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Treatment with mouse/rat C5-siRNA/LNP suppresses disease symptoms and loss of neuromuscular junctions in experimental MG model in rats Disease severity (A), body weight change (B), hemolysis (C), and complement C5 expression levels in the liver (D) of MG rats (n = 3/group) treated with PBS, Ctrl-siRNA/LNP, or 0.01, 0.03, or 0.1 mg/kg of mouse/rat (m/r) C5-siRNA/LNP are shown. (A) Rats received i.v. administration of PBS, 0.1 mg/kg Ctrl-siRNA/LNP, or 0.01–0.1 mg/kg of m/r C5-siRNA/LNP on day 0. On day 7, <t>anti-AChR</t> mAb <t>(mAb35,</t> 1 mg/kg) was i.v. administered to rats to induce the symptoms of MG. Disease severity was monitored on days 8 and 9. (B) Body weights of rats treated with PBS, Ctrl-siRNA/LNP, or rat C5-siRNA/LNP were expressed as a ratio of the initial body weight (day 0). (C and D) Blood samples were collected from the jugular vein on days 3, 7, 8, and 9 and livers were collected on day 9 to evaluate the complement hemolytic activity in the serum and liver complement C5 mRNA expression levels (rat C5/rat GAPDH). (E) Soleus muscle sections were stained with Alexa Fluor 488-labeled α-BTX, and fluorescence signal was quantified for at least 50 or more neuromuscular junctions in each treated rat. Data are presented as the mean ± SEM. n = 3 in each group; ∗∗p < 0.01 versus Ctrl-siRNA/LNP.
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Treatment with mouse/rat C5-siRNA/LNP suppresses disease symptoms and loss of neuromuscular junctions in experimental MG model in rats Disease severity (A), body weight change (B), hemolysis (C), and complement C5 expression levels in the liver (D) of MG rats (n = 3/group) treated with PBS, Ctrl-siRNA/LNP, or 0.01, 0.03, or 0.1 mg/kg of mouse/rat (m/r) C5-siRNA/LNP are shown. (A) Rats received i.v. administration of PBS, 0.1 mg/kg Ctrl-siRNA/LNP, or 0.01–0.1 mg/kg of m/r C5-siRNA/LNP on day 0. On day 7, anti-AChR mAb (mAb35, 1 mg/kg) was i.v. administered to rats to induce the symptoms of MG. Disease severity was monitored on days 8 and 9. (B) Body weights of rats treated with PBS, Ctrl-siRNA/LNP, or rat C5-siRNA/LNP were expressed as a ratio of the initial body weight (day 0). (C and D) Blood samples were collected from the jugular vein on days 3, 7, 8, and 9 and livers were collected on day 9 to evaluate the complement hemolytic activity in the serum and liver complement C5 mRNA expression levels (rat C5/rat GAPDH). (E) Soleus muscle sections were stained with Alexa Fluor 488-labeled α-BTX, and fluorescence signal was quantified for at least 50 or more neuromuscular junctions in each treated rat. Data are presented as the mean ± SEM. n = 3 in each group; ∗∗p < 0.01 versus Ctrl-siRNA/LNP.

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identification of potent siRNA targeting complement C5 and its robust activity in pre-clinical models of myasthenia gravis and collagen-induced arthritis

doi: 10.1016/j.omtn.2023.01.005

Figure Lengend Snippet: Treatment with mouse/rat C5-siRNA/LNP suppresses disease symptoms and loss of neuromuscular junctions in experimental MG model in rats Disease severity (A), body weight change (B), hemolysis (C), and complement C5 expression levels in the liver (D) of MG rats (n = 3/group) treated with PBS, Ctrl-siRNA/LNP, or 0.01, 0.03, or 0.1 mg/kg of mouse/rat (m/r) C5-siRNA/LNP are shown. (A) Rats received i.v. administration of PBS, 0.1 mg/kg Ctrl-siRNA/LNP, or 0.01–0.1 mg/kg of m/r C5-siRNA/LNP on day 0. On day 7, anti-AChR mAb (mAb35, 1 mg/kg) was i.v. administered to rats to induce the symptoms of MG. Disease severity was monitored on days 8 and 9. (B) Body weights of rats treated with PBS, Ctrl-siRNA/LNP, or rat C5-siRNA/LNP were expressed as a ratio of the initial body weight (day 0). (C and D) Blood samples were collected from the jugular vein on days 3, 7, 8, and 9 and livers were collected on day 9 to evaluate the complement hemolytic activity in the serum and liver complement C5 mRNA expression levels (rat C5/rat GAPDH). (E) Soleus muscle sections were stained with Alexa Fluor 488-labeled α-BTX, and fluorescence signal was quantified for at least 50 or more neuromuscular junctions in each treated rat. Data are presented as the mean ± SEM. n = 3 in each group; ∗∗p < 0.01 versus Ctrl-siRNA/LNP.

