title: CD30 Downregulation, MMAE Resistance, and MDR1 Upregulation Are All Associated with Resistance to Brentuximab Vedotin.
abstract: Brentuximab vedotin (BV) is an antibody-drug conjugate that specifically delivers the potent cytotoxic drug monomethyl auristatin E (MMAE) to CD30-positive cells. BV is FDA approved for treatment of relapsed/refractory Hodgkin lymphoma and anaplastic large cell lymphoma (ALCL); however, many patients do not achieve complete remission and develop BV-resistant disease. We selected for BV-resistant Hodgkin lymphoma (L428) and ALCL (Karpas-299) cell lines using either constant (ALCL) or pulsatile (Hodgkin lymphoma) exposure to BV. We confirmed drug resistance by MTS assay and analyzed CD30 expression in resistant cells by flow cytometry, qRT-PCR, and Western blotting. We also measured drug exporter expression, MMAE resistance, and intracellular MMAE concentrations in BV-resistant cells. In addition, tissue biopsy samples from 10 Hodgkin lymphoma and 5 ALCL patients who had relapsed or progressed after BV treatment were analyzed by immunohistocytochemistry for CD30 expression. The resistant ALCL cell line, but not the Hodgkin lymphoma cell line, demonstrated downregulated CD30 expression compared with the parental cell line. In contrast, the Hodgkin lymphoma cell line, but not the ALCL cell line, exhibited MMAE resistance and increased expression of the MDR1 drug exporter compared with the parental line. For both Hodgkin lymphoma and ALCL, samples from patients relapsed/resistant on BV persistently expressed CD30 by immunohistocytochemistry. One Hodgkin lymphoma patient sample expressed MDR1 by immunohistocytochemistry. Although loss of CD30 expression is a possible mode of BV resistance in ALCL in vitro models, this has not been confirmed in patients. MMAE resistance and MDR1 expression are possible modes of BV resistance for Hodgkin lymphoma both in vitro and in patients.
authors.affiliation: Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California. rchen@coh.org.
authors.forename: Robert
authors.initials: R
authors.lastname: Chen
authors.affiliation: Department of Cancer Biology, City of Hope, Duarte, California.
authors.forename: Jessie
authors.initials: J
authors.lastname: Hou
authors.affiliation: Department of Cancer Biology, City of Hope, Duarte, California.
authors.forename: Edward
authors.initials: E
authors.lastname: Newman
authors.affiliation: Department of Pathology, City of Hope, Duarte, California.
authors.forename: Young
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Cancer Biology, City of Hope, Duarte, California.
authors.forename: Cecile
authors.initials: C
authors.lastname: Donohue
authors.affiliation: Division of Biostatistics, Beckman Research Institute of City of Hope, Duarte, California.
authors.forename: Xueli
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California.
authors.forename: Sandra H
authors.initials: SH
authors.lastname: Thomas
authors.affiliation: Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, California.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Forman
authors.affiliation: Department of Cancer Biology, City of Hope, Duarte, California.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Kane
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D000079963:Brentuximab Vedotin; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006689:Hodgkin Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018796:Immunoconjugates / Q000494:pharmacology*; D007150:Immunohistochemistry; D020128:Inhibitory Concentration 50; D017730:Ki-1 Antigen / Q000235:genetics / Q000378:metabolism*; D017728:Lymphoma, Large-Cell, Anaplastic / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009842:Oligopeptides / Q000378:metabolism / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018796:Immunoconjugates; D017730:Ki-1 Antigen; D009842:Oligopeptides; D000079963:Brentuximab Vedotin; C495575:monomethyl auristatin E
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