Reprogramming the tumor microenvironment via self-amplified RNA (SafeR) circuits

通过自扩增 RNA (SafeR) 电路重新编程肿瘤微环境

• 批准号: 9206914
• 负责人: RON WEISS
• 金额: $ 55.02万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-12 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9206914
• 关键词: 4T1; Antigen-Presenting Cells; Behavior; Breast Cancer Cell; Cell physiology; Cells; Clinical; Complex; Cytotoxic T-Lymphocyte-Associated Protein 4; DNA; Data; Development; Disease; Disease Outcome; Disseminated Malignant Neoplasm; Dose; Effector Cell; Engineering; Environment; Equilibrium; FDA approved; Gene Expression; Genetic; Genetic Transcription; Genomics; Goals; Immune; Immune response; Immunity; Immunosuppression; Immunosuppressive Agents; In Vitro; Individual; Infiltration; Libraries; Mammalian Cell; Measurement; Mediating; Methods; MicroRNAs; Monitor; Mus; Myelogenous; PDCD1LG1 gene; Patients; Pharmaceutical Preparations; RNA; RNA Binding; RNA-Binding Proteins; Receptor Signaling; Regimen; Regulatory T-Lymphocyte; Replicon; Safety; Series; Serum; Signal Pathway; Signal Transduction; Suppressor-Effector T-Lymphocytes; Synthetic Genes; System; Systems Analysis; Systems Biology; T-Lymphocyte; Testing; Therapeutic; Tissues; Treatment Efficacy; Variant; base; cancer cell; cancer immunotherapy; cancer therapy; cancer type; cell type; chemokine; chemotherapy; cytokine; cytokine therapy; design; experience; genetic regulatory protein; immune checkpoint blockade; improved; in vivo; intercellular communication; melanoma; miRNA expression profiling; neoplastic cell; outcome forecast; programs; promoter; protein degradation; protein expression; public health relevance; receptor; response; sensor; small molecule; synthetic biology; systemic toxicity; targeted treatment; tool; tumor; tumor eradication; tumor microenvironment

 DESCRIPTION: Data from patients in diverse cancer types show that increased immune cell infiltration of tumors correlates with improved patient prognosis. It is also clear that a host of immunosuppressive signals are produced by tumors in order to block immune attack, and thus the endogenous immune response is in most cases unable to eliminate established tumors. Recently, efforts to alter the suppressive signaling in tumors through "checkpoint blockade" drugs that block individual suppressive signaling receptors on T-cells have shown promising clinical results, demonstrating tumor regression through shifting of the tumor microenvironment toward a pro-immunity state. However, it is likely that blocking individual signaling pathways will be insufficient for cancer immunotherapy to reach its full potential, because tumors create a complex network of suppressive signals. Here we propose an approach based on integration of synthetic biology and systems biology to reprogram the suppressive microenvironment in tumors through delivery of sophisticated multi-step genetic circuits to cells in the tumor microenvironment. This strategy will be enabled by the development of self-replicating RNA-based circuits that can (i) identify cell types specifically in the tumor through miRNA expression profiles, (ii) utilize small molecule-regulated induction of new gene expression programs that act in trans on surrounding cells in the tumor, and (iii) alter the function of identified cells in a coordinated fashion to tip the balance within the tumor from immune suppression to immune-mediated tumor destruction. Our specific aims are: (1) We will design a platform for in vivo RNA-based synthetic biology comprising RNA-binding regulatory proteins and small molecule induced protein degradation domains; (2) We will profile the miRNA signatures of 4T1 breast cancer cells and B16F10 melanoma cancer cells and create RNA-encoded multi-input classifier circuits that permit replicon expression only in target cell types, and (3) We will build a series f increasingly sophisticated programmed cancer therapies where cytokine secretion and necroptotic gene expression is restricted to tumor cells and can be precisely regulated by FDA approved small molecules, along with safety switch mechanisms to eliminate replicon-encoded circuits in healthy cells. We will deliver our therapeutic circuits into mouse tumors in vivo and monitor anticancer immune responses, using systems biology principles to analyze the resulting response of multi-cell networks in the tumor. A key goal of these studies will be to design RNA circuits which drive transfected tumor/immune cells to act in trans on surrounding cells in the microenvironment, to achieve a tumor-wide change in the tumor milieu without the need for successful circuit delivery to every cell in the tumor. Results from this project will provide a ne RNA-based toolkit for engineering mammalian cell function, demonstrate the utility of these approaches in vivo, and provide a framework for reprogramming tumors that overcomes limitations of traditional chemotherapy and targeted drug treatments.

 产品说明：来自不同癌症类型患者的数据显示，肿瘤的免疫细胞浸润增加与患者预后改善相关。同样清楚的是，肿瘤产生大量免疫抑制信号以阻断免疫攻击，因此内源性免疫应答在大多数情况下不能消除已建立的肿瘤。最近，通过阻断T细胞上的个体抑制性信号传导受体的“检查点阻断”药物来改变肿瘤中的抑制性信号传导的努力已经显示出有希望的临床结果，证明了通过肿瘤微环境向促免疫状态的转变来实现肿瘤消退。然而，阻断单个信号通路可能会 癌症免疫疗法不足以充分发挥其潜力，因为肿瘤会产生复杂的抑制信号网络。在这里，我们提出了一种基于合成生物学和系统生物学整合的方法，通过将复杂的多步遗传电路传递到肿瘤微环境中的细胞来重新编程肿瘤中的抑制性微环境。这种策略将通过开发基于自复制RNA的电路来实现，该电路可以（i）通过miRNA表达谱特异性地鉴定肿瘤中的细胞类型，（ii）利用小分子调节的新基因表达程序的诱导，该新基因表达程序反式作用于肿瘤中的周围细胞，和（iii）以协调的方式改变所鉴定细胞的功能，以使肿瘤内的平衡从免疫抑制向免疫介导的肿瘤破坏倾斜。我们的具体目标是：（1）我们将设计一个基于RNA的体内合成生物学平台，包括RNA结合调节蛋白和小分子诱导的蛋白降解结构域;（2）我们将分析4 T1乳腺癌细胞和B16 F10黑色素瘤癌细胞的miRNA特征，并创建RNA编码的多输入分类器电路，其允许复制子仅在靶细胞类型中表达，以及（3）我们将建立一系列日益复杂的程序化癌症疗法，其中细胞因子分泌和坏死性凋亡基因表达仅限于肿瘤细胞，并且可以由FDA批准的小分子精确调节，沿着安全开关机制以消除健康细胞中的复制子编码电路。我们将把我们的治疗回路输送到体内小鼠肿瘤中，并监测抗癌免疫反应，利用系统生物学原理分析肿瘤中多细胞网络的反应。这些研究的一个关键目标将是设计RNA电路，驱动转染的肿瘤/免疫细胞反式作用于微环境中的周围细胞，以实现肿瘤环境的肿瘤范围内的变化，而不需要将电路成功递送到肿瘤中的每个细胞。该项目的结果将为工程哺乳动物细胞功能提供一个基于新RNA的工具包，证明这些方法在体内的实用性，并为克服传统化疗和靶向药物治疗的局限性的肿瘤重编程提供一个框架。