Off-The-Shelf Dual Targeted NK Cells For NHL

用于 NHL 的现成双靶向 NK 细胞

• 批准号: 10172127
• 负责人: DANIEL J WEISDORF
• 金额: $ 29.4万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10172127
• 关键词: ATAC-seq; Adoptive Transfer; Affinity; Allogenic; Antigen Targeting; Antigens; Autologous; Autologous Transplantation; B-Cell Lymphomas; CD19 gene; CD34 gene; Cell Death; Cell Line; Cell physiology; Cell surface; Cells; Cellular Metabolic Process; ChIP-seq; Clinical; Clinical Trials; Clip; Clone Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Data; Development; Dose; Dose-Limiting; Engineering; Engraftment; Enzymes; Exhibits; FCGR3B gene; FDA approved; Failure; Funding; Future; Gene-Modified; Genes; Genetic Engineering; Goals; Half-Life; Heterogeneity; Immune; In complete remission; Individual; Infusion procedures; Ink; Interleukin-15; Investigation; Knock-out; Lead; Lymphoma; MS4A1 gene; Maintenance Therapy; Malignant - descriptor; Mediating; Membrane; Metabolic; Metabolism; Minnesota; Modification; NK cell therapy; Natural Killer Cells; Non-Hodgkin' s Lymphoma; Oral; Oxidative Stress; Patients; Phenotype; Prevention strategy; Process; Production; Relapse; Reporting; Resistance; Role; Running; Site; Specificity; T-Lymphocyte; Testing; Therapeutic; Toxic effect; Transgenic Organisms; Translations; Transplantation; Universities; Variant; Work; antibody-dependent cell cytotoxicity; base; cancer therapy; cellular transduction; chimeric antigen receptor; chimeric antigen receptor T cells; cytokine; cytokine release syndrome; disorder later incidence prevention; disorder risk; engineered NK cell; experience; gastrointestinal; high risk; improved; in vivo; induced pluripotent stem cell; inhibitor/antagonist; manufacturing process; metabolic abnormality assessment; metabolic fitness; metabolic profile; overexpression; patient derived xenograft model; phase 1 study; post-transplant; pre-clinical; preclinical trial; prevent; progenitor; programs; receptor; response; rituximab; safety study; scale up; small molecule; stem cell differentiation; synergism; targeted treatment; transcriptome sequencing

Limiting relapse is the biggest challenge following autologous transplantation for non-Hodgkin’s lymphoma. Antigen-directed CAR T cell products and other targeted therapies may be changing the field, yet for high-risk lymphomas, new and safer approaches are needed. We will study the safety and efficacy of antigen-directed, off-the-shelf, induced pluripotent stem cell (iPSC)-derived NK cells engineered for dual-antigen targeting. We have optimized the differentiation of iPSC-derived CD34+ cells to highly potent NK cells, scaled production at the University of Minnesota, and can now produce hundreds of doses in a manufacturing run with a clonal iPSC transduced with a high-affinity, non-cleavable CD16 (called hnCD16). In late 2019, we initiated clinical trials testing this engineered iPSC product (designated FT516). The overarching hypothesis for this project is that dual targeting of NK cells with a CAR and through hnCD16 will protect against relapse after auto-transplant for lymphoma and that future gene edits to alter metabolism will enhance adoptive transfer. This hypothesis fits with the themes of this Program, and Project 2 will inform the Program on the development of off-the-shelf NK cells, NK CAR, and the role of membrane IL-15, as well as pre-clinically on whether manipulating metabolism will enhance NK cell therapy. These investigations are supported by the following Specific Aims. Aim 1 will test dual targeting of NK cells to CD19 and CD20 using two mechanisms of action, ADCC and an NK CAR, to prevent relapse after autologous transplantation for lymphoma. We will test a triple gene-modified iPSC-derived NK cell product (FT596) transduced with hnCD16, an NK cell-optimized CD19 CAR, and membrane bound 15/IL-15Ra fusions. FT596 will be combined with rituximab as maintenance therapy to limit relapse after autologous HCT. This trial is FDA-approved and will first establish an MTD at day +30 after engraftment and then, if single dosing is safe, move to day +7 before engraftment with the goal of 3 doses in the first hundred days to prevent relapse. Aim 2 will test enhancement of NK cell metabolic fitness by ARID5B overexpression and CD38 knockout. Our preliminary data shows both adaptive NK cells and CD38 knockout iPSC-derived NK cells resist oxidative stress- induced cell death. ARID5B overexpression, naturally increased in adaptive NK cells, mediates similar effects. We will generate new iPSC lines to build on FT596 with transgenic expression of ARID5B or a CRISPR knockout of CD38. These clones will be evaluated to probe the mechanisms on how they drive NK cell metabolism and augment function. Aim 3 will test whether manipulating ARID5B and CD38 will drive metabolic fitness and enhance FT596 persistence and function in vivo. These pre-clinical trials will inform future clinical trial modifications—all to promote enhanced anti-lymphoma activity by extending transferred NK cell persistence and potency. These metabolic and potency findings will directly inform Project 1 to improve cell targeting and metabolism, Project 3 for cell persistence, and all 3 Projects to define the advantages of autonomous presentation of IL-15.

