Endogenous PAD4 in Breast Cancer Cells Mediates Cancer Extracellular Chromatin Network Formation and Promotes Lung Metastasis

• Published in: Molecular cancer research Vol. 18; no. 5; pp. 735 - 747
• Main Authors: Shi, Lai, Yao, Huanling, Liu, Zheng, Xu, Ming, Tsung, Allan, Wang, Yanming
• Format: Journal Article
• Language: English
• Published: United States 01.05.2020
• Subjects: Animals; Apoptosis; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cell Proliferation; Chromatin - genetics; Chromatin - metabolism; Extracellular Traps; Female; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms - genetics; Lung Neoplasms - metabolism; Lung Neoplasms - secondary; Mice; Mice, Inbred BALB C; Mice, Knockout; Mice, Nude; Prognosis; Protein-Arginine Deiminase Type 4 - genetics; Protein-Arginine Deiminase Type 4 - metabolism; Tumor Cells, Cultured; Xenograft Model Antitumor Assays
• DOI: 10.1158/1541-7786.MCR-19-0018
• ISSN: 1541-7786; 1557-3125

Peptidyl arginine deiminase 4 (PAD4/PADI4) is a posttranslational modification enzyme that converts protein arginine or mono-methylarginine to citrulline. The PAD4-mediated hypercitrullination reaction in neutrophils causes the release of nuclear chromatin to form a chromatin network termed neutrophil extracellular traps (NET). NETs were first described as antimicrobial fibers that bind and kill bacteria. However, it is not known whether PAD4 can mediate the release of chromatin DNA into the extracellular space of cancer cells. Here, we report that murine breast cancer 4T1 cells expressing high levels of PADI4 can release cancer extracellular chromatin networks (CECN) and . Deletion of using CRISPR/Cas9 abolished CECN formation in 4T1 cells. deletion from 4T1 cells also reduced the rate of tumor growth in an allograft model, and decreased lung metastasis by 4T1 breast cancers. DNase I treatment, which degrades extracellular DNA including CECNs, also reduced breast to lung metastasis of wild-type 4T1 cells in allograft experiments in the -knockout mice. We further demonstrated that DNase I treatment in this mouse model did not alter circulating tumor cells but decreased metastasis through steps after intravasation. Taken together, our genetic studies show that PAD4 plays a cell autonomous role in cancer metastasis, thus revealing a novel strategy for preventing cancer metastasis by inhibiting cancer cell endogenous PAD4. IMPLICATIONS: This study shows that PADI4 can mediate the formation of CECNs in 4T1 cells, and that endogenous PADI4 plays an essential role in breast cancer lung metastasis. VISUAL OVERVIEW: http://mcr.aacrjournals.org/content/molcanres/18/5/735/F1.large.jpg.