Heparanome-Mediated Rescue of Oligodendrocyte Progenitor Quiescence Following Inflammatory Demyelination
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Research Articles: Neurobiology of Disease Heparanome-mediated rescue of oligodendrocyte progenitor quiescence following inflammatory demyelination https://doi.org/10.1523/JNEUROSCI.0580-20.2021 Cite as: J. Neurosci 2021; 10.1523/JNEUROSCI.0580-20.2021 Received: 10 March 2020 Revised: 3 December 2020 Accepted: 5 January 2021 This Early Release article has been peer-reviewed and accepted, but has not been through the composition and copyediting processes. The final version may differ slightly in style or formatting and will contain links to any extended data. Alerts: Sign up at www.jneurosci.org/alerts to receive customized email alerts when the fully formatted version of this article is published. Copyright © 2021 the authors PI-88 inhibits IFN-Jsignaling in OPCs 1 Heparanome-mediated rescue of oligodendrocyte progenitor quiescence following inflammatory 2 demyelination 3 4 Darpan Saraswat1, R. Ross Welliver1, Roopa Ravichandar1, Ajai Tripathi3, Jessie J. Polanco2, 5 Jacqueline Broome1, Edward Hurley4, Ranjan Dutta3, M Laura Feltri4, Fraser J. Sim1, 2 6 7 1Department of Pharmacology and Toxicology, Jacob’s School of Medicine and Biomedical Sciences, 8 University at Buffalo, Buffalo, NY, USA, 2Neuroscience Program, Jacob’s School of Medicine and 9 Biomedical Sciences, University at Buffalo, Buffalo, NY, USA, 3Department of Neuroscience, Cleveland 10 Clinic, Cleveland, OH, USA, 4Hunter James Kelly Research Institute, Departments of Biochemistry & 11 Neurology, School of Medicine and Biomedical Sciences, University at Buffalo, Buffalo, NY, USA. 12 13 Address correspondence to: 14 Fraser Sim, Ph.D. 15 Department of Pharmacology and Toxicology 16 Jacob’s School of Medicine and Biomedical Sciences 17 University at Buffalo 18 955 High Street 19 Buffalo, NY 14203 20 716-829-2151 21 [email protected] 22 23 Keywords: quiescence, human, glia, RNA-seq, heparanase inhibitor 24 Running title: PI-88 inhibits IFN-Jsignaling in OPCs 25 26 Abstract, 237 words; Introduction, 680 words; Result: 3371; Discussion, 1812 words. 27 28 Tables: 0; Figures: 9 29 30 Extended Data Figures: 11 31 Page 1 of 40 PI-88 inhibits IFN-Jsignaling in OPCs 32 Acknowledgements: This work was supported by NINDS grant (R01NS104021), NCATS 33 (UL1TR001412), National Multiple Sclerosis Society (RG-1701-26750), the Kalec Multiple Sclerosis 34 Foundation, the Change MS Foundation, the Skarlow Memorial Trust. JJP received additional support 35 from NIGMS (R25GM09545902), NCATS (UL1TR001412-S1), and New York State Department of 36 Health (Empire State Stem Cell Fund) through the Stem Cells in Regenerative Medicine Fellowship 37 (NYSTEM C30290GG). We acknowledge the assistance of the Confocal Microscope and Flow 38 Cytometry Facility in the School of Medicine and Biomedical Sciences, University at Buffalo. 39 40 Conflict of Interest: The authors declare no competing financial interests. 41 Page 2 of 40 PI-88 inhibits IFN-Jsignaling in OPCs 42 ABSTRACT 43 The proinflammatory cytokine IFN-J, which is chronically elevated in multiple sclerosis, induces 44 pathological quiescence in human oligodendrocyte progenitor cells (OPCs) via upregulation of the 45 transcription factor PRRX1. In this study using animals of both sexes, we investigated the role of 46 heparan sulfate proteoglycans in the modulation of IFN-J signaling following demyelination. We found 47 that IFN-J profoundly impaired OPC proliferation and recruitment following adult spinal cord 48 demyelination. IFN-J-induced quiescence was mediated by direct signaling in OPCs as conditional 49 genetic ablation of IFNJR1 (Ifngr1) in adult NG2+ OPCs completely abrogated these inhibitory effects. 50 Intriguingly, OPC-specific IFN-Jsignaling contributed to failed oligodendrocyte differentiation, which 51 was associated with hyperactive Wnt/Bmp target gene expression in OPCs. We found that PI-88, a 52 heparan sulfate mimetic, directly antagonized IFN-J to rescue human OPC proliferation and 53 differentiation in vitro and blocked the IFN-J mediated inhibitory effects on OPC recruitment in vivo. 54 Importantly, heparanase modulation by PI-88 or OGT2155 in demyelinated lesion rescued IFN-J 55 mediated axonal damage and demyelination. In addition to OPC-specific effects, IFN-J augmented 56 lesions were characterized by increased size, reactive astrogliosis and proinflammatory 57 microglial/macrophage activation along with exacerbated axonal injury and cell death. Heparanase 58 inhibitor treatment rescued many of the negative IFN-J-induced sequalae suggesting a profound 59 modulation of the lesion environment. Together, these results suggest that the modulation of the 60 heparanome represents a rational approach to mitigate the negative effects of proinflammatory 61 signaling and rescuing pathological quiescence in the inflamed and demyelinated human brain. 