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

基本信息

批准号：
10548833
负责人：
Jayakar V Nayak
金额：
$ 46.99万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
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/10548833
关键词：
Air Airway Disease Animal Model Anion Transport Proteins Anions Architecture Autologous Basal Cell Behavior assessment Bicarbonates Biocompatible Materials Biological Assay Biological Models CRISPR correction CRISPR/Cas technology Cell Count Cell Differentiation process Cell Fraction Cell Proliferation 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 Dissociation Electrolytes Engineering Engraftment Epidermal Growth Factor Epithelial Cells Epithelium 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 Proliferating Proteins Publishing Regulator Genes Respiratory Failure Respiratory System Respiratory Tract Infections Rodent Rodent Model Scientist Sinus Site Small Intestinal Submucosa Sorting Stem cell transplant Stromal Cells Surgical Models System Technology Testing Time Tissues Translating Translations Transplantation Transplantation Surgery VX-770 Water airway epithelium autosome behavior in vitro bioluminescence imaging cell behavior cell replacement therapy constitutive expression cystic fibrosis airway 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 stem cell expansion stem cell proliferation stem cell replacement stem cell therapy stem cells success symptom treatment treatment strategy

项目摘要

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.

项目摘要/摘要囊性纤维化（CF）是由囊性纤维化突变引起的常染色体隐性单基因疾病跨膜电导调节剂（CFTR）基因。最常见的突变称为∆F508，发生在〜85％的CF患者。由于CFTR基因编码为阴离子转运蛋白，因此CFTR突变改变了电解质和水运输，导致致密，病理，粘液和其他分泌物。最多 CFTR功能障碍的有害影响发生在呼吸系统中，复发感染和上和下气道的炎症。尽管医疗疗法取得了长足的进展，但对于改善CF的改善，耐用疗法仍然是巨大的未满足的需求。在过去的三年中，我们的完整科学家和医生的合作小组已决定为患有CF的患者制定一种新型的基于干细胞的治疗策略。对于几个关键原因，我们将我们的努力指向基于细胞的CF上呼吸道疾病的疗法主要人类气道基本干细胞，称为ABC。第一个主要里程碑是利用CRISPR/CAS9 基因组编辑技术可有效纠正CF患者培养的ABC中的∆F508突变接受鼻窦手术。这种基因校正方法已导致氯化阴离子的显着恢复 ABC的运输量从0-3％到30-40％。这种令人鼓舞和新发表的开发现在提供用于将CFTR基因校正的ABC重新引入的临床前路线图，作为干细胞的体内环境替代疗法。在此提案中，我们将严格确定体内最有效，最有效的生物材料平台人类ABC的移植和锻炼，并评估实验室中基因校正的ABC的行为使用上呼吸道移植的显微外科模型系统的活动物模型发达。概述的实验是临床前步骤，以便将这种方法转化为CF 患者为CF患者产生一种创新的和可能的变革性疗法，第一个茎基于细胞的人类气道疾病疗法。