title: In vivo anti-tumor efficacy of afucosylated anti-CS1 monoclonal antibody produced in glycoengineered Pichia pastoris.
abstract: Monoclonal antibody (mAb) therapy has been successfully used for the treatment of B-cell lymphomas and is currently extended for the treatment of multiple myeloma (MM). New developments in MM therapeutics have achieved significant survival gains in patients but the disease still remains incurable. Elotuzumab (HuLuc63), an anti-CS1 monoclonal IgG1 antibody, is believed to induce anti-tumor activity and MM cytotoxicity through antibody dependent cellular cytotoxicity (ADCC) and inhibition of MM cell adhesion to bone marrow stromal cells (BMSCs). Modulations of the Fc glycan composition at the N297 site by selective mutations or afucosylation have been explored as strategies to develop bio-better therapeutics with enhanced ADCC activity. Afucosylated therapeutic antibodies with enhanced ADCC activity have been reported to possess greater efficacy in tumor growth inhibition at lower doses when compared to fucosylated therapeutic antibodies. The N-linked glycosylation pathway in Pichia pastoris has been engineered to produce human-like N-linked glycosylation with uniform afucosylated complex type glycans. The purpose of this study was to compare afucosylated anti-CS1 mAb expressed in glycoengineered Pichia pastoris with fucosylated anti-CS1 mAb expressed in mammalian HEK293 cells through in vitro ADCC and in vivo tumor inhibition models. Our results indicate that Fc glycosylation is critical for in vivo efficacy and afucosylated anti-CS1 mAb expressed in glycoengineered Pichia pastoris shows a better in vivo efficacy in tumor regression when compared to fucosylated anti-CS1 mAb expressed in HEK293 cells. Glycoengineered Pichia pastoris could provide an alternative platform for generating homogeneous afucosylated recombinant antibodies where Fc mediated immune effector function is important for efficacy.
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Sujatha
authors.initials: S
authors.lastname: Gomathinayagam
authors.affiliation: Biologics Discovery, Palo Alto, Merck Research Laboratories, 901 California Avenue, Palo Alto, CA 94304, United States.
authors.forename: Drake
authors.initials: D
authors.lastname: Laface
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Nga Rewa
authors.initials: NR
authors.lastname: Houston-Cummings
authors.affiliation: Biologics Discovery, Palo Alto, Merck Research Laboratories, 901 California Avenue, Palo Alto, CA 94304, United States.
authors.forename: Ruban
authors.initials: R
authors.lastname: Mangadu
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Renee
authors.initials: R
authors.lastname: Moore
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Ishaan
authors.initials: I
authors.lastname: Shandil
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Nathan
authors.initials: N
authors.lastname: Sharkey
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Li
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States.
authors.forename: Terrance A
authors.initials: TA
authors.lastname: Stadheim
authors.affiliation: GlycoFi Inc., A Wholly-Owned Subsidiary of Merck & Co Inc., 16 Cavendish Court, Lebanon, NH 03766, United States. Electronic address: dzha@mdanderson.org.
authors.forename: Dongxing
authors.initials: D
authors.lastname: Zha
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D000912:Antibodies, Neoplasm / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D060846:Cell Engineering; D045744:Cell Line, Tumor; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D016513:Mice, SCID; D009101:Multiple Myeloma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D009374:Neoplasms, Experimental / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D011994:Recombinant Proteins
keywords: ADCC; Afucosylation; Anti-CS1; HuLuc63; Monoclonal antibodies; Multiple myeloma; N-glycosylation; Pichia pastoris