CircSMARCA5 Regulates VEGFA mRNA Splicing and Angiogenesis in Glioblastoma Multiforme Through the Binding of SRSF1

Cancers - Tập 11 Số 2 - Trang 194
Davide Barbagallo1, A Caponnetto1, Duilia Brex1, Federica Mirabella1, Cristina Barbagallo1, Giovanni Lauretta1, Antonio Morrone2, Francesco Certo3, Giuseppe Broggi2, Rosario Caltabiano2, Giuseppe Barbagallo3, Vittoria Spina‐Purrello4, Marco Ragusa5, Cinzia Di Pietro1, Thomas B. Hansen6, Michele Purrello1
1Department of Biomedical and Biotechnological Sciences—Section of Biology and Genetics “Giovanni Sichel”, University of Catania, 95123 Catania, Italy
2Department of Medical, Surgical and Advanced Technological Sciences “G.F. Ingrassia”, University of Catania, 95123 Catania, Italy
3Multidisciplinary Research Center on Brain Tumors Diagnosis and Therapy, University of Catania, 95123 Catania, Italy
4Department of Biomedical and Biotechnological Sciences—Section of Medical Biochemistry, University of Catania, 95123 Catania, Italy
5Department of Biomedical and Biotechnological Sciences—Section of Biology and Genetics “Giovanni Sichel”, University of Catania, 95123 Catania, Italy; Oasi Research Institute-IRCCS, 94018 Troina, Italy
6Department of Molecular Biology and Genetics (MBG), Aarhus University, 8000 Aarhus C, Denmark

Tóm tắt

Circular RNAs are a large group of RNAs whose cellular functions are still being investigated. We recently proposed that circSMARCA5 acts as sponge for the splicing factor Serine and Arginine Rich Splicing Factor 1 (SRSF1) in glioblastoma multiforme (GBM). After demonstrating by RNA immunoprecipitation a physical interaction between SRFS1 and circSMARCA5, we assayed by real-time PCR in a cohort of 31 GBM biopsies and 20 unaffected brain parenchyma controls (UC) the expression of total, pro-angiogenic (Iso8a) and anti-angiogenic (Iso8b) mRNA isoforms of Vascular Endothelial Growth Factor A (VEGFA), a known splicing target of SRSF1. The Iso8a to Iso8b ratio: (i) increased in GBM biopsies with respect to UC (p-value < 0.00001); (ii) negatively correlated with the expression of circSMARCA5 (r-value = −0.46, p-value = 0.006); (iii) decreased in U87-MG overexpressing circSMARCA5 with respect to negative control (p-value = 0.0055). Blood vascular microvessel density, estimated within the same biopsies, negatively correlated with the expression of circSMARCA5 (r-value = −0.59, p-value = 0.00001), while positively correlated with that of SRSF1 (r-value = 0.38, p-value = 0.00663) and the Iso8a to Iso8b ratio (r-value = 0.41, p-value = 0.0259). Kaplan-Meier survival analysis showed that GBM patients with low circSMARCA5 expression had lower overall and progression free survival rates than those with higher circSMARCA5 expression (p-values = 0.033, 0.012, respectively). Our data convincingly suggest that circSMARCA5 is an upstream regulator of pro- to anti-angiogenic VEGFA isoforms ratio within GBM cells and a highly promising GBM prognostic and prospective anti-angiogenic molecule.

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Tài liệu tham khảo

Hansen, 2013, Natural RNA circles function as efficient microRNA sponges, Nature, 495, 384, 10.1038/nature11993

Memczak, 2013, Circular RNAs are a large class of animal RNAs with regulatory potency, Nature, 495, 333, 10.1038/nature11928

Ebbesen, 2017, Insights into circular RNA biology, RNA Biol., 14, 1035, 10.1080/15476286.2016.1271524

Zhang, 2018, Circular RNAs: Promising Biomarkers for Human Diseases, EBioMedicine, 34, 267, 10.1016/j.ebiom.2018.07.036

Dahl, 2018, Enzyme-free digital counting of endogenous circular RNA molecules in B-cell malignancies, Lab. Invest., 98, 1657, 10.1038/s41374-018-0108-6

