Abstract P2-16-04: MNX1 promotes tumor sensitivity to HER2-targeted therapy in HER2-positive breast cancer

Abstract

Abstract Background: Female breast cancer has been the world’s leading cancer incidence among women. HER2-positive breast cancer accounts for about 15%~20% of all breast cancer, with a high degree of malignancy and easy metastasis and recurrence. With the development of medical technology and the continuous innovation of HER2 targeting drugs, patients with HER2-positive breast cancer have more treatment options and their prognosis had been greatly improved. However, there is still a lack of biomarkers for HER2-positive early breast cancer in clinical practice. Methods: This study prospectively collected single-center (Fudan University Shanghai Cancer Center, FUSCC) preoperative core needle biopsy samples of breast cancer patients from July 2017 to July 2018 who received neoadjuvant paclitaxel, carboplatin plus with trastuzumab (PCH). The patients were divided into pCR and non-PCR groups, and 81 patients were enrolled. The differential expression of genes was screened and validated by RNA-seq. The gene expression data for GSE181574 (this database has a total of 105 cases of breast cancer tissue samples were collected by core needle biopsy before neoadjuvant treatment. 52 cases underwent Ado-trastuzumab emtansine plus pertuzumab, 9 cases of paclitaxel plus trastuzumab, and 44 cases of paclitaxel plus trastuzumab and pertuzumab), GSE52707 (SK-BR-3 Lapatinib resistance cell group vs control group), and GSE15043 (BT474 Herceptin-resistant cell group vs control group), were downloaded from the Gene Expression Omnibus (GEO) database. Cell cloning formation, proliferation assay, and drug sensitivity experiments were conducted in MNX1 ectopic and knockdown cell lines. Co-IP assay, RNA-Seq, and ChIP-Seq analysis were used to explore the downstream pathways that MNX1 might be involved. Results: High-throughput sequencing results of core needle biopsy samples from 81 HER2-positive breast cancer patients were divided into pCR and non-pCR groups. Using DEseq2 packet analysis to screen differentially expressed genes, P-value < 0.05, | log2FoldChange | > 1 as the filter, there were 620 up-regulated genes and 715 down-regulated genes in the pCR group. The same method was used for the analysis of the GSE181574 data set. Combined the two datasets used P values and log2FoldChange (P-value < 0.05, | log2FoldChange | > 1.2) as selection criteria, we found the MNX1 and PNMT expressions were significantly higher in the pCR group. Subsequently, differential analysis of GSE52707 and GSE15043 datasets showed that MNX1 and PNMT expression levels were reduced in drug-resistant cell lines. Considering the P-value and | log2FoldChange |, we ultimately selected motor neuron and pancreatic homeobox 1(MNX1) gene as the target factors for follow-up studies. In the cloning formation and proliferation assay, overexpression of MNX1 inhibits proliferation and clonal formation of HER2-positive breast cancer cells. In the drug sensitivity experiments overexpression of MNX1 enhances the sensitivity of HER2-positive breast cancer cells to tyrosine kinase inhibitors (TKI) such as lapatinib and pyrotinib. MNX1 knockdown reduces the sensitivity of HER2-positive breast cancer cells to TKI. Through RNA-seq and ChIP-seq, we found that CD-M6PR might be a downstream target gene regulated by MNX1. Verification by Dual-Luciferase Reporter Assay MNX1 was found to positively regulate CD-M6PR transcription. By Co-IP assay, it was found that MNX1 interacted with EEF1D, and EEF1D stabilized MNX1. MNX1 and CD-M6PR affect the proliferation and drug sensitivity of breast cancer cells by down-regulating the PI3K-AKT-mTOR pathway. Conclusion: Overexpression of MNX1 increases the sensitivity of HER-2 positive cells to TKI. MNX1 binds to EEF1D and is stabilized by EEF1D. MNX1 positively regulates M6PR and affects the proliferation of breast cancer cells and the sensitivity of breast cancer cells to TKI through the PI3K-AKT-mTOR pathway. Citation Format: Wei-Ru Chi, Bingqiu Xiu, Min Xiong, Pei Li, Qi Zhang, Lun Li, Jianjing Hou, Xujie Zhou, Yuting Sang, Ming Chen, Liyi Zhang, Jingyan Xue, Yayun Chi, Jiong Wu. MNX1 promotes tumor sensitivity to HER2-targeted therapy in HER2-positive breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-16-04.