Fig. 2: Kinetics of dual mRNAs-encoded hepatic PCC and comparison of dual mRNA therapy with carglumic acid in PA mice.
From: Dual mRNA therapy restores metabolic function in long-term studies in mice with propionic acidemia
a, b Kinetics of dual mRNAs-encoded hepatic PCC subunits and enzyme activity. PA hypomorphic mice of mixed gender were administered a single IV bolus injection (1 mg/kg) of dual mRNAs (n = 4 mice/time point) or Luc control mRNA (n = 3 mice/time point) encapsulated in LNPs and were sacrificed at various time points. a Absolute quantification of hPCCA and hPCCB proteins in livers from dual mRNAs-injected PA mice by LC-MS/MS using signature peptides for wild-type (WT) hPCCA and hPCCB protein sequences. hPCCA and hPCCB were below the LLOQ in all livers from Luc control mRNA-injected mice. b PCC enzyme activity in hepatic mitochondria was measured. Shaded bars represent the range of endogenous PCC enzyme activity levels from all Luc mRNA-injected mice across all time points (n = 21). c–e Substantial reductions in plasma ammonia and primary disease biomarkers due to dual mRNAs in contrast to carglumic acid. PA hypomorphic female mice were administered a single IV bolus dose of Tris-sucrose buffer (n = 11) or 1 mg/kg of dual mRNAs or Luc control mRNA (n = 12) encapsulated in LNPs and were sacrificed 7 days post injection. Additional PA hypomorphic female mice were administered repeat doses of 1% carboxymethyl cellulose (CMC, n = 11) or carglumic acid (1200 mg/kg/day, twice daily, n = 12) via oral gavage for 7 consecutive days and then sacrificed. Unaffected Pcca+/− mice (n = 9) were IV injected with Tris-sucrose buffer as control. Mice were bled at study days 0 (pre-treatment), 2, and 7. Plasma ammonia (c) and liver 2MC (e) at day 7 upon sacrifice, and plasma 2MC (d) were assessed. Data are shown as mean ± SEM. Plasma and liver 2MC levels in unaffected mice were <LLOQ. P values were obtained from Tukey’s post hoc pairwise multiple comparison test following a one-way or two-way ANOVA, and are provided in the source data. ***P < 0.001 comparing dual mRNA therapy with carglumic acid. +++P < 0.001 comparing dual mRNAs or carglumic acid group with control groups (1% CMC, Tris-sucrose, and Luc mRNA). Pre, pre-treatment on day 0.
