Optimizing Surgical Transplant of CFTR Gene-Corrected Human Basal Stem Cells to the Upper Airway

优化 CFTR 基因校正的人类基底干细胞至上呼吸道的手术移植

• 批准号: 10361467
• 负责人: Jayakar V Nayak
• 金额: $ 47.01万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10361467
• 关键词: Air; Airway Disease; Animal Model; Anion Transport Proteins; Anions; Architecture; Autologous; Behavior; Bicarbonates; Biocompatible Materials; Biological Assay; Biological Models; CRISPR correction; CRISPR/Cas technology; Cell Count; Cell Differentiation process; Cell Fraction; Cell Survival; Cell Therapy; Cell Transplantation; Cells; Chimerism; Chlorides; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Collagen; Collagen Type I; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Delta F508 mutation; Development; Differentiated Gene; Discrimination; Electrolytes; Engineering; Engraftment; Epidermal Growth Factor; Epithelial; Epithelial Cells; FDA approved; Family suidae; Functional disorder; Green Fluorescent Proteins; Growth Factor; Heparin Binding; Human; Immune; Immunohistochemistry; In Vitro; Infiltration; Inflammation; Life Expectancy; Liquid substance; Luciferases; Lung; Maxillary Sinus; Measures; Mediating; Medical; Membrane; Mendelian disorder; Methods; Microsurgery; Modality; Modeling; Mucous body substance; Mus; Mutation; Operative Surgical Procedures; Outcome; Pathologic; Patients; Pattern; Physicians; Proteins; Publishing; Regulator Genes; Respiratory Failure; Respiratory System; Respiratory Tract Infections; Rodent; Rodent Model; Scientist; Sinus; Site; Small Intestinal Submucosa; Stem cell transplant; Stromal Cells; Surgical Models; System; Technology; Testing; Time; Tissues; Translating; Translations; Transplantation; VX-770; Water; airway epithelium; base; behavior in vitro; bioluminescence imaging; cell behavior; cell replacement therapy; cystic fibrosis airway epithelia; cystic fibrosis patients; disease-causing mutation; experimental study; gene correction; genome editing; human stem cells; immunosuppressed; improved; in vivo; in vivo engraftment; innovation; migration; mouse model; novel; personalized strategies; pre-clinical; protein expression; recruit; recurrent infection; replacement tissue; restoration; scaffold; stem cell expansion; stem cell proliferation; stem cell replacement; stem cell therapy; stem cells; success; symptom treatment; treatment strategy

PROJECT SUMMARY/ABSTRACT Cystic fibrosis (CF) is an autosomal recessive single-gene disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. The most common mutation, termed ∆F508, occurs in ~85% of CF patients. Because the CFTR gene encodes for an anion transport protein, CFTR mutations alter electrolyte and water transport, resulting in dense, pathologic, mucous and other secretions. The most harmful effects of CFTR dysfunction occur in the respiratory system, with recurrent infections and inflammation of the upper and lower airways. Despite substantial progress with medical therapies, there remains a tremendous unmet need for improved, durable therapies for CF. For the past 3 years, our collaborative group of complementary scientists and physicians has determined to develop a novel, stem cell-based treatment strategy for patients suffering from CF. For several critical reasons, we directed our efforts to cell-based therapy of CF upper airway disease using ex vivo-expanded, primary human airway basal stem cells, termed ABCs. The first major milestone was to utilize CRISPR/Cas9 genome editing technology to efficiently correct the ∆F508 mutation in ABCs cultured from CF patients undergoing sinus surgery. This gene correction approach has led to significant restoration of chloride anion transport from 0-3% to 30-40% in ABCs. This encouraging, and newly published, development now provides a pre-clinical roadmap for re-introducing CFTR gene-corrected ABCs into in vivo contexts as a stem cell replacement therapy. In this proposal, we will rigorously determine the most efficient, biomaterial platform for ex vivo-to-in vivo transplant and engraftment of human ABCs, and assess the behavior of gene-corrected ABCs in the lab and live animal model using a microsurgical model system of upper airway transplantation that we have developed. The experiments outlined are essential pre-clinical steps in order to translate this approach to CF patients to generate an innovative and possibly transformative therapy for patients with CF, and the first stem cell-based therapy for human airway disease.

项目概要/摘要 囊性纤维化（CF）是一种常染色体隐性遗传单基因疾病，由囊性纤维化突变引起 跨膜电导调节（CFTR）基因。最常见的突变称为 ΔF508，发生在 约 85% 的 CF 患者。由于 CFTR 基因编码阴离子转运蛋白，CFTR 突变会改变 电解质和水的运输，产生致密的、病理性的、粘液和其他分泌物。最 CFTR 功能障碍的有害影响发生在呼吸系统中，伴有反复感染和 上呼吸道和下呼吸道炎症。尽管医学治疗取得了重大进展，但 对于改进的、持久的 CF 疗法仍然存在巨大的未满足的需求。 在过去的三年里，我们由互补的科学家和医生组成的合作小组决定 为 CF 患者开发一种基于干细胞的新型治疗策略。对于几个关键的 由于这些原因，我们将努力方向转向使用离体扩增、基于细胞的治疗 CF 上呼吸道疾病。 原代人气道基底干细胞，称为 ABC。第一个重要里程碑是利用 CRISPR/Cas9 基因组编辑技术可有效纠正 CF 患者培养的 ABC 中的 ΔF508 突变 正在接受鼻窦手术。这种基因校正方法显着恢复了氯阴离子 ABC 中的运输从 0-3% 增加到 30-40%。这一令人鼓舞的新发布的发展现在提供了 将 CFTR 基因校正的 ABC 作为干细胞重新引入体内环境的临床前路线图 替代疗法。 在本提案中，我们将严格确定离体到体内最有效的生物材料平台 人类 ABC 的移植和植入，并评估基因校正 ABC 在实验室中的行为 使用我们拥有的上呼吸道移植显微外科模型系统的活体动物模型 发达。概述的实验是将这种方法转化为 CF 的重要临床前步骤 为 CF 患者提供一种创新的、可能具有变革性的治疗方法，以及第一个干细胞疗法 针对人类气道疾病的细胞疗法。