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（由申请人提供）：遗传性肺泡蛋白沉积症（hPAP）是一种表面活性剂积累增加导致呼吸衰竭的疾病，目前尚无药物治疗方法。它是由于 CSF2RA 或 CSF2RB 突变破坏了 GM-CSF 向肺泡巨噬细胞 (AM) 发出的信号而引起的，CSF2RA 或 CSF2RB 分别编码 GM-CSF 受体 (GM-R) α 和 β 亚基。长期目标是开发基因疗法来恢复 AM 中的 GM-CSF 信号传导，从而恢复对表面活性剂清除、肺泡稳态、肺功能和宿主防御至关重要的 GM-CSF 依赖性 AM 功能。这里的目的是评估一种新的治疗方法，即肺巨噬细胞移植（PMT）。初步数据显示，GM-R 缺陷 (GM-RKO) 小鼠会患上与人类 hPAP 相同的 hPAP 肺部疾病，包括肺部 GM-CSF 水平升高，这赋予具有功能性 GM-R 的 AM 选择性生存优势。核心假设是，安全性增强、慢病毒载体介导的造血干细胞/前体细胞 (HSPC) 中 GM-R 表达、扩增至巨噬细胞以及基因校正细胞的自体 PMT（无需清髓）将是有效且安全的 hPAP 治疗方法。初步数据显示，单次 PMT 治疗可以纠正 GM-RKO 小鼠中的 hPAP 至少一年（评估的最长时间），而不会出现相关不良事件，并且可以使用申请人实验室中成熟的方法轻松转导人类 HSPC，并扩展到表达功能性 GM-R 的巨噬细胞。理由是预期结果将为未来临床试验的设计提供信息，并提供临床前安全性和有效性数据以及 GMP 获得监管部门批准以在人体上测试这种方法所需的制造程序和验证数据。该假设将在四个具体目标上进行测试：1）确定 GM-RKO 小鼠中 hPAP 基因转移/PMT 疗法的效率和动力学； 2) 优化人类 HSPC 衍生的基因校正巨噬细胞的扩增，使其具有最大的 PMT 植入潜力； 3) 确定基因转移/PMT 在 GM-RKO 小鼠中与基因转移、HSPC 巨噬细胞扩增、PMT 和转导细胞药理耗竭相关的临床前研究中的安全性； 4) 开发和验证 HSPC 衍生的基因校正巨噬细胞的生产方案，并编写 hPAP 基因转移/PMT 治疗的临床方案和研究性新药申请。该方法具有创新性，因为它明显不同于目前通过全肺灌洗物理去除表面活性剂的低效、高侵入性方法，而是使用一种新方法来恢复 AM 功能。拟议的研究意义重大，因为它有望确定针对 hPAP 儿童的特定疗法的可行性以及可能对其他疾病有用的新型疗法 (PMT)，并评估多项安全性改进以降低基因疗法的风险。结果将揭示GM-CSF调节AM群体大小的基本机制，提供AM寿命的估计，并为基于巨噬细胞的治疗的发展奠定基础。