A Human iPSC-based Cell Therapy for Canavan Disease

项目摘要 卡纳万病（CD）是一种罕见的常染色体隐性遗传神经发育障碍， 婴儿期。大多数儿童与发病CD，最普遍的形式的疾病，将死亡的第一个 十年的生活。这种疾病既没有治愈方法，也没有标准治疗方法。CD是由遗传 乙酰化酶（ASPA）基因中的突变，该基因编码通过以下途径合成的代谢酶： 大脑中的少突胶质细胞ASPA分解N-乙酰基-天冬氨酸（NAA），一种在 个脑袋NAA的产生和分解周期似乎对于维持白色物质至关重要 大脑由髓鞘覆盖的神经纤维组成。CD的适应症包括缺乏ASPA活性， NAA的积累、髓鞘形成缺陷和脑中的海绵状变性（空泡化）。临床 症状包括运动功能受损和智力迟钝。目前尚无获批的治疗方法 这个条件。因此，CD的有效治疗存在明确的未满足的医学需求。 人类诱导多能干细胞（hiPSC）技术的发展为人类的免疫治疗开辟了令人兴奋的途径。 细胞疗法在我们的初步研究中，我们已经使用hiPSC技术来产生CD患者iPSC， 将这些iPSC分化成神经祖细胞（CD iNPC）。然后我们引入了一个功能性ASPA基因 通过慢病毒转导进入CD iNPC，以产生基因工程化的功能性含ASPA的 iNPC，称为ASPA iNPC。为了将ASPA iNPC细胞产品推向临床，我们开发了电流 生产ASPA iNPC的药品生产质量管理规范（cGMP）兼容工艺。结果ASPA 在CD小鼠模型中测试从三个不同CD患者产生的iNPC的功效和安全性。 在移植到CD小鼠的脑中后，ASPA iNPC提供持续的ASPA活性，导致 显著降低NAA水平，显著挽救脑中的海绵变性和髓鞘形成缺陷， 并显著改善移植CD小鼠的运动功能。重要的是，没有肿瘤发生或其他 在移植小鼠中观察到副作用。这些可靠的临床前数据为以下方面提供了有力的依据： 建议的研究。 本研究的目的是建立一种基于hiPSC的CD细胞疗法。细胞产品具有 在临床前研究中被证明是持久有效的，具有良好的安全性。我们建议 以下具体目标：目标1：进行IND启用确认运行并进行最终生产 ASPA iNPC细胞产品。目的2：进行明确的临床前有效性和安全性/致瘤性研究 ASPA iNPC细胞产品。目标3：获得IND批准。目标4：进行I期临床试验， 确定向CD患者施用ASPA iNPC细胞产品的安全性和可行性。这项研究可能 导致CD的新型细胞疗法的发展，并证明使用基于hiPSC的细胞的可行性 用于治疗类似疾病的产品。

Project Summary Canavan disease (CD) is a rare, autosomal recessive neurodevelopmental disorder that affects children from infancy. Most children with infantile-onset CD, the most prevalent form of the disease, will die within the first decade of life. There is neither a cure nor a standard treatment for this disease. CD is caused by genetic mutations in the aspartoacylase (ASPA) gene, which encodes a metabolic enzyme synthesized by oligodendrocytes in the brain. ASPA breaks down N-acetyl-aspartate (NAA), an amino acid derivative in the brain. The cycle of production and breakdown of NAA appears to be critical for maintaining the white matter of the brain, which consists of nerve fibers covered by myelin. Indications of CD include lack of ASPA activity, accumulation of NAA, myelination defect, and spongy degeneration (vacuolation) in the brain. The clinical symptoms include impaired motor function and mental retardation. There is currently no approved therapy for this condition. Therefore, there is a clear, unmet medical need for an effective therapy for CD. The development of human induced pluripotent stem cell (hiPSC) technology has opened exciting avenues for cell therapy. In our preliminary studies, we have used hiPSC technology to generate CD patient iPSCs and differentiated these iPSCs into neural progenitor cells (CD iNPCs). We then introduced a functional ASPA gene into CD iNPCs through lentiviral transduction to generate genetically engineered functional ASPA-containing iNPCs, termed ASPA iNPCs. In order to move the ASPA iNPC cell product to the clinic, we developed current Good Manufacturing Practice (cGMP)-compatible process to manufacture the ASPA iNPCs. The resultant ASPA iNPCs generated from three different CD patients were tested in a CD mouse model for efficacy and safety. After being transplanted into brains of CD mice, the ASPA iNPCs provided sustained ASPA activity, led to significantly lower NAA level, considerable rescue of spongy degeneration and myelination defect in the brain, and substantially improved motor function in the transplanted CD mice. Importantly, no tumorigenesis or other adverse effect was observed in the transplanted mice. These robust preclinical data provide strong rationale for the proposed study. The object of the proposed research is to establish a hiPSC-based cell therapy for CD. The cell products have proven in preclinical studies to be long lasting and efficacious with a favorable safety profile. We propose the following Specific Aims: Aim 1: To conduct IND-enabling qualification runs and perform final manufacturing of the ASPA iNPC cell products. Aim 2: To perform definitive preclinical efficacy and safety/tumorigenicity studies of the ASPA iNPC cell products. Aim 3: To obtain IND approval. Aim 4: To conduct a phase I clinical trial to establish the safety and feasibility of administering the ASPA iNPC cell products to CD patients. This study could lead to the development of a novel cell therapy for CD and demonstrate the feasibility of using hiPSC-based cell products for the treatment of similar diseases.