrosirius red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.

Journal: Cell Reports Medicine

Article Title: Cross-species tropism of AAV.CPP.16 in the respiratory tract and its gene therapies against pulmonary fibrosis and viral infection

doi: 10.1016/j.xcrm.2025.102144

Figure Lengend Snippet: An AAV.CPP.16-mediated gene therapy for idiopathic pulmonary fibrosis (A) Schematic diagram illustrating design of a bifunctional fusion protein that inhibits VEGF and TGF-β1 and construction of an AAV vector expressing such a protein. (B) Experimental design for testing the AAV.CPP.16 gene therapy (CPP.16-IPF trap) in a bleomycin-induced IPF mouse model. i.t., intratracheal; inhal., Inhaled; i.g., intragastric. (C) Representative H&E staining on lung sections in mice with different treatments. Scale bar: 200 μm for low-magnification images and 50 μm for enlarged images. (D) Quantification of lung cell areas in H&E-stained lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. (E) Representative Masson, picrosirius red, and αSMA immunohistochemical staining of lung sections in mice with different treatments. Masson (blue) and picrosirius red (red) stainings label collagen expression. Scale bar: 200 μm. (F) Quantification of fibrotic areas on lung sections. n = 3–6 per group, mean ± SEM, one-way ANOVA with Tukey’s post-test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. PBS, phosphate-buffered saline; BLM, bleomycin; IPF, idiopathic pulmonary fibrosis; PFD, pirfenidone; H&E, hematoxylin and eosin.

Article Snippet: Picrosirius red staining , Servicebio , Cat# G1078.

Techniques: Plasmid Preparation, Expressing, Staining, Immunohistochemical staining, Saline