title: The Characterization of GSDMB Splicing and Backsplicing Profiles Identifies Novel Isoforms and a Circular RNA That Are Dysregulated in Multiple Sclerosis.
abstract: Abnormalities in alternative splicing (AS) are emerging as recurrent features in autoimmune diseases (AIDs). In particular, a growing body of evidence suggests the existence of a pathogenic association between a generalized defect in splicing regulatory genes and multiple sclerosis (MS). Moreover, several studies have documented an unbalance in alternatively-spliced isoforms in MS patients possibly contributing to the disease etiology. In this work, using a combination of PCR-based techniques (reverse-transcription (RT)-PCR, fluorescent-competitive, real-time, and digital RT-PCR assays), we investigated the alternatively-spliced gene encoding Gasdermin B,GSDMB, which was repeatedly associated with susceptibility to asthma and AIDs. The in-depth characterization ofGSDMBAS and backsplicing profiles led us to the identification of an exonic circular RNA (ecircRNA) as well as of novelGSDMBin-frame and out-of-frame isoforms. The non-productive splicing variants were shown to be downregulated by the nonsense-mediated mRNA decay (NMD) in human cell lines, suggesting thatGSDMBlevels are significantly modulated by NMD. Importantly, both AS isoforms and the identified ecircRNA were significantly dysregulated in peripheral blood mononuclear cells of relapsing-remitting MS patients compared to controls, further supporting the notion that aberrant RNA metabolism is a characteristic feature of the disease.
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. giulia.cardamone@st.hunimed.eu.
authors.forename: Giulia
authors.initials: G
authors.lastname: Cardamone
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. elvezia_maria.paraboschi@humanitasresearch.it.
authors.forename: Elvezia Maria
authors.initials: EM
authors.lastname: Paraboschi
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. valeria.rimoldi@humanitasresearch.it.|Humanitas Clinical and Research Center, Via Manzoni 56, 20089 Rozzano, Milan, Italy. valeria.rimoldi@humanitasresearch.it.
authors.forename: Valeria
authors.initials: V
authors.lastname: Rimoldi
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. stefano.duga@hunimed.eu.|Humanitas Clinical and Research Center, Via Manzoni 56, 20089 Rozzano, Milan, Italy. stefano.duga@hunimed.eu.
authors.forename: Stefano
authors.initials: S
authors.lastname: Duga
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. giulia.solda@hunimed.eu.|Humanitas Clinical and Research Center, Via Manzoni 56, 20089 Rozzano, Milan, Italy. giulia.solda@hunimed.eu.
authors.forename: Giulia
authors.initials: G
authors.lastname: Soldà
authors.affiliation: Department of Biomedical Sciences, Humanitas University, Via Manzoni 113, 20089 Rozzano, Milan, Italy. rosanna.asselta@hunimed.eu.|Humanitas Clinical and Research Center, Via Manzoni 56, 20089 Rozzano, Milan, Italy. rosanna.asselta@hunimed.eu.
authors.forename: Rosanna
authors.initials: R
authors.lastname: Asselta
mesh_terms: D017398:Alternative Splicing; D016022:Case-Control Studies; D005091:Exons; D005260:Female; D005786:Gene Expression Regulation; D023061:Gene Order; D006801:Humans; D008297:Male; D009103:Multiple Sclerosis / Q000097:blood / Q000235:genetics*; D020529:Multiple Sclerosis, Relapsing-Remitting / Q000097:blood / Q000235:genetics; D009363:Neoplasm Proteins / Q000235:genetics*; D059365:Nonsense Mediated mRNA Decay; D012313:RNA; D012326:RNA Splicing; D000079962:RNA, Circular; D000094524:Gasdermins
chemical_list: D009363:Neoplasm Proteins; D012313:RNA; D000079962:RNA, Circular; C527978:GSDMB protein, human; D000094524:Gasdermins
keywords: GSDMB; alternative splicing; circRNA; multiple sclerosis; nonsense-mediated mRNA decay