Mechanical regulation of T cell receptor and co-receptor responses in cancer immunotherapy

项目总结 在获得性免疫反应中，细胞毒性T淋巴细胞(CTL)不断“爬行”寻找外源证据 其他细胞表面的多肽碎片。一旦T细胞遇到带有外源或突变的靶细胞 多肽，然后它被激活，释放出强大的免疫反应。新出现的证据表明，细胞 传递到T细胞受体(TCR)的机械力有助于其在抗原识别中的高度特异性 并促进T细胞活化。这并不令人惊讶，因为TCR和其他T细胞辅助受体结合了它们的同源物 只有当两个动态细胞在物理上“接触”时，才有配基。作为了解分子的作用的第一步 在调节T细胞反应时，重要的是我们要测量传递给配体的力的大小- 受体复合体，然后将机械事件与信号和功能反应联系起来。 我的博士研究(F99阶段)一直专注于开发方法来测量和阐明 免疫反应中的机械力。我已经设计了一种微粒子张力传感器，它可以让人 高吞吐量的感受器作用力，也用于测量弯曲细胞连接处的作用力。另外，我还用了这个 筛选调节细胞力学的药物的剂量-反应功能的试验。因为T细胞反应 通过一系列辅助受体进行微调，我测试了力学在LFA-1功能中的作用。在这项工作中，我 证明了LFA-1整合素的大小迫使微调TCR触发了激活和抗原 歧视。此外，我揭示了机械活性的LFA-1定义了细胞毒的允许区 分泌，抑制LFA-1的力量显著消除细胞毒性。 我的研究表明，受体协同调节T细胞的反应。在我F99阶段的剩余时间里， 我将研究感受力之间的机械通讯。具体地说，我将开发一种DNA折纸 用于构图配体和测量力学事件的时空共局部化的纳米设备。之后，我 将通过在细胞膜表面设计张力探头来检验这一假说 活细胞。这将使人们能够控制和测量真实细胞-细胞连接处的TCR力，这些细胞连接处模仿 免疫突触的化学和物理特性。 对于我的博士后工作(K00阶段)，我的目标是通过利用T 细胞力学在增强免疫反应特异性中的作用。在过继细胞治疗(ACT)中，治疗后 T细胞回输时，需要使用细胞因子等辅助药物来促进免疫重建。 然而，非特异性药物释放会导致副作用和T细胞耗尽。为了应对这一挑战，我将 用DNA笼子装饰T细胞，这种笼子可以机械地触发肿瘤区域内被包裹的药物的释放。如果 这项工作的成功将显著提高ACT的效率，并提供第一个链接的范例 从机械生物学到癌症免疫治疗。

PROJECT SUMMARY In the adaptive immune response, cytotoxic T lymphocyte (CTL) continuously “crawl” seeking evidence of foreign peptide fragments on the surface of other cells. Once the T cell encounters a target cell with foreign or mutant peptides, then it is activated unleashing a potent immune response. Emerging evidence suggests that cell mechanical forces transmitted to the T cell receptor (TCR) contribute to its high specificity in antigen recognition and promote T-cell activation. This is not surprising, as the TCR and other T cell co-receptors bind their cognate ligands only when two dynamic cells physically “touch”. As a first step toward understanding the role of molecular forces in tuning T cell response, it is important that we measure the magnitude of forces transmitted to ligand- receptor complexes and then to relate mechanical events to signaling and functional responses. My PhD research (F99 phase) has focused on developing methods to measure and elucidate the role of mechanical forces in immune response. I have designed a microparticle tension senor that allows one to quantify receptor forces in high throughput and also to measure forces at curved cell junctions. Additionally, I used this assay to screen the dose-response function of drugs that modulate cell mechanics. Because T cell responses are fine tuned by an array of co-receptors, I tested the role of mechanics in LFA-1 function. In this work, I demonstrated that the magnitude of LFA-1 integrin forces fine tunes TCR triggered activation and antigen discrimination. In addition, I revealed mechanically active LFA-1 defines the permissive zones for cytotoxic secretion, and suppression of LFA-1 forces significantly abrogates cytotoxicity. My work suggests that receptors cooperate to tune T-cell responses. For the remainder of my F99 phase, I will investigate the mechano-communication between receptor forces. Specifically, I will develop a DNA origami nano device to pattern ligands and measure spatiotemporal colocalization of mechanical events. Afterwards, I will proceed to test this hypothesis on cell plasma membrane by engineering tension probes on the surface of living cells. This will enable one to control and measure TCR-forces at authentic cell-cell junctions that mimic the chemical and physical properties of the immune synapse. For my postdoctoral work (K00 phase), I aim to improve upon current cancer therapies by leveraging T cell mechanics in boosting the specificity of immune response. In adoptive cell therapy (ACT), after therapeutic T-cell reinfusion, adjuvant drugs such as cytokines need to be administered to boost immune reconstitution. However, nonspecific drug release causes side effects and T-cell exhaustion. To address this challenge, I will decorate T-cells with DNA cages that mechanically trigger the release of encapsuled drugs at the tumor zone. If successful, this work will significantly enhance the ACT efficiency and offer the first example that links mechanobiology to cancer immunotherapy.