Targeted cell-to-cell mRNA delivery of therapeutic circuits

项目摘要 信使核糖核酸作为一种诱导细胞内蛋白质表达的试剂具有很大的治疗潜力。精准交付 从通过基因纠正致病突变，到病态细胞类型的mrna可能有很多应用。 编辑以诱导病态细胞类型中的杀伤到细胞状态重编程。尽管有这种潜力，但它仍然 向特定的细胞亚群传递信使核糖核酸具有挑战性。一种安全运送信使核糖核酸的通用方法 患者体内的靶细胞类型将释放出mRNA的全部治疗潜力。活细胞是理想的 因为它们可以被编程为使用传感和逻辑来专门将mRNA递送到 目标细胞群。 为了实现这一目标，我们实验室最近开发出了能够在合成出口中输出mrna的递送细胞。 车辆(同步电动汽车)。这些颗粒可以将信使核糖核酸转移到表达信使核糖核酸的细胞。在这 提案，我们的目标是创建、优化和整合元素，以生成理想的基于细胞的mRNA传递 站台。在目标1中，我们将设计原代人类T细胞，一种与临床相关的细胞类型，作为交付细胞。在AIM 2、我们将开发通过工程条件发送和伪分型来控制靶向特异性的策略。 在目标3中，我们将通过传递一个电路来演示活体中的细胞-细胞递送和mRNA的功能 诱导肿瘤细胞死亡，这应该绕过限制传统免疫的免疫抑制 基于细胞的疗法。 总之，这项工作将扩大我们在细胞-细胞mRNA传递方面的能力，并使一种新的、以细胞为基础的肿瘤成为可能 避开免疫抑制的杀戮策略。此外，当我们专注于与癌症相关的应用时 在这项提案中，这项工作将建立一个在生物医学领域具有广泛实用性的平台，包括体内细胞 重新编程和基因组编辑。

Project Summary mRNA holds great therapeutic potential as an agent to induce expression of proteins in cells. Precise delivery of mRNA to diseased cell types could have numerous applications, from correcting pathogenic mutations by gene editing to inducing killing in diseased cell types to cell state reprogramming. Despite this potential, it remains challenging to deliver mRNA to specific subsets of cells. A general method to safely deliver mRNA cargo to target cell types within a patient’s body would unlock the full therapeutic potential of mRNA. Living cells are ideal delivery vehicles because they can be programmed to use sensing and logic to specifically deliver mRNA to target cell populations. Towards this goal, our lab has recently developed delivery cells capable of exporting mRNA in synthetic export vehicles (synEVs). These particles can transfer mRNA cargo to receiver cells, where it is expressed. In this proposal, we aim to create, optimize, and integrate elements to generate an ideal cell-based mRNA delivery platform. In Aim 1, we will engineer primary human T cells, a clinically relevant cell type, as delivery cells. In Aim 2, we will develop strategies to control targeting specificity by engineering conditional sending and pseudotyping. In Aim 3, we will demonstrate cell-cell delivery and functionality of mRNA in vivo by delivering a circuit that induces cell death to tumor cells, which should bypass the immunosuppression that limits traditional immune cell-based therapies. Together, this work will expand our capabilities in cell-cell mRNA delivery and enable a novel, cell-based tumor killing strategy that circumvents immunosuppression. Further, while we focus on applications related to cancer in this proposal, this work will establish a platform with broad utility across biomedicine, including for in vivo cell reprogramming and genome editing.