title: Anti-nicastrin monoclonal antibodies elicit pleiotropic anti-tumour pharmacological effects in invasive breast cancer cells.
abstract: The goal of targeted cancer therapies is to specifically block oncogenic signalling, thus maximising efficacy, while reducing side-effects to patients. The gamma-secretase (GS) complex is an attractive therapeutic target in haematological malignancies and solid tumours with major pharmaceutical activity to identify optimal inhibitors. Within GS, nicastrin (NCSTN) offers an opportunity for therapeutic intervention using blocking monoclonal antibodies (mAbs). Here we explore the role of anti-nicastrin monoclonal antibodies, which we have developed as specific, multi-faceted inhibitors of proliferation and invasive traits of triple-negative breast cancer cells. We use 3D in vitro proliferation and invasion assays as well as an orthotopic and tail vail injection triple-negative breast cancer in vivo xenograft model systems. RNAScope assessed nicastrin in patient samples. Anti-NCSTN mAb clone-2H6 demonstrated a superior anti-tumour efficacy than clone-10C11 and the RO4929097 small molecule GS inhibitor, acting by inhibiting GS enzymatic activity and Notch signalling in vitro and in vivo. Confirming clinical relevance of nicastrin as a target, we report evidence of increased NCSTN mRNA levels by RNA in situ hybridization (RNAScope) in a large cohort of oestrogen receptor negative breast cancers, conferring independent prognostic significance for disease-free survival, in multivariate analysis. We demonstrate here that targeting NCSTN using specific mAbs may represent a novel mode of treatment for invasive triple-negative breast cancer, for which there are few targeted therapeutic options. Furthermore, we propose that measuring NCSTN in patient samples using RNAScope technology may serve as companion diagnostic for anti-NCSTN therapy in the clinic.
authors.affiliation: Division of Surgery and Cancer, Department of Oncology, ICTEM Hammersmith Hospital Campus, Imperial College London, Du Cane Road, W12 0NN, London, UK, a.filipovic@imperial.ac.uk.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Filipović
authors.forename: Ylenia
authors.initials: Y
authors.lastname: Lombardo
authors.forename: Monica
authors.initials: M
authors.lastname: Faronato
authors.forename: Monica
authors.initials: M
authors.lastname: Fronato
authors.forename: Joel
authors.initials: J
authors.lastname: Abrahams
authors.forename: Eric
authors.initials: E
authors.lastname: Aboagye
authors.forename: Quang-De
authors.initials: QD
authors.lastname: Nguyen
authors.forename: Barbara Borda
authors.initials: BB
authors.lastname: d'Aqua
authors.forename: Anne
authors.initials: A
authors.lastname: Ridley
authors.forename: Andrew
authors.initials: A
authors.lastname: Green
authors.forename: Emad
authors.initials: E
authors.lastname: Rahka
authors.forename: Ian
authors.initials: I
authors.lastname: Ellis
authors.forename: Chiara
authors.initials: C
authors.lastname: Recchi
authors.forename: Natasa
authors.initials: N
authors.lastname: Przulj
authors.forename: Anida
authors.initials: A
authors.lastname: Sarajlić
authors.forename: Jean-Rene
authors.initials: JR
authors.lastname: Alattia
authors.forename: Patrick
authors.initials: P
authors.lastname: Fraering
authors.forename: Mahendra
authors.initials: M
authors.lastname: Deonarain
authors.forename: R Charles
authors.initials: RC
authors.lastname: Coombes
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D064726:Triple Negative Breast Neoplasms / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; C413604:nicastrin protein; D053829:Amyloid Precursor Protein Secretases