title: Development of human serine protease-based therapeutics targeting Fn14 and identification of Fn14 as a new target overexpressed in TNBC.
abstract: The cytokine TWEAK and its receptor, Fn14, have emerged as potentially valuable targets for cancer therapy. Granzyme B (GrB)-containing Fn14-targeted constructs were generated containing either the Fn14 ligand TWEAK (GrB-TWEAK) or an anti-Fn14 humanized single-chain antibody (GrB-Fc-IT4) as the targeting moieties. Both constructs showed high affinity and selective cytotoxicity against a panel of Fn14-expressing human tumor cells including triple-negative breast cancer (TNBC) lines. Cellular expression of the GrB inhibitor PI-9 in target cells had no impact on the cytotoxic effect of either construct. Cellular expression of MDR1 showed no cross-resistance to the fusion constructs. GrB-TWEAK and GrB-Fc-IT4 activated intracellular caspase cascades and cytochrome c-related proapoptotic pathways consistent with the known intracellular functions of GrB in target cells. Treatment of mice bearing established HT-29 xenografts with GrB-TWEAK showed significant tumor growth inhibition compared with vehicle alone (P < 0.05). Both GrB-TWEAK and GrB-Fc-IT4 displayed significant tumor growth inhibition when administered to mice bearing orthotopic MDA-MB-231 (TNBC) tumor xenografts. The Cancer Genome Atlas analysis revealed that Fn14 mRNA expression was significantly higher in TNBC and in HER2-positive disease (P < 0.0001) compared with hormone receptor-positive breast cancer, and in basal-like 2 tumors (P = 0.01) compared with other TNBC molecular subtypes. IHC analysis of a 101 patient TNBC tumor microarray showed that 55 of 101 (54%) of tumors stained positive for Fn14, suggesting that this may be an excellent potential target for precision therapeutic approaches. Targeting Fn14 using fully human, GrB-containing fusion constructs may form the basis for a new class of novel, potent, and highly effective constructs for targeted therapeutic applications.
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas.
authors.forename: Khalid A
authors.initials: KA
authors.lastname: Mohamedali
authors.affiliation: Department of Breast Medical Oncology, MD Anderson Cancer Center, Houston, Texas. Department of Systems Biology, MD Anderson Cancer Center, Houston, Texas.
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Gonzalez-Angulo
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas.
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Surgery, Center for Vascular and Inflammatory Diseases, and the Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland.
authors.forename: Mary
authors.initials: M
authors.lastname: Migliorini
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas.
authors.forename: Lawrence H
authors.initials: LH
authors.lastname: Cheung
authors.affiliation: Integrated Cancer Genomics Division, Translational Genomics Research Institute, Phoenix, Arizona.
authors.forename: Janine
authors.initials: J
authors.lastname: LoBello
authors.affiliation: Department of Biostatistics, MD Anderson Cancer Center, Houston, Texas.
authors.forename: Xiudong
authors.initials: X
authors.lastname: Lei
authors.affiliation: Department of Bioinformatics, MD Anderson Cancer Center, Houston, Texas.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qi
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas.
authors.forename: Walter N
authors.initials: WN
authors.lastname: Hittelman
authors.affiliation: Department of Surgery, Center for Vascular and Inflammatory Diseases, and the Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Winkles
authors.affiliation: Cancer and Cell Biology Division, Translational Genomics Research Institute, Phoenix, Arizona.
authors.forename: Nhan L
authors.initials: NL
authors.lastname: Tran
authors.affiliation: Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, Texas. mrosenbl@mdanderson.org.
authors.forename: Michael G
authors.initials: MG
authors.lastname: Rosenblum
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D005260:Female; D053804:Granzymes / Q000235:genetics / Q000494:pharmacology; D057809:HEK293 Cells; D019073:HT29 Cells; D006801:Humans; D019169:Jurkat Cells; D061986:MCF-7 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D058990:Molecular Targeted Therapy; D018124:Receptors, Tumor Necrosis Factor / Q000096:biosynthesis / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D000074050:TWEAK Receptor; D064726:Triple Negative Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D023041:Xenograft Model Antitumor Assays
chemical_list: D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C000613297:TNFRSF12A protein, human; D000074050:TWEAK Receptor; C000613299:Tnfrsf12a protein, mouse; D053804:Granzymes