Luzp1 Is An Understudied Transcriptional Regulator In Triple Negative Breast Cancer (Tnbc).

Abstract Introduction: TNBC is associated with poorer prognosis compared to other types of breast cancer due to poorly understood aggressive metastatic behavior. Proteomic profiling prioritized Leucine-Zipper Protein 1 (LUZP1) as an understudied TNBC-specific protein from an analysis of breast cancer patient-derived xenografts (PDXs) [PMC5379071]. Approach: RNA-seq data and isobaric tags for relative and absolute quantitation (iTRAQ) data from 16 PDXs were used to identify differentially expressed genes in basal-like and claudin-low breast cancers, compared to luminal and HER2-enriched subtypes. Multiple datasets (DepMap, METABRIC, and TCGA) were used to validate the enrichment of LUZP1 in TNBC. T-test was applied to identify differential genes/proteins. Pearson correlation analysis was used to identify RNAs that are correlated with LUZP1 protein level in CTPAC proteogenomic clinical sample data [PMC5102256 and Cell in press]. Gene Set Enrichment Analysis (GSEA) was used to identify enriched pathways. Cell fractionation was used to determine the subcellular expression of LUZP1. qPCR was used to validate differential expression observed in RNA-seq. Results: Leucine-Zipper Protein 1 (LUZP1) was differentially elevated in basal-like and claudin-low PDX models versus luminal and HER2-enriched at the mRNA and protein levels across 16 PDXs. Furthermore, LUZP1 mRNA was significantly enriched in TNBCs, relative to luminal and HER2-enriched subtypes, in breast cancer cell lines (data from DepMap), and human breast tumors (data from METABRIC and TCGA). LUZP1 has three putative nuclear localization signals and three leucine-zipper motifs, thought to mediate its protein dimerization and DNA-binding properties. Fractionation of TNBC cells showed that LUZP1 was predominately present in the nucleus. RNA-sequencing of control and MDA-MB-231 LUZP1 knockdown cells revealed that several extracellular matrix (ECM)-related pathways and epithelial-to-mesenchymal transition (EMT) were regulated by LUZP1. In particular, expression of migration, invasion, and metastasis genes such as matrix metalloproteinases (MMPs), Bone Morphogenetic Protein 2 (BMP2), and Transforming Growth Factor Beta 2 (TGFB2) were found to be LUZP1 regulated. Additionally, in CPTAC breast cancer proteogenomic data, LUZP1-correlated proteins and transcripts were enriched in ECM-related pathways and EMT, confirming the pathophysiological relevance of the cell-line knock-down data. Conclusion: LUZP1 is a novel TNBC-enriched protein in breast cancer PDXs, cell lines, and human tumors data. Gene silencing experiments demonstrated that LUZP1 transcriptionally regulates metastasis-related genes, and thus may drive a transcriptional program responsible for invasion, metastasis and EMT in TNBC. Functional data will be presented. Citation Format: Junkai Wang, Doug Chan, Beom-Jun Kim, Gloria V. Echeverria, Eric Jaehnig, Bing Zhang, Matthew J. Ellis. LUZP1 is an understudied transcriptional regulator in triple negative breast cancer (TNBC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2877.