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Anti-prion Protein Antibody 6D11 Restores Cellular Proteostasis of Prion Protein Through Disrupting Recycling Propagation of PrPSc and Targeting PrPSc for Lysosomal Degradation

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Abstract

PrPSc is an infectious and disease-specific conformer of the prion protein, which accumulation in the CNS underlies the pathology of prion diseases. PrPSc replicates by binding to the cellular conformer of the prion protein (PrPC) expressed by host cells and rendering its secondary structure a likeness of itself. PrPC is a plasma membrane anchored protein, which constitutively recirculates between the cell surface and the endocytic compartment. Since PrPSc engages PrPC along this trafficking pathway, its replication process is often referred to as “recycling propagation.” Certain monoclonal antibodies (mAbs) directed against prion protein can abrogate the presence of PrPSc from prion-infected cells. However, the precise mechanism(s) underlying their therapeutic propensities remains obscure. Using N2A murine neuroblastoma cell line stably infected with 22L mouse-adapted scrapie strain (N2A/22L), we investigated here the modus operandi of the 6D11 clone, which was raised against the PrPSc conformer and has been shown to permanently clear prion-infected cells from PrPSc presence. We determined that 6D11 mAb engages and sequesters PrPC and PrPSc at the cell surface. PrPC/6D11 and PrPSc/6D11 complexes are then endocytosed from the plasma membrane and are directed to lysosomes, therefore precluding recirculation of nascent PrPSc back to the cell surface. Targeting PrPSc by 6D11 mAb to the lysosomal compartment facilitates its proteolysis and eventually shifts the balance between PrPSc formation and degradation. Ongoing translation of PrPC allows maintaining the steady-state level of prion protein within the cells, which was not depleted under 6D11 mAb treatment. Our findings demonstrate that through disrupting recycling propagation of PrPSc and promoting its degradation, 6D11 mAb restores cellular proteostasis of prion protein.

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Funding

This work was supported by the following awards from the National Institute on Aging R01 AG029635 (MJS), R01 AG031221 (MJS), and R01 AG053990 (MJS), and by the following award from the National Science Foundation CHE-1507946 (KK).

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Authors and Affiliations

  1. Department of Neurology, New York University School of Medicine, 550 First Avenue, Science Building, Room 1007, New York, NY, 10016, USA

    Joanna E. Pankiewicz, Sandrine Sanchez & Martin J. Sadowski

  2. Department of Psychiatry, New York University School of Medicine, New York, NY, 10016, USA

    Martin J. Sadowski

  3. Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY, 10016, USA

    Joanna E. Pankiewicz & Martin J. Sadowski

  4. Department of Chemistry, New York University, New York, NY, 10003, USA

    Kent Kirshenbaum

  5. New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY, 10314, USA

    Regina B. Kascsak & Richard J. Kascsak

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  1. Joanna E. Pankiewicz
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Correspondence to Martin J. Sadowski.

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All mouse care and experimental procedures were approved by the Institutional Animal Care and Use Committee of the New York University School of Medicine.

Conflict of Interest

JEP, SS, KK, and MJS declare no competing interests. RBK and RJK receive royalties for licensing the 6D11 clones, which currently is marketed by BioLegend (San Diego, CA).

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Pankiewicz, J.E., Sanchez, S., Kirshenbaum, K. et al. Anti-prion Protein Antibody 6D11 Restores Cellular Proteostasis of Prion Protein Through Disrupting Recycling Propagation of PrPSc and Targeting PrPSc for Lysosomal Degradation. Mol Neurobiol 56, 2073–2091 (2019). https://doi.org/10.1007/s12035-018-1208-4

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