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.

 描述：来自不同癌症类型患者的数据表明，肿瘤免疫细胞浸润的增加与患者预后的改善相关。同样清楚的是，肿瘤会产生大量免疫抑制信号，以阻止免疫攻击，从而阻止内源性免疫反应。最近，通过“检查点阻断”药物阻断 T 细胞上的单个抑制信号受体来改变肿瘤中的抑制信号传导，已显示出有希望的临床结果，证明肿瘤通过以下方式消退。然而，阻断单个信号通路可能会导致肿瘤微环境向免疫状态转变。 不足以让癌症免疫疗法充分发挥其潜力，因为肿瘤会产生复杂的抑制信号网络，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，通过传递复杂的多步骤遗传来重新编程肿瘤中的抑制微环境。该策略将通过开发基于 RNA 的自我复制电路来实现，该电路可以（i）通过 miRNA 表达谱特异性识别肿瘤中的细胞类型，（ii）利用小分子调节的诱导。新的基因表达程序反式作用于肿瘤周围的细胞，并且（iii）以协调的方式改变已识别细胞的功能，以改变肿瘤内的平衡，从免疫抑制转向免疫介导的肿瘤破坏。我们将设计一个基于 RNA 的体内合成生物学平台，包含 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 4T1 乳腺癌细胞和 B16F10 黑色素瘤细胞的 miRNA 特征，并创建 RNA-编码的多输入分类器电路，仅允许在靶细胞类型中表达复制子，以及（3）我们将构建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，并且可以通过 FDA 批准进行精确调节小分子以及安全开关机制消除了健康细胞中的复制子编码电路，我们将把我们的治疗电路传递到小鼠体内肿瘤中并监测抗癌免疫反应，利用系统生物学原理来分析多细胞的反应。这些研究的一个关键目标是设计RNA回路，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，以实现肿瘤范围内的肿瘤环境变化，而不需要该项目的结果将为设计哺乳动物细胞功能提供一个新的基于RNA的工具包，证明这些方法在体内的实用性，并提供一个克服传统化疗局限性的肿瘤重编程框架。以及靶向药物治疗。