Inhibition of TNF-alpha Signaling to Reduce Intervertebral Disc Inflammation

摘要 与椎间盘（IVD）退变相关的慢性背痛是一个重大问题， 仅在美国就有数十亿。我们的建议将探索导致椎间盘退变的因素的新方面 和炎症，从而加速对IVD变性和相关背痛的研究。 TNF-α诱导蛋白8（TNFAIP 8，称为TIPE）家族由四个成员（TNFAIP 8和TIPE）组成。 技巧1 -3）。它们是最近发现的新型细胞质蛋白，是调节炎症的关键因子。我们 初步数据显示TNFAIP 8家族蛋白调节巨噬细胞浸润到损伤的IVD中。 我们提出的前提是TNFAIP 8家族成员可能是IVD炎症的关键调节因子， 一种新的肿瘤坏死因子受体（TNFR）-1的小分子抑制剂可能代表一种治疗方法， 减少炎症的工具。 在目标1中，我们假设TNFAIP 8/TIPE 2的基因失活导致了炎症的减少， 受伤的老鼠尾巴IVD。我们将确定TNFAIP 8/TIPE 2的基因失活是否会改善IVD 炎症和变性。我们已经获得并正在维持突变小鼠的群体， TNFAIP 8家族的两个成员（TNFAIP 8-/-、TIPE 2-/-和TNFAIP 8/TIPE 2双敲除）。小鼠尾部 IVD将受到针刺损伤的挑战。巨噬细胞浸润和细胞因子基因的变化 将在突变小鼠之间以及与它们的野生型同窝对照比较损伤后的表达。 在目标2中，我们进一步假设一种新的TNFR 1小分子抑制剂（SGT 11）可以 代表了一种减轻炎症的治疗工具。小分子特别有吸引力，因为它们可以 被修改用于口服药物递送。SGT 11已被证明通过改变构象来抑制炎症 TNFR1。与目前抑制TNFR 1和TNFR 2活化的抗TNF治疗不同，SGT 11抑制TNFR 1和TNFR 2活化。 TNFR 1，而不是TNFR 2。如果成功，这将是第一个调节IVD炎症的小分子。我们将 确定SGT 11是否可以改善IVD炎症并降低野生型中TNFAIP 8/TIPE 2的表达， 老鼠.将用针刺穿小鼠尾部IVD，伴或不伴SGT 11处理。变化 IVD中的巨噬细胞浸润、细胞因子和TNFAIP 8家族基因表达将在 SGT 11处理组和对照组。 这项探索性研究开辟了一个新的方向，因为调节TNF-TNFAIP 8 以前没有在IVD中研究过遗传方法和小分子抑制剂对axia的影响。发现 调节IVD炎症的药物将导致从目前的侵入性手术到晚期的范式转变。 将疾病分期为早期非侵入性治疗，从而使患者受益。

Abstract Chronic back pain related to intervertebral disc (IVD) degeneration is a significant problem, costing billions in the U.S. alone. Our proposal will explore new aspects of the factors that lead to disc degeneration and inflammation, thus accelerating research on IVD degeneration and related back pain. The TNF-α-induced protein-8 (TNFAIP8, known as TIPE) family consists of four members (TNFAIP8 and TIPE1-3). They are novel cytoplasmic proteins recently found to be key factors regulating inflammation. Our preliminary data have shown that TNFAIP8 family proteins regulate macrophage infiltration into the injured IVD. The premise of our proposal is that TNFAIP8 family members may be key regulators in IVD inflammation, and that a novel small molecule inhibitor of tumor necrosis factor receptor (TNFR)-1 could represent a therapeutic tool to reduce inflammation. In Aim 1, we hypothesize that genetic inactivation of TNFAIP8/TIPE2 results in reduced inflammation in the injured mouse tail IVD. We will determine if genetic inactivation of TNFAIP8/TIPE2 will ameliorate IVD inflammation and degeneration. We have acquired and are currently maintaining colonies of mutant mice lacking two members of the TNFAIP8 family (TNFAIP8-/-, TIPE2-/-, and TNFAIP8/TIPE2 double knockout). Mouse tail IVDs will be challenged with a needle puncture injury. Changes in macrophage infiltration and cytokine gene expression post-injury will be compared among the mutant mice and with their wild type littermate controls. In Aim 2, we further hypothesize that a novel small molecule inhibitor of TNFR1 (SGT11) could represent a therapeutic tool to reduce inflammation. Small molecules are especially attractive since they could be modified for oral drug delivery. SGT11 has been shown to inhibit inflammation by changing the conformation of TNFR1. Unlike current anti-TNF therapy that inhibits both TNFR1 and TNFR2 activation, SGT11 inhibits TNFR1 but not TNFR2. If successful, this will be the first small molecule to modulate IVD inflammation. We will determine if SGT11 could ameliorate IVD inflammation and reduce TNFAIP8/TIPE2 expression in the wild-type mouse. Mouse tail IVDs will be punctured with a needle, with or without SGT11 treatment. Changes in macrophage infiltration, cytokine and TNFAIP8 family gene expression in the IVD will be compared between SGT11-treated and control groups. The proposed exploratory study breaks ground toward a novel direction since modulating TNF-TNFAIP8 axia with genetic methods and small molecule inhibitors has not been previously studied in the IVD. Discovery of drugs that modulate IVD inflammation would result in a paradigm shift from current invasive surgeries on late stage disease to early noninvasive therapies, thus benefiting patients.