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

项目摘要 在获得性免疫应答中，细胞毒性T淋巴细胞（CTL）不断“爬行”，寻找外源性免疫应答的证据， 其他细胞表面的肽片段。一旦T细胞遇到具有外源或突变的靶细胞， 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， 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特异性 促进T细胞活化这并不令人惊讶，因为TCR和其他T细胞共受体结合它们的同源受体。 配体只有当两个动态细胞物理“接触”。作为了解分子生物学作用的第一步， 调节T细胞反应的力量，重要的是我们测量传递给配体的力量的大小， 受体复合物，然后将机械事件与信号传导和功能反应联系起来。 我的博士研究（F99阶段）专注于开发方法来测量和阐明 免疫反应中的机械力我设计了一个微粒张力传感器， 高通量中的受体力，并且还测量弯曲细胞连接处的力。另外，我用这个 测定以筛选调节细胞力学的药物的剂量反应功能。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 表明LFA-1整联蛋白的大小迫使TCR触发的活化和抗原的微调， 歧视此外，我发现机械活性LFA-1定义了细胞毒性的允许区域， 分泌和抑制LFA-1力显著消除细胞毒性。 我的工作表明受体合作调节T细胞反应。在我的F99阶段的剩余时间里， 我将研究感受器力之间的机械通信。具体来说，我会发明一种DNA“折纸” 纳米器件图案配体和测量时空共定位的机械事件。后来我 将继续测试这一假设的细胞质膜上的工程张力探针的表面上， 活细胞这将使人们能够控制和测量真实的细胞-细胞连接处的TCR-力，所述细胞-细胞连接模拟细胞-细胞连接。 免疫突触的化学和物理特性。 对于我的博士后工作（K 00阶段），我的目标是通过利用T 增强免疫反应特异性的细胞机制。在过继性细胞疗法（ACT）中，治疗后 T细胞回输，需要给予细胞因子等辅助药物以促进免疫重建。 然而，非特异性药物释放引起副作用和T细胞耗竭。为了应对这一挑战，我将 用DNA笼子装饰T细胞，机械地触发肿瘤区域的药物释放。如果 如果成功，这项工作将大大提高ACT的效率，并提供第一个将 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.