Article Snippet: After 7 days, rats were inoculated intravenously with 1 mg/kg of antibodies against the AChR (clone Mab35 no. BE0123, BioXCell).

Techniques: Expressing, Activity Assay, Staining, Labeling, Fluorescence

Mouse/rat C6-siRNA/LNP treatment suppresses MG symptoms in rats Disease severity (A), body weight change (B), hemolysis (C), and complement C6 expression levels in the liver (D) of MG rats (n = 3/group) treated with PBS, Ctrl-siRNA/LNP, or 0.1 and 1 mg/kg mouse/rat (m/r) C6-siRNA/LNP are shown. (A) Rats received i.v. administration of PBS, 0.1 mg/kg of Ctrl-siRNA/LNP, or 0.01 or 0.1 mg/kg m/r C6-siRNA/LNP on day 0. On day 7, anti-AChR mAb (Mab35, 1 mg/kg) was i.v. administered to rats to induce the symptoms of MG. Disease severity was monitored on days 8 and 9 (B). Body weights of rats treated with PBS, Ctrl-siRNA/LNP, or rat C6-siRNA/LNP were expressed as a ratio of the initial body weight (day 0). (C and D) Blood samples were collected from the jugular vein on days 3, 7, 8, and 9 and livers were collected on day 9 to evaluate the complement hemolytic activity in serum and liver complement C6 mRNA expression levels (rat C6/rat GAPDH). Data are presented as the mean ± SEM. n = 4 in each group; ∗p < 0.05 and ∗∗p < 0.01 versus Ctrl-siRNA/LNP.

Journal: Molecular Therapy. Nucleic Acids

Article Title: Identification of potent siRNA targeting complement C5 and its robust activity in pre-clinical models of myasthenia gravis and collagen-induced arthritis

doi: 10.1016/j.omtn.2023.01.005

Figure Lengend Snippet: Mouse/rat C6-siRNA/LNP treatment suppresses MG symptoms in rats Disease severity (A), body weight change (B), hemolysis (C), and complement C6 expression levels in the liver (D) of MG rats (n = 3/group) treated with PBS, Ctrl-siRNA/LNP, or 0.1 and 1 mg/kg mouse/rat (m/r) C6-siRNA/LNP are shown. (A) Rats received i.v. administration of PBS, 0.1 mg/kg of Ctrl-siRNA/LNP, or 0.01 or 0.1 mg/kg m/r C6-siRNA/LNP on day 0. On day 7, anti-AChR mAb (Mab35, 1 mg/kg) was i.v. administered to rats to induce the symptoms of MG. Disease severity was monitored on days 8 and 9 (B). Body weights of rats treated with PBS, Ctrl-siRNA/LNP, or rat C6-siRNA/LNP were expressed as a ratio of the initial body weight (day 0). (C and D) Blood samples were collected from the jugular vein on days 3, 7, 8, and 9 and livers were collected on day 9 to evaluate the complement hemolytic activity in serum and liver complement C6 mRNA expression levels (rat C6/rat GAPDH). Data are presented as the mean ± SEM. n = 4 in each group; ∗p < 0.05 and ∗∗p < 0.01 versus Ctrl-siRNA/LNP.

Article Snippet: After 7 days, rats were inoculated intravenously with 1 mg/kg of antibodies against the AChR (clone Mab35 no. BE0123, BioXCell).

Techniques: Expressing, Activity Assay