We targeted NMI based on our previous proteomics analysis using FFPE breast cancer samples with neoadjuvant settings and text-mining analytics to verify NMI as a target agent to overcome chemoresistance through in vitro and in vivo studies.
Quantitative proteomics was analyzed for three different breast cancer cell lines with NMI gene silencing. We performed computation pathway enrichment based on domain knowledge for text-mining and bioinformatics tools to identify critical pathways related to chemotherapeutic sensitivity. A total of 8 breast cancer cell lines with or without endogenous chemoresistance were enrolled to define the important pathways and molecules in determining chemosensitivity through cell-titer glo assay, FACS, 3D spheroid and invasion assay, ROS assay by DCFDA and Mitotracker. Interaction network analyses were investigated to define the signaling pathway landscapes with public network databases and bioinformatic network evaluations. To verify the chemosensitive roles of NMI in vivo setting, we are conducting animal tests and immunostaining in human breast cancer samples where the patients received neoadjuvant chemotherapy.
A total of 972 were confirmed to be significantly altered proteins after NMI gene silencing. A vast number of cell cycle-related proteins, which were downregulated considerably in NMI suppressed group led us to verify NMI's biological function on chemosensitivity through molecular biology-driven assays. Cell-titer glo assay and FACS revealed significantly induced cytotoxicity and apoptosis in both hormone receptor positive and negative groups without NMI gene after treatment with three different chemotherapeutic agents. The 3D-spheroid assay demonstrated a reduced spheroid formation in the case group. DCFDA and Mitotracker assay revealed increased intracellular and intramitochondrial ROS levels. We built biological network models based on in-silico and biology-driven assays. Currently, we are conducting in vivo validation using an animal test model and human samples.
Our biological evidence for NMI can provide novel insights to overcome chemo-resistance in breast cancer.
Clinical trial identification
Legal entity responsible for the study
Han Suk Ryu.
This research was suppoorted by a grant of the Korea Health technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea.
All authors have declared no conflicts of interest.