Epigenetic modulation of macrophage polarization prevents lumbar disc degeneration

Inflammation plays an essential role in the development of lumbar disc degeneration (LDD), although the exact effects of macrophage subtypes on LDD remain unclear. Based on previous studies, we hypothesized that M2-polarization of local macrophages and simultaneous suppression of their production of fibrotic transforming growth factor beta 1 (TGFβ1) could inhibit progression of LDD. Thus, we applied an orthotopic injection of adeno-associated virus (AAV) carrying shRNA for DNA Methyltransferase 1 (DNMT1) and/or shRNA for TGFβ1 under a macrophage-specific CD68 promoter to specifically target local macrophages in a mouse model for LDD. We found that shDNMT1 significantly reduced levels of the pro-inflammatory cytokines TNFα, IL-1β and IL-6, significantly increased levels of the anti-inflammatory cytokines IL-4 and IL-10, significantly increased M2 macrophage polarization, significantly reduced cell apoptosis in the disc degeneration zone and significantly reduced LDD-associated pain. The anti-apoptotic and anti-pain effects were further strengthened by co-application of shTGFβ1. Together, these data suggest that M2 polarization of macrophages induced by both epigenetic modulation and suppressed production and release of TGFβ1 from polarized M2 macrophages, may have a demonstrable therapeutic effect on LDD.

[1]  B. V. D. van der Kallen,et al.  Disc inflammation and Modic changes show an interaction effect on recovery after surgery for lumbar disc herniation , 2019, European Spine Journal.

[2]  M. Barbosa,et al.  The inflammatory response in the regression of lumbar disc herniation , 2018, Arthritis Research & Therapy.

[3]  Chunyang Meng,et al.  Changes of proteoglycan and collagen II of the adjacent intervertebral disc in the cervical instability models. , 2016, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[4]  Huidong Shi,et al.  Epigenetic regulation of macrophage polarization and inflammation by DNA methylation in obesity. , 2016, JCI insight.

[5]  Ming Wu,et al.  Differential expression of p38 MAPK α, β, γ, δ isoforms in nucleus pulposus modulates macrophage polarization in intervertebral disc degeneration , 2016, Scientific Reports.

[6]  Yue Zhou,et al.  Growth and Differentiation Factor-5 Contributes to the Structural and Functional Maintenance of the Intervertebral Disc , 2015, Cellular Physiology and Biochemistry.

[7]  J. Lu,et al.  Cell and molecular biology of intervertebral disc degeneration: current understanding and implications for potential therapeutic strategies , 2014, Cell proliferation.

[8]  D. Samartzis,et al.  Clarifying the nomenclature of intervertebral disc degeneration and displacement: from bench to bedside. , 2014, International journal of clinical and experimental pathology.

[9]  G. Natoli,et al.  Transcriptional regulation of macrophage polarization: enabling diversity with identity , 2011, Nature Reviews Immunology.

[10]  Liwei Wang,et al.  The Fas–FADD death domain complex structure reveals the basis of DISC assembly and disease mutations , 2010, Nature Structural &Molecular Biology.

[11]  Richard A. Flavell,et al.  The polarization of immune cells in the tumour environment by TGFβ , 2010, Nature Reviews Immunology.

[12]  L. Setton,et al.  Proinflammatory cytokine expression profile in degenerated and herniated human intervertebral disc tissues. , 2010, Arthritis and rheumatism.

[13]  J. Pollard.  Trophic macrophages in development and disease , 2009, Nature Reviews Immunology.

[14]  S. Gordon,et al.  Alternative activation of macrophages: an immunologic functional perspective. , 2009, Annual review of immunology.

[15]  H. Gruber,et al.  Senescence in Cells of the Aging and Degenerating Intervertebral Disc: Immunolocalization of Senescence-Associated &bgr;-Galactosidase in Human and Sand Rat Discs , 2007, Spine.

[16]  Rinat Abramovitch,et al.  VEGF-Induced Adult Neovascularization: Recruitment, Retention, and Role of Accessory Cells , 2006, Cell.

[17]  S. Gordon,et al.  Monocyte and macrophage heterogeneity , 2005, Nature Reviews Immunology.

[18]  C. Christiansen,et al.  Association between spine disc degeneration and type II collagen degradation in postmenopausal women: the OFELY study. , 2004, Arthritis and rheumatism.

[19]  S. Kokubun,et al.  Angiogenesis and Inflammatory Cell Infiltration in Lumbar Disc Herniation , 2003, Spine.

[20]  D. Greaves,et al.  Autocrine Deactivation of Macrophages in Transgenic Mice Constitutively Overexpressing IL-10 Under Control of the Human CD68 Promoter1 , 2002, The Journal of Immunology.

[21]  Jong-Beom Park,et al.  Expression of Fas Ligand and Apoptosis of Disc Cells in Herniated Lumbar Disc Tissue , 2001, Spine.

[22]  T. Shimizu,et al.  Roles of tumour necrosis factor‐alpha (TNF‐α), transforming growth factor‐beta (TGF‐β), and IL‐10 in the modulation of intercellular adhesion molecule‐1 (ICAM‐1) expression by macrophages during mycobacterial infection , 2000, Clinical and experimental immunology.

[23]  H. Gruber,et al.  Anti-Apoptotic Effects of IGF-1 and PDGF on Human Intervertebral Disc Cells In Vitro , 2000, Spine.

[24]  H. Iwata,et al.  The Involvement of Matrix Metalloproteinases and Inflammation in Lumbar Disc Herniation , 1998, Spine.

[25]  H E Gruber,et al.  Analysis of Aging and Degeneration of the Human Intervertebral Disc: Comparison of Surgical Specimens With Normal Controls , 1998, Spine.

[26]  H. Maeda,et al.  TGF-beta enhances macrophage ability to produce IL-10 in normal and tumor-bearing mice. , 1995, Journal of immunology.

[27]  H. Nick,et al.  Regulation of inducible nitric oxide synthase mRNA levels by LPS, INF-γ, TGF-β, and IL-10 in murine macrophage cell lines and rat peritoneal macrophages , 1994 .

[28]  R. Dubner,et al.  A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia , 1987, Pain.

[29]  J. Scott,et al.  Proteoglycan-collagen interactions in intervertebral disc. A chondroitin sulphate proteoglycan associates with collagen fibrils in rabbit annulus fibrosus at the d-e bands , 1986, Bioscience reports.

[30]  Henson Pm.  Mechanisms of exocytosis in phagocytic inflammatory cells. Parke-Davis Award Lecture. , 1980 .

[31]  P. Gerdhem,et al.  Markers of inflammation and fibrinolysis in relation to outcome after surgery for lumbar disc herniation. A prospective study on 177 patients , 2015, European Spine Journal.

[32]  I. Shapiro,et al.  Role of cytokines in intervertebral disc degeneration: pain and disc content , 2014, Nature Reviews Rheumatology.

[33]  H. Nick,et al.  Regulation of inducible nitric oxide synthase mRNA levels by LPS, INF-gamma, TGF-beta, and IL-10 in murine macrophage cell lines and rat peritoneal macrophages. , 1994, Biochemical and biophysical research communications.

[34]  H. Muir,et al.  Experimental intervertebral disc degeneration: morphologic and proteoglycan changes over time. , 1981, Arthritis and rheumatism.

[35]  P. Henson,et al.  Mechanisms of exocytosis in phagocytic inflammatory cells. Parke-Davis Award Lecture. , 1980, The American journal of pathology.