Novel non-genotoxic ligand-based CD117-directed CAR T conditioning in the context of hematopoietic stem cell transplantation and leukemia treatment

造血干细胞移植和白血病治疗背景下新型非基因毒性配体 CD117 定向 CAR T 调理

• 批准号: 10613307
• 负责人: Gianna Branella
• 金额: $ 3.86万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10613307
• 关键词: Acute; Acute Myelocytic Leukemia; Adverse event; Antigen Targeting; Antigens; Bone Marrow; Bone Marrow Ablation; Bone Marrow Cells; Busulfan; Cancer Relapse; Cell Line; Cell Therapy; Cells; Cellular Stress; Child; Childhood Acute Myeloid Leukemia; Chimeric Proteins; Clinical; Coculture Techniques; Cytotoxic T-Lymphocytes; Data; Development; Disease; Dose; Drops; Effector Cell; Engraftment; FDA approved; Future; Goals; Hematinics; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histocompatibility; Immune; In Vitro; Incidence; Infertility; Knowledge; Leukemic Cell; Ligands; Major Histocompatibility Complex; Malignant - descriptor; Malignant Neoplasms; Memory; Modification; Monoclonal Antibodies; Myelogenous; Myeloid Progenitor Cells; Organ failure; Pathologist; Patients; Phenotype; Prognosis; Proteins; Proto-Oncogene Protein c-kit; Protocols documentation; Receptor Signaling; Recombinants; Refractory Disease; Regimen; Relapse; Resistance; Second Primary Cancers; Serum; Signal Transduction; Specificity; Stains; Stem Cell Factor; Stem cell transplant; Stress; Structure; Survival Rate; T memory cell; T-Lymphocyte; Therapeutic; Therapeutic Uses; Tissues; Toxic effect; Transfection; Translating; Transplantation; Transplantation Conditioning; Tumor Antigens; Tumor Burden; Whole-Body Irradiation; acute myeloid leukemia cell; antigen binding; cancer cell; chemotherapeutic agent; chemotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; conditioning; cytotoxic; cytotoxicity; design; effective therapy; effectiveness evaluation; exhaustion; experimental study; genotoxicity; graft vs host disease; graft vs leukemia effect; hematopoietic cell transplantation; improved; in vivo; leukemia; leukemia relapse; leukemia treatment; leukemic stem cell; mouse model; neoplastic cell; novel; pediatric patients; preconditioning; receptor; receptor expression; side effect; stem cells; targeted treatment; transgene expression; transplantation therapy; treatment strategy; tumor; γδ T cells; Acute; Acute Myelocytic Leukemia; Adverse event; Antigen Targeting; Antigens; Bone Marrow; Bone Marrow Ablation; Bone Marrow Cells; Busulfan; Cancer Relapse; Cell Line; Cell Therapy; Cells; Cellular Stress; Child; Childhood Acute Myeloid Leukemia; Chimeric Proteins; Clinical; Coculture Techniques; Cytotoxic T-Lymphocytes; Data; Development; Disease; Dose; Drops; Effector Cell; Engraftment; FDA approved; Future; Goals; Hematinics; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Histocompatibility; Immune; In Vitro; Incidence; Infertility; Knowledge; Leukemic Cell; Ligands; Major Histocompatibility Complex; Malignant - descriptor; Malignant Neoplasms; Memory; Modification; Monoclonal Antibodies; Myelogenous; Myeloid Progenitor Cells; Organ failure; Pathologist; Patients; Phenotype; Prognosis; Proteins; Proto-Oncogene Protein c-kit; Protocols documentation; Receptor Signaling; Recombinants; Refractory Disease; Regimen; Relapse; Resistance; Second Primary Cancers; Serum; Signal Transduction; Specificity; Stains; Stem Cell Factor; Stem cell transplant; Stress; Structure; Survival Rate; T memory cell; T-Lymphocyte; Therapeutic; Therapeutic Uses; Tissues; Toxic effect; Transfection; Translating; Transplantation; Transplantation Conditioning; Tumor Antigens; Tumor Burden; Whole-Body Irradiation; acute myeloid leukemia cell; antigen binding; cancer cell; chemotherapeutic agent; chemotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; conditioning; cytotoxic; cytotoxicity; design; effective therapy; effectiveness evaluation; exhaustion; experimental study; genotoxicity; graft vs host disease; graft vs leukemia effect; hematopoietic cell transplantation; improved; in vivo; leukemia; leukemia relapse; leukemia treatment; leukemic stem cell; mouse model; neoplastic cell; novel; pediatric patients; preconditioning; receptor; receptor expression; side effect; stem cells; targeted treatment; transgene expression; transplantation therapy; treatment strategy; tumor; γδ T cells

