Novel cell therapy for anemia of CKD

治疗 CKD 贫血的新型细胞疗法

• 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
• 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

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）：慢性肾脏疾病（CKD）估计影响了美国人口的7％，并导致产生促红细胞生成素（EPO）的周围和周围成纤维细胞的损失。目前，CKD的Epo缺陷贫血已接受重组EPO类似注射治疗，这些注射最近与不希望的副作用有关，例如中风的风险增加，心脏病发作和深静脉血栓形成，这可能无法进一步使用该疗法。尽管这些副作用的机制尚不清楚，但很明显，每周或每月的EPO类似物的推注剂量不会概括这种重要的激素的生理调节，而剂量给药可能会改变EPO信号传导途径。因此，迫切需要开发CKD贫血的替代疗法。本文中，我们使用非病毒转座子介导的基因转移来描述一种创新的实验设计，从而制定了CKD贫血治疗的新策略。基因修饰的T淋巴细胞的特异性针对持续性（潜在）病毒（例如爱泼斯坦 - 巴尔病毒（EBV））的长期（> 8年）在体内长期（> 8年）在体内生存，这是由于慢性病毒抗原刺激。此外，临床前和近期临床研究表明，通过激活共转移的自杀基因，可以很容易地将T细胞诱导到凋亡中，从而提供了额外的安全性和控制层。因此，我们假设病毒特异性T细胞在遗传上修饰至诱导 Express EPO和单独诱导的自杀基因代表了CKD贫血的持续和安全治疗的理想候选细胞群体。在特定的目标1中，我们建议修改特定病毒的鼠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，以测量它们在调节体内血细胞比容水平的有效性。具体目标2的重点是将这些遗传修饰扩展到人T细胞，并在体外测试它们，以通过慢性病毒抗原刺激长期传播它们的能力，并在需要时诱导地表达EPO并在需要时进行选择性诱导的细胞消融。在特定的目标3中，我们将评估转基因的人T细胞的功能 来自CKD的患者并确定EBV特异性T细胞的频率及其在存在和不存在转基因EPO EPO的情况下对EBV抗原的反应。 公共卫生相关性：该项目的重点是开发用于慢性肾脏疾病贫血的高效，安全和新颖的细胞疗法。提出的策略也可以用于治疗其他各种人类疾病。