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

基本信息

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
项目类别：
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

项目摘要

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：潜水癌症患者的数据表明，肿瘤的免疫剂浸润增加与改善患者提示相关。同样很明显，肿瘤会产生许多免疫抑制信号以阻止免疫攻击，因此内源性免疫响应在大多数情况下无法消除已建立的肿瘤。最近，通过“检查点阻断”药物改变肿瘤中抑制性信号的努力，这些药物阻断了T细胞上个体抑制信号受体的抑制性信号受体，已显示出令人鼓舞的临床结果，这表明通过将肿瘤微环境转移到促疾病状态，这表明了肿瘤的回归。但是，阻止单个信号通路可能会由于肿瘤会产生复杂的抑制信号网络，因此不足以使癌症免疫疗法发挥其全部潜力。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，以通过将复杂的多步遗传回路传递到肿瘤微环境中的细胞中，以重新编程肿瘤中抑制性微环境。该策略将通过开发自我复制的基于RNA的电路来启用（i）（i）通过miRNA表达谱在肿瘤中特别识别细胞类型，（（ii）使用小分子调节的小分子调节的新基因表达程序，这些诱导的新基因表达程序在肿瘤中的周围细胞中作用于周围细胞上的trans中，以及（iii）在肿瘤中的pive液中的pive液中的pive液在pive中，以使tamied tymient themient themient tymied tymied tymied tymied tymor tymor的功能在平衡的功能中，以使tamore的功能保持在肿瘤中，以使tamore的功能保持在较小的状态。破坏。我们的具体目的是：（1）我们将设计一个基于体内RNA的合成生物学的平台，以完成RNA结合调节蛋白和小分子诱导的蛋白质降解结构域； (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 replican 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批准的小分子，以及安全开关机制，以消除健康细胞中复制编码的电路。我们将使用系统生物学原理来分析肿瘤中多细胞网络的产生响应，将治疗回路输送到体内小鼠肿瘤中并监测抗癌免疫调查。这些研究的一个关键目标是设计RNA电路，这些RNA电路驱动翻译肿瘤/免疫细胞以在微环境中的周围细胞上作用，以实现肿瘤环境中肿瘤环境的变化，而无需成功将电路传递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基于NE RNA的工具包，证明了这些方法在体内的实用性，并为重新编程肿瘤提供了一个框架，从而克服了传统化学疗法和靶向药物治疗的限制。