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：
10428544
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 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10428544
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 α-galactosidase A (α-gal A). α-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 α-gal A in a Fabry disease model. We will extend our mouse studies with EPO to α-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 α-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 α-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 α-gal A in a mouse model of Fabry disease. We propose to deliver optimized human CAR-T cells expressing human α-gal A in a NOD/SCID/Fabry mouse model. We will evaluate for engraftment, vaccination response, α-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.

能够在体内持续治疗蛋白递送的细胞疗法有可能使这两个肾脏受益疾病及其并发症。在上一个赠款周期中，我们使用转座子修饰的抗原特异性T细胞使用促红细胞生成素（EPO）在体内递送治疗蛋白的特异性T细胞（EPO）作为模型系统。我们证明了慢性肾脏疾病贫血的鼠EPO和治疗在小鼠作为模型系统中，我们证明了抗原特异性人的诱导人EPO表达体外T细胞。我们建议通过将研究扩展到人体T细胞在体内环境中并测试一种创新方法来增强长期治疗酶 Fabry疾病的递送是由于α-半乳糖苷酶A（α-GAL A）丧失而导致的。 α-gal a - / - 小鼠代表人类Fabry疾病的动物模型，这是由于缺乏代谢脂肪所需的酶而导致的全身性疾病，包括肾脏疾病。在AIM 1中，我们将测试转座子修饰的抗原特异性T细胞用于在Fabry病模型中表达α-GAL A。我们将使用EPO扩展小鼠研究到A中的α-GAL A Fabry病模型使用抗原特异性小鼠T细胞和疫苗接种。我们将基因修饰人类T细胞表达嵌合抗原受体（CAR）以及荧光素酶或人α-galA。点头/scid/fabry小鼠评估工程抗原特异性人T细胞植入的能力在体内模型中进行疫苗接种和短期表达α-GAL A。虽然穿孔在T细胞中很重要为了清除恶性细胞，穿孔蛋白途径也有助于清除抗原表达细胞疫苗接种后。在AIM 2中，我们将测试T细胞中的穿孔蛋白敲除作为增强长期治疗的一种方式蛋白质从抗原特异性T细胞中递送。我们建议在抗原 - 特定的T细胞以确定它是否会增强输送治疗蛋白的细胞的长期持久性。目标 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 法布里病。我们建议在A中传递表达人α-GAL A的优化的人类T细胞点头/scid/fabry鼠标模型。我们将评估植入，疫苗接种反应，α-GAL A活性和组织中的球形三方糖酰胺水平。拟议的研究将导致新细胞疗法的发展对于肾脏疾病及其并发症，并具有远远超出肾脏的热撞击的潜力。

项目成果

期刊论文数量（18）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Temporal self-regulation of transposition through host-independent transposase rodlet formation.

DOI：
10.1093/nar/gkw1115
发表时间：
2017-01-09
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Woodard LE;Downes LM;Lee YC;Kaja A;Terefe ES;Wilson MH
通讯作者：
Wilson MH

Gene therapy for kidney disease: targeting cystinuria.

DOI：
10.1097/mnh.0000000000000768
发表时间：
2022-03-01
期刊：
Current opinion in nephrology and hypertension
影响因子：
3.2
作者：
Peek JL;Wilson MH
通讯作者：
Wilson MH

Targeting piggyBac transposon integrations in the human genome.

针对人类基因组中的 piggyBac 转座子整合。

DOI：
10.1007/978-1-62703-761-7_9
发表时间：
2014
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Galvan,DanielL;Kettlun,ClaudiaS;Wilson,MatthewH
通讯作者：
Wilson,MatthewH

Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney.

肾盂水动力注射用于肾脏中蛋白质的非病毒表达。

DOI：
10.3791/56324
发表时间：
2018
期刊：
Journal of visualized experiments : JoVE
影响因子：
0
作者：
Woodard,LaurenE;Welch,RichardC;Williams,FelishaM;Luo,Wentian;Cheng,Jizhong;Wilson,MatthewH
通讯作者：
Wilson,MatthewH

CRISPR/Cas9 engineering of a KIM-1 reporter human proximal tubule cell line.