Targeted cell-to-cell mRNA delivery of therapeutic circuits

项目摘要 mRNA作为诱导细胞中蛋白质表达的试剂具有巨大的治疗潜力。精确输送 mRNA到病变细胞类型可以有许多应用，从通过基因纠正致病突变， 编辑到诱导杀死患病细胞类型到细胞状态重编程。尽管有这种潜力， 将mRNA递送到特定的细胞亚群具有挑战性。将mRNA货物安全地递送至 患者体内的靶细胞类型将释放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特异性地将mRNA递送到 靶细胞群。 为了实现这一目标，我们的实验室最近开发了能够以合成输出的方式输出mRNA的递送细胞 车辆（synEV）。这些颗粒可以将mRNA货物转移到受体细胞，在那里它被表达。在这 我们的目标是创建，优化和整合元件，以产生理想的基于细胞的mRNA递送 平台在目标1中，我们将设计原代人类T细胞，一种临床相关的细胞类型，作为递送细胞。在aim中 2、我们将通过工程条件发送和假型化来开发控制靶向特异性的策略。 在目标3中，我们将通过递送一个电路来证明mRNA在体内的细胞-细胞递送和功能，该电路 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间mRNA传递方面的能力，并使一种新的基于细胞的肿瘤成为可能。 避免免疫抑制的杀死策略。此外，当我们专注于与癌症相关的应用时， 在这项提议中，这项工作将建立一个在生物医学上具有广泛用途平台，包括用于体内细胞 重编程和基因组编辑。

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.