Kristensen, 2018, Circular RNAs in cancer: Opportunities and challenges in the field, Oncogene, 37, 555, 10.1038/onc.2017.361

Dragomir, 2018, Circular RNAs in Cancer—Lessons Learned From microRNAs, Front. Oncol., 8, 179, 10.3389/fonc.2018.00179

Floris, 2017, Regulatory Role of Circular RNAs and Neurological Disorders, Mol. Neurobiol., 54, 5156, 10.1007/s12035-016-0055-4

Altesha, M.A., Ni, T., Khan, A., Liu, K., and Zheng, X. (2018). Circular RNA in cardiovascular disease. J. Cell Physiol.

Barbagallo, 2016, Dysregulated miR-671-5p/CDR1-AS/CDR1/VSNL1 axis is involved in glioblastoma multiforme, Oncotarget, 7, 4746, 10.18632/oncotarget.6621

Pajares, 2007, Alternative splicing: An emerging topic in molecular and clinical oncology, Lancet Oncol., 8, 349, 10.1016/S1470-2045(07)70104-3

Wang, B.D., and Lee, N.H. (2018). Aberrant RNA Splicing in Cancer and Drug Resistance. Cancers (Basel), 10.

Montes, 2019, RNA Splicing and Disease: Animal Models to Therapies, Trends Genet., 35, 68, 10.1016/j.tig.2018.10.002

Ostrom, 2017, CBTRUS Statistical Report: Primary brain and other central nervous system tumors diagnosed in the United States in 2010-2014, Neuro-Oncology, 19, v1, 10.1093/neuonc/nox158

Eger, 2018, Circular RNA Splicing, Adv. Exp. Med. Biol., 1087, 41, 10.1007/978-981-13-1426-1_4

Qin, 2018, Circ-UBR5: An exonic circular RNA and novel small nuclear RNA involved in RNA splicing, Biochem. Biophys. Res. Commun., 503, 1027, 10.1016/j.bbrc.2018.06.112

Meyer, 2014, circRNA biogenesis competes with pre-mRNA splicing, Mol. Cell, 56, 55, 10.1016/j.molcel.2014.08.019

Conn, 2015, The RNA binding protein quaking regulates formation of circRNAs, Cell, 160, 1125, 10.1016/j.cell.2015.02.014

Barbagallo, D., Caponnetto, A., Cirnigliaro, M., Brex, D., Barbagallo, C., D’Angeli, F., Morrone, A., Caltabiano, R., Barbagallo, G.M., and Ragusa, M. (2018). CircSMARCA5 Inhibits Migration of Glioblastoma Multiforme Cells by Regulating a Molecular Axis Involving Splicing Factors SRSF1/SRSF3/PTB. Int. J. Mol. Sci., 19.

Das, 2014, Emerging functions of SRSF1, splicing factor and oncoprotein, in RNA metabolism and cancer, Mol. Cancer Res., 12, 1195, 10.1158/1541-7786.MCR-14-0131

Karni, 2007, The gene encoding the splicing factor SF2/ASF is a proto-oncogene, Nat. Struct. Mol. Biol., 14, 185, 10.1038/nsmb1209

Ratnadiwakara, M., Archer, S.K., Dent, C.I., Ruiz De Los Mozos, I., Beilharz, T.H., Knaupp, A.S., Nefzger, C.M., Polo, J.M., and Anko, M.L. (2018). SRSF3 promotes pluripotency through Nanog mRNA export and coordination of the pluripotency gene expression program. Elife, 7.

Madhavan, 2009, Rembrandt: helping personalized medicine become a reality through integrative translational research, Mol. Cancer Res., 7, 157, 10.1158/1541-7786.MCR-08-0435

Tomczak, 2015, The Cancer Genome Atlas (TCGA): an immeasurable source of knowledge, Contemp. Oncol. (Pozn), 19, A68

Pratt, 2016, Robust transcriptome-wide discovery of RNA-binding protein binding sites with enhanced CLIP (eCLIP), Nat. Methods, 13, 508, 10.1038/nmeth.3810

Zang, J., Lu, D., and Xu, A. (2018). The interaction of circRNAs and RNA binding proteins: An important part of circRNA maintenance and function. J. Neurosci. Res.

