Macrophage-based Human Gene Therapy for Hereditary PAP

基于巨噬细胞的遗传性 PAP 人类基因治疗

基本信息

批准号：
8206634
负责人：
Bruce C Trapnell
金额：
$ 21.88万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-12-15 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8206634
关键词：
Address Adult Affect Alternative Therapies Alveolar Macrophages Alveolus Animals Autoantibodies Autoimmune Process Autologous Autologous Bone Marrow Transplantation Bacteriophages Biochemistry Biological Assay Biological Markers Bone Marrow Bone Marrow Purging Bone Marrow Transplantation Breathing Bronchoalveolar Lavage Bronchoalveolar Lavage Fluid Bronchoscopes CD34 gene CSF2RA gene CSF2RB gene CSF3 gene Catabolism Cell Therapy Cells Child Childhood Cyclophosphamide Cytology Data Diagnosis Disease Engraftment Figs - dietary Flow Cytometry Functional disorder Future General Anesthesia Genes Granulocyte-Macrophage Colony-Stimulating Factor Granulocyte-Macrophage Colony-Stimulating Factor Receptors Growth Factor Hereditary Disease Histologic Histopathology Human ITGAM gene Immune response Implant Individual Infection Inherited Lentivirus Vector Lobe Lung Lung diseases Measures Mechanics Mediating Methods Molecular Diagnosis Mus Mutation Myeloid Cells Pathogenesis Patients Phosphorylation Procedures Proteins Pulmonary Alveolar Proteinosis Pulmonary Surfactants Receptor Gene Reporting Respiratory Failure Respiratory Insufficiency Retroviridae Risk STAT5A gene Saline Serum Signal Transduction Sodium Chloride Testing Therapeutic Transduction Gene Unconscious State Virus Water base cellular transduction cohort effective therapy functional restoration gene correction gene therapy gene therapy clinical trial improved in vivo macrophage molecular pathology monocyte mouse model nonhuman primate novel novel strategies peripheral blood pre-clinical receptor receptor expression receptor function response restoration rituximab success surfactant uptake

项目摘要

ABSTRACT Recessive mutations in CSF2RA, the gene encoding the ¿-chain of the granulocyte/macrophage-colony stimulating factor receptor (GM-R), were recently reported by the applicant as a newly-identified genetic disorder causing pulmonary alveolar proteinosis (PAP) in children. Recognizing increased serum GM-CSF as a disease biomarker, the applicant identified a cohort of affected individuals and defined the pathogenesis, presentation, diagnosis, molecular pathophysiology, and therapy response of hereditary PAP. The disease is characterized by the progressive accumulation of surfactant in alveolar macrophages (AM) and alveoli, resulting in respiratory insufficiency and, in severe cases, respiratory failure. Current therapy is whole lung lavage, a procedure performed under general anesthesia in which one lung is mechanical ventilated while the other is repeatedly filled with warmed saline, mechanically percussed to emulsify the accumulated surfactant, and then drained to physically remove it. The procedure is repeated as required, which in some children is every two months. PAP occurs in genetically modified mice deficient in the ¿-chain of the GM-R (GM-R¿KO mice) or in GM-CSF (GMKO mice), and in humans with neutralizing GM-CSF autoantibodies (autoimmune PAP), all of which result in pulmonary histopathology and molecular pathology similar to hereditary PAP in children. Without stimulation by GM-CSF, surfactant catabolism in AM is impaired and results in reduced pulmonary clearance and progressive accumulation of pulmonary surfactant. Bone marrow transplantation (BMT) is a therapeutic option for these children with PAP and was attempted in one child who died of a lung infection before engraftment was complete. The accumulated surfactant in PAP likely increases the infection risk associated with myeloablation, which is required for BMT. PAP in GM-R¿KO mice was 'cured' by the applicant by autologous transplantation of bone marrow after retrovirus-mediated gene transduction to correct GM-R function. The central hypothesis of this proposal is that ex-vivo lentiviral vector-mediated restoration of functional GM-R expression in autologous monocytes or G-CSF-mobilized CD34+ cells followed by intrapulmonary administration into non-myeloablated recipients will be safe, well-tolerated, and effective therapy of hPAP. This hypothesis will be tested in 3 Specific Aims: (1) macrophage-mediated cell therapy of hPAP in mice; (2) macrophage-mediated gene therapy of hPAP in mice; (3) preclinical correction of CSF2RA expression and surfactant catabolism in macrophages from children with hPAP, evaluated in vivo in the lungs of non-human primates. A novel approach avoiding known impediments to lung gene therapy will take advantage of the natural survival advantage of GM-R gene-corrected cells conferred by the elevated levels of GM-CSF, a potent growth factor for both human and murine AM. This approach gives hereditary PAP an outstanding chance of being the first human lung disease to be successfully treated by gene therapy and provides a feasible and potential therapeutic alternative for a devastating disease in children.

