NanoOptogenetic immunotherapy for B cell lymphoma

项目摘要/摘要： 嵌合抗原受体(Cars)是一种工程重组受体，由 T细胞受体(TCR)和共刺激受体的关键信号模块 以建立有效的抗肿瘤免疫。基因工程的CAR-T细胞被重新注入到 患者识别和攻击癌细胞。CAR-T细胞疗法已经显示出非常 临床试验取得了令人振奋的结果。然而，由于缺乏对剂量的精确控制， T细胞活动的位置和时间，这种方法涉及到一些重要的安全性 需要克服的挑战。例如，发生了细胞因子释放综合征(CRS) 由于反应CAR-T的大量和不受控制的细胞因子释放，可能会导致 症状从发烧到潜在的致命器官破坏。此外, 传统的CAR T细胞也与正常的靶向破坏有关 组织，这被称为“靶点上，肿瘤外”的效应，因为肿瘤抗原也 在几种正常组织中有一定水平的表达。因此，不受控制的 积极扩增的CAR-T细胞与心脏、肺或肝脏中的细胞发生交叉反应 会给病人带来毁灭性的后果。为了解决这一具有挑战性的问题，我们 建议开发允许时空控制的非侵入性方法 B细胞治疗肿瘤嵌合抗原受体(CAR)T细胞的研究 以淋巴瘤为测试案例。这项建议是基于两项关键发现 团队成员。首先，我们已经创造了一种通过光基因来调节 钙信号在可见光对基质相互作用中的作用 分子1(STIM1)，激活T细胞ORAI1钙通道以安装效应器 免疫反应。第二个是我们对上转换纳米粒子的开发 (UCNPs)，更具体地说，它们用作体内中继节点以捕获和转换低电平 将深层组织穿透性和近红外辐射(NIR)转化为可见光，用于 活体光电应用。在我们的初步结果中，我们已经证明 使用体外UCNPs和视神经双重工程方法确实可以 光遗传学指示免疫细胞(即树突状细胞)攻击小鼠的黑色素瘤， 近红外光疗有效地抑制了黑色素瘤的生长和转移 肺部。我们提出了三个具体目标。对于目标1，我们将开发可调光的汽车 在工程治疗性T细胞中。在目标2中，我们将设计新一代的UCNP 改进了与OptoCAR的兼容性。在目标3中，我们将确定 纳米光遗传CAR-T平台的体内外研究。这一新战略将克服 许多目前基于CAR-T的方法的局限性，并将使新的 在基础科学和人类健康方面的应用。

Project Summary/Abstract: Chimeric antigen receptors (CARs) are engineered recombinant receptors composed of key signaling modules from both the T cell receptor (TCR) and co-stimulatory receptors to mount effective anti-tumor immunity. Engineered CAR-T cells be reinfused into the patient to recognize and attack cancer cells. CAR-T cell therapy has shown very promising results in clinical trials. However, owing to a lack of precise control of the dose, location, and timing of T cell activity, this method involves some significant safety challenges to be overcome. For example, cytokine release syndrome (CRS), occurred due to a large and uncontrolled release of cytokines in response to CAR-T, may cause symptoms ranging from fever to potentially fatal organ destructions. In addition, conventional CAR T cells are also associated with the targeted destruction of normal tissue, which is known as “on-target, off-tumor” effects since tumor antigens are also expressed at a certain level in several normal tissues. Therefore, the uncontrolled aggressively amplified CAR-T cells cross-react with cells in the heart, lung or liver and cause devastating consequences in patients. To address this challenging issue, we propose to develop non-invasive methodologies that allow for the spatiotemporal control of chimeric antigen receptor (CAR) T cells for cancer treatment by using B cell lymphoma as a test case. This proposal is based on two key discoveries made by the members of the team. First, we have created a way to optogenetically modulate the function of calcium signaling by conferring visible light sensitivity to stromal interaction molecule 1 (STIM1), activating the ORAI1 calcium channel in T cells to mount effector immune responses. The second is our development of upconversion nanoparticles (UCNPs), more specifically their use as in vivo relay nodes to capture and convert low power, deep tissue-penetrant, and near infrared radiation (NIR) into visible light for in vivo optogentic applications. In our preliminary results, we have demonstrated that the use of ex vivo UCNPs and optogentic dual engineering approach can indeed optogenetically instruct immune cells (i.e,, dentritic cells) to attack melanoma in mice, and that NIR light therapy effectively suppresses melanoma growth and metastasis to lungs. We propose three specific aims. For Aim 1, we will develop photo-tunable CARs in engineered therapeutic T cells. In Aim 2, we will devise new generations of UCNPs with improved compatibility with OptoCARs. In Aim 3, we will determine the efficacy of nano-optogenetic CAR-T platforms in vitro and in vivo. This new strategy will overcome many of the limitations of current CAR-T based approaches, and will enable new applications in both fundamental science and human health.