Dataset Information

DAPK3 modulates migration and invasion of triple negative breast cancers

ABSTRACT: A mass spectrometry (MS)-based kinase inhibitor pulldown assay (KIPA) was applied to analyze 16 patient-derived xenografts (PDXs) leading to the discovery that Death-Associated Protein Kinase 3 (DAPK3) is significantly and specifically overexpressed in triple negative breast cancer (TNBC). Validation studies confirmed the enrichment of DAPK3 at the protein level, independent of RNA expression, in both TNBC cell lines and tumors. Genomic knockout of DAPK3 in TNBC cell lines inhibited in vitro migration and invasion, along with downregulation of an epithelial-mesenchymal transition (EMT) signature. The kinase and leucine-zipper domains within DAPK3 were shown by mutational analysis to be essential for functionality. Notably, DAPK3 was found to inhibit the levels of desmoplakin (DSP), a crucial component of the desmosome complex, thereby explaining TNBC migration and invasion effects. Further exploration with immunoprecipitation-mass spectrometry (IP-MS) identified Leucine-zipper protein 1 (LUZP1) as a strong binding partner of DAPK3, engaging in a leucine-zipper-domain-mediated interaction that protects DAPK3 from degradation. Thus, DAPK3 emerges as a novel regulator of EMT components in TNBC.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Breast Cancer Cell

DISEASE(S): Breast Cancer

SUBMITTER: Matthew Holt

LAB HEAD: Matthew V Holt

PROVIDER: PXD049187 | Pride | 2025-05-06

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	48653_1_ME480_DAPK3_IP_CONT_1_2_3_5_half.raw	Raw
	48653_1_ME480_DAPK3_IP_CONT_4_6_onefourth.raw	Raw
	48654_1_ME480_DAPK3_IP_D3_1_2_3_5_half.raw	Raw
	48654_1_ME480_DAPK3_IP_D3_4_6_onefourth.raw	Raw
	Junkai_2663_PTUM_WT_1_1.raw	Raw

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Publications

Death-associated protein kinase 3 modulates migration and invasion of triple-negative breast cancer cells.

Wang Junkai J Tran-Huynh Anh M AM Kim Beom-Jun BJ Chan Doug W DW Holt Matthew V MV Fandino Diana D Yu Xin X Qi Xiaoli X Wang Jin J Zhang Weijie W Wu Yi-Hsuan YH Anurag Meenakshi M Zhang Xiang H F XHF Zhang Bing B Cheng Chonghui C Foulds Charles E CE Ellis Matthew J MJ

PNAS nexus 20240912 9

Sixteen patient-derived xenografts (PDXs) were analyzed using a mass spectrometry (MS)-based kinase inhibitor pull-down assay (KIPA), leading to the observation that death-associated protein kinase 3 (DAPK3) is significantly and specifically overexpressed in the triple-negative breast cancer (TNBC) models. Validation studies confirmed enrichment of DAPK3 protein, in both TNBC cell lines and tumors, independent of mRNA levels. Genomic knockout of <i>DAPK3</i> in TNBC cell lines inhibited in vitro ...[more]

PMID: 39319326

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Project description:In this study we performed a systematic evaluation of functional miRNA-mRNA interactions associated with the aggressiveness of breast cancer cells using a combination of integrated miRNA and mRNA expression profiling, bioinformatics prediction, and functional assays. Analysis of the miRNA expression identified 11 miRNAs that were differentially expressed, including 7 down-regulated (miR-200c, miR-205, miR-203, miR-141, miR-34a, miR-183, and miR-375) and 4 up-regulated miRNAs (miR-146a, miR-138, miR-125b1 and miR-100), in aggressive cell lines when compared to normal and less aggressive cell lines. Transient overexpression of miR-200c, miR-205, and miR-375 in MDA-MB-231 cells led to the inhibition of cell migration and invasion. The integrated analysis of miRNA and mRNA expression identified 35 known and novel target genes of miR-200c, miR-205, and mir-375, including CFL2, LAMC1, TIMP2, ZEB1, CDH11, PRKCA, PTPRJ, PTPRM, LDHB, and SEC23A. Surprisingly, the majority of these genes (27 genes) were target genes of miR-200c, suggesting that it plays a more important role in regulating the aggressiveness of breast cancer cells. We characterized one of the target genes of miR-200c, CFL2, and demonstrated that CFL2 is overexpressed in aggressive breast cancer cell lines and can be significantly down-regulated by exogenous miR-200c. Tissue microarray analysis further revealed that CFL2 expression in primary breast cancer tissue correlated with tumor grade. To our knowledge, this study is the first systematic screening of functional miRNA target genes in aggressive breast cancer cells. The results obtained from this study may improve our understanding of the role of these candidate miRNAs and their target genes in relation to breast cancer aggressiveness and ultimately lead to the identification of novel biomarkers associated with prognosis. Affymetrix miRNA 1.0 arrays were performed according to the manufacturer's directions on small RNA extracted from 12 breast cancer cell lines including 4 aggressive cell lines, 6 less aggressive cell lines and 2 imortalized breast epithelium cell lines