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

基本信息

批准号：
9029117
负责人：
Stephanie Cherqui
金额：
$ 34.88万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-01-01 至 2020-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9029117
关键词：
Ablation Address Affect Age-Years Autologous Autologous Transplantation Basement membrane Biochemical Bone Marrow Brain Carbohydrates Cell Death Cell membrane Cells Clinical Clinical Trials Complementary DNA Cysteamine Cystine Cystinosis Data Disease Disease Progression End stage renal failure Epithelium European Eye Family Fanconi Syndrome Functional disorder Funding Future Gene-Modified Genes Grant Healed Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Hereditary Disease Homologous Transplantation Homozygote Human In Vitro Individual Investigation Investigational New Drug Application Kidney Knowledge Laboratories Lead Lentivirus Vector Life Liver Lysosomes Measures Mediating Metabolic Metabolic Diseases Mus Muscle Mutation Nanotubes Organ Pathogenesis Patients Pharmacology and Toxicology Pharmacotherapy Phase I Clinical Trials Phenotype Phosphotransferases Play Process Protein Isoforms Protocols documentation RNA Interference Renal function Research Personnel Risk Role Stem cell transplant Stem cells Symptoms Testing Therapeutic Tissue Preservation Tissues Transplantation Tubular formation United States Food and Drug Administration Vesicle Work apical membrane base cell type cellular transduction clinical application disease phenotype gene therapy genetic profiling healing improved in vivo insight kidney preservation knock-down lentivirally transduced macrophage mortality mouse model novel therapeutics prevent progenitor public health relevance stem cell therapy tissue repair trafficking treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Cystinosis is a metabolic hereditary disease characterized by intracellular accumulation of cystine. Affected individuals typically present with proximal tubulopathy before one year of age and eventually progress to end- stage renal failure. Cystine accumulation also leads to multi-organ dysfunction. Using the mouse model for cystinosis, the Ctns-/- mice, we showed that transplantation of wildtype Sca1+ HSC expressing Ctns resulted in dramatic reductions of tissue cystine content and long-term of kidney preservation. Within the scope of the original R01-DK090058 grant, we optimized a protocol to obtain efficient HSC transduction and showed that ex vivo transduced HSCs using our lentiviral vector construct, pCCL-CTNS, were also capable of decreasing cystine content in all tissues and improved kidney function in Ctns-/- mice. After submitting a pre-Investigator New Drug (IND) application to Food and Drug Administration (FDA), we are now conducting the pharmacology/toxicology studies required for inclusion in an IND for a phase I clinical trial for autologous gene- modified-HSC transplantation for cystinosis. While this work offers new hope for the treatment of cystinosis, we are proposing to investigate two critical questions for the future clinical application of this strategy. In Specific Aim 1, we will investigate if the second CTNS isoform, CTNSLKG, would improve the actual gene therapy strategy. Cystinosin-LKG is found in the lysosomes and at the plasma membrane but its function is unknown. However, CTNSLKG has been found highly expressed in PTCs and other cell types that depend on vesicular trafficking and that correlate with cystinosis clinical features including the Fanconi syndrome. We also recently showed that vesicular trafficking was impaired in cystinosis cells. Thus, our hypothesis is that cystinosinLKG is involved in vesicular trafficking of transporters at the apical membrane of the PTCs and that introduction of the two CTNS isoforms via stem cell therapy would augment the therapeutic impact especially the kidney function. If so, these data will serve as the basis for modifying subsequent clinical trial strategies submitted to the FDA but may also elucidate the pathogenesis of the Fanconi syndrome in cystinosis that has remained a mystery all these years despite considerable scientific investigation. In Specific Aim 2, we will investigate if the patients homozygote for the 57-kb deletion, the most common mutation in cystinosis, can be treated with autologous HSC transplantation. Indeed, this large deletion also removes the adjacent Carbohydrate kinase-like (CARKL) gene encoding the sedoheptulokinase (SHPK). While the absence of this gene does not have an obvious consequence on disease phenotype, SHPK has been recently shown to control macrophage differentiation. Since we recently showed that macrophages play a key role in tissue repair in cystinosis after HSC transplantation, it is critical to verify that the absence of CARKL gene will not impact our stem cell treatment strategy. This work s critical for the future clinical trial for cystinosis but alsowill advance the understanding on macrophage-mediated tissue repair and the role of CARKL in this process.

描述（由申请人提供）：胱氨酸病是一种代谢性遗传性疾病，其特征是受影响的个体通常存在胱氨酸的细胞内积累。一岁前的近端肾小管病变并最终进展为终末期肾衰竭也会导致多器官功能障碍，使用胱氨酸中毒小鼠模型，我们表明移植表达野生型 Sca1+ HSC。 Ctns 导致组织胱氨酸含量显着降低并实现肾脏的长期保存。在最初的 R01-DK090058 资助范围内，我们优化了方案以获得高效的 HSC。转导并表明，在提交预研究新药 (IND) 后，使用我们的慢病毒载体构建体 pCCL-CTNS 体外转导的 HSC 也能够降低所有组织中的胱氨酸含量并改善 Ctns-/- 小鼠的肾功能。目前，我们正在向美国食品药品监督管理局 (FDA) 提交申请，正在进行自体基因修饰 HSC 移植治疗胱氨酸病的 I 期临床试验 IND 所需的药理学/毒理学研究。这项工作为胱氨酸病的治疗带来了新的希望，我们建议研究该策略未来临床应用的两个关键问题，在具体目标 1 中，我们将研究第二种 CTNS 异构体 CTNSLKG 是否会改善实际的基因治疗。 Cystinosin-LKG 存在于溶酶体和质膜中，但其功能尚不清楚，但已发现 CTNSLKG 在 PTC 和其他依赖囊泡运输的细胞类型中高度表达。这与包括 Fanconi 综合征在内的胱氨酸病临床特征相关。因此，我们的假设是胱氨酸蛋白 LKG 参与了 PTC 顶膜的囊泡运输。通过干细胞治疗的两种 CTNS 异构体将增强治疗效果，尤其是肾功能。如果是这样，这些数据将作为修改提交给 FDA 的后续临床试验策略的基础，但。还可能阐明胱氨酸中毒范科尼综合征的发病机制，尽管进行了大量的科学研究，但多年来这仍然是一个谜。在具体目标 2 中，我们将调查患者是否存在 57-kb 缺失（胱氨酸中毒最常见的突变）的纯合子。事实上，这种大的缺失也去除了邻近的编码景天庚酮激酶的碳水化合物激酶样（CARKL）基因。 (SHPK)。虽然该基因的缺失不会对疾病表型产生明显的影响，但最近已证明 SHPK 可以控制巨噬细胞分化，因为我们最近表明巨噬细胞在 HSC 移植后的组织修复中发挥着关键作用。验证 CARKL 基因的缺失不会影响我们的干细胞治疗策略至关重要。这项工作对于未来的胱氨酸病临床试验至关重要，而且也将增进对巨噬细胞介导的组织修复以及 CARKL 在其中的作用的理解。过程。