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

基本信息

批准号：
10630917
负责人：
Caleb Bashor
金额：
$ 34.1万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
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/10630917
关键词：
Acute Graft Versus Host Disease Address Anti-Inflammatory Agents Autoimmune Diseases Behavior Behavioral Cell Shape Cell Therapy Cell physiology Cells Classification Coupling Cues Custom Data Detection Development Devices Diagnostic Dimerization Discrimination Disease Engineering Environment Event Excision Foundations G-Protein-Coupled Receptors Genetic Transcription Goals Growth Factor Human Immune Infection Inflammation Inflammatory Intelligence 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 Time Tissues Translations Work cellular engineering clinically relevant cytokine design environmental change established cell line extracellular flexibility improved mesenchymal stromal cell nervous system disorder next generation non-Native novel programs rapid detection receptor response small molecule synthetic biology synthetic construct technology platform tissue trauma tool treatment response

项目摘要

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.

概括细胞使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。工程人工信号网络，使膜受体介导传感与疾病相关的信号对治疗反应可能会导致发展的突破能够自主检测和治疗疾病的基于动态细胞的治疗装置的。相比之下到依赖磷酸化来传递外部信号的天然信号网络，当前方法用于构建人类中的合成信号网络，依靠非本地调节机制并运行在缓慢的时间尺度上或通过一次性事件。结果，建立感官和 - 响应这些程序，使环境波动能够伴随着输出响应，或者可以灵活地进行响应公司多样化的受体介导的投入。设计基于磷酸化的感官和响应的能力程序可以实现类似于本地途径的功能行为，包括快速检测和集成细胞外信号。通过在不同的细胞外环境之间进行微调的歧视，例如程序可以增强基于细胞的疗法的安全性和功效概况。在这个项目中，我们将建立一个在人类细胞中基于工程合成磷酸化的工程合成磷酸化的可概括方法，放置了一个下一代细胞疗法设备，能够感应与疾病相关的分子提示和将它们转换为定量定义的治疗反应。为了实现我们的目标，我们将利用我们的实验室最近开发了合成生物学平台，该平台可以自下而上的合成phos- 使用工程信号蛋白的伪造电路。正如我们的初步工作所表明的那样，这个平台可用于创建连接受体介导的细胞外分子检测的合成信号通路向下游细胞过程激活（例如，转录）。在这里，我们将调查这个平台是否可用于设计和反应计划以治疗炎症性疾病。具体来说，我们将：1）证明合成途径与受体有多种生物分子提示的受体连接的能力与炎症有关； 2）工程师信号网络，这些信号网络集成多个信号并响应不可在存在炎症提示的定义组合的情况下； 3）间充质的测试途径基质细胞（MSC）评估我们平台的可翻译性。我们的工作将阐明工程合成信号电路，并提供一个强大的技术平台，用于创建定制的可以准确区分健康组织和患病组织特征的感官和响应程序。在除了疾病监测和诊断应用外，这些基于细胞的基于细胞的治疗设备可能是使用的疾病从炎症和自身免疫性疾病到组织创伤和癌症等疾病。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Engineering the next generation of cell-based therapeutics.

DOI：
10.1038/s41573-022-00476-6
发表时间：
2022-09
期刊：
Nature reviews. Drug discovery
影响因子：
0
作者：
通讯作者：

Engineering synthetic phosphorylation signaling networks in human cells.

在人体细胞中工程合成磷酸化信号网络。

DOI：
10.1101/2023.09.11.557100
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Yang,Xiaoyu;Rocks,JasonW;Jiang,Kaiyi;Walters,AndrewJ;Rai,Kshitij;Liu,Jing;Nguyen,Jason;Olson,ScottD;Mehta,Pankaj;Collins,JamesJ;Daringer,NicholeM;Bashor,CalebJ
通讯作者：
Bashor,CalebJ