CAREER: Manipulating the Innate Immune Response to Improve Gene Therapy

PI: Elmer， Jacob提案号：1651837该项目的目标是通过识别和修饰几种癌细胞系中涉及对外来DNA免疫反应的基因来改善基因治疗。这种方法有望通过用小分子抑制剂和抑制剂蛋白抑制靶基因来增强基因传递。体细胞抵御病毒感染的第一道防线是先天免疫反应（IIR），它由几种酶和受体组成，主动寻找细胞内的致病分子。细胞质DNA激活IIR可引起炎症，抑制蛋白质翻译，甚至导致宿主细胞自毁。虽然所有这些反应都能保护细胞免受病毒的侵害，但它们也会严重阻碍基因治疗。事实上，大多数基因治疗将治疗性DNA传递到细胞质中，在那里它无意中激活了IIR。这个项目的目标是通过解决这个问题来改进基因治疗。我们将首先确定在各种癌细胞系中对细胞质DNA的先天免疫反应中涉及的确切蛋白质和基因。然后用小分子抑制剂和/或病毒抑制剂蛋白抑制这些基因/蛋白质，从而增强基因传递。作为抑制IIR的另一种选择，转基因表达将通过使用已知被IIR激活的启动子来增加。总之，这些实验将同时改善基因治疗和增加我们对细胞质DNA的先天免疫反应的理解。此外，该计划还将包括一系列全年的外展活动，这些活动将直接惠及社区。例如，PI将组织一个扩展项目，使代表性不足的高中生能够在维拉诺瓦大学进行基因传递和生化工程实验。PI还将为高中教师开设暑期研究体验项目，让他们在PI进行基因治疗实验。然后在秋季学期与他们的班级分享这些经验。最后，在这个项目中进行的每一个实验也将被记录下来，并用于制作教育/培训视频。

PI: Elmer, Jacob Proposal No: 1651837The goal of this project is to improve gene therapy by identifying and modifying the genes involved in the immune response to extraneous DNA in several cancer cell lines. Such an approach is expected to enhance gene delivery by inhibiting the target genes with both small molecule inhibitors and inhibitor proteins.A somatic cell's first line of defense against viral infection is the innate immune response (IIR) consisting of several enzymes and receptors that actively seek out pathogenic molecules within the cell. Activation of the IIR by cytoplasmic DNA can then induces inflammation, inhibits protein translation, and even cause the host cell to self-destruct. While all of these reactions defend the cell from viruses, they can significantly hinder gene therapy. Indeed, most gene therapy treatments deliver therapeutic DNA to the cytoplasm, where it unintentionally activates the IIR. The goal of this project is to improve gene therapy by addressing this problem. We will begin by identifying the exact proteins and genes involved in the innate immune response to cytoplasmic DNA in a variety of cancer cell lines. Gene delivery will then be enhanced by inhibiting those genes/proteins with small molecule inhibitors and/or viral inhibitor proteins. As an alternative to inhibition of the IIR, transgene expression will be increased by using promoters that are known to be activated by the IIR. Altogether, these experiments will simultaneously improve gene therapy and increase our understanding of the innate immune response to cytoplasmic DNA. In addition, this project will also include a series of year-round outreach activities that will directly benefit the community. For example, the PI will organize an outreach program that enables underrepresented high school students to perform gene delivery and biochemical engineering experiments at Villanova. The PI will also create a summer research experience program for high school teachers that allows them to perform gene therapy experiments in the PI?s lab and then share those experiences with their classes in the fall semester. Finally, each of the experiments conducted in this project will also be recorded and used to create educational/training videos.