Next‐generation sequencing reveals a novel role of lysine‐specific demethylase 1 in adhesion of rhabdomyosarcoma cells. Issue 12 (19th December 2019)

• Record Type: Journal Article
• Authors: Haydn, Tinka; Kehr, Sarah; Willmann, Dominica; Metzger, Eric; Schüle, Roland; Fulda, Simone
• Is Part Of: International journal of cancer. Volume 146:Issue 12(2020)
• Page Start: 3435
• Page End: 3449
• Publication Date: 2019-12-19
• Subjects: LSD1 -- rhabdomyosarcoma -- cancer; Cancer -- Periodicals; Cancer -- Prevention -- Periodicals; 616.994
• DOI: 10.1002/ijc.32806
• Languages: English
• ISSNs: 0020-7136

Abstract

Abstract : Lysine‐specific demethylase 1 (LSD1), a histone lysine demethylase with the main specificity for H3K4me2, has been shown to be overexpressed in rhabdomyosarcoma (RMS) tumor samples. However, its role in RMS biology is not yet well understood. Here, we identified a new role of LSD1 in regulating adhesion of RMS cells. Genetic knockdown of LSD1 profoundly suppressed clonogenic growth in a panel of RMS cell lines, whereas LSD1 proved to be largely dispensable for regulating cell death and short‐term survival. Combined RNA and ChIP‐sequencing performed to analyze RNA expression and histone methylation at promoter regions revealed a gene set enrichment for adhesion‐associated terms upon LSD1 knockdown. Consistently, LSD1 knockdown significantly reduced adhesion to untreated surfaces. Importantly, precoating of the plates with the adhesives collagen I or fibronectin rescued this reduced adhesion of LSD1 knockdown cells back to levels of control cells. Using KEGG pathway analysis, we identified 17 differentially expressed genes (DEGs) in LSD1 knockdown cells related to adhesion processes, which were validated by qRT‐PCR. Combining RNA and ChIP‐sequencing results revealed that, within this set of genes, SPP1, C3AR1, ITGA10 and SERPINE1 also exhibited increased H3K4me2 levels at their promoter regions in LSD1 knockdown compared to control cells. Indeed, LSD1 ChIP experiments confirmed enrichment of LSD1 at their promoter regions, suggesting a direct transcriptional regulation by LSD1. By identifying a new role of LSD1 in the modulation of cell adhesion and clonogenic growth of RMS cells, these findings highlight the importance of LSD1 in RMS. Abstract : What's new? Rhabdomyosarcoma is a soft‐tissue tumor in children with poor outcome in advanced cases. Using various next‐generation sequencing methods, the authors identified a novel role of lysine‐specific demethylase (LSD1), an epigenetic regulator involved in stem cell maintenance and carcinogenesis, in rhabdomyosarcoma. LSD1 knockdown reduced clonogenic growth and adhesion of tumor cells, while being largely without effect for cell death and short‐term survival, underscoring the nuances of LSD1 action in rhabdomyosarcoma biology.