title: Quantitative Proteomic Verification of Membrane Proteins as Potential Therapeutic Targets Located in the 11q13 Amplicon in Cancers.
abstract: Tumor types can be defined cytologically by their regions of chromosomal amplification, which often results in the high expression of both mRNA and proteins of certain genes contained within the amplicon. An important strategy for defining therapeutically relevant targets in these situations is to ascertain which genes are amplified at the protein level and, concomitantly, are key drivers for tumor growth or maintenance. Furthermore, so-called passenger genes that are amplified with driver genes and a manifest on the cell surface can be attractive targets for an antibody-drug conjugate approach (ADC). We employed a tandem mass spectrometry proteomics approach using tumor cell lines to identify the cell surface proteins whose expression correlates with the 11q13 amplicon. The 11q13 amplicon is one of the most frequently amplified chromosomal regions in human cancer, being present in 45% of head and neck and oral squamous cell carcinoma (OSCC) and 13-21% of breast and liver carcinomas. Using a panel of tumor cell lines with defined 11q13 genomic amplification, we identified the membrane proteins that are differentially expressed in an 11q13 amplified cell line panel using membrane-enriched proteomic profiling. We found that DSG3, CD109, and CD14 were differentially overexpressed in head and neck and breast tumor cells with 11q13 amplification. The level of protein expression of each gene was confirmed by Western blot and FACS analysis. Because proteins with high cell surface expression on selected tumor cells could be potential antibody drug conjugate targets, we tested DSG3 and CD109 in antibody piggyback assays and validated that DSG3 and CD109 expression was sufficient to induce antibody internalization and cell killing in 11q13-amplified cell lines. Our results suggest that proteomic profiling using genetically stratified tumors can identify candidate antibody drug conjugate targets. Data are available via ProteomeXchange with the identifier PXD002486.
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Heather
authors.initials: H
authors.lastname: Hoover
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Jason
authors.initials: J
authors.lastname: Marchese
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Christopher
authors.initials: C
authors.lastname: Rothwell
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Jason
authors.initials: J
authors.lastname: Borawoski
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Jeffery
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: L Alex
authors.initials: LA
authors.lastname: Gaither
authors.affiliation: Novartis Institutes for Biomedical Research , Cambridge, Massachusetts 02139, United States.
authors.forename: Nancy
authors.initials: N
authors.lastname: Finkel
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D002880:Chromosomes, Human, Pair 11; D006258:Head and Neck Neoplasms / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D040901:Proteomics; D053719:Tandem Mass Spectrometry
chemical_list: D008565:Membrane Proteins
keywords: antibody-drug conjugate; head and neck carcinoma; mass spectrometry; tandem mass tag