62 63 SIGNIFICANCE STATEMENT 64 The failure of remyelination in multiple sclerosis contributes to neurological dysfunction and 65 neurodegeneration. The activation and proliferation of oligodendrocyte progenitor cells (OPCs) is a 66 necessary step in the recruitment phase of remyelination. Here, we show that the proinflammatory 67 cytokine interferon-gamma directly acts on OPCs to induce pathological quiescence and thereby limit 68 recruitment following demyelination. Heparan sulfate is a highly structured sulfated carbohydrate 69 polymer that is present on the cell surface and regulates several aspects of the signaling 70 microenvironment. We find that pathological interferon-gamma can be blocked by modulation of the 71 heparanome following demyelination using either a heparan mimetic or by treatment with heparanase 72 inhibitor. These studies establish the potential for modulation of heparanome as a regenerative 73 approach in demyelinating disease. 74 Page 3 of 40 PI-88 inhibits IFN-Jsignaling in OPCs 75 INTRODUCTION 76 Multiple sclerosis (MS), an autoimmune disease of the central nervous system, is characterized 77 by demyelination, neurological dysfunction, and neurodegeneration (Lassmann, 2014). 78 Oligodendrocyte progenitor cells (OPCs) are vital for the endogenous regenerative process known as 79 remyelination that restores myelin and prevents ongoing neurodegeneration that typify progressive 80 forms of MS (Franklin and Ffrench-Constant, 2017). The failure of remyelination in MS has been largely 81 attributed to a failure of the generation of new oligodendrocytes. This is likely due to impaired 82 oligodendrocyte differentiation along with a combination of insufficient migration, death, or pathological 83 quiescence of OPCs. The inhibitory tissue environment is characterized by reactive astrogliosis and 84 astroglial scar formation (Ponath et al., 2018), as well as proinflammatory microglial/macrophage 85 infiltration (Lassmann et al., 2001; Wang et al., 2019). Together, the environment acts to limit the 86 recruitment of OPCs into demyelinated lesions and may thereby prevent proliferation, differentiation 87 and remyelination. 88 Interferon gamma (IFN-J) is a pleotropic pro-inflammatory cytokine implicated in MS 89 pathogenesis (Popko et al., 1997; Steinman, 2001). IFN-J induces microglial M1 polarization (Martinez 90 and Gordon, 2014), up-regulation of class I/II MHC, and increased expression of pro-inflammatory 91 cytokines. In MS patients, IFN-J levels are frequently elevated prior to onset of clinical symptoms (Beck 92 et al., 1988). Indeed, IFN-J treatment in MS patients resulted in severe disease exacerbation (Panitch 93 et al., 1987). However, the precise roles of IFN-J are more complex as studies in animals have also 94 implicated IFN-J in neuroprotection (reviewed in Arellano et al., 2015). IFN-J signals via the ubiquitously 95 expressed IFNJR1 that is coupled to the JAK/STAT-1 pathway and results in the up-regulation of 96 interferon regulatory factor (IRF-1). Interestingly, single nucleotide polymorphisms in IRF-1 are 97 associated with progressive MS (Fortunato et al., 2008) and IRF-1-expressing OL lineage cells are 98 present in MS lesions (Ren et al., 2011; Loda and Balabanov, 2012). In vitro high doses of IFN-J induce 99 OPC apoptosis (Vartanian et al., 1995; Baerwald and Popko, 1998; Horiuchi et al., 2006; Wang et al., 100 2010). However, at lower doses and in the absence of serum, IFN-J instead induces reversible cell- 101 cycle quiescence in rodent (Agresti et al., 1996; Tanner et al., 2011) and human OPCs (Wang et al., 102 2018). In vivo excessive IFN-J during CNS development induces hypomyelination (Corbin et al., 1996; 103 LaFerla et al., 2000) and demyelination (Horwitz et al., 1997). Following demyelination, IFN-J impairs 104 remyelination in both EAE and cuprizone mice models (Lin et al., 2006). Oligodendrocyte-specific 105 SOCS1 overexpression, a negative regulator of IFN-J receptor, antagonized the hypomyelinating 106 effects of IFN-J supporting a direct cell autonomous role following IFN-J signaling (Balabanov et al., 107 2006). Page 4 of 40 PI-88 inhibits IFN-Jsignaling in OPCs 108 Recently, we identified PRRX1 as a transcriptional regulator of pathological quiescence in 109 hOPCs that is upregulated by IFN-J and Bmp signaling (Wang et al., 2018). Moreover, IFN-J coinjection 110 with lysolecithin results in a model of proinflammatory focal demyelination that recapitulates PRRX1 111 upregulation and OPC quiescence (Wang et al., 2018). In this study, we further examined the role of 112 endogenous IFN-J signaling on OPCs and the potential for rescuing the negative effects of IFN-J by 113 modulating the local OPC heparanome that regulates remyelination