Abstract | BACKGROUND: METHODS: Pre-chemotherapeutic needle biopsy of 143 invasive ductal carcinomas undergoing the same chemotherapeutic regimen was subjected to proteomic analysis. Four different machine learning algorithms were employed to determine signature protein combinations. Immunoreactive markers were selected using three common candidate proteins from the machine-learning algorithms and verified by immunohistochemistry using 123 cases of independent needle biopsy FFPE samples. The regulation of chemotherapeutic response and necroptotic cell death was assessed using lentiviral YARS overexpression and depletion 3D spheroid formation assay, viability assays, LDH release assay, flow cytometry analysis, and transmission electron microscopy. The ROS-induced metabolic dysregulation and phosphorylation of necrosome complex by YARS were assessed using oxygen consumption rate analysis, flow cytometry analysis, and 3D cell viability assay. The therapeutic roles of SMAC mimetics ( LCL161) and a pan-BCL2 inhibitor (ABT-263) were determined by 3D cell viability assay and flow cytometry analysis. Additional biologic process and protein- protein interaction pathway analysis were performed using Gene Ontology annotation and Cytoscape databases. RESULTS: YARS was selected as a potential biomarker by proteomics-based machine-learning algorithms and was exclusively associated with good response to chemotherapy by subsequent immunohistochemical validation. In 3D spheroid models of breast cancer cell lines, YARS overexpression significantly improved chemotherapy response via phosphorylation of the necrosome complex. YARS-induced necroptosis sequentially mediated mitochondrial dysfunction through the overproduction of ROS in breast cancer cell lines. Combination treatment with necroptosis-inducing small molecules, including a SMAC mimetic ( LCL161) and a pan-BCL2 inhibitor (ABT-263), showed therapeutic efficacy in YARS-overexpressing breast cancer cells. CONCLUSIONS: Our results indicate that, before chemotherapy, an initial screening of YARS protein expression should be performed, and YARS-positive breast cancer patients might consider the combined treatment with LCL161 and ABT-263; this could be a novel stepwise clinical approach to apply new targeted therapy in breast cancer patients in the future.
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Authors | Kyung-Min Lee, Hyebin Lee, Dohyun Han, Woo Kyung Moon, Kwangsoo Kim, Hyeon Jeong Oh, Jinwoo Choi, Eun Hye Hwang, Seong Eun Kang, Seock-Ah Im, Kyung-Hun Lee, Han Suk Ryu |
Journal | Breast cancer research : BCR
(Breast Cancer Res)
Vol. 22
Issue 1
Pg. 130
(11 25 2020)
ISSN: 1465-542X [Electronic] England |
PMID | 33239070
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Aniline Compounds
- Apoptosis Regulatory Proteins
- BCL2 protein, human
- DIABLO protein, human
- LCL161
- Mitochondrial Proteins
- Proto-Oncogene Proteins c-bcl-2
- Sulfonamides
- Thiazoles
- Tyrosine-tRNA Ligase
- navitoclax
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Topics |
- Aniline Compounds
(pharmacology, therapeutic use)
- Antineoplastic Combined Chemotherapy Protocols
(pharmacology, therapeutic use)
- Apoptosis Regulatory Proteins
(agonists, metabolism)
- Biopsy
- Breast
(pathology)
- Breast Neoplasms
(pathology, therapy)
- Carcinoma, Ductal, Breast
(pathology, therapy)
- Cell Line, Tumor
- Clinical Decision-Making
(methods)
- Drug Synergism
- Female
- Humans
- Mastectomy
- Mitochondrial Proteins
(agonists, metabolism)
- Necroptosis
(drug effects)
- Neoadjuvant Therapy
(methods)
- Proto-Oncogene Proteins c-bcl-2
(antagonists & inhibitors, metabolism)
- Sulfonamides
(pharmacology, therapeutic use)
- Thiazoles
(pharmacology, therapeutic use)
- Tyrosine-tRNA Ligase
(analysis, metabolism)
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