PROJECT SUMMARY It is essential to study improved therapies for the treatment of advanced acute myeloid leukemia (AML), as nearly 30% of children will relapse or have refractory disease, bringing the survival down to a mere 20%. Hematopoietic stem cell transplantation (HSCT) is curative for relapsed AML; however, the genotoxicity of bone marrow (BM) conditioning represents a substantial barrier in its use. Current BM conditioning regimens consist of a combination of alkylating chemotherapeutic agents such as busulfan and high doses of total body irradiation (TBI). Similar to the non-targeted chemotherapeutics used as first-line treatment for AML, these conditioning agents are also highly genotoxic and have several harmful toxicities outside of the hematopoietic compartment, such as organ failure and secondary malignancies. In contrast, cellular therapies provide strong support for the use of targeted therapies in the relapsed setting, as evident by five total FDA approved chimeric antigen receptor (CAR) products for the treatment of relapsed cancers, three of which have been approved within the last year. Therefore, we hypothesize that the development of non-genotoxic conditioning regimens that specifically target hematopoietic stem cells (HSCs) and AML will be transformative to the treatment of the disease. The use of c- kit (CD117) as a target for non-genotoxic conditioning has been explored by our lab and others due to its expression on HSCs. Up to 90% of AML patients have c-kit expression, and this expression correlates with poor prognosis and resistance to chemotherapy. We hypothesize that specifically targeting c-kit for AML treatment will reduce tumor burden in addition to serving as a non-genotoxic conditioning regimen prior to HSCT. This proposal seeks to explore a novel ligand-based c-kit directed CAR (SCF CAR) to target HSCs (Specific Aim 1) and AML (Specific Aim 2) by utilizing the c-kit receptor’s natural ligand stem cell factor (SCF) as the recombinant antigen binding domain of our CAR. Importantly, ligands offer a greater understanding of receptor-ligand interactions and can potentially reduce tonic CAR signaling through enhanced protein stability, leading to less T cell exhaustion, among other key advantages. Furthermore, our lab has optimized the use of γδ T cells as a cytotoxic alternative to ⍺β T cells. γδ T cells are uniquely beneficial for this setting since they i) are innate immune cells that do not form memory T cell phenotypes to the same extent as ⍺β T cells, ii) do not cause graft-vs-host disease (GvHD) when transplanted across major histocompatibility (MHC) barriers, iii) contribute to the innate killing of leukemia cells via graft-vs-leukemia (GvL) interactions, as γδ T cells can recognize cancer cell stress antigens, and iv) have been shown to enhance survival and decrease leukemia relapse post-HSCT. We therefore propose utilizing the cytotoxic abilities of γδ T cells to capitalize on these effects, as our lab has optimized the ex vivo expansion and modification of γδ T cells via the development of a serum-free protocol, making our lab uniquely suited to perform these experiments. Our goal is to advance the use of CAR T therapy to patients with AML and to elucidate the broader application of a combination cancer and bone marrow conditioning treatment.

项目摘要 研究改进治疗晚期急性髓样白血病（AML）的疗法至关重要，因为 30％的儿童将退休或患有难治性疾病，使生存率仅为20％。造血 干细胞移植（HSCT）可用于复发的AML。但是，骨髓（BM）的遗传毒性 条件代表其使用的实质性障碍。当前的BM调节方案由 绿色化学治疗剂（例如busulfan和高剂量的全身照射）的组合 （TBI）。类似于用作AML的一线治疗的非靶向化学治疗剂，这些条件 药物也具有高度的遗传毒性，并且在造血区外有几种有害毒性， 例如器官衰竭和继发性恶性肿瘤。相反，细胞疗法为 在继电器环境中使用靶向疗法，这是由五个FDA批准的嵌合抗原受体证明的 （汽车）用于处理继电器癌的产品，其中三种在去年已获得批准。 因此，我们假设非生物毒性调节方案的发展，该方案专门针对 造血干细胞（HSC）和AML将转化为疾病的治疗。使用C- 套件（CD117）作为非生物毒性调节的目标，我们的实验室和其他因素探索了它 在HSC上的表达。多达90％的AML患者具有C-KIT表达，并且该表达与较差 预后和对化学疗法的抵抗力。我们假设该专门针对C-KIT进行AML治疗 除了在HSCT之前用作非生物毒性调节方案外，还将减少肿瘤燃烧。这 提案旨在探索一种新型的基于配体的C-KIT定向汽车（SCF CAR）来瞄准HSC（特定AIM 1） 和AML（特定目标2）通过使用C-KIT接收器的天然配体干细胞因子（SCF）作为重组 我们汽车的抗原结合域。重要的是，配体对接收器配体有更多的了解 相互作用，可以通过增强的蛋白质稳定性来减少强直性汽车信号传导，从而导致T较少 细胞耗尽，除其他关键优势。此外，我们的实验室优化了γδT细胞的使用 ⍺βT细胞的细胞毒性替代品。 γδT细胞在这种情况下具有独特的有益，因为它们是先天免疫 不形成记忆T细胞表型的细胞与⍺βT细胞的程度相同，ii）不会引起移植物-VS宿主 疾病（GVHD）在主要组织相容性（MHC）障碍物（III）中移植时，有助于先天 由于γδT细胞可以识别癌细胞应激 抗原和IV）已显示可增强HSCT后血症并降低白血病继电器。因此，我们 提出利用γδT细胞的细胞毒性能力来利用这些影响，因为我们的实验室优化了 通过开发无血清方案的开发γδT细胞的离体扩展和修饰，使我们的实验室 独特地适合执行这些实验。我们的目标是推进对患者的使用CAR T治疗 AML并阐明癌症组合和骨髓调理治疗的更广泛应用。