Gene Therapy Clinical Candidate Development for Carbamoyl Phosphate Synthetase Deficiency

氨基甲酰磷酸合成酶缺乏症的基因治疗临床候选药物开发

基本信息

批准号：
10339836
负责人：
Gerald S Lipshutz
金额：
$ 38.92万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10339836
关键词：
Address Affect Ammonia Animal Model Anorexia Biotechnology Birth Brain Carbamyl Phosphate Cerebral Edema Clinical Clinical Protocols Codon Nucleotides Coma Complementary DNA Cryopreservation DNA cassette Data Defect Dependovirus Development Diet Disease Disease model Dose Drug Metabolic Detoxication Encephalopathies Enhancers Enzymatic Biochemistry Enzymes Gene Delivery Gene Transduction Agent Genes Genetic Genetic Diseases Genome Glutamine Goals Hepatic Hepatocyte Hereditary Disease Human Hybrids Hyperammonemia Hyperargininemia Hypoventilation Impairment Incidence Infant Knockout Mice Laboratories Lethargies Life Ligase Liver Mammals Mediating Medical Mendelian disorder Metabolic Pathway Mission Mitochondrial Proteins Morbidity - disease rate Mus Mutation N acetyl L glutamate Neonatal Neonatal Mortality Nervous System Trauma Neurologic Nitrogen Organ Pathway interactions Patients Periodicity Phase Phenotype Plasma Posture Pre-Clinical Model Prevention strategy Production Proteins Public Health Publishing Regulator Genes Regulatory Element Research Risk Seizures Serotyping Site Survivors Technology Testing Therapeutic Transgenes Transgenic Mice Transplantation United States National Institutes of Health Urea cycle disorders Vectorial capacity Viral Vector Vulnerable Populations adeno-associated viral vector base clinical application clinical candidate clinical development clinically translatable cost critical period disability effective therapy enzyme deficiency experience gene product gene replacement gene therapy genetic approach high risk homologous recombination human disease humanized mouse immunosuppressed in vivo liver transplantation liver xenograft model development mortality mortality risk mouse model natural hypothermia neonate novel novel therapeutic intervention novel therapeutics postnatal development pre-clinical pre-clinical therapy programs promoter severe intellectual disability treatment strategy urea cycle vector wasting

Address Affect Ammonia Animal Model Anorexia Biotechnology Birth Brain Carbamyl Phosphate Cerebral Edema Clinical Clinical Protocols Codon Nucleotides Coma Complementary DNA Cryopreservation DNA cassette Data Defect Dependovirus Development Diet Disease Disease model Dose Drug Metabolic Detoxication Encephalopathies Enhancers Enzymatic Biochemistry Enzymes Gene Delivery Gene Transduction Agent Genes Genetic Genetic Diseases Genome Glutamine Goals Hepatic Hepatocyte Hereditary Disease Human Hybrids Hyperammonemia Hyperargininemia Hypoventilation Impairment Incidence Infant Knockout Mice Laboratories Lethargies Life Ligase Liver Mammals Mediating Medical Mendelian disorder Metabolic Pathway Mission Mitochondrial Proteins Morbidity - disease rate Mus Mutation N acetyl L glutamate Neonatal Neonatal Mortality Nervous System Trauma Neurologic Nitrogen Organ Pathway interactions Patients Periodicity Phase Phenotype Plasma Posture Pre-Clinical Model Prevention strategy Production Proteins Public Health Publishing Regulator Genes Regulatory Element Research Risk Seizures Serotyping Site Survivors Technology Testing Therapeutic Transgenes Transgenic Mice Transplantation United States National Institutes of Health Urea cycle disorders Vectorial capacity Viral Vector Vulnerable Populations adeno-associated viral vector base clinical application clinical candidate clinical development clinically translatable cost critical period disability effective therapy enzyme deficiency experience gene product gene replacement gene therapy genetic approach high risk homologous recombination human disease humanized mouse immunosuppressed in vivo liver transplantation liver xenograft model development mortality mortality risk mouse model natural hypothermia neonate novel novel therapeutic intervention novel therapeutics postnatal development pre-clinical pre-clinical therapy programs promoter severe intellectual disability treatment strategy urea cycle vector wasting

