Fig. 2: Silencing FcRn binding increases tissue retention and reduces peripheral exposure of IL-12Fc. | Nature Communications

Fig. 2: Silencing FcRn binding increases tissue retention and reduces peripheral exposure of IL-12Fc.

From: FcRn-silencing of IL-12Fc prevents toxicity of local IL-12 therapy and prolongs survival in experimental glioblastoma

Fig. 2: Silencing FcRn binding increases tissue retention and reduces peripheral exposure of IL-12Fc.

A Binding of modified IL-12Fc to FcRn at pH = 6.0 was determined by surface plasmon resonance (SPR) with hFcRn in the immobilized phase. Signal response (RU) over time for IL-12Fc WT (left) or IL-12Fc NHQ (right). Curves relate to concentrations (nM, top to bottom): IL-12Fc WT: 365 – 122 – 41 – 14 – 5; IL-12Fc NHQ: 3645 – 1215 – 405 – 135 – 45. B Dissociation constant KD (nM) of IL-12Fc WT (IHH) and modified variants as well as control antibodies, calculated from SPR equilibrium analysis (Fig. S2C). C IL-12 levels in the ipsilateral hemisphere 6 h after intraparenchymal administration of 1 μg of the indicated IL-12Fc variants via CED into hFcRn Tg32 mice. Mean ± SD. Outlier removal based on Grubbs’ test. One-way ANOVA with Tukey’s multiple comparison test. IL-12Fc WT: n = 9, IL-12Fc IAQ: n = 8, IL-12Fc AAA: n = 9, IL-12Fc NHQ: n = 9 animals. D IL-12 levels in the ipsilateral hemisphere 6 h after intraparenchymal administration of 1 μg of the indicated IL-12Fc variants via CED into hFcRn Tg32 mice treated intravenously (i.v.) with 100 μg of the FcRn inhibitor MST-HN Fc. Mean ± SD. Unpaired two-tailed t-test of groups IL-12Fc NHQ to IL-12Fc WT group with (p = 0.0177) and without FcRn inhibitor (non-significant, p = 0.8663). IL-12Fc WT: n = 5, IL-12Fc NHQ: n = 5, IL-12Fc WT + MST-HN Fc: n = 8, IL-12Fc NHQ + MST-HN Fc: n = 8 animals. E Extended pharmacokinetic (PK) analysis. Indicated IL-12 variants were administered to hFcRn Tg32 mice (n = 7–12 animals/timepoint/group; detailed group sizes in Table S2) and analyzed as in C). Mean concentration of IL-12 in brain (top) and plasma (bottom) across the next 7 days. Outlier removal based on Grubbs’ test. Separate curves with individual data points in Fig. S2I, J. One-way ANOVA with Dunnett’s test IL-12Fc NHQ vs IL-12Fc WT and rIL-12. Statistically significant difference only to rIL-12. IL-12Fc WT: green; IL-12Fc NHQ: orange; rIL-12: black. F Overall exposure (area under curve, AUC) analysis for brain (top) and plasma (bottom) in E) until day 3. Mean ± SEM. Unpaired two-tailed t-test IL-12Fc NHQ vs IL-12Fc WT and rIL-12 corrected for multiple testing using Bonferroni correction. Source data are provided as a Source Data file.

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