Session: 651. Multiple Myeloma and Plasma Cell Dyscrasias: Basic and Translational: Poster III
Hematology Disease Topics & Pathways:
Research, Translational Research
Methods: AT-02 was produced from a CHO pool using a perfusion cell culture production process, purified by Protein A, anion exchange, and cation exchange chromatographies, and characterized by mass spectrometry and SDS-capillary electrophoresis. AT-02 was biotinylated and used for immunohistochemical detection AL amyloid in formalin fixed tissues. Binding to heparin (as a surrogate for amyloid-associated hypersulfated heparan sulfate proteoglycans) and synthetic AL fibrils was studied using both ELISA and surface plasmon resonance assays. Activation of complement following binding to amyloid substrates was determined using a C5b9 liberation assay in the presence of human plasma. AT-02 labelled with the near infra-red fluorophore Dylight800 was administered to mice bearing subcutaneous human AL amyloid and the uptake monitored over ten (10) days using optical imaging.
Results: AT-02 bound cardiac AL amyloid in tissue sections with high specificity and intensity. The binding potency for synthetic rVl6WIL fibrils, as well as human AL amyloid extracts, was sub-nanomolar in the ELISA assay. Incubation of AT-02 with AL amyloid extract induced phagocytosis by activated human THP-1 cells via FcR-mediated uptake and induced complement activation in the presence of human plasma. Fluorophore labelled AT-02 co-localized with human AL amyloidomas in mice and remained bound to the amyloid for ten days post-injection. Pretreatment of human AL amyloid extract with AT-02 prior to subcutaneous injection enhanced phagocytosis and clearance of amyloid in the amyloidoma mouse model.
Conclusion: Clearance of AL amyloid from affected organs is a significant clinical unmet need for patients. Removing amyloid could positively impact organ function and reverse dysfunction, as seen in some patients where AL amyloid spontaneously regresses coincident with a complete hematologic remission. The humanized IgG1-peptide fusion, AT-02, potently binds AL amyloid and may serve as an opsonin to induce macrophage-mediated phagocytosis and clearance of tissue amyloid deposits.
Acknowledgments: This study is supported by a grant from Attralus Inc, CA, USA. AT-02 was provided by Attralus Inc.
Disclosures: Wall: Attralus, Inc.: Current equity holder in private company, Current holder of stock options in a privately-held company, Patents & Royalties, Research Funding. Kennel: Attralus, Inc.: Current equity holder in private company, Patents & Royalties. Stuckey: Attralus, Inc.: Current equity holder in private company. Richey: Attralus, Inc.: Current equity holder in private company. Martin: Attralus, Inc.: Current equity holder in private company, Patents & Royalties.