Development of senolytic CAR T cells as new therapeutic agents

项目摘要/摘要 衰老是一种细胞程序，导致应激反应中不可逆转的细胞周期停滞。虽然在 短期内促进组织动态平衡的恢复，缺乏对衰老细胞的免疫清除 因此，它们的慢性积累导致了慢性促炎环境，这已被证明 促进肿瘤的发展并促进与慢性组织相关的各种病理 损害，如肺或肝纤维化，糖尿病和动脉粥样硬化等。上一次感冒药 这些方法依赖于非选择性化学抑制剂的使用，这些化学抑制剂具有非靶标毒性，并且 对于衰老细胞之间的异质性，都是低效的。我论文项目的长期目标是 首次开发嵌合抗原受体(CAR)T细胞作为活性感受剂。作为第一步 为了开发衰老CAR T，我们将识别衰老特定的表面分子，以更好地识别， 确定衰老细胞的特征并将其作为目标。同时，我们将利用这些分子的选择性特征。 目的是制造能分解感觉神经的CAR T细胞，然后我们对其有效性和安全性进行研究 临床前衰老模型。我研究生学习最初几年的初步数据有力地支持了 拟议工作的可行性：我们已经确定了一个主要的细胞表面分子 在衰老细胞上表达，并形成靶向它的CAR T细胞。我们进一步提供了初步数据 表明这些CAR T细胞是真正的感觉剂，能够在培养和培养中消除衰老细胞 老鼠。在我剩下的博士工作中，我们继续验证蛋白质和分解衰老的CAR T细胞，并获得 更好地了解他们的作用机制和安全概况，并利用我们的发现来开发增强 版本以及具有新确定的因素的组合目标策略。完成拟议的 这项工作将导致有效的CAR T细胞的开发，并为其临床提供进一步的数据 开发作为癌症和其他一系列衰老的新治疗选择- 相关疾病。我的博士后研究将继续研究衰老和CAR T细胞，但 焦点的细微变化试图了解衰老是如何与分泌表型相关的 (SASP)影响CAR T细胞的活性。该提案旨在阐明确切的机制(S) 衰老刺激CAR T活性：通过微环境和/或CAR T活动本身的调节 并利用这一知识来开发组合方法的增强CAR T结构以改进 CAR-T对实体瘤的疗效。总体而言，这两个项目将首次揭示 将衰老和细胞治疗领域相结合，将导致高效的发展 增感剂以及提高CAR T细胞在实体瘤中的疗效的新策略。

PROJECT SUMMARY/ABSTRACT Senescence is a cellular program that leads to irreversible cell cycle arrest in response to stress. Although in the short term it promotes restoration of tissue homeostasis, the lack of immune clearance of senescent cells and thus their chronic accumulation leads to a chronic pro-inflammatory environment that has been shown to promote tumor development and to contribute to a variety of pathologies associated with chronic tissue damage such as lung or liver fibrosis, diabetes and atherosclerosis among others. Previous senolytic approaches have relied on the use of non-selective chemical inhibitors that have off-target toxicities and, owing to heterogeneity between senescent cells, are inefficient. The long term objective of my thesis project is to develop, for the first time, Chimeric Antigen Receptor (CAR) T cells as living senolytic agents. As a first step towards developing senolytic CAR T we will identify senescence-specific surface molecules to better identify, characterize and target senescent cells. In parallel, we will exploit the selective features of these molecules with the aim of producing senolytic CAR T cells, which we then characterize their efficacy and safety profile in preclinical senescence models. Preliminary data from the initial years of my graduate studies strongly supports the feasibility of the proposed work: we have already identified one cell surface molecule predominantly expressed on senescent cells and developed CAR T cells targeting it. We further provide preliminary data showing that these CAR T cells are bona fide senolytics capable of eliminating senescent cells in culture and in mice. For the rest of my doctoral work we continue to validate the protein and senolytic CAR T cells and gain better understanding of their mechanism of action and safety profile and use our findings to develop enhanced versions as well as combinatorial targeting strategies with newly identified factors. Completion of the proposed work will lead to the development of effective senolytic CAR T cells and provide further data for their clinical development as novel therapeutic options in cancer as well as for a wide range of other senescence- associated diseases. My postdoctoral research will continue to study senescence and CAR T cells but with a slight change of focus in order to try to understand how the Senescence Associated Secretory Phenotype (SASP) influences the activity of CAR T cells. The proposal aims to elucidate the exact mechanism(s) whereby senescence stimulates CAR T activity: through modulation of microenvironment and/or CAR T activity itself and exploit this knowledge to develop enhanced CAR T constructs of combinatorial approaches to improve CAR T efficacy in solid tumors. Overall these two projects will shed light for the first time into the possibility of combining the fields of senescence and cellular therapy and will lead to the development of highly effective senolytic agents as well as new strategies to improve the efficacy of CAR T cells in solid tumors.