Nongenotoxic conditioning for gene therapy and allogeneic transplantation in Fanconi anemia

范可尼贫血基因治疗和同种异体移植的非基因毒性调理

• 批准号: 10305635
• 负责人: Bruce R Blazar
• 金额: $ 6.75万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-20 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10305635
• 关键词: Affect; Alloantigen; Allogeneic Bone Marrow Transplantation; Allogenic; Amanitins; Antibodies; Antibody-drug conjugates; Autologous; Biological Assay; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Bone marrow failure; Candidate Disease Gene; Cell Count; Cell Transplantation; Cell physiology; Cells; Child; Consensus; Cyclophosphamide; DNA Damage; DNA Repair; DNA Repair Disorder; Defect; Development; Disease; Dose; Engineering; Engraftment; Environment; Fanconi Anemia Complementation Group A Protein; Fanconi' s Anemia; Future; Gene-Modified; Genetic; Graft Rejection; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histopathology; Homologous Transplantation; Immune; Immunotoxins; Infusion procedures; Inherited; Knockout Mice; Lentivirus Vector; Leukemic Cell; Life; Measures; Mediating; Methods; Mus; Myeloproliferative disease; Outcome; PTPRC gene; Patients; Peripheral; Pharmaceutical Preparations; Process; Proto-Oncogene Protein c-kit; Protocols documentation; Radiation therapy; Regimen; Residual Tumors; Residual state; Ribosomes; Risk; Second Primary Cancers; Siblings; Squamous cell carcinoma; T-Lymphocyte; Therapeutic; Tissues; Toxic effect; Toxin; acute myeloid leukemia cell; antibody conjugate; antigen-specific T cells; base; cancer risk; cell transformation; chemotherapy; conditioning; cytotoxic; gene therapy; gene transplantation for gene therapy; genotoxicity; in vivo; innovation; irradiation; knock-down; leukemia; leukemic transformation; mouse model; novel; novel strategies; post-transplant; prevent; risk minimization; small hairpin RNA; standard of care; stem cell engraftment; stem cell gene therapy; stem cell population; stem cells

ABSTRACT Fanconi anemia (FA) is an inherited bone marrow (BM) failure disorder resulting from an intrinsic defect in DNA repair leading to an increased risk of cancers such as acute myeloid leukemia and squamous cell carcinoma. Approximately 130,000 children born worldwide each year are affected by FA. Currently, the only cure for the hematologic complications of FA is an allogeneic BM or hematopoietic stem cell transplant (HSCT) from a suitable HLA donor. A key component is preparing the recipient BM using some form of conditioning to both eliminate diseased cells and promote engraftment of donor product. All of the currently used conditioning regimens for FA rely on the use of alkylating chemotherapy drugs and/or irradiation, both of which are associated with an increased risk of developing secondary malignancies, especially in DNA repair disorders like FA. As an alternative strategy, antibody drug conjugates (ADCs) targeting hematopoietic stem cells (HSCs) are a promising nongenotoxic method of facilitating engraftment of gene-modified autologous or allogeneic grafts. Recent studies have shown the effective use of ADCs with either CD45 or CD117 (c-Kit) antibodies conjugated to the immunotoxin saporin (SAP). Since the general consensus is that genotoxic conditioning should be avoided in FA and other diseases with DNA repair defects, we propose to develop novel approaches to overcome these critical limitations for current gene therapy and HSCT protocols. Thus, in Aim 1, we will develop nongenotoxic conditioning regimens for FA using a FANCA knockout mouse model to optimally deplete residual HSCs and facilitate engraftment of gene-modified or allogeneic cells. Despite eliminating as many host HSCs as safely possible, there will be a risk of remaining host HSCs, which can result in residual disease-related hematopoiesis after transplantation of gene-modified cells and also in the setting of nonmyeloablative, T-cell depleted allogeneic HSCT. Aim 2 will pursue a novel approach to eliminate residual FA cells after gene therapy or allogeneic HSCT. While Aim 1 seeks to avoid allo-HSCT complications, not all FA patients will be good candidates for gene therapy. Thus, in Aim 3, we will determine whether our novel nongenotoxic conditioning approach can deplete host HSCs and prevent host immune-mediated BM graft rejection and thus permit allogeneic HSC engraftment in Fanca-/- mice. The proposed studies will develop an entirely novel approach of nongenotoxic conditioning for autologous HSC gene therapy and as a key component of a novel regimen for allogeneic HSC transplantation. In addition, we describe an innovative strategy, applicable to both gene therapy and allogeneic transplantation, to eliminate residual and uncorrected FA hematopoietic cells that may develop into leukemic cells post-transplant.

抽象的 Fanconi贫血（FA）是由DNA中固有缺陷导致的遗传骨髓（BM）衰竭障碍 修复导致癌症急性髓样白血病和鳞状细胞癌等癌症的风险增加。 每年出生的大约130,000名儿童受FA的影响。目前，唯一的治疗方法 Fa的血液学并发症是一种同种异体BM或造血干细胞移植（HSCT） HLA捐助者。一个关键组件是使用某种形式的调理来准备收件人BM以消除 患病的细胞并促进供体产品的植入。所有当前使用的适用于FA的调节方案 依靠使用烷基化化疗药物和/或辐照，这两者都与 增加了继发性恶性肿瘤的风险，尤其是在FA等DNA修复障碍中。作为 替代策略，靶向造血干细胞（HSC）的抗体药物缀合物（ADC）是一个有希望的 促进基因修饰自体或同种异体移植物促进植入的非核毒性方法。最近的研究 已经显示了与CD45或CD117（C-KIT）抗体相结合的ADC的有效使用 Saporin（SAP）。由于一般共识是应避免在FA中避免遗传毒性调节 患有DNA修复缺陷的疾病，我们建议开发新的方法来克服这些临界局限性 用于当前的基因治疗和HSCT方案。因此，在AIM 1中，我们将开发非核调理 使用FANCA敲除鼠标模型的FA方案，以最佳耗尽残留HSC并促进 基因修饰或同种异体细胞的植入。尽管尽可能安全地消除了HOST HSC，但 将有剩余宿主HSC的风险，这可能导致与疾病相关的残留造血 基因改性细胞的移植以及在非乳清，T细胞耗尽同种异体的情况下 HSCT。 AIM 2将采用一种新的方法来消除基因治疗或同种异体HSCT后消除残留的FA细胞。 虽然AIM 1试图避免Allo-HSCT并发症，但并非所有FA患者都将成为基因治疗的良好候选者。 因此，在AIM 3中，我们将确定我们的新型非核调理方法是否可以耗尽宿主HSC 并防止宿主免疫介导的BM移植物排斥反应，从而允许FANCA中的同种异体HSC植入 - / - 老鼠。拟议的研究将开发一种完全新颖的非核调节方法的方法 HSC基因疗法，是同种异体HSC移植的新型方案的关键组成部分。此外， 我们描述了一种适用于基因治疗和同种异体移植的创新策略，以消除 移植后可能发展成白血病细胞的残留和未矫正的FA造血细胞。