Prenatal Cell and Gene Therapy for Hemophilia A

A 型血友病的产前细胞和基因治疗

• 批准号: 10014648
• 负责人: Graca Duarte Almeida-Porada
• 金额: $ 69.89万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-14 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10014648
• 关键词: AFP gene; Adjuvant; Allogenic; Amniotic Fluid; Animal Model; Antibody Formation; Autologous; B-Lymphocytes; Binding; Biological Assay; Biomedical Engineering; Birth; Blood Circulation; Blood Coagulation Factor; Bone Marrow; Cell Therapy; Cells; Childhood; Clinical; Coagulation Process; Complication; Congenital Disorders; Country; Development; Diagnosis; Disease; Dose; Engraftment; Exposure to; F8 gene; Family health status; Family history of; Fetus; Frequencies; Fright; Genetic Diseases; Healthcare Systems; Hemophilia A; Human; IgG4; Immune; Immune Tolerance; Immune response; Immunologics; Individual; Infant; Infusion procedures; Life; Location; Marrow; Modeling; Molecular Chaperones; Mothers; Patients; Pattern; Phenotype; Physical activity; Population; Prenatal Diagnosis; Procedures; Production; Proteins; Replacement Therapy; Reporting; Resistance development; Risk; Safety; Sheep; Source; Stem cell transplant; Stromal Cells; Symptoms; T-Lymphocyte; Testing; Therapeutic; Therapeutic Intervention; Transgenes; Transplant Recipients; Transplantation; Ultrasonography; Work; base; boys; cell type; curative treatments; efficacy testing; engineered stem cells; fetal; gene therapy; genetic inhibitor; improved; in utero transplantation; in vivo; inhibitor/antagonist; minimal risk; minimally invasive; mortality risk; postnatal; pre-clinical; prenatal; prenatal exposure; prenatal testing; prevent; psychologic; response; stem cells; stromal progenitor; therapeutic gene; therapeutic protein; trafficking; transgene expression; vector

Project Summary Intrauterine stem cell transplantation (IUTx) using a minimally invasive, ultrasound-guided approach has been performed in ~50 human patients for 14 different genetic disorders, proving that this procedure poses minimal risk to both the fetus and the mother. Hemophilia A (HA) is an ideal disease to be treated by IUTx, since 75% of HA cases can be diagnosed through prenatal screening. IUTx could be curative, or at least convert severe HA to a moderate/mild phenotype, significantly reducing the need for FVIII infusions. Importantly, exposure to vector-encoded FVIII during early immunologic development would induce tolerance FVIII and avoid formation of inhibitors, the most serious and threatening complication in HA treatment. Indeed, recent studies showed in both severe and non-severe HA patients, that the development of FVIII inhibitors is associated with increased risk of death. Using the same preclinical animal model, that allowed the delineation of the conditions used in clinical IUTx, we showed that prenatal infusion of transduced bone marrow stromal progenitor cells (MSPs) resulted in robust long-term, multi-organ integration into both parenchyma and vasculature, long-term release of secreted transgene products into the circulation, and development of immune tolerance. Using the same sheep model, similar results were obtained after IUTx of amniotic fluid-derived stromal progenitors (AFPs). Utilizing a line of sheep that emulates the genetics, inhibitor formation, and clinical symptoms of the severe form of human HA, we showed that the postnatal transplantation of paternal (haploidentical) MSPs engineered to express high levels of FVIII led to complete phenotypic correction of two pediatric HA sheep, reversal of existing hemarthroses, and return to normal physical activity. However, like with FVIII infusions in human patients, these two sheep developed FVIII inhibitors following postnatal treatment. This work thus provides compelling evidence that the transplantation of FVIII-expressing stromal cells could be curative for HA, if inhibitor formation could be avoided. Therefore, we hypothesize that using an IUTx-based strategy to treat HA will overcome the two major shortcomings that have been observed following factor replacement therapy and current gene therapy approaches to treat HA, namely a lack of sustained FVIII expression and immunologic responses to the therapeutic protein. We propose to: 1) determine the ideal cell source to obtain the highest long-term engraftment levels and the in vivo distribution patterns that maximize release of FVIII into the circulation; 2) use HA sheep to demonstrate that an IUTx approach using the optimal cell source and FVIII transgene construct, can be curative, or at least permanently convert severe HA to a mild phenotype; 3) test whether receiving IUTx precludes inhibitor induction following postnatal FVIII infusion and/or induces tolerance that persists even after postnatal challenge with FVIII in adjuvant. Upon completion, these studies will prove the safety and efficacy of using cells as a FVIII delivery platform, and demonstrate the ability of IUTx to cure or improve HA phenotype, and defeat the immune-related hurdles that currently hinder clinical HA treatment.

