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 使用微创，超声引导的方法已经是宫内干细胞移植（IUTX） 在约50名人类患者中，患有14种不同的遗传疾病，证明此过程呈最少 胎儿和母亲的风险。血友病A（HA）是一种理想的疾病，因为75％ 可以通过产前筛查诊断出HA病例。 IUTX可以治愈，或者至少转化为严重 HA达到中等/轻度的表型，大大减少了对FVIII输注的需求。重要的是，接触 早期免疫发育过程中载体编码的FVIII会诱导耐受性FVIII并避免形成 抑制剂，HA治疗中最严重和威胁性的并发症。确实，最近的研究表明 严重和非严重HA患者，FVIII抑制剂的发展与增加有关 死亡风险。使用相同的临床前动物模型，允许划定使用中的条件 临床IUTX，我们表明了转导的骨髓基质祖细胞（MSP）的产前输注 导致长期稳健的多器官整合到实质和脉管系统中，长期释放 分泌的转基因产物进入循环和免疫耐受性的发展。使用相同 绵羊模型，在IUTX的羊水衍生的基质祖细胞（AFP）之后获得了类似的结果。 利用一系列模仿遗传学，抑制剂形成和严重的临床症状 人类HA的形式，我们表明父亲（单倍型）MSP的产后移植 表达高水平的FVIII导致两只小儿HA绵羊的完全表型校正，逆转 现有的呼吸道，并恢复正常的体育锻炼。但是，就像人类的FVIII输注一样 患者，这两只绵羊在产后治疗后产生了FVIII抑制剂。因此，这项工作提供了 令人信服的证据表明，如果FVIII表达基质细胞的移植对于HA可以治愈，如果 可以避免抑制剂形成。因此，我们假设使用基于IUTX的策略来治疗HA 将克服因素替代疗法和 当前治疗HA的基因疗法方法，即缺乏持续的FVIII表达和免疫学 对治疗蛋白的反应。我们建议：1）确定理想的细胞源以获得最高 长期植入水平和体内分布模式，使FVIII最大化 循环; 2）使用HA绵羊证明使用最佳单元格和FVIII的IUTX方法 转基因构建体可以是治愈的，也可以至少永久将严重的HA转化为轻度表型； 3）测试 在产后FVIII输注和/或诱导公差后，是否接受IUTX排除抑制剂诱导 即使在佐剂中对FVIII的产后挑战之后，这种情况仍然存在。完成后，这些研究将证明 将细胞用作FVIII输送平台的安全性和功效，并证明IUTX治愈或 改善HA表型，并击败目前阻碍临床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