Defective Integrator activity shapes the transcriptome of patients with multiple sclerosis: mouse model

While the role of the immune system in multiple sclerosis (MS) is widely acknowledged, our comprehension of the transcriptional foundations of this prevalent neurological disorder remains largely incomplete. Here, we conducted high-depth RNA sequencing on monocytes from a cohort of patients with MS to explore rare RNA species associated with the disease. In a subset of the patients, a markedly altered transcriptome was observed, coinciding with a decreased expression of the epigenetic silencer HP1α/CBX5. These patients also exhibited diminished expression of multiple genes encoding subunits of the Integrator complex. Alongside, there were clear indications of decreased Integrator activity, including impaired U snRNA and enhancer-RNA maturation/degradation and dysregulated RNA polymerase II (RNAPII) pause-release. Inactivation of Cbx5 in the mouse recapitulated the defects in Integrator activity and resulted in hypersensitivity to Experimental Autoimmune Encephalomyelitis (EAE). While these data implied an unexpected function for HP1α in regulating RNAPII pause-release, they also showed that the dysregulation of numerous genes in MS patients could be attributed to either the biased distribution of transcription toward the 5' end of genes or the heightened elongation and stability of enhancer RNAs. An impaired Integrator activity, which has been hinted at by mutations within Integrator subunits previously linked to an increased risk of MS, provides a well-defined mechanistic understanding of the transcriptional anomalies associated with the disease, while introducing new criteria to categorize MS patients. C57BL/6N-Atm1Brd Cbx5tm1a(EUCOMM)Wtsi/WtsiOrl strain inactivated for Cbx5 was received from the EUCOMM Consortium. Either control mice or Cbx5 KO mice were included or not in an experimental autoimmune encephalomyelitis (EAE) protocol, as indicated. For each mouse, CD4+ T cells were purified from spleenocytes using the Miltenyi kit 130-104-454. Total RNA was extracted by Trizol LS (Thermo Fisher Scientific, ref. 10296028), according to the manufacturer's protocol. Total RNA library preparation and sequencing were performed by Novogene Co., Ltd, as a lncRNA sequencing service, including lncRNA directional library preparation with rRNA depletion (Ribo-Zero Magnetic Kit), quantitation, pooling and Paired End 150 sequencing (30G raw data) on Illumina HiSeq 2500 platform.