Prenatal pulmonary cell gene editing to cure monogenic lung diseases

产前肺细胞基因编辑治疗单基因肺部疾病

• 批准号: 10447104
• 负责人: William H. Peranteau
• 金额: $ 39.43万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10447104
• 关键词: Adult; Albumins; Alleles; Alveolar; Birth; CRISPR/Cas technology; Cause of Death; Cells; Cessation of life; Chronic Obstructive Pulmonary Disease; Chronic lung disease; Clustered Regularly Interspaced Short Palindromic Repeats; Cystic Fibrosis; DNA Double Strand Break; DNA Repair; Data; Development; Diagnosis; Disease; Epithelial Cells; Fetal Weight; Fetus; Foundations; GTP-Binding Protein alpha Subunits, Gs; Genes; Genetic; Genetic Diseases; Goals; Growth; Health; Hepatocyte; Homeostasis; Human; Immune; Immune Tolerance; Inherited; Injury; Knowledge; Leukocyte Elastase; Life; Liver; Long-Term Effects; Lung; Lung Transplantation; Lung diseases; Mediating; Medical; Mission; Modeling; Morbidity - disease rate; Mus; Mutation; Neonatal Hyperoxic Injury; Nonhomologous DNA End Joining; Organ; Pathology; Patients; Perinatal; Perinatal mortality demographics; Population; Proliferating; Protein Secretion; Proteins; Public Health; Publishing; Reporter; Research; Respiratory Failure; Risk; System; Technology; Testing; Therapeutic; Transplantation; United States National Institutes of Health; Work; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; base; cell type; disability; disease-causing mutation; endonuclease; epithelial stem cell; fetal; gene correction; human disease; human model; hyperoxia induced lung injury; in utero; in vivo; injured airway; innovation; mortality; mouse model; neonatal injury; novel; novel therapeutics; perinatal period; postnatal; prenatal; repaired; stem cells; success; surfactant; therapeutic gene

PROJECT SUMMARY Congenital monogenic lung diseases, including cystic fibrosis, surfactant protein disorders, and alpha-1 antitrypsin deficiency (A1ATD), can cause perinatal respiratory failure and death or chronic lung disease. Despite medical advances, therapy options are limited, often focused on treating disease complications, and culminating in the need for a lung transplant for many patients. Thus, there is a critical need for novel therapies for monogenic lung diseases. Many monogenic lung disease-causing mutations are well known, can be diagnosed before birth, and, often, one mutation is responsible for the majority of cases. A G→A mutation (Glu342Lys, the PiZ allele) in the SERPINA1 gene accounts for 90% of A1ATD mutations and results in severe disease, increasing the risk of developing chronic obstructive pulmonary disease. Advances in CRISPR gene editing technology provide an unprecedented opportunity to permanently correct disease-causing mutations in monogenic lung diseases after a single treatment. Although encouraging, in vivo CRISPR gene editing studies targeting other organs in adult mouse models of human diseases highlight limitations to the postnatal approach including low-levels of homology directed repair (HDR) due to inaccessible and nonproliferative target cells and a mature immune barrier. These obstacles are even more daunting in the postnatal lung, a barrier organ with immune and physical barriers in which only 1% of epithelial progenitor cells, the target cell population for most lung diseases, are cycling at homeostasis. In utero gene editing has the potential to overcome these barriers and treat perinatal lethal diseases before the onset of irreversible pathology. The fetus is immunologically tolerant and progenitor cells of multiple organs, including the lung, are highly proliferative and accessible during development. The objective of this proposal is to establish the feasibility of prenatal lung gene editing and use prenatal gene editing to treat a mouse A1ATD model as a model for monogenic lung diseases. Our central hypotheses are that prenatal pulmonary cell gene editing is more efficient than postnatal editing and prenatal gene editing will not have a detrimental effect on edited pulmonary progenitor cell fate. We hypothesize that prenatal HDR can provide therapeutic levels of circulating alpha-1 antitrypsin protein and pulmonary cell gene correction in the A1ATD mouse model. Our hypotheses are based on our published data demonstrating efficient liver and pulmonary epithelial cell editing via prenatal CRISPR-nonhomologous end joining (NHEJ). To attain our objective, we will pursue the following aims: 1) evaluate prenatal pulmonary cell gene editing in normal and neonatal injury states, 2) evaluate prenatal HDR targeting the lung and compare it to postnatal HDR, and 3) correct A1ATD by prenatal HDR and compare it to postnatal HDR. Our research is innovative in the prenatal timing and targeting of the lung for therapeutic gene editing. The significant contribution of this work will be to provide the foundation for a one- shot, long-term therapy that cures A1ATD and is applicable to other monogenic lung diseases.

