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: 9888096
• 负责人: Bruce R Blazar
• 金额: $ 84.95万
• 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 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9888096
• 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; 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; Pancytopenia; 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 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.

摘要 范可尼贫血（FA）是一种遗传性骨髓（BM）衰竭性疾病，由DNA的内在缺陷引起 修复导致癌症风险增加，如急性髓性白血病和鳞状细胞癌。 全世界每年约有13万名新生儿受到FA的影响。目前，唯一的治疗方法 FA的血液学并发症是来自合适的造血干细胞移植（HSCT）的同种异体BM或造血干细胞移植。 HLA供体。一个关键的组成部分是准备受体BM使用某种形式的条件反射，以消除 病变细胞并促进供体产物的植入。目前使用的所有FA预处理方案 依赖于使用烷基化化疗药物和/或辐射，这两者都与癌症相关。 发生继发性恶性肿瘤的风险增加，特别是在DNA修复疾病如FA中。作为 作为一种替代策略，靶向造血干细胞（HSC）的抗体药物缀合物（ADC）是一种有前途的治疗方法。 促进基因修饰的自体或同种异体移植物植入的非基因毒性方法。最近的研究 已经显示ADC与缀合至免疫毒素的CD 45或CD 117（c-Kit）抗体的有效使用 皂草素（SAP）。由于普遍的共识是遗传毒性条件应避免在FA和其他 我们建议开发新的方法来克服这些关键的限制， 基因治疗和造血干细胞移植方案因此，在目标1中，我们将开发非遗传毒性条件作用 使用FANCA敲除小鼠模型的FA方案，以最佳地耗尽残留的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尽可能多的宿主HSC， 将存在残留宿主HSC的风险，这可能导致在移植后残留疾病相关的造血作用。 基因修饰的细胞移植，以及非清髓性、T细胞耗竭的同种异体 HSCT。目的二是寻找一种新的方法来消除基因治疗或异基因造血干细胞移植后残留的FA细胞。 虽然目标1旨在避免allo-HSCT并发症，但并非所有FA患者都是基因治疗的良好候选人。 因此，在目标3中，我们将确定我们的新的非遗传毒性条件处理方法是否可以耗尽宿主HSC 并防止宿主免疫介导的骨髓移植排斥，从而允许同种异体HSC在Fanca-/- 小鼠拟议的研究将开发一种全新的非遗传毒性的自体预处理方法， HSC基因治疗是异基因HSC移植新方案的关键组成部分。此外，本发明还提供了一种方法， 我们描述了一种创新的策略，适用于基因治疗和同种异体移植，以消除 移植后可能发展成白血病细胞的残留和未校正的FA造血细胞。