Macrophage Based Gene Therapy for Hereditary Pulmonary Alveolar Proteinosis

基于巨噬细胞的遗传性肺泡蛋白沉积症基因治疗

• 批准号: 8725410
• 负责人: Bruce C Trapnell
• 金额: $ 66.83万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725410
• 关键词: Ablation; Adverse event; Alveolar Macrophages; Autologous; Bone Marrow Purging; Bronchoalveolar Lavage; CD34 gene; CSF2RA gene; CSF2RB gene; Catabolism; Cells; Child; Clinical; Clinical Protocols; Clinical Trials; Data; Data Reporting; Development; Diagnosis; Disease; Dose; Engraftment; Event; Foundations; Functional disorder; Future; Gene Transfer; Generations; Genes; Genetic; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Health; Hematopoietic; Hematopoietic stem cells; Homeostasis; Host Defense; Human; Inherited; Investigational New Drug Application; Kinetics; Knowledge; Laboratories; Lentivirus Vector; Lipids; Longevity; Lung; Lung diseases; Mediating; Methods; Mission; Mus; Mutation; Outcome; Pathogenesis; Patients; Phase; Pluripotent Stem Cells; Population Sizes; Preparation; Procedures; Production; Proteins; Protocols documentation; Public Health; Pulmonary Alveolar Proteinosis; Receptor Gene; Research; Research Methodology; Respiratory Failure; Respiratory physiology; Safety; Saline; Signal Transduction; Testing; Therapeutic immunosuppression; Time; Transgenes; Transplantation; Validation; Work; Writing; alveolar homeostasis; base; caspase-9; cellular transduction; design; gene correction; gene therapy; in vivo; innovation; macrophage; man; novel strategies; novel therapeutic intervention; pre-clinical; preclinical efficacy; preclinical safety; preclinical study; precursor cell; promoter; receptor; receptor expression; receptor function; risk variant; stem; surfactant; transgene expression

DESCRIPTION (provided by applicant): Hereditary pulmonary alveolar proteinosis (hPAP) is a disorder of increased surfactant accumulation resulting in respiratory failure for which no pharmacologic therapy exists. It is caused by disruption of GM-CSF signaling to alveolar macrophages (AMs) by mutations in CSF2RA or CSF2RB, which encode GM-CSF receptor (GM-R) α and β subunits, respectively. The long-term goal is to develop gene therapy to restore GM-CSF signaling in AMs and, thereby, GM-CSF-dependent AM functions critical to surfactant clearance, alveolar homeostasis, lung function and host defense. The objective here is to evaluate a novel therapeutic approach, pulmonary macrophage transplantation (PMT). Preliminary data show that GM-R deficient (GM-RKO) mice develop hPAP lung disease that is identical to hPAP in humans including an increased level of pulmonary GM-CSF, which confers a selective survival advantage to AMs with functional GM-Rs. The central hypothesis is that safety-enhanced, lentiviral vector-mediated GM-R expression in hematopoietic stem/precursor cells (HSPCs), expansion into macrophages, and autologous PMT of gene-corrected cells without myeloablation will be effective and safe as therapy of hPAP. Preliminary data show that a single PMT treatment can correct hPAP in GM-RKO mice for at least one year (the longest time evaluated) without associated adverse events, and that human HSPCs can be readily transduced using well-established methods in the applicants' laboratory and expanded into macrophages expressing functional GM-Rs. The rationale is that anticipated results will inform the design of future a clinical trial and provide the preclinical safety and efficacy data, and GMP manufacturing procedures and validation data required to obtain regulatory approval to test this approach in humans. The hypothesis will be tested in four specific aims: 1) determine the efficiency and kinetics of gene transfer/PMT therapy of hPAP in GM-RKO mice; 2) optimize the expansion of human HSPC-derived, gene- corrected macrophages with maximum PMT engraftment potential; 3) determine the safety of gene transfer/PMT in preclinical studies related to gene transfer, macrophage expansion from HSPCs, PMT, and pharmacologic depletion of transduced cells in GM-RKO mice; 4) develop and validate protocols for the manufacture of HSPC-derived, gene-corrected macrophages, and write a clinical protocol and investigational new drug application for gene transfer/PMT therapy of hPAP. The approach is innovative because it departs markedly from the current inefficient, highly invasive method of physically removing surfactant by whole lung lavage and in- stead uses a novel approach to restore AM function. The proposed research is significant because it is expected to establish the feasibility of a specific therapy for children with hPAP and a new type of therapy (PMT) that may be useful for other diseases, and evaluate multiple safety improvements to reduce the risks of gene therapy. Results will inform a fundamental mechanism by which GM-CSF regulates AM population size, pro- vide an estimate of AM lifespan, and lay the foundation for the development of macrophage-based therapy.

描述（通过应用提供）：遗传性肺肺泡蛋白质病（HPAP）是一种生存积累增加的疾病，导致呼吸衰竭，不存在药物治疗。它是由GM-CSF信号传导对肺泡巨噬细胞（AMS）的破坏引起的，该突变分别编码GM-CSF受体（GM-R）α和β亚基的CSF2RA或CSF2RB中的突变。长期目标是开发基因疗法以恢复AMS中的GM-CSF信号传导，从而恢复GM-CSF依赖性AM的功能，对生存清除，肺泡稳态，肺功能和宿主防御至关重要。这里的目的是评估一种新型的治疗方法，肺巨噬细胞移植（PMT）。初步数据表明，GM-R缺乏症（GM-RKO）小鼠患有HPAP肺疾病，与人类的HPAP相同，包括肺部GM-CSF水平升高，这使功能性GM-RS具有选择性的生存优势。中心假设是，在造血词干/前体细胞（HSPC）中，安全增强的慢病毒介导介导的GM-R表达，在没有骨髓的基因校正细胞的自体PMT中扩张到巨噬细胞中，将有效，并且是HPAP的治疗。初步数据表明，单一的PMT治疗可以纠正GM-RKO小鼠的HPAP至少一年（评估时间为最长的时间）而没有相关的不良事件，并且可以在应用实验室中使用良好的方法来轻松地翻译人类HSPC，并扩展到表达功能GM-RS的巨噬细胞中。理由是，预期的结果将为未来的临床试验的设计提供信息，并提供临床前的安全性和效率数据，以及GMP 获得监管机构批准所需的制造程序和验证数据，以在人类中测试这种方法。该假设将以四个具体目的进行检验：1）确定GM-RKO小鼠中HPAP基因转移/PMT治疗的效率和动力学； 2）优化具有最大PMT植入潜力的人类HSPC衍生的基因校正巨噬细胞的扩展； 3）确定基因转移/PMT在与基因转移，巨噬细胞扩张相关的临床前研究中的安全性。 4）制定和验证用于制造HSPC衍生的，基因校正的巨噬细胞的方案，并为HPAP的基因转移/PMT治疗撰写临床方案和研究新药物应用。这种方法具有创新性，因为它明显偏离了当前效率低下，高度侵入性的方法，即通过整个肺灌洗，并使用一种新颖的方法来恢复AM功能。拟议的研究很重要，因为预计它将确定针对HPAP儿童和一种新型疗法（PMT）的特定疗法的可行性，该治疗可能对其他疾病有用，并评估多种安全性改善以降低基因疗法的风险。结果将为GM-CSF调节AM人口规模的基本机制提供依据，从而估算了AM寿命，并为基于巨噬细胞的治疗的发展奠定了基础。