项目摘要 先天性单基因肺疾病，包括囊性纤维化、表面活性蛋白紊乱和α-1 抗胰蛋白酶缺乏症（A1 ATD）可导致围产期呼吸衰竭和死亡或慢性肺病。尽管 医学进步，治疗选择有限，往往集中在治疗疾病并发症，并最终 许多病人需要肺移植。因此，迫切需要用于单基因治疗的新疗法。 肺部疾病许多单基因肺病致病突变是众所周知的，可以在出生前诊断， 而且通常一个突变导致了大多数病例一个G→A突变（Glu 342 Lys，PiZ等位基因）， SERPINA 1基因占A1 ATD突变的90%，并导致严重疾病，增加了 慢性阻塞性肺病CRISPR基因编辑技术的进展提供了 这是一个前所未有的机会，可以永久纠正单基因肺病中的致病突变， 一次治疗。尽管令人鼓舞，但针对成人其他器官的体内CRISPR基因编辑研究 人类疾病的小鼠模型突出了出生后方法的局限性，包括低水平的 同源性定向修复（HDR）由于不可接近和非增殖性靶细胞和成熟的免疫 屏障这些障碍在出生后的肺中更令人生畏，肺是一个具有免疫和生理功能的屏障器官。 屏障，其中只有1%的上皮祖细胞，大多数肺部疾病的靶细胞群， 在体内平衡循环。子宫内基因编辑有可能克服这些障碍， 在不可逆转的病理学发生之前，胎儿是免疫耐受的， 包括肺在内的多个器官的细胞在发育过程中高度增殖和可接近。的 该提案的目的是建立产前肺基因编辑的可行性，并使用产前基因编辑 将小鼠A1 ATD模型作为单基因肺疾病的模型进行治疗。我们的主要假设是， 产前肺细胞基因编辑比产后编辑更有效，而产前基因编辑不会 对编辑的肺祖细胞命运具有有害影响。我们假设产前HDR可以提供 A1 ATD中循环α-1抗胰蛋白酶蛋白和肺细胞基因校正的治疗水平 小鼠模型我们的假设是基于我们发表的数据，表明有效的肝脏和肺 通过产前CRISPR-非同源末端连接（NHEJ）进行上皮细胞编辑。为了实现我们的目标，我们将 追求以下目标：1）评估正常和新生儿损伤状态下的产前肺细胞基因编辑， 2)评估靶向肺的产前HDR并将其与产后HDR进行比较，以及3）通过产前HDR校正A1 ATD。 HDR，并将其与出生后的HDR进行比较。我们的研究在产前时机和肺靶向方面具有创新性 用于治疗性基因编辑这项工作的重大贡献将是为一个- 一次注射，长期治疗，治愈A1 ATD，适用于其他单基因肺病。

项目成果

Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.

• DOI: 10.1016/j.jconrel.2021.10.031
• 发表时间: 2022-01
• 期刊: Journal of controlled release : official journal of the Controlled Release Society
• 作者: Swingle KL;Billingsley MM;Bose SK;White B;Palanki R;Dave A;Patel SK;Gong N;Hamilton AG;Alameh MG;Weissman D;Peranteau WH;Mitchell MJ
• 通讯作者: Mitchell MJ

InUtero Gene Therapy: Progress and Challenges.

• DOI: 10.1016/j.molmed.2021.05.007
• 发表时间: 2021-08
• 期刊: Trends in molecular medicine
• 影响因子: 13.6
• 作者: Bose SK;Menon P;Peranteau WH
• 通讯作者: Peranteau WH

Foetal genome editing.

胎儿基因组编辑。

• DOI: 10.1097/gco.0000000000000854
• 发表时间: 2023
• 期刊: Current opinion in obstetrics & gynecology
• 影响因子: 2.1
• 作者: Bose,SouravK;Kennedy,Kara;Peranteau,WilliamH
• 通讯作者: Peranteau,WilliamH

Prenatal Gene Therapy for Metabolic Disorders.

• DOI: 10.1097/grf.0000000000000662
• 发表时间: 2021-12-01
• 期刊: Clinical obstetrics and gynecology
• 影响因子: 1.5
• 作者: Coons B;Peranteau WH
• 通讯作者: Peranteau WH

The Future of In Utero Gene Therapy.

子宫基因疗法的未来。

• DOI: 10.1007/s40291-020-00445-y
• 发表时间: 2020-04
• 期刊: MOLECULAR DIAGNOSIS & THERAPY
• 影响因子: 4
• 作者: Peranteau, William H.;Flake, Alan W.
• 通讯作者: Flake, Alan W.