展开

项目摘要

Project Summary/Abstract The urea cycle is the major pathway for detoxification of ammonia in mammals. Carbamoyl phosphate synthetase 1 (CPS1) deficiency is a neurologically-devastating condition that, while late- onset does occur, the disorder primarily is a condition affecting neonates. The deficiency of this enzyme is characterized clinically by periodic episodes of hyperammonemia resulting in progressive irreversible neurological injury and severe CNS impairment, particularly during a period of critical postnatal development; the condition is associated with a high likelihood of early neonatal mortality. The long-term goal of this program is to develop a clinical candidate gene therapy vector that will result in ammonia control and restore ureagenesis in CPS1 deficiency. Recent advances in gene therapy have led to the concept of using adeno-associated virus (AAV)-based biotechnology to treat CPS1 deficiency; at present, this monogenic disorder has no completely effective therapy except liver transplantation, though often not occurring until the infant has reached a size where successful transplantation is more likely but multiple episodes of neurological injury have been sustained. In addition to being one of the most difficult urea cycle disorders to treat clinically, the development of a gene therapy approach has been hampered by several challenges. First, until recently, there were no animal models of the disorder. Second, CPS1, the most abundant mitochondrial protein in the human liver, must be expressed at high level in hepatocytes. And third, the size of the CPS1 cDNA at 4.5kb places limitations on the expression cassette size; this may be the greatest hindrance for a clinically translatable vector that can be produced at high titer. This proposal seeks to advance a gene therapy approach using small hepatocyte-specific gene regulatory elements to produce a compact CPS1-expressing AAV and a hybrid/dual vector AAV as approaches for this poorly treated disorder. The Lipshutz laboratory recently developed two murine models of CPS1 deficiency and has published and preliminary data on the efficacy of an AAV approach; this proposal is to complete the development and bring forth a clinical candidate vector. Preliminary and published data: The research group has: 1) developed a conditional Cps1-deficient knockout mouse, replicating neonatal onset CPS1 deficiency; 2) demonstrated hepatocyte-specific expression of CPS1 from single vector AAV that has led to plasma ammonia control; 3) developed a dual (or split) AAV approach to express CPS1 in the liver; and 4) has produced humanized mice where CPS1-deficient hepatocytes have repopulated the murine liver; the latter is essential for testing these vector approaches. Specific Aim 1: Optimize the AAV constructs for hepatic CPS1 expression and identify the optimal candidate to advance. Specific Aim 2: Test different hepatotropic serotypes of AAV vector- based CPS1 expression utilizing the optimized transgene cassette in a Cps1-deficient humanized mouse model. The proposed research is significant as it is expected that at completion a final clinical candidate and serotype will have been chosen for establishing hepatic gene replacement for CPS1 deficiency.

项目摘要/摘要尿素周期是哺乳动物氨解毒的主要途径。氨基甲酰磷酸合成酶1（CPS1）缺乏症是一种神经蒸发的疾病，虽然迟到确实发生了，该疾病主要是影响新生儿的疾病。该酶的不足是通过周期性发作在临床上的特征，导致渐进性不可逆神经系统损伤和严重的中枢神经系统损害，特别是在产后关键的时期；该条件与早期新生儿死亡率的高可能性有关。该计划的长期目标是开发临床候选基因治疗载体，该载体将导致氨控制并恢复尿素。在CPS1缺乏症中。基因疗法的最新进展导致了使用腺相关病毒的概念（AAV）基于治疗CPS1缺乏症的生物技术；目前，这种单基因疾病还没有完全除了肝移植以外的有效疗法，尽管通常不会发生在婴儿达到大小之前在成功的移植更有可能的地方，但神经损伤的多个发作已经持续。除了是临床治疗最困难的尿素周期疾病之一之外，基因的发展治疗方法受到了一些挑战的阻碍。首先，直到最近，还没有动物模型疾病。其次，必须表达人肝脏中最丰富的线粒体蛋白CPS1 在肝细胞中高水平。第三，在4.5kb处的CPS1 cDNA的大小使表达式限制盒式大小；这可能是可在高处生产的临床可翻译矢量的最大障碍滴度。该建议旨在使用小肝细胞特异性基因调节来推进基因治疗方法出现紧凑的CPS1表达AAV和混合/双向矢量AAV的元素治疗障碍不佳。 Lipshutz实验室最近开发了两种CPS1缺乏症的鼠模型和已经发布了有关AAV方法功效的初步数据；该建议是完成开发并提出临床候选媒介。初步和已发布的数据：研究小组 HES：1）开发了有条件的CPS1缺陷敲除小鼠，复制新生儿发作CPS1缺乏症； 2）从单个矢量AAV中证明了CPS1的肝细胞特异性表达，导致血浆氨控制; 3）开发了一种双重（或分裂）AAV方法，以在肝脏中表达CPS1； 4）已经产生人源化小鼠CPS1缺陷肝细胞已重新吞噬鼠肝；后者对于测试这些向量方法。特定目标1：优化肝CPS1表达式AAV构建体和确定最佳候选人的进步。特定目标2：测试AAV载体的不同肝脏血清型在CPS1缺陷的人源化小鼠模型中利用优化的转基因盒的基于CPS1的表达。拟议的研究非常重要，因为预计完成最终的临床候选和血清型将选择用于建立CPS1缺乏症的肝基因替代。