Human Donor Stem/Progenitor Cells with Dual Gene Editing (CD33 and CLL-1) Enable "Two Hit" targeting of Acute Myeloid Leukemia"

具有双基因编辑（CD33 和 CLL-1）的人类供体干细胞/祖细胞能够实现急性髓系白血病的“两次打击”

• 批准号: 10112414
• 负责人: Abdullah M Ali
• 金额: $ 18.93万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-03 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10112414
• 关键词: Ablation; Activities of Daily Living; Acute Myelocytic Leukemia; Alleles; Allogenic; Antibody-drug conjugates; Antigen Targeting; Antigens; Biological Assay; Blast Cell; Bone Marrow; CD34 gene; CRISPR/Cas technology; Cell Line; Cells; Clinical; Clinical Research; Clinical Trials; Clonal Expansion; Colony-Forming Units Assay; Disease; Disease model; Disease remission; Dose; Drug Efflux; Dysmyelopoietic Syndromes; Engineering; Engraftment; Flow Cytometry; Funding; Gemtuzumab Ozogamicin; Genes; Genetic; Genomics; Goals; Grant; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; IL3RA gene; IL6 gene; IL8 gene; Immunotherapeutic agent; Immunotherapy; In Vitro; Injections; Institutional Review Boards; Legal patent; Letters; Leukemic Cell; Lymphoid; Malignant Neoplasms; Measures; Mediating; Methods; Modeling; Monitor; Morbidity - disease rate; Mus; Mutation; Myelogenous; Myeloid Cells; Myeloproliferative disease; Myelosuppression; PTPRC gene; Patients; Phagocytosis; Phase; Postremission Therapy; Pre-Clinical Model; Preparation; Production; Publishing; Refractory; Relapse; Remission Induction; Residual Neoplasm; Resistance; Resistance development; Site; Stem cell transplant; TNF gene; Testing; Therapeutic; Toxic effect; Transplantation; Tumor Antigens; Veno-Occlusive Disease; Work; acute myeloid leukemia cell; bi-specific T cell engager; chimeric antigen receptor T cells; chronic T-cell leukemia; curative treatments; cytokine; cytokine release syndrome; deep sequencing; experimental study; high risk; improved outcome; in vitro Assay; in vivo; innovation; leukemia; macrophage; monocyte; mouse model; novel; novel strategies; novel therapeutic intervention; overexpression; patient derived xenograft model; peripheral blood; phase 1 study; pre-clinical; preservation; progenitor; relapse patients; relapse risk; resistance mechanism; stem; stem cells; therapeutic gene; translational study

PROJECT SUMMARY/ABSTRACT: The goal of this study is to develop a novel therapeutic strategy for AML, a malignancy of myeloid lineage, that represents a disease with enormous unmet therapeutic need. Hematopoietic stem cell transplant (HSCT) is the only curative treatment option for patients with relapsed/refractory AML but up to 40% patients further relapse after HSCT. Targeting of AML cells using agents directed against a lineage specific antigen such as CD33 using the antibody drug conjugate Gemtuzimab Ozagamycin (GO; Mylotarg) has improved outcomes, but the use of Mylotarg is associated with severe myelosuppression. This is due to targeting of both the leukemia cells and normal myeloid cells (including stem, progenitor and myeloid cells in the donor graft) that also express CD33 (the normal myeloid cells are concomitantly killed by Mylotarg, which also has other toxicities, including veno- occlusive disease at high doses). We reasoned that by ablating CD33 expression using genomic engineering methods in donor stem/progenitor cells, we could generate stem/progenitor cells for transplant that are resistant to Mylotarg treatment, while rendering the AML cells uniquely sensitive to anti-CD33 therapy such as low- dose Mylotarg, anti-CD33 CAR-T or CD33 bi-specific T cell engagers (BiTEs). Indeed, in a recently published “proof of concept study”, we demonstrated that this strategy (either Mylotarg alone, CART-33 or both) enabled the complete killing of an engrafted human CD33+ AML cell line, while allowing fully functional hematopoietic repopulation by CD33- HSPCs. In preliminary experiments with primary human AML cells, however, we found that despite AML clearance with Mylotarg, a small fraction of AML persisted in the bone marrow after 16 weeks of treatment. These AML cells are CD33+ but also express a second lineage antigen, CLL-1+ (Clec12a). We reasoned that targeting multiple antigens will result reduce the chances of relapse. In this proposal, we plan to test the hypothesis that long-term leukemia remission is achieved when two antigens on AML cells are targeted, either serially or simultaneously. To test this hypothesis, we will measure the engraftment, differentiation, and functional potential of HSPCs that are gene-edited to ablate expression of CLL-1 antigen alone or in combination with CD33 (Aim 1). We will test this two-hit treatment approach using patient derived xenograft (PDX) models of primary human AML and also test the engraftment, differentiation, and functional potential of single (CLL-1) and double (CLL-1 and CD33) deletion HSPC in this context (Aim 2). The long-term objective of this study is to develop novel cell-based gene therapeutic strategies in combination with immunotherapeutic approaches for the treatment of AML and related cancers such myelodysplastic syndromes.

项目摘要/摘要： 这项研究的目的是为AML制定一种新型的治疗策略，即髓样谱系的恶性肿瘤，该策略是 代表一种巨大未满足治疗需求的疾病。造血干细胞移植（HSCT）是 接力/难治性AML的患者唯一的治疗方法，但多达40％的患者进一步缓解 HSCT之后。使用针对谱系特异性抗原（例如CD33）靶向AML细胞的靶向AML细胞 抗体药物共轭gemtuzimab ozagamycin（GO; mylotarg）有改善的预后 mylotarg与严重的骨髓抑制有关。这是由于两个白血病细胞和 正常的髓样细胞（包括供体移植中的茎，祖细胞和髓样细胞）也表达CD33 （正常的髓样细胞同时被mylotarg杀死，Mylotarg也具有其他毒性，包括静脉 高剂量的闭塞性疾病）。我们认为，通过使用基因组工程消融CD33表达 供体干/祖细胞中的方法，我们可以生成具有抗性的移植的茎/祖细胞 进行mylotarg治疗，同时使AML细胞对抗CD33疗法的独特敏感，例如低 - 剂量mylotarg，抗CD33 CAR-T或CD33 BI特异性T细胞Endagers（位）。确实，在最近出版的 “概念验证研究”，我们证明了这种策略（单独的mylotarg，Cart-33或两者都启用） 完全杀死了植入的人CD33+ AML细胞系，同时允许完全功能型造血 CD33-HSPC的重生。 但是，在针对原代人AML细胞的初步实验中，我们发现目的地AML清除率 经过16周的治疗后，骨髓中的一小部分AML持续存在。这些AML细胞 是CD33+，但也表达第二个谱系抗原Cll-1+（CLEC12A）。我们认为针对多个 抗原将导致减少退休的机会。在该提议中，我们计划检验以下假设 当AML细胞上的两种抗原靶向串行或简单地靶向时，就可以减轻白血病。 为了检验这一假设，我们将测量HSPC的植入，分化和功能潜力 基因编辑单独或与CD33结合的Cll-1抗原的灌溉表达（AIM 1）。我们将测试 这种使用原代人AML的患者衍生型（PDX）模型的两次打击治疗方法以及 测试单个（CLL-1）和双（CLL-1和CD33）的植入，分化和功能潜力 在此上下文中删除HSPC（AIM 2）。这项研究的长期目的是开发基于细胞的新基因 治疗策略与AML治疗的免疫治疗方法结合使用 癌症症状综合征。