Charley, 2014, Sponging of cellular proteins by viral RNAs, Curr. Opin. Virol., 9, 14, 10.1016/j.coviro.2014.09.001

Barbagallo, 2018, LncRNA UCA1, Upregulated in CRC Biopsies and Downregulated in Serum Exosomes, Controls mRNA Expression by RNA-RNA Interactions, Mol. Ther. Nucleic Acids, 12, 229, 10.1016/j.omtn.2018.05.009

Kim, 2016, LncRNA OIP5-AS1/cyrano sponges RNA-binding protein HuR, Nucleic Acids Res., 44, 2378, 10.1093/nar/gkw017

Anczukow, 2015, SRSF1-Regulated Alternative Splicing in Breast Cancer, Mol. Cell., 60, 105, 10.1016/j.molcel.2015.09.005

Ezponda, 2018, The oncogenic RNA-binding protein SRSF1 regulates LIG1 in non-small cell lung cancer, Lab. Invest., 98, 1562, 10.1038/s41374-018-0128-2

Amin, 2011, WT1 mutants reveal SRPK1 to be a downstream angiogenesis target by altering VEGF splicing, Cancer Cell, 20, 768, 10.1016/j.ccr.2011.10.016

Mavrou, 2015, Serine-arginine protein kinase 1 (SRPK1) inhibition as a potential novel targeted therapeutic strategy in prostate cancer, Oncogene, 34, 4311, 10.1038/onc.2014.360

Ye, 2016, Altered ratios of pro- and anti-angiogenic VEGF-A variants and pericyte expression of DLL4 disrupt vascular maturation in infantile haemangioma, J. Pathol., 239, 139, 10.1002/path.4715

Amin, 2015, Alternative splicing of TIA-1 in human colon cancer regulates VEGF isoform expression, angiogenesis, tumour growth and bevacizumab resistance, Mol. Oncol., 9, 167, 10.1016/j.molonc.2014.07.017

Gilbert, 2014, A randomized trial of bevacizumab for newly diagnosed glioblastoma, N. Engl. J. Med., 370, 699, 10.1056/NEJMoa1308573

Holdt, 2018, Circular RNAs as Therapeutic Agents and Targets, Front. Physiol., 9, 1262, 10.3389/fphys.2018.01262

Fukuhara, 2006, Utilization of host SR protein kinases and RNA-splicing machinery during viral replication, Proc. Natl. Acad. Sci. USA, 103, 11329, 10.1073/pnas.0604616103

Peritz, 2006, Immunoprecipitation of mRNA-protein complexes, Nat. Protoc., 1, 577, 10.1038/nprot.2006.82

Vento, 2010, Molecular profiling of human oocytes after vitrification strongly suggests that they are biologically comparable with freshly isolated gametes, Fertil. Steril., 94, 2804, 10.1016/j.fertnstert.2010.04.060

Livak, 2001, Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method, Methods, 25, 402, 10.1006/meth.2001.1262

Urbano, 2016, Altered expression of uncoupling protein 2 in GLP-1-producing cells after chronic high glucose exposure: Implications for the pathogenesis of diabetes mellitus, Am. J. Physiol. Cell Physiol., 310, C558, 10.1152/ajpcell.00148.2015

Caltabiano, 2016, ADAM 10 expression in primary uveal melanoma as prognostic factor for risk of metastasis, Pathol. Res. Pract., 212, 980, 10.1016/j.prp.2016.08.003

Weidner, 1991, Tumor angiogenesis and metastasis—Correlation in invasive breast carcinoma, N. Engl. J. Med., 324, 1, 10.1056/NEJM199101033240101

Mikkelsen, V.E., Stensjoen, A.L., Granli, U.S., Berntsen, E.M., Salvesen, O., Solheim, O., and Torp, S.H. (2018). Angiogenesis and radiological tumor growth in patients with glioblastoma. BMC Cancer, 18.

Burton, 2012, Assessment of mitotic rate reporting in melanoma, Am. J. Surg., 204, 969, 10.1016/j.amjsurg.2012.05.021

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