cGMP Manufacture Of FVIII-Expressing Placental Cells For Hemophilia A

用于治疗 A 型血友病的表达 FVIII 的胎盘细胞的 cGMP 生产

• 批准号: 9811291
• 负责人: Graca Duarte Almeida-Porada
• 金额: $ 38.57万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9811291
• 关键词: Address; Adult; Animal Model; Animals; Antibodies; Antibody Formation; Biodistribution; Biomedical Engineering; Birth; Cell Therapy; Cells; Chronic; Clinic; Clinical; Clinical Trials; Coagulation Process; Codon Nucleotides; Connective Tissue; Cyclic GMP; Development; Engraftment; Enrollment; Evaluation; Exposure to; F8 gene; Factor VIII; Family health status; Fright; Future; Gene Delivery; Gene-Modified; General Population; Healthcare Systems; Hematoma; Hemophilia A; Hemorrhage; Hemostatic function; Human; Immune; Immunophenotyping; In Vitro; Infant; Infusion procedures; Inherited; Injections; Karyotype determination procedure; Lead; Lentivirus Vector; Modeling; Mothers; Mus; Muscle; Patients; Phase I Clinical Trials; Placenta; Plasma; Population; Production; Proteins; Quality of life; Replacement Therapy; Resistance development; Risk; Safety; Sheep; Site; Stem cells; Symptoms; Therapeutic; Transgenes; Treatment Efficacy; Tumorigenicity; Viral Vector; adeno-associated viral vector; arthropathies; base; cell bank; cellular transduction; clinically relevant; cost; curative treatments; gene product; gene therapy; improved; in vitro Assay; in vivo; inhibiting antibody; inhibitor/antagonist; integration site; meetings; neutralizing antibody; potency testing; prenatal; psychologic; systemic toxicity; therapeutic evaluation; vector

PROJECT SUMMARY Current treatments for hemophilia A consist of frequent FVIII infusions to maintain hemostasis. Replacement therapy is lifelong, expensive, unavailable for ~75% of the world's hemophiliacs, and >30% of patients develop inhibitors to Factor VIII (FVIII). There is thus a critical need to develop affordable HA therapies with longer- lasting benefit or permanent cure. Clinical trials directly infusing AAV vectors for in vivo FVIII gene delivery have yielded promising results in adults, but pre-existing neutralizing antibodies (NAbs) to AAV are present in up to 50% of the population and inhibit target cell transduction when AAV is delivered systemically. As a result, clinical trials using AAV can currently only enroll subjects with no- or low-titer pre-existing AAV NAbs. Prenatal treatment (PNTx) for hemophilia A would circumvent the immune barriers present in adult patients, and could induce lifelong tolerance to FVIII, and thereby eliminate the risk of FVIII inhibitor formation, the most feared problem in treatment/management of hemophilia A. Although direct injection of viral vectors into prenatal recipients would likely be successful, numerous safety concerns need to be addressed before this approach can enter the clinic. A cell and gene therapy-based approach, in which cells are transduced in vitro, would enable multiple safeguards to be implemented in production, e.g., copy number and integration site analyses, that are not possible with direct vector injection. Such an approach would also eliminate the possibility of off-target gene-modification. Using a large animal model of PNTx, we showed that an off-the-shelf therapy comprised of human placental cells (PLCs) transduced with a lentiviral vector (LV) encoding a bioengineered, codon-optimized FVIII transgene, designated mcoET3, resulted in plasma FVIII activity levels that exceeded that of control non-transplanted animals by over 20%, and that these levels were maintained for at least 1 year after birth. Based on these results, a Pre-IND meeting with the FDA was held to discuss the path to a clinical trial. It was agreed that while it was vital to consider safety of the product to safeguard the patient, it was also imperative to protect the mother, particularly with respect to possible exposure to the gene- modified cells and/or gene product. To directly address this FDA guidance, we herein propose to: Aim 1: Manufacture, characterize, and authenticate, in vitro, a gene-modified PLC-based hemophilia A therapeutic product under cGMP conditions; and Aim 2: Confirm the therapeutic potential and safety of mcoET3- expressing human PLCs in vivo, in mice and in a large animal model of PNTx, determine whether there is transfer of the product across the placenta to the mother following PNTx, and if so, define the immune consequences of such transfer. At the completion of this project, we expect to have demonstrated the innocuity of the PNTx approach, and to have developed a safe and effective PLC-based hemophilia A therapeutic that could be used in a PNTx Phase-I clinical trial.

项目摘要 血友病的当前治疗方法由频繁的FVIII输注来维持止血。替代品 治疗是终生的，昂贵的，无法获得约75％的世界血友病，> 30％的患者发展 VIII因子（FVIII）的抑制剂。因此，迫切需要开发负担得起的HA疗法， 持久的好处或永久治愈。临床试验直接注入体内FVIII基因递送的AAV载体 在成年人中产生了有希望的结果，但是对AAV的中和抗体（NAB）存在于 当AAV系统地交付时，多达50％的人群并抑制靶细胞转导。作为 结果，使用AAV的临床试验目前只能招募具有无现或低位剂的AAV NAB的受试者。 血友病A的产前治疗（PNTX）将规避成年患者中存在的免疫屏障， 并可能引起对FVIII的终身耐受性，从而消除FVIII抑制剂形成的风险，最多 令人担心的血友病治疗/治疗中的问题。尽管将病毒向量直接注射到 产前接收者可能会成功，在此之前需要解决许多安全问题 方法可以进入诊所。细胞和基因治疗方法，其中细胞在体外转导， 将使多个保障措施在生产中实施，例如副本编号和集成站点 分析，直接向量注射是不可能的。这样的方法也将消除 脱靶基因修饰的可能性。使用大型的PNTX动物模型，我们证明了一个现成的 由用慢病毒载体（LV）转导的人类胎盘细胞（PLC）的治疗 指定的MCOET3的生物工程，密码子优化的FVIII转基因导致血浆FVIII活性水平 这超过了对控制非移植动物的动物的超过20％以上，并且这些水平得到了维持 出生后至少1年。基于这些结果，举行了与FDA的预定会议，讨论 临床试验的途径。同意的是，考虑到产品的安全以保护该产品至关重要 患者，必须保护母亲，特别是在可能暴露于基因的情况下 修饰的细胞和/或基因产物。为了直接解决此FDA指南，我们在此建议：AIM 1： 在体外制造，表征和身份验证基因改性PLC的基因性血友病A治疗 CGMP条件下的产品；和目标2：确认MCOET3-的治疗潜力和安全性 在体内表达人体PLC，在小鼠和大型动物模型中，确定是否存在 在PNTX之后，产品在胎盘上转移到母亲，如果是，则定义免疫 这种转移的后果。该项目完成时，我们希望已经证明 PNTX方法的无害性，并开发出一种安全有效的基于PLC的血友病A 可以在PNTX I期临床试验中使用的治疗性。