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: 8209009
• 负责人: Stephanie Cherqui
• 金额: $ 28.91万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 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 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209009
• 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. PUBLIC HEALTH RELEVANCE: Cystinosis is a hereditary disease characterized by the accumulation of cystine in all the cells of the body leading to cell death and tissue damage to kidneys, liver, eyes, muscle and brain. The long-term objective of this project is to develop a new treatment for cystinosis, the strategy is to use the patient's own bone marrow stem cells for transplantation and to genetically modify them ex vivo to introduce a functional version of the defective gene (CTNS). As pre-clinical studies, we will use the mouse model for cystinosis that accumulates cystine in all the tissues and develops similar defects to those of the human children.

描述（由申请人提供）：胱氨酸病是一种代谢性遗传性疾病，其特征为胱氨酸在细胞内蓄积。受影响的个体通常在一岁之前出现近端小管病变（范可尼综合征），并且在第一个十年结束时没有特定的治疗进展到终末期肾衰竭。胱氨酸蓄积最终导致多器官功能障碍。药物半胱胺降低胱氨酸的细胞内浓度。然而，需要规律间隔的剂量和许多不期望的副作用使得其施用困难。半胱胺不能预防近端肾小管病变或终末期肾功能衰竭.该项目的长期目标是通过移植经遗传修饰的自体造血干细胞以表达功能性CTNS基因来开发胱氨酸病的新治疗方法。作为临床前研究，我们将使用Ctns-/-鼠模型用于胱氨酸病。这些动物在所有受试器官中积累胱氨酸和胱氨酸晶体，并出现与受影响人类中观察到的相似的眼部变化、神经缺陷和肾损伤。我们的初步数据显示，移植表达Ctns的同系全骨髓细胞（BMC）或纯化的造血干细胞（HSC）导致BMC或HSC衍生的细胞的组织植入，并且在所有测试的组织中胱氨酸含量显著降低。这种治疗还防止了肾功能障碍的发展和进展。我们现在建议使用Sca 1 + HSC分离自Ctns-/-小鼠和慢病毒载体（LV）用于递送CTNS基因离体。LV已经证明了其在小鼠和人类中长期HSC转导的功效。在具体目标1中，我们提出使用LV表达报告基因优化鼠Sca 1 + HSC的转导，以提高临床试验的安全性和效率，并显著降低成本。在具体目标2中，我们将测试LV转导的、表达CTNS的HSC是否可以预防年轻小鼠中的胱氨酸病介导的组织损伤，并潜在地逆转老年小鼠中的胱氨酸病介导的损伤。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，以测试肾功能障碍、眼异常、骨异常和神经缺陷的预防或治疗。还将测试该策略的免疫应答和安全性以及药物介导的骨髓消融在Ctns-/-小鼠中的毒性。这项工作代表了胱氨酸病的第一个干细胞和基因治疗策略，并为未来的临床试验奠定了基础。它还代表了自体造血干细胞移植策略治疗其他溶酶体贮积症的概念证明。 公共卫生关系：胱氨酸病是一种遗传性疾病，其特征是胱氨酸在身体所有细胞中积累，导致细胞死亡和肾、肝、眼、肌肉和脑的组织损伤。该项目的长期目标是开发一种新的胱氨酸病治疗方法，其策略是使用患者自身的骨髓干细胞进行移植，并对其进行体外遗传修饰，以引入缺陷基因的功能版本（CTNS）。作为临床前研究，我们将使用胱氨酸病的小鼠模型，该模型在所有组织中积累胱氨酸，并产生与人类儿童相似的缺陷。