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Functional assay development for tafazzin enzyme replacement candidate selection

用于 tafazzin 酶替代候选选择的功能测定开发

基本信息

批准号：
10722856
负责人：
MICHAEL T CHIN
金额：
$ 62.3万
依托单位：
TUFTS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10722856
关键词：
3-Methylglutaconic aciduria type 2 Acyltransferase Affect Algorithms Antibody Formation Autopsy Biochemical Biological Biological Assay Cardiolipins Cardiomyopathies Cell membrane Cells Cessation of life Child Childhood Congestive Heart Failure Cultured Cells Cyclic Neutropenia Defect Development Disease Dose Drug Kinetics Drug or chemical Tissue Distribution Endosomes Endotoxins Enzyme Tests Enzyme-Linked Immunosorbent Assay Enzymes Evaluation Freedom Functional disorder Genes Genetic Diseases Heart failure Hereditary Disease Human In Vitro Intravenous Knockout Mice Label Left Ventricular Function Life Link Lipids Live Birth Longitudinal Studies Measurement Measures Mitochondria Modeling Monitor Morphology Mus Muscle Muscle hypotonia Mutation Organ Oxygen Consumption Patients Penetration Peptides Pharmacology Phase Physiological Plasma Population Primary idiopathic dilated cardiomyopathy Production Proteins Protocols documentation Quality Control Reactive Oxygen Species Reagent Recombinants Research Project Grants Respiration Serum Standardization Therapeutic Tissues Toxic effect Toxicology assay development candidate identification candidate selection clinical development cohort commercialization detection assay effective therapy effectiveness measure efficacious treatment efficacy evaluation efficacy study enzyme activity enzyme deficiency enzyme replacement therapy immunogenicity in vitro activity in vivo in vivo evaluation infancy intraperitoneal lead candidate manufacture manufacturing scale-up middle age mitochondrial dysfunction mitochondrial membrane mouse model peptide drug premature process evaluation process optimization therapeutic enzyme uptake

项目摘要

Abstract: Functional assay development for tafazzin enzyme replacement candidate selection In the proposed research project, we will perform key biochemical, cellular and physiological studies to evaluate the potential of various modified recombinant tafazzin enzyme therapeutics to become the first effective treatment for Barth Syndrome, a rare, life-threatening disorder with no efficacious therapy. Barth Syndrome is a rare X-linked disorder affecting 1:300,000 live births, resulting from defects in the gene encoding Tafazzin, an acyltransferase that modifies cardiolipin to the tetralinoleoyl form and is essential for mitochondrial respiration. Patients with Barth Syndrome develop cardiomyopathy, muscular hypotonia and cyclic neutropenia during childhood, rarely surviving to middle age. At present, no effective therapy exists for these patients. We are developing potential enzyme replacement therapeutics in which recombinant tafazzin is modified to contain a cellular penetrating peptide that promotes uptake into tafazzin-deficient cells, sometimes in conjunction with an endosomal escape peptide to facilitate escape from lysosomal degradation. We have found that these recombinant tafazzin enzyme replacement therapeutics (rTERTs) can enter tafazzin-deficient cells, localize to mitochondria and correct both cardiolipin remodeling and mitochondrial respiration defects, in vitro and in vivo to varying degrees. To facilitate the commercialization of these reagents, we will develop standardized biochemical, cellular and physiological assays to directly compare enzymatic function, cellular uptake, rescue of cardiolipin remodeling defects, and rescue of defective mitochondrial respiration in cultured cells. Based on these studies, we have developed a selection algorithm to identify a cohort of potential lead candidates that demonstrate the greatest biochemical and biological activity in vitro. The top candidates identified by this algorithm will undergo production process optimization and evaluation for activity in vivo. The in vivo evaluation will consist of pharmacokinetics, tissue distribution, toxicology and efficacy studies in tafazzin knockout mice using an in house tafazzin LC-MS peptide detection assay, an in house ELISA assay to measure antibody formation, in conjunction with measurements of tafazzin function and left ventricular function. We believe that our studies will establish standardized assays to characterize potential enzyme replacement therapies for Barth Syndrome in the R61 phase, thereby facilitating evaluation of potential lead candidates in a mouse model of Barth Syndrome in the R33 phase, ultimately leading to the identification of a lead candidate for further clinical development.

翻译后摘要：塔法津酶替代候选选择功能测定的发展在拟议的研究项目中，我们将进行关键的生化，细胞和生理研究，评估各种修饰的重组tafazzin酶治疗剂成为第一个有效治疗巴斯综合征，一种罕见的，危及生命的疾病，没有有效的治疗。Barth 综合征是一种罕见的X连锁疾病，影响1：300，000活产，由基因缺陷引起编码Tafazzin，一种将心磷脂修饰为四氢萘油酰形式的酰基转移酶，线粒体呼吸患有巴特综合征的患者会出现心肌病、肌张力减退和儿童期周期性中性粒细胞减少症，很少能活到中年。目前还没有有效的治疗方法，这些病人。我们正在开发潜在的酶替代疗法，其中重组tafazzin是修饰为含有细胞穿透肽，其促进tafazzin缺陷细胞的摄取，有时与内体逃逸肽结合以促进从溶酶体降解中逃逸。我们有发现这些重组tafazzin酶替代疗法（rTERTs）可以进入tafazzin缺陷型细胞，定位于线粒体并纠正心磷脂重塑和线粒体呼吸缺陷，体外和体内不同程度地。为了促进这些试剂的商业化，我们将开发标准化的生物化学、细胞和生理学测定，以直接比较酶功能、细胞摄取，挽救心磷脂重塑缺陷，并挽救有缺陷的线粒体呼吸，细胞基于这些研究，我们开发了一种选择算法，以确定潜在的铅队列在体外表现出最大生化和生物活性的候选物。的最佳候选者通过该算法鉴定的化合物将进行生产工艺优化和体内活性评价。的体内评价将包括他法津的药代动力学、组织分布、毒理学和功效研究使用内部tafazzin LC-MS肽检测测定、内部ELISA测定测量抗体形成，结合tafazzin功能和左心室功能的测量，功能我们相信，我们的研究将建立标准化的检测方法来表征潜在的酶，在R61阶段对Barth综合征进行替代治疗，从而促进潜在的电极导线评估在R33期的Barth综合征小鼠模型中的候选人，最终导致鉴定出进一步临床开发的主要候选人。