Novel cell therapy for sustained therapeutic protein delivery in vivo

用于体内持续治疗性蛋白质递送的新型细胞疗法

基本信息

批准号：
10011826
负责人：
MATTHEW H WILSON
金额：
$ 41.67万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10011826
关键词：
Affect Alpha-galactosidase Anemia Animal Model Animals Antigens Autopsy Biological Models CAR T cell therapy Cell Therapy Cells Chronic Kidney Failure Clinical Data Communication Cytotoxic T-Lymphocytes Development Disease Disease model Engineering Engraftment Enzymes Erythropoietin Fabry Disease Fatty acid glycerol esters Future Gene-Modified Grant Half-Life Heart Diseases Hematocrit procedure Histology Human In Vitro Infusion procedures Kidney Kidney Diseases Knock-out Long-Term Effects Longevity Luciferases Mediating Modeling Morbidity - disease rate Mus Nature Organ Pathway interactions Patients Production Proteins Stroke System Systemic disease T-Cell Receptor T-Lymphocyte Technology Testing Therapeutic Time Tissues Toxic effect Toxicology Transgenic Organisms Translating Vaccination antigen-specific T cells cancer cell cancer therapy chimeric antigen receptor chimeric antigen receptor T cells clinical efficacy cost enzyme replacement therapy experimental study globotriaosylceramide human disease human model in vivo in vivo Model in vivo imaging inducible gene expression innovation mouse model mouse perforin novel novel therapeutics perforin pre-clinical response therapeutic enzyme therapeutic protein vaccination strategy

项目摘要

A cell therapy capable of sustained therapeutic protein delivery in vivo has the potential to benefit both kidney disease and its complications. During our previous grant cycle, we developed and validated technology using transposon-modified antigen specific T cells for therapeutic protein delivery in vivo using erythropoietin (EPO) as a model system. We demonstrated delivery of murine EPO and therapy for anemia of chronic kidney disease in mice as a model system, and we demonstrated inducible human EPO expression from antigen-specific human T cells in vitro. We propose to significantly advance beyond our previous grant by extending our studies to human T cells in an in vivo context and testing an innovative approach to enhance long-term therapeutic enzyme delivery for Fabry disease that results from loss of -galactosidase A (-gal A). -gal A -/- mice represent an animal model of human Fabry disease, which results from lack of an enzyme needed to metabolize fats leading systemic disease including kidney disease. In aim 1, we will test transposon-modified antigen-specific T cells for expression of -gal A in a Fabry disease model. We will extend our mouse studies with EPO to -gal A in a Fabry disease model using antigen-specific mouse T cells and vaccination. We will gene modify human T cells to express a chimeric antigen receptor (CAR) along with luciferase or human -gal A. Cells will be infused into NOD/SCID/Fabry mice to evaluate the ability of engineered antigen-specific human T cells to engraft, respond to vaccination, and short-term expression -gal A in an in vivo model. Although perforin is important in T cells for clearance of malignant cells, the perforin pathway also contributes to clearance of antigen expressing cells post vaccination. In aim 2, we will test perforin knockout in T cells as a way of enhancing long-term therapeutic protein delivery from antigen-specific T cells. We propose to test perforin knockout in the setting of antigen- specific T cells to determine if it will boost long-term persistence of cells delivering therapeutic proteins. In aim 3, we will test transposon-modified antigen-specific T cells for long-term expression of -gal A in a mouse model of Fabry disease. We propose to deliver optimized human CAR-T cells expressing human -gal A in a NOD/SCID/Fabry mouse model. We will evaluate for engraftment, vaccination response, -gal A activity, and globotriaosylceramide levels in tissues. The proposed studies will lead to the development of new cell therapies for kidney disease and its complications and have the potential for therapeutic impact well beyond the kidney.

能够在体内持续输送治疗性蛋白质的细胞疗法有可能使双肾受益在我们之前的资助周期中，我们使用开发和验证了技术。转座子修饰的抗原特异性 T 细胞，使用促红细胞生成素 (EPO) 在体内递送治疗性蛋白作为模型系统，我们展示了小鼠 EPO 的输送和慢性肾病贫血的治疗。以小鼠为模型系统，我们证明了抗原特异性人类可诱导人类 EPO 表达我们建议通过将我们的研究扩展到体外，从而显着超越我们之前的资助。在体内环境中研究人类 T 细胞并测试增强长期治疗酶的创新方法因α-半乳糖苷酶 A (α-gal A) 缺失而导致的法布里病的分娩代表了一种小鼠。人类法布里病的动物模型，该病是由于缺乏代谢脂肪所需的酶而导致的在目标 1 中，我们将测试转座子修饰的抗原特异性 T 细胞。法布里病模型中 α-gal A 的表达我们将把 EPO 的小鼠研究扩展到 a 中的 α-gal A。法布里病模型使用抗原特异性小鼠 T 细胞和疫苗接种，我们将对人类 T 细胞进行基因修饰。表达嵌合抗原受体 (CAR) 以及荧光素酶或人 α-gal A。细胞将被注入 NOD/SCID/Fabry 小鼠评估工程化抗原特异性人类 T 细胞移植、应答的能力尽管穿孔素在 T 细胞中很重要，但在体内模型中，疫苗接种和短期表达 α-gal A 仍发挥着重要作用。对于恶性细胞的清除，穿孔素途径也有助于抗原表达细胞的清除在目标 2 中，我们将测试 T 细胞中的穿孔素敲除，作为增强长期治疗的一种方法。我们建议在抗原特异性 T 细胞中测试穿孔敲除。特定的 T 细胞，以确定它是否会促进细胞递送治疗蛋白的长期持续性。 3、我们将测试转座子修饰的抗原特异性T细胞在小鼠模型中长期表达α-gal A 我们建议在一个细胞中提供表达人类 α-gal A 的优化人类 CAR-T 细胞。我们将评估 NOD/SCID/Fabry 小鼠模型的植入、疫苗接种反应、α-gal A 活性和拟议的研究将导致新细胞疗法的开发。用于肾脏疾病及其并发症，并且具有远远超出肾脏范围的潜在治疗作用。