Input encoding in T-cell receptor signaling

T 细胞受体信号传导中的输入编码

• 批准号: 9903295
• 负责人: Hao Yuan Kueh
• 金额: $ 19.44万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9903295
• 关键词: Address; Adopted; Affect; Affinity; Antigens; Architecture; Biology; Cell Line; Cells; Chronic; Color; Detection; Devices; Discrimination; Disease; Dose; Engineering; Environment; Exhibits; Feedback; Functional disorder; Generations; Image; Image Analysis; Imaging Techniques; Ligands; Malignant Neoplasms; Measurement; Measures; Mediating; Modeling; Monitor; Mus; Output; Pathway interactions; Peptides; Performance; Physiological; Receptor Signaling; Reporter; Role; Series; Signal Pathway; Signal Transduction; Source; Surface; System; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Time; Viral; Virus; Virus Diseases; Work; automated image analysis; cancer cell; cancer therapy; cellular engineering; combinatorial; design; detector; engineered T cells; exhaust; exhaustion; experimental study; individual response; insight; live cell imaging; man; mathematical model; mouse model; novel; operation; pathogen; response

ABSTRACT Signal sensing circuits in cells often perform at a level that rivals or even exceeds that of man-made detection devices. Understanding how these circuits perform optimally and robustly in a noisy cellular environment will yield fundamental insights into their underlying design principles, and will enable us to re-purpose these circuits for cell engineering. The signaling circuit that mediates antigen detection by T-cells exhibits particularly striking sensing capabilities – it can selectively respond to even a few copies of antigenic ligand, while retaining an ability to distinguish ligand levels over a five order of magnitude range. How T-cell receptor signaling circuits can achieve such remarkable selectivity, sensitivity and dynamic range in their operation is not understood. To address this question, we have developed a novel multi-pathway fluorescent reporter system that enables, for the first time, simultaneous live tracking of the activity of the three primary signaling pathways downstream of the T-cell receptor at the single-cell level. Here, we combine this multi-pathway reporter with quantitative live- cell imaging, mathematical modeling and perturbation analysis to elucidate the control strategies underlying T- cell ligand sensing, and determine how they are disrupted in T-cell dysfunction. Firstly, we will (I) perform a systematic, quantitative characterization of input/output responses of individual pathways and T-cell receptor engagement. This characterization will be performed at the single-cell level, using a combination of high throughput live imaging and computational image analysis. Next, we will (II) elucidate regulatory feedback loops underlying these responses. To do so, we will perform mathematical modeling of candidate feedback architectures, followed by iterative experimental testing. Finally, we will (III) determine how these input/output states are perturbed upon T-cell dysfunction, using live-cell imaging techniques in conjunction with mouse models. These studies will generate fundamental insights into antigen sensing mechanisms by T-cells, yielding guiding principles for engineering T-cells to treat cancer and other diseases. More broadly, this work will also elucidate principles underlying architecture and design of mammalian signaling circuits, impacting systems and signaling biology studies across diverse fields.

抽象的 单元格中的信号传感电路通常以障碍甚至超过人造检测的水平执行 设备。了解这些电路在噪声蜂窝环境中的最佳性能表现 产生对其基本设计原则的基本见解，并将使我们能够重新使用这些电路 用于细胞工程。通过T细胞介导抗原检测的信号传导电路特别引人注目 感知功能 - 它可以选择性地响应几份抗原配体，同时保留 在五个数量级范围内区分配体水平的能力。 T细胞接收器信号传导电路如何 无法理解其操作中的显着选择性，灵敏度和动态范围。到 解决这个问题，我们已经开发了一种新型的多条纹荧光记者系统，可以使 第一次，简单的实时跟踪对下游的三个主要信号通路的活动 单细胞水平的T细胞接收器。在这里，我们将此多条道路记者与定量实时结合 - 细胞成像，数学建模和扰动分析，以阐明T-的控制策略 细胞配体感测，并确定如何在T细胞功能障碍中破坏它们。首先，我们将（i）执行 单个途径和T细胞接收器的输入/输出响应的系统定量表征 订婚。这种表征将在单细胞水平上进行，使用高 吞吐量实时成像和计算图像分析。接下来，我们将（ii）阐明监管反馈 这些响应的循环。为此，我们将对候选反馈进行数学建模 结构，然后进行迭代实验测试。最后，我们将（iii）确定这些输入/输出如何 使用与小鼠结合使用的活细胞成像技术，状态在T细胞功能障碍时受到干扰 型号。这些研究将产生对T细胞的抗原感测机制的基本见解， 产生用于治疗癌症和其他疾病的工程T细胞的指导原则。更广泛地说，这项工作 还将阐明哺乳动物信号电路的建筑和设计的原则，影响 潜水领域的系统和信号生物学研究。