抽象的 CSF2RA 的隐性突变，编码 ¿ -粒细胞/巨噬细胞集落链刺激因子受体（GM-R），最近被申请人报告为新鉴定的遗传基因导致儿童肺泡蛋白沉积症 (PAP) 的疾病应识别为血清 GM-CSF 增加。疾病生物标志物，申请人确定了一组受影响的个体并确定了发病机制，遗传性 PAP 的表现、诊断、分子病理生理学和治疗反应。其特征是表面活性剂在肺泡巨噬细胞（AM）和肺泡中逐渐积累，导致呼吸功能不全，严重时会导致呼吸衰竭。目前的治疗方法是全肺治疗。灌洗，一种在全身麻醉下进行的手术，其中一侧肺进行机械通气，同时另一侧肺进行机械通气。另一部分反复注入温热的生理盐水，机械敲击使积累的表面活性剂乳化，然后根据需要重复进行排水以物理去除，这对于某些儿童来说是必需的。缺乏 ¿ 的转基因小鼠每两个月就会发生一次 PAP。 - GM-R 链条 (GM-R¿KO 小鼠）或 GM-CSF（GMKO 小鼠），以及具有中和 GM-CSF 自身抗体（自身免疫性 PAP），所有这些都会导致肺部组织病理学和分子病理学类似于遗传性 PAP 儿童如果没有 GM-CSF 的刺激，AM 中的表面活性剂分解代谢就会受到损害并导致减少。肺清除率和肺表面活性物质的逐渐积累。骨髓移植。 (BMT) 是这些患有 PAP 的儿童的一种治疗选择，曾在一名死于肺病的儿童身上进行过尝试 PAP 中积累的表面活性剂可能会增加感染。与骨髓清除相关的风险，这是 GM-R 中 PAP 所需的。 KO小鼠被“治愈” 申请人通过自体骨髓移植后逆转录病毒介导的基因转导来纠正该提案的中心假设是离体慢病毒载体介导的恢复。自体单核细胞或 G-CSF 动员的 CD34+ 细胞中功能性 GM-R 表达，然后对非清髓受体进行肺内给药将是安全、耐受性良好且有效的 hPAP 的治疗将在 3 个具体目标中得到检验：(1) 巨噬细胞介导的细胞治疗。小鼠中的 hPAP；(2) 小鼠中巨噬细胞介导的 hPAP 基因治疗；(3) CSF2RA 的临床前校正 hPAP 儿童巨噬细胞的表达和表面活性剂分解代谢，在肺体内进行评估需要一种新的方法来避免肺部基因治疗的已知障碍。利用 GM-R 基因校正细胞的自然生存优势，这是由于 GM-R 水平升高所赋予的 GM-CSF，一种针对人类和小鼠 AM 的有效生长因子。这种方法为遗传性 PAP 提供了一种可能。成为第一个通过基因疗法成功治疗的人类肺部疾病的绝佳机会为儿童的毁灭性疾病提供了可行且潜在的治疗替代方案。