SBIR Phase II: Formulation of a mRNA-Based Therapy for CTLN1 by Inverse Flash NanoPrecipitation

这个小企业创新研究（SBIR）二期项目的更广泛的影响/商业潜力将是通过使基因疗法的临床转化成为可能的新技术来增强美国和全球的医疗保健。生物技术尚未满足的主要需求之一是一种递送技术，使基因疗法能够到达体内所需的器官，而不会引起免疫反应。最近的进展为遗传病提供了许多潜在的治疗方法，但由于缺乏输送技术，它们在临床中的应用受到阻碍。该项目将为此目的验证一种新的非病毒平台，并将其应用于罕见疾病适应症的商业应用。该项目为尿素循环障碍患者提供了一种基因替代疗法，这些患者目前使用药物和饮食相结合的治疗方法，效果有限。他们的治疗方案需要每天服用40片，同时还要严格控制饮食，避免摄入过多的蛋白质。尽管如此，血氨升高仍会导致神经损伤和新生儿高死亡率。护理人员面临着监测饮食、补充剂和药物的重大负担。一种提供真正疾病矫正的基因替代疗法将对患者和护理人员产生重大影响。拟议中的项目将导致基因替代疗法的发展，这种疗法可以安全地反复给患有尿素循环障碍这类罕见疾病的患者服用。这些患者缺乏一种尿素循环酶，不能外源性输送。另一种治疗方法是以mRNA等核酸的形式传递指令，让体内的细胞制造缺失的酶。商业上已证实的方法——脂质纳米颗粒和病毒载体——由于毒性、免疫原性和剂量挑战，在这一适应症中失败。在这个项目中开发的平台提供了一种方法来克服这些限制，使用非病毒，聚合物-脂质杂交配方。该项目的范围包括尿素循环紊乱瓜氨酸血症I型啮齿动物模型的药理学和毒理学研究，这将产生支持进一步发展的临床前数据包。成功的项目执行将包括配方优化和疾病纠正（降低血氨水平）的临床前演示，并具有很强的安全性。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project will be to enhance American and global healthcare through new technologies enabling clinical translation of gene therapies. One of the major unmet needs within biotechnology is a delivery technology that enables gene therapies to reach the desired organ within the body without raising an immune response. Recent advances have unlocked many potential treatments for genetic diseases, but their use in the clinic is hampered by a lack of delivery technologies. This project will validate a new non-viral platform for this purpose and apply it to commercial use in a rare disease indication. The project provides a gene replacement therapy for Urea Cycle Disorder patients, who are currently treated using a combination of drugs and diet that shows limited effectiveness. Their regimens require up to 40 pills per day in combination with incredibly strict dietary control to avoid consuming too much protein. Even still, elevated blood ammonia results in neurological damage and high neonatal mortality. Caregivers face significant burdens of care to monitor diet, supplements, and medications. A gene replacement therapy that provides true disease correction would be transformational for patients and caregivers. The proposed project will result in the development of a gene replacement therapy that can be safely and repeatedly dosed to patients suffering from the class of rare diseases known as Urea Cycle Disorders. These patients lack an enzyme of the urea cycle that cannot be delivered exogenously. An alternative therapeutic approach is to deliver instructions, in the form of nucleic acids such as mRNA, for cells in the body to make the missing enzyme. The commercially proven methods of doing this – lipid nanoparticles and viral vectors – fail in this indication due to toxicity, immunogenicity, and dosing challenges. The platform developed in this project provides a means to overcome these limitations using a non-viral, polymer-lipid hybrid formulation. The scope of the project includes both pharmacology and toxicology studies in rodent models of the Urea Cycle Disorder Citrullinemia Type I, that will produce a pre-clinical data package supporting further development. Successful project execution will include formulation optimization and pre-clinical demonstration of disease correction (reduced blood ammonia levels) with a strong safety profile.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.