Ionizable lipid nanoparticles for the delivery of mRNA for CAR T cell engineering

用于 CAR T 细胞工程 mRNA 递送的可电离脂质纳米粒子

• 批准号: 10231910
• 负责人: Margaret M. Billingsley
• 金额: $ 4.6万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10231910
• 关键词: Acute Lymphocytic Leukemia; Adverse effects; Adverse event; Affect; Applied Research; Artificial nanoparticles; Autologous; B-Cell Lymphomas; B-Lymphocytes; Biocompatible Materials; Biological Assay; Biomedical Engineering; CAR T cell therapy; CTLL-2 Assay; Cancerous; Case Study; Cell Line; Cell Therapy; Cell membrane; Cell physiology; Cells; Cessation of life; Clinical; Clinical Engineering; Clinical Trials; Collaborations; Development; Disease remission; Dose; Electroporation; Encapsulated; Engineering; Event; FDA approved; Flow Cytometry; Fluorescence; Formulation; Future; Generations; Genomics; Gills; Gold; Harvest; Immune; Immune system; Immunology; Immunotherapy; Interferons; Interleukin-2; Investigation; Libraries; Life; Literature; Luciferases; Malignant Neoplasms; Mediating; Membrane; Messenger RNA; Methods; Modification; Mus; Patients; Pennsylvania; Property; Risk; Schools; Serum; Severities; System; T-Lymphocyte; TNF gene; Toxic effect; Training; Transfection; Translating; Translations; Treatment Efficacy; Universities; Viral; Virus; Work; Xenograft Model; acute lymphoblastic leukemia cell; base; burden of illness; cancer cell; cancer immunotherapy; cancer type; cell killing; cell type; cellular engineering; chimeric antigen receptor; chimeric antigen receptor T cells; clinical translation; clinically relevant; cytokine; cytokine release syndrome; cytotoxic; cytotoxicity; efficacy evaluation; engineered T cells; experience; hospital readmission; hypogammaglobulinemia; improved; in vivo; lipid nanoparticle; mRNA delivery; medical schools; mouse model; nanoparticle delivery; neurotoxicity; novel; novel strategies; nucleic acid delivery; protein expression; receptor expression; screening; side effect; success; tool; tumor; uptake

PROJECT SUMMARY CAR T cell immunotherapy is FDA approved for the treatment of acute lymphoblastic leukemia (ALL) and large B cell lymphoma and has shown success in inducing durable remission. However, the therapy is also associated with causing severe, life-threatening side effects—including cytokine release syndrome, B cell aplasia, and neurotoxicity—in 70% of patients receiving the treatment. Thus, there is a need to develop CAR T cells that maintain their therapeutic efficacy while minimizing adverse effects. Currently, CAR T cells are engineered using viruses that induce permanent CAR expression, but investigations into mRNA-based CAR T cells—which result in transient CAR expression—have been utilized in clinical trials and shown potential for mitigating long-term side effects of the immunotherapy. To create these mRNA CAR T cells, electroporation is utilized for T cell transfection, but it is cytotoxic and has no potential for translation to in vivo T cell delivery. Thus, this investigation aims to explore ionizable lipid nanoparticles (LNPs) as a delivery tool for the ex vivo engineering of T cells. LNPs have shown potent mRNA delivery in various cell types and can be easily modified to alter the physicochemical properties that impact delivery, which will allow for their optimization as a delivery platform for T cells specifically. In Aim 1, 24 novel LNPs will be screened for their ability to functionally deliver mRNA with low toxicity, and the top-performing LNP will be further optimized to determine the best formulation for delivery to primary T cells. In Aim 2, the LNP formulation selected in Aim 1 will be used to encapsulate CAR mRNA with different modifications to determine the best mRNA cargo for LNP-based delivery to T cells. With the top LNP and CAR mRNA cargo selected, Aim 3 will validate LNPs as a method for CAR T cell engineering as compared to electroporated mRNA- CAR T cells and virus-based CAR T cells via a survival study using an ALL mouse model. The completion of these aims will identify and optimize a mRNA delivery platform for T cells that, in future investigations, can be utilized for the screening of new CAR constructs or in vivo delivery. Ultimately, this work—conducted as an interdisciplinary project between sponsors in the Bioengineering Department and Medical School at University of Pennsylvania—will allow for the development of a novel LNP delivery platform for immune cell engineering.

项目摘要 CAR T细胞免疫疗法已批准用于治疗急性淋巴细胞白血病（ALL）和大型 B细胞淋巴瘤，并在诱导的持久缓解方面表现出了成功。但是，该疗法也与 引起严重，威胁生命的副作用 - 包括细胞因子释放综合征，B细胞性疾病和 神经毒性 - 在接受治疗的70％的患者中。那是需要开发汽车T细胞的 保持其治疗效果，同时最大程度地减少不良影响。目前，使用汽车T单元使用 影响永久性汽车表达的病毒，但对基于mRNA的CAR T细胞进行了研究 - 这是结果 在瞬态汽车表达中 - 已在临床试验中使用，并显示出缓解长期的潜力 免疫疗法的副作用。为了创建这些mRNA CAR T细胞，电穿孔用于T细胞 转染，但它具有细胞毒性，并且没有转化为体内T细胞递送的潜力。那是这个调查 旨在探索可离子脂质纳米颗粒（LNP）作为T细胞实体工程的输送工具。 LNP 已经显示出各种细胞类型的有效mRNA传递，并且可以轻松修改以改变物理 影响输送的属性，这将允许其优化作为T细胞的输送平台。 在AIM 1中，将筛选24个新型LNP，以使其功能性低毒性的mRNA的能力，并且 表现最佳的LNP将进一步优化，以确定向原代T细胞递送的最佳公式。 AIM 2，AIM 1中选择的LNP公式将用于封装具有不同修饰的CAR mRNA 确定基于LNP的T细胞递送的最佳mRNA货物。带有顶级LNP和汽车mRNA货物 选择，AIM 3将验证LNP作为CAR T细胞工程的方法，与电穿孔mRNA-相比 CAR T细胞和基于病毒的CAR T细胞通过使用所有小鼠模型的生存研究。完成 这些目标将识别并优化T细胞的mRNA输送平台，在以后的研究中，可以是 用于筛选新汽车构造或体内输送。最终，这项工作 - 生物工程系和大学医学院的赞助商之间的跨学科项目 宾夕法尼亚州 - 将允许开发一个新型的LNP输送平台用于免疫细胞工程。