Regulation of NGF by macrophage in intervertebral discs

• Project/Area Number: 20K22984
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0906:Surgery related to the biological and sensory functions and related fields
• Research Institution: Kitasato University
• Principal Investigator: Nakawaki Mitsufumi 北里大学, 医学部, 助教 (00623175)
• Project Period (FY): 2020-09-11 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 神経成長因子 / 椎間板性腰痛 / TGF-β / マクロファージ / 椎間板

Outline of Research at the Start

マウス椎間板傷害モデル、KOマウスおよび臨床検体を駆使してマクロファージおよびマクロファージ産生サイトカインを介したNGF制御機構の解明と椎間板性腰痛メカニズムを明らかにする。TNF-alpha KOマウスおよびTGF-β阻害剤を用いて椎間板におけるNGFの制御を検討する。マクロファージ除去剤、ケモカイン受容体KOマウスを用いてマクロファージによるNGF制御機構を検討する。手術時に採取したヒト椎間板組織を用いたトランスレーショナルスタディを展開し、TNF-alpha, TGF-beta, M1, M2 marker, NGF発現と腰痛の関連性を検討する。

Outline of Final Research Achievements

We studied NGF regulation in a mouse model of IVD injury. We examined mRNA levels of Tnfa, Tgfb, and Ngf in IVDs from control and IVD-injured mice.We used magnetic cell separation to isolate macrophage-rich and IVD cell-richfractions from injured IVDs. We also examined Ngf expression in injured IVDs from C57BL/6 J and Tnfa-knockout (KO) mice. To study the effect of TGF-β on Ngf expression, C57/BL6J mice were given an intraperitoneal injection of TGF-β inhibitor, SB431542 one and two days before harvesting IVDs. Tnfa, Tgfb, and Ngf expression was significantly increased in injured IVDs. Tnfa was predominantly expressed in the CD11b ( +) fraction, and Tgfb in the CD11b (-) fraction. Ngf expression was comparable between CD11b ( +) and CD11b (-) fractions, and between wild-type and Tnfa-KO mice. SB431542 suppressed TGF-β-mediated Ngf expression and NGF production in vitro. Further, SB431542 significantly reduced Ngf expression in IVDs.TGF-β may regulate NGF expression in vivo.

Academic Significance and Societal Importance of the Research Achievements

腰痛の生涯罹患率は85％と報告され、超高齢者社会を迎えた我が国における腰痛患者は
2,800万人にものぼる。腰痛による経済損失は年間7,000億円と推定されている。本研究で異化はNGFおよびNGF制御因子を標的とした腰痛治療の開発に役立つものと考えられる。

Research Products

• [Journal Article] TGF-β regulates nerve growth factor expression in a mouse intervertebral disc injury model (2021)
  • Author(s): Yokozeki Y, Uchida K, Kawakubo A, Nakawaki M, Okubo T, Miaygi M, Inoue G, Itakura M, Sekiguchi H, Takaso M
  • Journal Title: BMC Musculoskelet Disord Volume: 22 Issue: 1 Pages: 634-634
  • DOI: 10.1186/s12891-021-04509-w
  • Peer Reviewed / Open Access
• [Journal Article] Role of CD14-positive cells in inflammatory cytokine and pain-related molecule expression in human degenerated intervertebral discs (2020)
  • Author(s): Miyagi M, Uchida K, Takano S, Nakawaki M, Sekiguchi H, Nakazawa T, Imura T, Saito W, Shirasawa E, Kawakubo A, Akazawa T, Inoue G, Takaso M
  • Journal Title: J Orthop Res Volume: - Issue: 8 Pages: 1755-1762
  • DOI: 10.1002/jor.24839
  • Peer Reviewed
• [Presentation] TNF-α は椎間板組織内のiNOS発現を制御している (2020)
  • Author(s): 中脇充章、 内田健太郎、川久保歩、宮城正行、大久保 直、横関雄司、関口裕之、井上 玄、齋藤 亘、高相晶士
  • Organizer: 第35回日本整形外科学会基礎学術集会