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IgG-cleavage protein allows therapeutic AAV gene delivery in passively immunized MPS IIIA mice

Abstract

The widespread pre-existing αAAV-Abs in humans pose a critical challenge in translation of AAV gene therapy. The IgG degrading enzyme of Streptococci (IdeS) is demonstrated to specifically cleave IgG of humans and other species (not mouse). This study developed a modified new modified IdeS protein product (IdeSop). When incubated in vitro, IdeSop was shown to completely cleave human and rabbit IgGs within 6 h. To test IdeSop in a disease setting, we established a rabbitized αAAV9-Ab+ mouse by an IV infusion of purified acute αAAV9-Ab+ rabbit IgG into MPS IIIA mice, resulting in serum αAAV9-IgG at 1:6,400 and αAAV9-nAbs at 1:800. IdeSop-Ab-cleavage was shown to be dose-dependent. An IV IdeSop infusion at the effective doses resulted in rapid IgG depletion and clearance of pre-existing αAAV9-IgG and αAAV9-nAbs in rabbitized αAAV9-Abs+ MPS IIIA mice. Importantly, an IV injection of a high dose AAV9-hSGSHop vector (5 × 1013vg/kg) at 24 h post IdeSop treatment led to transduction as effective in αAAV9-Abs+ MPS IIIA mice, as in αAAV9-Abs-negative controls. We believe that transient IdeSop administration may offer a great tool to address the pre-existing-αAAV-Abs for the translation of rAAV gene therapy to treat diseases in humans, making effective rAAV gene therapy available to all patients in need.

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Fig. 1: Expression of functional recombinant IdeS.
Fig. 2: Dose response IgG cleavage of IdeSop.
Fig. 3: Depletion of αAAV9-IgG by IdeSop in vivo in rabbitized mice wt C57BL/6 mice were given an IV injection of 100 μl of purified αAAV9-Ab-positive Rb IgG via tail vein.
Fig. 4: IdeSop αAAV9-IgG depletion retained the transduction efficiency of IV-delivered scAAV9-hSGSH vector in rabbitized αAAV9-IgG+ MPS IIIA mice MPS IIIA mice (n = 8) were given an IV injection of Rb-IgG, of which half (n = 4) were then given an IV injection of IdeSop (0.45 mg/kg).
Fig. 5: IdeSop αAAV9-IgG depletion allow efficient rSGSH expression and clearance of GAG contents in the CNS and periphery tissues in rabbitized αAAV9-IgG+ MPS IIIA mice following an IV AAV9-hSGSH vector delivery MPS IIIA mice (n = 4/group) were given an IV injection of Rb-IgG, of which four were then given an IV injection of IdeSop (0.45 mg/kg).

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Acknowledgements

We thank the UNC Protein Production & Purification Core for manufacturing of the AbC product, and UNC Vector Core for vector production services. We appreciate Dr. Douglas M. McCarty’s constructive comments and editing assistance. We also thank Dr. Charles Askew for kindly sharing Huh7 cells. This work was supported by a research grant from the National Institute of Health (NIAID R21AI146653) and donations from the Sanfilippo syndrome patient community through Aislinne’s Wish Foundation and the Abby Grace Foundation. Bobo was also supported by a Gene Therapy Fellowship from North Carolina Biotechnology Center (NCBC Grant # 2020-GTF-6903). HF was also supported as site PI by a STTR grant from NIH/NINDS (R41NS115232).

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TB designed and performed the experiments, data analysis and wrote the paper; PS performed the experiments, data collection and analysis, and paper preparation; MR performed tissue collection and ELISA assays; LM performed PAGE, ELISA and NAb assays and data analysis; LF, RF, and NN performed protein production, purification and analysis; HF designed the project and wrote the paper.

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Correspondence to Haiyan Fu.

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HF is the founder and president of NeuroGT, Inc. Other authors declare no competing interests.

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Bobo, T.A., Samowitz, P.N., Robinson, M.I. et al. IgG-cleavage protein allows therapeutic AAV gene delivery in passively immunized MPS IIIA mice. Gene Ther (2022). https://doi.org/10.1038/s41434-022-00368-9

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