Lentiviral-transduced hematopoictic stem cell transplantation for cystinosis

慢病毒转导的造血干细胞移植治疗胱氨酸病

• 批准号: 8627162
• 负责人: Stephanie Cherqui
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8627162
• 关键词: 12 year old; Adolescent; Adult; Adverse effects; Affect; Age-Years; Animals; Autologous Transplantation; Blindness; Bone Marrow; Bone Marrow Cells; Bone Marrow Purging; Bone Marrow Stem Cell; Bone Marrow Stem Cell Transplantation; Brain; Cell Death; Cells; Child; Childhood; Chronic; Clinical; Clinical Trials; Confocal Microscopy; Cysteamine; Cystine; Cystinosis; Data; Defect; Development; Diabetes Mellitus; Disease; Disease Progression; Dose; Drug toxicity; Electrolytes; End stage renal failure; Engraftment; Eye; Eyedrops; Family; Fanconi Syndrome; Foundations; Functional disorder; Future; Gene Delivery; Genes; Graft Rejection; Green Fluorescent Proteins; Growth; Growth Factor; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hereditary Disease; Hour; Human; Hypogonadism; Hypothyroidism; Image; Immune response; Immunosuppression; Individual; Injury; Isogenic transplantation; Kidney; Lentivirus Vector; Life; Liquid substance; Liver; Luciferases; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolic Diseases; Modeling; Mus; Muscle; Myopathy; Neuraxis; Neurologic; Odors; Organ; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Photophobia; Pluripotent Bone Marrow Stem Cell; Population; Prevention; Protocols documentation; Reporter Genes; Rickets; Risk; Safety; Source; Stem cell transplant; Stem cells; Symptoms; Testing; Time; Tissues; Toxic effect; Transplantation; Viral Vector; Vomiting; Work; base; bone; cellular transduction; cost; gene delivery system; gene therapy; infancy; injured; mouse model; nephrogenesis; neuronal cell body; peripheral blood; pre-clinical; preclinical study; prevent; public health relevance; stem cell therapy; transduction efficiency; transgene expression; treatment strategy; vector; viral gene delivery; young adult

DESCRIPTION (provided by applicant): Cystinosis is a metabolic hereditary disease characterized by intracellular accumulation of cystine. Affected individuals typically present with proximal tubulopathy (Fanconi syndrome) before one year of age and without specific treatment progress to end-stage renal failure by the end of the first decade. Cystine accumulation eventually leads to multi-organ dysfunction. The drug cysteamine reduces the intracellular concentration of cystine. However, the need for regularly spaced doses and a number of undesirable side effects render its administration difficult. Moreover, cysteamine does not prevent the proximal renal tubulopathy or the end- stage renal failure. The long-term objective of this project is to develop a new treatment for cystinosis by transplantation of autologous hematopoietic stem cells genetically modified ex vivo to express a functional CTNS gene. As pre- clinical studies, we will use the Ctns-/- murine model for cystinosis. These animals accumulate cystine and cystine crystals in all organs tested and develop ocular changes, neurological defects and kidney injuries similar to those observed in affected humans. Our preliminary data showed that transplantation of syngeneic whole bone marrow cells (BMC) or purified hematopoietic stem cells (HSC) expressing Ctns resulted in tissue engraftment of BMC or HSC-derived cells and significant reductions of cystine content in all the tissues tested. This treatment also prevented the development and progression of kidney dysfunction. We now propose to use Sca1+ HSC isolated from Ctns-/- mice and lentiviral vector (LV) for delivering the CTNS gene ex vivo. LV has proven its efficacy for long-term HSC transduction in mice but also in humans. In Specific aim 1, we propose to optimize the transduction of murine Sca1+ HSC using LV expressing reporter genes to enhance the safety and efficiency of a clinical trial as well as significantly reduce costs. In Specific aim 2, we will test if LV- transduced, CTNS-expressing HSC can prevent cystinosis-mediated tissue injury in young mice and potentially reverse cystinosis-mediated injury in older mice. The efficiency of these strategies will be tested by measuring CTNS expression and cystine content in different tissue compartments and by well-established functional studies to test the prevention or treatment of the kidney dysfunction, eye anomalies, bone anomalies and neurological defects. The immune response and safety of this strategy will be also tested as well as the toxicity of drug-mediated myeloablation in Ctns-/- mice. This work represents the first stem cell and gene therapy treatment strategies for cystinosis and builds the foundations for a future clinical trial. It also represents a proof of concept for autotologous HSC transplantation strategy to treat other lysosomal storage disorders.

描述（由申请人提供）：胱氨酸病是一种以细胞内胱氨酸积累为特征的代谢遗传性疾病。受影响的个体通常在一岁之前出现近端肾小管病变（范可尼综合征），并且在第一个十年结束时没有进行特殊治疗，进展为终末期肾衰竭。胱氨酸积累最终导致多器官功能障碍。半胱胺药物可降低细胞内胱氨酸的浓度。然而，需要定期间隔剂量和许多不良副作用使其给药变得困难。此外，半胱胺不能预防近端肾小管病变或终末期肾衰竭。该项目的长期目标是通过移植经过离体基因改造表达功能性 CTNS 基因的自体造血干细胞来开发一种新的胱氨酸病治疗方法。作为临床前研究，我们将使用 Ctns-/- 小鼠胱氨酸病模型。这些动物在所有测试的器官中积累胱氨酸和胱氨酸晶体，并出现与受影响人类相似的眼部变化、神经缺陷和肾损伤。我们的初步数据表明，移植表达 Ctns 的同基因全骨髓细胞 (BMC) 或纯化的造血干细胞 (HSC) 导致 BMC 或 HSC 衍生细胞的组织植入，并且所有测试组织中胱氨酸含量显着降低。这种治疗还可以防止肾功能障碍的发生和进展。我们现在建议使用从 Ctns-/- 小鼠中分离的 Sca1+ HSC 和慢病毒载体 (LV) 来离体传递 CTNS 基因。 LV 已在小鼠和人类中证明了其对 HSC 长期转导的功效。在具体目标 1 中，我们建议使用 LV 表达报告基因优化小鼠 Sca1+ HSC 的转导，以提高临床试验的安全性和效率，并显着降低成本。在具体目标 2 中，我们将测试 LV 转导、表达 CTNS 的 HSC 是否可以预防年轻小鼠中胱氨酸介导的组织损伤，并可能逆转老年小鼠中胱氨酸介导的损伤。这些策略的有效性将通过测量不同组织区室中的 CTNS 表达和胱氨酸含量以及通过完善的功能研究来测试对肾功能障碍、眼睛异常、骨骼异常和神经缺陷的预防或治疗。该策略的免疫反应和安全性以及药物介导的清髓术在 Ctns-/- 小鼠中的毒性也将得到测试。这项工作代表了第一个针对胱氨酸病的干细胞和基因疗法治疗策略，并为未来的临床试验奠定了基础。它还代表了治疗其他溶酶体贮积症的自体造血干细胞移植策略的概念证明。