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细胞免疫疗法被FDA批准用于治疗急性淋巴细胞白血病（ALL）和大细胞白血病（ALL）。 B细胞淋巴瘤，并已成功诱导持久缓解。然而，这种疗法也与 导致严重的、危及生命的副作用，包括细胞因子释放综合征、B细胞发育不全， 神经毒性-在70%的患者接受治疗。因此，需要开发CAR T细胞， 保持其治疗功效，同时最小化副作用。目前，CAR T细胞是使用 病毒诱导永久性CAR表达，但对基于mRNA的CAR T细胞的研究， 在瞬时CAR表达中-已用于临床试验，并显示出减轻长期 免疫疗法的副作用为了产生这些mRNA CAR T细胞，将电穿孔用于T细胞 转染，但它是细胞毒性的，并且没有转化为体内T细胞递送的潜力。因此，本次调查 目的是探索可电离脂质纳米颗粒（LNP）作为T细胞体外工程的递送工具。的lnp 已经在各种细胞类型中显示出有效的mRNA递送，并且可以容易地进行修饰以改变其理化性质。 这些特性影响递送，这将允许它们作为T细胞特异性的递送平台进行优化。 在目标1中，将筛选24种新型LNP，以确定它们以低毒性功能性递送mRNA的能力，并且将其用于治疗癌症。 将进一步优化表现最好的LNP以确定递送至原代T细胞的最佳制剂。在 目的2，目的1中选择的LNP制剂将用于包封具有不同修饰的CAR mRNA 以确定用于基于LNP递送至T细胞的最佳mRNA货物。在LNP和CAR mRNA货物中， 选择，目标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ALL小鼠模型的存活研究。完成 这些目标将确定和优化T细胞的mRNA递送平台，在未来的研究中， 用于筛选新的CAR构建体或体内递送。最终，这项工作作为一项 生物工程系和大学医学院赞助者之间的跨学科项目 将允许开发用于免疫细胞工程的新型LNP递送平台。