Development of CM-CS1 CAR Treg to Treat Amyotrophic Lateral Sclerosis (ALS)

摘要 肌萎缩侧索硬化症(ALS)患者由于进行性运动神经元丢失而发生致命瘫痪 在大脑和脊髓中。美国每年新增肌萎缩侧索硬化症病例5000多例， 典型的发病年龄在40至70岁之间。虽然SOD1是一种细胞质蛋白，但错误折叠的SOD1可以 分泌并形成胞外低聚物和聚集体。超氧化物歧化酶-1(MSOD1)突变 导致SOD1的错误折叠和聚集，并在部分家族性ALS病例中发现。然而， 在许多散发的ALS病例的脊髓样本中也发现了错误折叠的SOD1。监管 T细胞(Tregs)具有耐受性和抗炎功能，目前正作为细胞基础被研究 阻断自体炎症免疫细胞的治疗方法。ALS患者Tregs(CD4+CD25hiCD127lo)数量较多 患者与疾病进展较慢有关。我们已经开发出新的嵌合抗原受体 (CARS)识别聚合SOD1并触发Treg函数。通过这种方式，我们的目标是提供一个大型 针对疾病相关蛋白的Tregs数量，并将在 折叠错误，聚合的SOD1。我们进一步加强了汽车树的活动，通过改造它们来 产生BNDF，一种关键的神经元生存因子。我们通过育种开发了一种新的ALS小鼠模型 将G93A SOD1转基因到NSG小鼠背景中，建立mSOD1-NSG小鼠。这些老鼠允许 植入人体细胞后会发展成一种进行性疾病，导致脊椎发炎 类似于肌萎缩侧索硬化症的脊髓和肢体瘫痪。该项目的目的是进行支持IND的研究 这是将这种疗法转化为临床所需的。

SUMMARY Amyotrophic lateral sclerosis (ALS) patients develop fatal paralysis as a result of progressive motor neuron loss in the brain and spinal cord. There are more than 5,000 new cases of ALS per year in the United States, with typical age of onset between 40 and 70 years of age. While SOD1 is a cytoplasmic protein, misfolded SOD1 can be secreted and form extracellular oligomers and aggregates. Mutations in superoxide dismutase-1 (mSOD1) result in misfolding and aggregation of SOD1 and are found in a subset of familial ALS cases. However, misfolded SOD1 has also been identified in spinal cord samples from many sporadic cases of ALS. Regulatory T cells (Tregs) have tolerogenic and anti-inflammatory functions and are being pursued as cell-based therapeutics to block auto-inflammatory immune cells. Higher numbers of Tregs (CD4+CD25hiCD127lo) in ALS patients are associated with a slower disease progression. We have developed novel chimeric antigen receptors (CARs) that recognize aggregated SOD1 and trigger Treg function. In this manner, we aim to provide a large number of Tregs that are specific for a disease-associated protein and will become activated at the site of misfolded, aggregated SOD1. We have further enhanced the activity of CAR Tregs by engineering them to produce BNDF, a key neuronal survival factor. We have developed a novel mouse model for ALS by breeding the G93A SOD1 transgene onto the NSG mouse background to create mSOD1-NSG mice. These mice allow the engraftment of human cells and they develop a progressive disease resulting in inflammation in the spinal cord and limb paralysis that mimic findings in ALS. The aim of this project is to perform IND-enabling studies required for translation of this therapy into the clinic.