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FAS-antisense 1 lncRNA and production of soluble versus membrane Fas in B-cell lymphoma

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Abstract

Impaired Fas-mediated apoptosis is associated with poor clinical outcomes and cancer chemoresistance. Soluble Fas receptor (sFas), produced by skipping of exon 6, inhibits apoptosis by sequestering Fas ligand. Serum sFas is associated with poor prognosis of non-Hodgkin’s lymphomas. We found that the alternative splicing of Fas in lymphomas is tightly regulated by a long-noncoding RNA corresponding to an antisense transcript of Fas (FAS-AS1). Levels of FAS-AS1 correlate inversely with production of sFas, and FAS-AS1 binding to the RBM5 inhibits RBM5-mediated exon 6 skipping. EZH2, often mutated or overexpressed in lymphomas, hyper-methylates the FAS-AS1 promoter and represses the FAS-AS1 expression. EZH2-mediated repression of FAS-AS1 promoter can be released by DZNeP (3-Deazaneplanocin A) or overcome by ectopic expression of FAS-AS1, both of which increase levels of FAS-AS1 and correspondingly decrease expression of sFas. Treatment with Bruton’s tyrosine kinase inhibitor or EZH2 knockdown decreases the levels of EZH2, RBM5 and sFas, thereby enhancing Fas-mediated apoptosis. This is the first report showing functional regulation of Fas repression by its antisense RNA. Our results reveal new therapeutic targets in lymphomas and provide a rationale for the use of EZH2 inhibitors or ibrutinib in combination with chemotherapeutic agents that recruit Fas for effective cell killing.

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Acknowledgements

This work was supported by Grants from the NCI/NIH(CA158692), NIDDK(DK091490), LLS(R6187-09), ACS(MSRG-10-052-01-LIB), the Richard Spencer Lewis Memorial Foundation and patients’ families. MDACC Flow Cytometry and Cellular Imaging Facility are funded by a Cancer Center Support Grant from the NCI (P30C16672). The Lymphoma Tissue Bank and the DNA Analysis Core Facility are supported by The University of Texas MD Anderson Cancer Center Support Grant CA16672 (National Institutes of Health). FAS-AS1 plasmids and plasmid encoding RBM5 and FAS-WT were provided by Suan Chuang, Taiwan and Juan Valcarcel, Spain, respectively. Since the time of submission of the manuscript to Leukemia, the FDA has approved the use of ibrutinib for relapsed mantle cell lymphoma and relapsed chronic lymphocytic leukemia, further underscoring the importance of analyzing apoptosis mechanisms that would apply to small lymphocytic lymphoma and other B-cell lymphomas.

Author contributions

LS designed, performed the majority of the experiments and analyzed the data. RM, FKB and JW performed some of the experiments. FS, SN and LWK reviewed work, provided critical suggestions and tumor specimens. LS, ZB and FS contributed to the research design, data analysis and writing of the manuscript.

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Correspondence to F Samaniego.

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Sehgal, L., Mathur, R., Braun, F. et al. FAS-antisense 1 lncRNA and production of soluble versus membrane Fas in B-cell lymphoma. Leukemia 28, 2376–2387 (2014). https://doi.org/10.1038/leu.2014.126

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