项目摘要 子宫内干细胞移植（IUTx）采用微创，超声引导的方法， 在约50名患有14种不同遗传疾病的人类患者中进行，证明该程序对人类的影响最小。 对胎儿和母亲都有风险。甲型血友病（Hemophilia A，HA）是IUTx治疗的理想疾病， 的HA病例可通过产前筛查确诊。IUTx可以治愈，或至少将严重的 HA至中度/轻度表型，显著减少FVIII输注的需要。重要的是，暴露于 在免疫发育早期，载体编码的FVIII可诱导耐受性FVIII， 抑制剂的产生是HA治疗中最严重和最具威胁性的并发症。事实上，最近的研究表明， 重度和非重度HA患者，FVIII抑制物的发生与增加 死亡的风险。使用相同的临床前动物模型，这允许描述用于治疗的条件。 在临床IUTx中，我们发现产前输注转导的骨髓基质祖细胞（MSP） 导致稳健的长期、多器官整合到实质和血管系统中，长期释放 分泌的转基因产物进入循环，以及免疫耐受的发展。使用相同的 绵羊模型，羊水来源的基质祖细胞（AFPs）的IUTx后获得了类似的结果。 利用一系列的羊，模仿遗传，抑制剂的形成，和临床症状的严重 人HA的形式，我们表明，出生后移植的父亲（单倍性）MSP工程， 表达高水平的FVIII导致两只儿科HA羊的表型完全纠正，逆转了 存在关节积血，并恢复正常的体力活动。然而，与人体中的FVIII输注一样， 在这些患者中，这两只绵羊在产后治疗后产生了FVIII抑制剂。这项工作提供了 令人信服的证据表明，移植FVIII表达基质细胞可以治愈HA，如果 可以避免抑制剂的形成。因此，我们假设使用基于IUTx的策略治疗HA 将克服因子替代疗法后观察到的两个主要缺点， 目前治疗HA的基因治疗方法，即缺乏持续的FVIII表达和免疫抑制， 对治疗性蛋白质的反应。我们建议：1）确定理想的细胞来源，以获得最高的 长期植入水平和体内分布模式，使FVIII最大限度地释放到 2）使用HA绵羊来证明使用最佳细胞来源和FVIII的IUTx方法 转基因构建体，可以是治愈性的，或至少永久地将重度HA转化为轻度表型; 3）测试 接受IUTx是否排除了出生后FVIII输注后的抑制剂诱导和/或诱导耐受性 即使在出生后用佐剂中的凝血因子VIII激发后，这种情况仍然存在。一旦完成，这些研究将证明 使用细胞作为FVIII递送平台的安全性和有效性，并证明IUTx治愈或 改善HA表型，并克服目前阻碍临床HA治疗的免疫相关障碍。

项目成果

Acceptability of prenatal diagnosis and prenatal treatment of haemophilia using cell and gene therapies within US haemophilia community.

美国血友病社区使用细胞和基因疗法对血友病进行产前诊断和产前治疗的可接受性。

• DOI: 10.1111/hae.14805
• 发表时间: 2023
• 期刊: Haemophilia : the official journal of the World Federation of Hemophilia
• 作者: Pham,QuanDM;Thomson,SharonM;Schaible,BurkN;Mills,KevinD;Atala,Anthony;Porada,ChristopherD;Almeida-Porada,Graça
• 通讯作者: Almeida-Porada,Graça

Comparison of different gene addition strategies to modify placental derived-mesenchymal stromal cells to produce FVIII.

• DOI: 10.3389/fimmu.2022.954984
• 发表时间: 2022
• 期刊: Frontiers in immunology
• 影响因子: 7.3