A synthetic signaling pathway engineering platform for creating precision cell-based sense-and-response devices

用于创建基于细胞的精密传感和响应设备的合成信号通路工程平台

• 批准号: 10280787
• 负责人: Caleb Bashor
• 金额: $ 34.49万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10280787
• 关键词: Acute Graft Versus Host Disease; Address; Anti-Inflammatory Agents; Autoimmune Diseases; Behavior; Behavioral; Cell Therapy; Cell physiology; Cells; Coupling; Cues; Custom; Data; Detection; Development; Devices; Diagnostic; Discrimination; Disease; Engineering; Environment; Event; Excision; Foundations; G-Protein-Coupled Receptors; Genetic Transcription; Goals; Growth Factor; Human; Immune; Infection; Inflammation; Inflammatory; Intelligence; Lead; Malignant Neoplasms; Mediating; Membrane; Mesenchymal Stem Cells; Modeling; Molecular; Monitor; Output; Pathway interactions; Patients; Phosphorylation; Phosphorylation Site; Process; Proteins; Proteolysis; Regulation; Research Personnel; Safety; Scheme; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Specificity; Surface; System; Technology; Testing; Therapeutic; Tissues; Translations; Work; base; cell transformation; cellular engineering; clinically relevant; cytokine; design; dimer; environmental change; established cell line; extracellular; flexibility; improved; mesenchymal stromal cell; nervous system disorder; next generation; non-Native; novel; programs; receptor; response; small molecule; synthetic biology; synthetic construct; time use; tissue trauma; tool; treatment response

SUMMARY Protein signaling networks are used by cells to sense, process, and respond to physical and molecular features in their external environment. Engineering artificial signaling networks that couple membrane receptor-mediated sensing of disease-associated signals to therapeutic responses could lead to breakthroughs in the development of dynamic cell-based therapeutic devices capable of autonomously detecting and treating disease. In contrast to native signaling networks, which rely on phosphorylation to transduce external signals, current approaches for constructing synthetic signaling networks in humans rely on nonnative regulatory mechanisms and operate on slow timescales or via single-turnover events. As a consequence, it is challenging to construct sense-and- respond programs that accurately couple environmental fluctuations to output response, or that can flexibly in- corporate diverse receptor-mediated inputs. The ability to engineer phosphorylation-based sense-and-response programs could enable functional behavior resembling native pathways, including rapid detecting and integration of extracellular signals. By enabling fine-tuned discrimination between different extracellular environments, such programs could enhance safety and efficacy profiles of cell-based therapies. In this project, we will establish a generalizable approach for engineering synthetic phosphorylation-based signaling in human cells, laying a foun- dation for next-generation cell therapy devices capable of sensing molecular cues associated with disease, and converting them into quantitatively defined therapeutic responses. To accomplish our goals, we will leverage a synthetic biology platform recently developed by our lab that enables bottom-up construction of synthetic phos- phorylation circuitry using engineered signaling proteins. As our preliminary work demonstrates, this platform can be used to create synthetic signaling pathways connecting receptor-mediated detection of extracellular mol- ecules to activation of downstream cellular processes (e.g., transcription). Here, we will investigate if this platform can be used to engineer sense-and respond program to treat inflammatory disease. Specifically, we will: 1) demonstrate the ability of synthetic pathways to be wired to receptors that sense diverse biomolecular cues associated with inflammation; 2) engineer signaling networks that integrate multiple signals and respond exclu- sively in the presence of defined combinations of inflammatory cues and; 3) test pathways in mesenchymal stromal cells (MSCs) to assess translatability of our platform. Our work will illuminate foundational principles for engineering synthetic signaling circuits and deliver a powerful technology platform for creating customized sense-and-respond programs that can precisely distinguish between features of healthy and diseased tissue. In addition to disease monitoring and diagnostic applications, these precision cell-based therapy devices could be used treat diseases ranging from inflammatory and autoimmune disorders, to tissue trauma and cancer.

总结 细胞利用蛋白质信号网络来感知、处理和响应物理和分子特征 在他们的外部环境中。工程化人工信号网络，其耦合膜受体介导的 检测疾病相关信号对治疗反应的影响可能会导致开发 基于细胞的动态治疗设备能够自主检测和治疗疾病。相比之下 对于依赖磷酸化来抑制外部信号的天然信号传导网络， 在人类中构建合成信号网络依赖于非天然调节机制， 在缓慢的时间尺度上或通过单一的周转事件。因此，构建意义和 响应程序，准确地耦合环境波动的输出响应，或可以灵活地在- 企业多样的受体介导的输入。设计基于磷酸化的感知和反应的能力 程序可以使功能行为类似于天然途径，包括快速检测和整合 细胞外信号。通过在不同的细胞外环境之间进行微调区分， 这些程序可以增强基于细胞的疗法的安全性和有效性。在这个项目中，我们将建立一个 在人类细胞中工程化合成的基于磷酸化的信号传导的可推广的方法，奠定了基础， 用于能够感测与疾病相关的分子线索的下一代细胞治疗设备的开发，以及 将其转化为定量定义的治疗反应。为了实现我们的目标，我们将利用 我们实验室最近开发的合成生物学平台，可以自下而上构建合成磷酸盐， 使用工程化信号蛋白的磷酸化电路。正如我们的初步工作表明，这个平台 可用于创建连接受体介导的细胞外摩尔检测的合成信号通路， 导致下游细胞过程的激活（例如，转录）。在这里，我们将调查，如果这个平台 可用于设计感觉和应答程序以治疗炎症性疾病。具体而言，我们将：1） 证明了合成途径与感受不同生物分子信号的受体相连的能力 与炎症相关; 2）工程信号网络，整合多种信号和响应， 在存在确定的炎症线索组合的情况下; 3）间充质细胞中的测试途径 基质细胞（MSC）来评估我们平台的可翻译性。我们的工作将阐明基本原则， 设计合成信号电路，并提供强大的技术平台， 传感和响应程序，可以精确区分健康和患病组织的特征。在 除了疾病监测和诊断应用外，这些精确的基于细胞的治疗设备还可以 用于治疗从炎症和自身免疫性疾病到组织创伤和癌症的疾病。