Brandon Nemieboka1, Sai Kiran Sharma2, Thapi Dharma Rao3, Kimberly J Edwards2, Su Yan4, Pei Wang4, Ashwin Ragupathi2, Alessandra Piersigilli5, David R Spriggs6, Jason S Lewis7. 1. Gerstner Sloan Kettering Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center, NY, USA; Department of Radiology, Memorial Sloan Kettering Cancer Center, NY, USA. 2. Department of Radiology, Memorial Sloan Kettering Cancer Center, NY, USA. 3. Department of Medicine, Memorial Sloan Kettering Cancer Center, NY, USA. 4. Eureka Therapeutics Inc., CA, USA. 5. Tri-Institutional Laboratory of Comparative Pathology, Memorial Sloan Kettering Cancer Center, Weill Cornell Medical College and The Rockefeller University, NY, USA. 6. Department of Medicine, Memorial Sloan Kettering Cancer Center, NY, USA; Department of Pharmacology, Weill Cornell Medical College, New York, NY, USA. 7. Gerstner Sloan Kettering Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center, NY, USA; Department of Radiology, Memorial Sloan Kettering Cancer Center, NY, USA; Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA; Department of Pharmacology, Weill Cornell Medical College, New York, NY, USA; Department of Radiology, Weill Cornell Medical College, New York, NY, USA; Radiochemistry and Molecular Imaging Probes Core, Memorial Sloan Kettering Cancer Center, New York, NY, USA. Electronic address: lewisj2@mskcc.org.
Abstract
INTRODUCTION: Despite its limitations, CA125 remains the most widely used biomarker for the diagnosis and treatment monitoring of ovarian cancer. Targeting the unshed portion of serum biomarkers such as CA125/MUC16 may afford more specific imaging and targeting of MUC16-positive tumors in High Grade Serous Ovarian Cancer (HGSOC) patients. METHODS: Six monoclonal antibodies raised against the 58 amino acid sequence between the extracellular cleavage site and the transmembrane region of MUC16 were radiolabeled with [89Zr]Zr4+. The radioimmunoconjugates were evaluated in vitro for molar activities, target binding affinity, cellular internalization and serum stability. In vivo characterization was performed via longitudinal positron emission tomography (PET) imaging and ex vivo biodistribution studies in mice bearing subcutaneous xenografts of SKOV3 cells transfected with the proximal 114 amino-acids of MUC16 carboxy-terminus (SKOV3+). RESULTS: In vitro screening identified 9C9 and 4H11 as the lead antibody candidates based on their comparable binding affinities, serum stability and cellular internalization profiles. Despite an identical molecular footprint for binding to MUC16, [89Zr]Zr-DFO-4H11 yielded a more favorable in vivo radiopharmacologic profile. Furthermore, a humanized variant of 4H11 capable of binding MUC16 in vitro also yielded excellent in vivo profile in subcutaneous xenograft models of SKOV3+, OVCAR3 tumors and a patient-derived xenograft model representative of HGSOC. CONCLUSION: Radiopharmacologic screening of antibodies early during their development can provide crucial information pertinent to the in vitro characterization and in vivo pharmacokinetics. The favorable in vivo profile demonstrated by humanized 4H11 combined with the use of its murine predecessor for immunohistochemical staining of biopsied tumor tissues from HGSOC patients makes a unique pair of antibodies that is poised for clinical translation.
INTRODUCTION: Despite its limitations, CA125 remains the most widely used biomarker for the diagnosis and treatment monitoring of ovarian cancer. Targeting the unshed portion of serum biomarkers such as CA125/MUC16 may afford more specific imaging and targeting of MUC16-positive tumors in High Grade Serous Ovarian Cancer (HGSOC) patients. METHODS: Six monoclonal antibodies raised against the 58 amino acid sequence between the extracellular cleavage site and the transmembrane region of MUC16 were radiolabeled with [89Zr]Zr4+. The radioimmunoconjugates were evaluated in vitro for molar activities, target binding affinity, cellular internalization and serum stability. In vivo characterization was performed via longitudinal positron emission tomography (PET) imaging and ex vivo biodistribution studies in mice bearing subcutaneous xenografts of SKOV3 cells transfected with the proximal 114 amino-acids of MUC16 carboxy-terminus (SKOV3+). RESULTS: In vitro screening identified 9C9 and 4H11 as the lead antibody candidates based on their comparable binding affinities, serum stability and cellular internalization profiles. Despite an identical molecular footprint for binding to MUC16, [89Zr]Zr-DFO-4H11 yielded a more favorable in vivo radiopharmacologic profile. Furthermore, a humanized variant of 4H11 capable of binding MUC16 in vitro also yielded excellent in vivo profile in subcutaneous xenograft models of SKOV3+, OVCAR3tumors and a patient-derived xenograft model representative of HGSOC. CONCLUSION: Radiopharmacologic screening of antibodies early during their development can provide crucial information pertinent to the in vitro characterization and in vivo pharmacokinetics. The favorable in vivo profile demonstrated by humanized 4H11 combined with the use of its murine predecessor for immunohistochemical staining of biopsied tumor tissues from HGSOC patients makes a unique pair of antibodies that is poised for clinical translation.
Authors: K Nustad; R C Bast; T J Brien; O Nilsson; P Seguin; M R Suresh; T Saga; S Nozawa; O P Børmer; H W de Bruijn; M Nap; A Vitali; M Gadnell; J Clark; K Shigemasa; B Karlsson; F T Kreutz; D Jette; H Sakahara; K Endo; E Paus; D Warren; S Hammarström; P Kenemans; J Hilgers Journal: Tumour Biol Date: 1996
Authors: Sai Kiran Sharma; Serge K Lyashchenko; Hijin A Park; Nagavarakishore Pillarsetty; Yorann Roux; Jiong Wu; Sophie Poty; Kathryn M Tully; John T Poirier; Jason S Lewis Journal: Nucl Med Biol Date: 2019-05-03 Impact factor: 2.408
Authors: Lindsey A Torre; Britton Trabert; Carol E DeSantis; Kimberly D Miller; Goli Samimi; Carolyn D Runowicz; Mia M Gaudet; Ahmedin Jemal; Rebecca L Siegel Journal: CA Cancer J Clin Date: 2018-05-29 Impact factor: 508.702
Authors: Ahmedin Jemal; Ram C Tiwari; Taylor Murray; Asma Ghafoor; Alicia Samuels; Elizabeth Ward; Eric J Feuer; Michael J Thun Journal: CA Cancer J Clin Date: 2004 Jan-Feb Impact factor: 508.702
Authors: R C Bast; T L Klug; E St John; E Jenison; J M Niloff; H Lazarus; R S Berkowitz; T Leavitt; C T Griffiths; L Parker; V R Zurawski; R C Knapp Journal: N Engl J Med Date: 1983-10-13 Impact factor: 91.245
Authors: Sai Kiran Sharma; Kyeara N Mack; Alessandra Piersigilli; Jacob Pourat; Kimberly J Edwards; Outi Keinänen; Maria S Jiao; Huiyong Zhao; Brandy White; Cory L Brooks; Elisa de Stanchina; Madi R Madiyalakan; Michael A Hollingsworth; Prakash Radhakrishnan; Jason S Lewis; Brian M Zeglis Journal: Clin Cancer Res Date: 2022-03-01 Impact factor: 12.531