A Human iPSC-based Cell Therapy for Canavan Disease

项目摘要 Canavan病(CD)是一种罕见的常染色体隐性遗传性神经发育障碍，影响儿童从 婴儿期。大多数患有婴儿期CD的儿童将在第一个月内死亡。 十年的生命。这种疾病既没有治愈的方法，也没有标准的治疗方法。Cd是由遗传因素引起的 天冬氨酸酯酶(ASPA)基因突变，该基因编码一种由 大脑中的少突胶质细胞。ASPA分解N-乙酰-天冬氨酸(NAA)，这是一种在体内的氨基酸衍生物 大脑。NAA的产生和分解的周期似乎对维持脑白质至关重要 大脑，由被髓鞘覆盖的神经纤维组成。CD的症状包括缺乏天冬氨酸氨基转移酶的活性， 脑内NAA积聚、髓鞘形成缺陷和海绵状变性(空泡化)。临床部 症状包括运动功能受损和智力低下。目前还没有批准的治疗方法 这种情况。因此，对CD的有效治疗存在明显的、未得到满足的医学需求。 人类诱导多能干细胞(HiPSC)技术的发展为 细胞疗法。在我们的初步研究中，我们使用HiPSC技术来生成CD患者的IPSC和 将这些IPSCs分化为神经前体细胞(CD INPC)。然后我们引入了一个有功能的aspa基因。 慢病毒转导CD-iNPC产生含ASPA的基因工程功能 INPC，称为ASPA iNPC。为了将aspa inpc细胞产品推向临床，我们开发了 符合良好制造规范(CGMP)的流程，以生产ASPA iNPC。由此产生的ASPA 在CD小鼠模型中，对三种不同CD患者产生的iNPC进行了有效性和安全性测试。 在被移植到CD小鼠的大脑中后，ASPA iNPC提供了持续的ASPA活性，导致 显著降低NAA水平，相当大程度上挽救了脑部海绵状变性和髓鞘缺陷， 并显著改善了移植CD小鼠的运动功能。重要的是，没有肿瘤形成或其他 在移植的小鼠中观察到不良反应。这些强大的临床前数据提供了强有力的理由 拟议的研究。 本研究的目的是建立一种基于HiPSC的CD细胞治疗方法。细胞产品有 在临床前研究中证明是持久和有效的，具有良好的安全性。我们建议 以下具体目标：目标1：开展支持IND的资质运行并进行最终制造 ASPA INPC细胞产品。目的2：进行明确的临床前疗效和安全性/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，以 建立给CD患者使用ASPA INPC细胞产品的安全性和可行性。这项研究可能 导致了CD的一种新的细胞疗法的发展，并证明了使用基于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.