Novel cell therapy for anemia of CKD

治疗 CKD 贫血的新型细胞疗法

基本信息

批准号：
8305209
负责人：
MATTHEW H WILSON
金额：
$ 34.04万
依托单位：
BAYLOR COLLEGE OF MEDICINE
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 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8305209
关键词：
Ablation Adverse effects Affect Alternative Therapies Anemia Animals Antigens Bolus Infusion Cell Therapy Cells Chronic Chronic Kidney Failure Cicatrix Clinical Research Deep Vein Thrombosis Dose Effectiveness Erythropoietin Experimental Designs Fibroblasts Frequencies Gene Activation Gene Expression Gene Transfer Gene-Modified Genes Genetic Genomics Hematocrit procedure Hormones Human Human Herpesvirus 4 Immunophenotyping In Vitro Injection of therapeutic agent Latent Virus Life Maintenance Measures Mediating Modeling Modification Mus Myocardial Infarction Patients Physiological Population Property Recombinant Erythropoietin Red Blood Cell Count Regulation Risk Safety Signal Pathway Site Specificity Stroke System T-Lymphocyte Testing Tetracyclines Thrombosis Toxic effect Transgenes Vaccination Viral Antigens Virus Wild Type Mouse analog genotoxicity human disease in vivo innovation mouse model novel patient population pre-clinical preclinical evaluation response suicide gene therapeutic protein

项目摘要

DESCRIPTION (provided by applicant): Chronic kidney disease (CKD) affects an estimated 7% of the US population and results in scarring and loss of peritubular fibroblasts which produce erythropoietin (EPO). EPO-deficient anemia of CKD is currently treated with recombinant EPO analog injections that have recently been associated with undesired side effects such as increased risk of stroke, heart attacks, and deep vein thrombosis which may preclude further use of this therapy. Although the mechanisms of these side effects are unclear, it is clear that bolus dosing of EPO analogs either weekly or monthly does not recapitulate the physiologic regulation of this important hormone and bolus dosing may alter EPO signaling pathways. Thus, there is a critical need to develop alternative therapies for anemia of CKD. Herein we describe an innovative experimental design using non-viral transposon-mediated gene transfer to develop a new strategy for therapy of anemia of CKD. Genetically modified T lymphocytes whose specificity is directed to persistent (latent) viruses such as Epstein-Barr virus (EBV) survive long-term (>8 years) in stable numbers in vivo due to chronic viral antigen stimulation. Moreover, preclinical and recent clinical studies have shown T cells can be readily induced to apoptose by activation of a co-transferred suicide gene, providing an additional layer of safety and control. We therefore hypothesize that virus specific T cells genetically modified to inducibly express EPO and a separately inducible suicide gene represent an ideal candidate cell population for sustained and safe treatment of anemia of CKD. In specific aim 1, we propose to modify virus specific murine T cells to inducibly express EPO and a suicide gene and we will infuse them into wild type and CKD mice to measure their effectiveness in regulating hematocrit levels in vivo. Specific aim 2 focuses on extending these genetic modifications to human T cells and testing them in vitro for their ability to be propagated long-term via chronic viral antigen stimulation, as well as inducibly express EPO and undergo selectively induced cell ablation if needed. In specific aim 3, we will evaluate the functionality of genetically modified human T cells from patients with CKD and determine the frequency of EBV-specific T cells and their response to EBV antigen in the presence and absence of transgenically expressed EPO ex vivo. PUBLIC HEALTH RELEVANCE: This project is focused on developing an efficient, safe, and novel cell therapy for anemia of chronic kidney disease. The proposed strategy could also be used for therapy for a variety of other human diseases.

描述（由申请人提供）：慢性肾病 (CKD) 影响了大约 7% 的美国人口，并导致疤痕形成和产生促红细胞生成素 (EPO) 的肾小管周围成纤维细胞的损失。 CKD 的 EPO 缺乏性贫血目前采用重组 EPO 类似物注射剂进行治疗，最近发现注射剂会产生不良副作用，例如中风、心脏病和深静脉血栓形成的风险增加，这可能会阻碍该疗法的进一步使用。尽管这些副作用的机制尚不清楚，但很明显，每周或每月推注 EPO 类似物并不能概括这种重要激素的生理调节，并且推注剂量可能会改变 EPO 信号通路。因此，迫切需要开发 CKD 贫血的替代疗法。在此，我们描述了一种创新的实验设计，利用非病毒转座子介导的基因转移来开发治疗 CKD 贫血的新策略。转基因 T 淋巴细胞的特异性针对持久性（潜伏）病毒，如 Epstein-Barr 病毒 (EBV)，由于慢性病毒抗原刺激，可在体内以稳定数量长期（>8 年）存活。此外，临床前和最近的临床研究表明，通过激活共转移的自杀基因，可以很容易地诱导 T 细胞凋亡，从而提供额外的安全和控制层。因此，我们假设病毒特异性 T 细胞经过基因修饰，可诱导表达 EPO 和单独诱导的自杀基因代表了持续、安全治疗 CKD 贫血的理想候选细胞群。在具体目标 1 中，我们建议修饰病毒特异性鼠 T 细胞以诱导表达 EPO 和自杀基因，并将它们注入野生型和 CKD 小鼠中，以测量它们在体内调节血细胞比容水平的有效性。具体目标 2 侧重于将这些基因修饰扩展到人类 T 细胞，并在体外测试它们通过慢性病毒抗原刺激长期繁殖的能力，以及诱导表达 EPO 并在需要时进行选择性诱导细胞消融的能力。在具体目标 3 中，我们将评估转基因人类 T 细胞的功能来自 CKD 患者的 EBV 特异性 T 细胞的频率及其在体外转基因表达的 EPO 存在和不存在的情况下对 EBV 抗原的反应。公共健康相关性：该项目的重点是开发一种有效、安全和新颖的细胞疗法来治疗慢性肾病贫血。所提出的策略还可用于治疗多种其他人类疾病。