sc-rAAV8 via Lumbar Puncture as Spinal Analgesic Drug Delivery Systems

腰椎穿刺术是一种很有吸引力的神经系统新疗法。 因为它在患者身上是安全的；在床边进行；FDA批准用于传统药物。 然而，到目前为止，它在基因传递方面的应用一直是无效的，不符合安全原则，并且 未能瞄准神经元。我们已经开发出高效的基因转移到初级感官 背根神经节(DRGs)的神经元通过LP，即鞘内(IT)，利用自身互补 重组腺相关病毒血清8型(sc-rAAV8)。单次给药sc-rAAV8 表达镇痛基因前内啡肽原(pp？EP)或大鼠抗-EP的突变体 炎症基因白介素10(rIL10/F129S)可显著(P&lt；0.0001)缓解症状 在E3个月的大鼠慢性神经病理性疼痛模型中，这是一个重要的功能结果。这个 这一应用的假设是sc-rAAV8通过lp是一种可行的治疗难治性疾病的基因递送产品。 慢性疼痛可供专业研究中心外的临床医生使用。为了 为了调查/建立临床翻译的潜力，我们提出了以下具体目标： 目的1.在封闭与开放的IT空间中用体视学方法量化DRG的转导，表征 对IT载体的血清学免疫，并决定器官分布。目标2.确定 神经病理性疼痛模型中最强的候选治疗基因并确定是否存在 与传统疼痛治疗的药理协同作用。目的3.检测抗伤害感受剂 该方法在一种新的反映不治之症癌症患者疼痛的大鼠模型中的有效性 未来可能进行的临床试验-情景。目的4.检测IT sc-rAAV8是否有效地转导DRG 大型动物的神经元。意义和未来：慢性疼痛影响着5000万美国人 仅在美国就产生了1000亿美元/年的成本(如PA-07-282所述)。此应用程序 提出一种基因治疗方法，如果研究计划成功，该方法将成为候选方法 用于晚期恶性肿瘤患者的严重疼痛的临床开发。IT sc-rAAV8可能 作为机制验证试验的工具，例如验证作为治疗靶点的小胶质细胞激活 在人类身上，也可能成为治疗难治性慢性疼痛的新药。

Lumbar puncture (LP) is an attractive route for delivering novel therapies to the nervous system because it is safe in patients; performed at the bedside; and FDA-approved for conventional drugs. However, its use for gene delivery has to date been ineffective; has not met safety principles; and has failed to target neurons. We have developed highly effective gene transfer to the primary sensory neurons of the dorsal root ganglia (DRGs) via LP, i.e. intrathecally (IT), using self-complementary recombinant adeno-associated virus serotype 8 (sc-rAAV8). A single administration of sc-rAAV8 expressing the analgesic gene prepro-¿-endorphin (pp¿EP) or a mutant form of the rat anti- inflammatory gene interleukin-10 (rIL10/F129S) led to highly significant (p<0.0001) relief of symptoms for e3 months in a rat chronic neuropathic pain model, an important functional outcome. The hypothesis of this application is that sc-rAAV8 via LP is a viable gene delivery product for intractable chronic pain usable by clinicians outside of specialized research centers. In order to investigate/establish the potential for clinical translation, we propose the following Specific Aims: Aim 1. To quantify DRG transduction by stereology in the closed vs. open IT space, characterize serological immunity to the IT vector, and determine organ distribution. Aim 2. To identify the strongest candidate therapeutic gene in the neuropathic pain model and to define if there is pharmacological synergy with conventional pain treatments. Aim 3. To test the antinociceptive efficacy of the approach in a new rat model reflecting pain in patients with incurable cancer, a possible future clinical trial-scenario. Aim 4. To test if IT sc-rAAV8 effectively transduces DRG neurons in large animals. Significance & future perspective: Chronic pain affects 50 million Americans and incurs costs of >$100 billion/year in the US alone (as stated in PA-07-282). This application proposes a gene therapy approach, which, if the Research Plan succeeds, will become a candidate for clinical development in patients with severe pain from advanced malignancies. IT sc-rAAV8 may serve as a tool for proof-of-mechanism trials, e.g. to validate microglial activation as therapeutic target in humans, and may also become a new drug for otherwise intractable chronic pain.