限制复发是非霍奇金淋巴瘤自体移植后的最大挑战。 抗原导向的 CAR T 细胞产品和其他靶向疗法可能正在改变这个领域，但对于高风险 淋巴瘤，需要新的、更安全的方法。我们将研究抗原导向的安全性和有效性， 现成的诱导多能干细胞 (iPSC) 衍生的 NK 细胞，专为双抗原靶向而设计。我们 优化了 iPSC 衍生的 CD34+ 细胞向高效 NK 细胞的分化，并在 明尼苏达大学，现在可以使用克隆 iPSC 在生产过程中生产数百剂药物 用高亲和力、不可切割的 CD16（称为 hnCD16）转导。 2019年底，我们启动了临床试验 测试该工程 iPSC 产品（指定为 FT516）。该项目的总体假设是 使用 CAR 和 hnCD16 双重靶向 NK 细胞将防止自体移植后复发 淋巴瘤以及未来改变新陈代谢的基因编辑将增强过继转移。这个假设符合 该计划的主题，项目 2 将为该计划提供有关现成 NK 细胞开发的信息， NK CAR、膜 IL-15 的作用，以及临床前操纵代谢是否会 加强NK细胞治疗。这些调查得到以下具体目标的支持。目标 1 将测试双重 使用 ADCC 和 NK CAR 两种作用机制将 NK 细胞靶向 CD19 和 CD20，以预防 淋巴瘤自体移植后复发。我们将测试三重基因修饰的 iPSC 衍生 NK 细胞 用 hnCD16（一种 NK 细胞优化的 CD19 CAR）和膜结合的 15/IL-15Ra 转导的产品 (FT596) 融合。 FT596 将与利妥昔单抗联合作为维持治疗，以限制自体 HCT 后的复发。 该试验已获得 FDA 批准，将首先在植入后第 30 天建立 MTD，然后，如果单次给药 是安全的，请移至植入前+7天，目标是在前一百天内注射3剂以防止复发。 目标 2 将测试 ARID5B 过表达和 CD38 敲除对 NK 细胞代谢适应性的增强作用。我们的 初步数据显示适应性 NK 细胞和 CD38 敲除 iPSC 衍生的 NK 细胞都能抵抗氧化应激 诱导细胞死亡。 ARID5B 过度表达在适应性 NK 细胞中自然增加，介导类似的效果。 我们将生成新的 iPSC 系，以 FT596 为基础，转基因表达 ARID5B 或 CRISPR 敲除 CD38。这些克隆将被评估，以探究它们如何驱动 NK 细胞代谢和 增强功能。目标 3 将测试操纵 ARID5B 和 CD38 是否会促进代谢健康和 增强 FT596 的体内持久性和功能。这些临床前试验将为未来的临床试验提供信息 修改——所有这些都是为了通过延长转移的 NK 细胞持久性和 效力。这些代谢和效力研究结果将直接为项目 1 提供信息，以改善细胞靶向和 新陈代谢、细胞持久性项目 3 以及定义自主优势的所有 3 个项目 IL-15 的介绍。