Harnessing CAR T cells to deconstruct the interconnectivity among hallmarks of aging

总结 到目前为止，已经描述了老化过程的九个特征，可以分为三大类 类别：主要标志或损伤原因（基因组不稳定性，端粒磨损，表观遗传改变 和蛋白质稳态的丧失）、拮抗剂标志或对损伤的反应（细胞衰老、线粒体 功能障碍和失调的营养感应）和老化表型的综合标志或表现 (stem细胞耗竭和改变的细胞间通讯）。尽管对每一个人进行了深入的研究， 作为一个标志，人们对它们之间的相互联系以及支配这些相互作用的等级制度知之甚少。限制 这些研究中的一个因素是开发、繁殖和老化小鼠模型所需的时间和资源， 将允许解开相互作用，以及缺乏高度选择性的小分子， 体细胞方法。在这方面，我们最近的工作首次表明了采用细胞免疫的可行性。 治疗，以消除已经进入特定细胞程序（如细胞衰老）的细胞。作为证据- 在一项概念研究中，我们将uPAR确定为在细胞衰老中上调的表面分子，并开发了 CAR T细胞能够以高效力和特异性消除衰老细胞。CAR T细胞具有优势， 它们可以体细胞地施用，并且在专门靶向表达 特异性抗原此外，它们提供了空间和时间控制的可能性，使其具有高度的通用性 工具.在此，我们利用我们团队在衰老生物学和细胞工程技术方面的专业知识， 了解衰老的标志之间的相互作用。我们的目标是深入描述 衰老清除CAR T细胞对衰老的拮抗和整合标志;开发组织特异性衰老清除 CAR-T细胞可以让我们解开生理寿命中的标志之间的层次关系， 研究调节相互作用以治疗年龄相关病理的治疗潜力。 初步数据有力地支持了拟议工作的可行性：我们成功地安全地 在自然衰老的小鼠中使用了衰老清除CAR T细胞，并显示出它们在消除靶细胞方面的高效力。 在很长一段时间内组织中的阳性细胞。重要的是，用衰老清除性CAR T细胞治疗导致 健康状况的显著改善。在我们的应用中，我们继续以系统的方式表征 跨标志的效果，并利用细胞治疗的灵活性来解开管理 交互.总的来说，我们希望我们的研究将导致多功能的工具来询问衰老生物学，以及 为一系列与年龄有关的疾病开发新的疗法。

SUMMARY To date nine hallmarks of the aging process have been described, which can be grouped into three major categories: primary hallmarks or cause of damage (genomic instability, telomere attrition, epigenetic alterations and loss of proteostasis), antagonist hallmarks or responses to damage (cellular senescence, mitochondrial dysfunction and deregulated nutrient sensing) and integrative hallmarks or manifestations of the aging phenotype (stem cell exhaustion and altered intercellular communication). Despite intensive research on each individual hallmark, little is known about their interconnectivity and the hierarchies governing those interactions. A limiting factor in these studies is the time and resources needed for developing, breeding and aging mouse models that would allow to untangle the interactions as well as the lack of highly selective small molecules that would enable somatic approaches. In this regard, our recent work showed for the first time the feasibility of employing cellular therapy to eliminate cells that have entered a specific cellular program such as cellular senescence. In a proof- of-concept study we identified uPAR as a surface molecule upregulated in cellular senescence and developed CAR T cells able to eliminate senescent cells with high potency and specificity. CAR T cells have the advantage that they can be administered somatically and are highly effective at exclusively targeting cells expressing a specific antigen. In addition, they offer the possibility of spatial and temporal control making them highly versatile tools. Herein we leverage our team’s expertise in senescence biology and cell engineering technology to shed light into the interactions among the hallmarks of aging. Our goals are to characterize in depth the impact of senolytic CAR T cells on the antagonistic and integrative hallmarks of aging; develop tissue specific senolytic CAR T cells that would allow us to untangle hierarchies among the hallmarks across physiological lifespan and study the therapeutic potential of modulating the interactions for the treatment of age-related pathologies. Preliminary data strongly supports the feasibility of the proposed work: we have successfully and safely employed senolytic CAR T cells in naturally aged mice and showed their high potency at eliminating target positive cells in the tissues over long periods of time. Importantly, treatment with senolytic CAR T cells results in significant improvements in healthspan. In our application, we continue to characterize in a systematic way the effects across hallmarks and harness the flexibility of cellular therapy to untangle the hierarchies governing the interactions. Overall, we expect our studies will lead to versatile tools to interrogate aging biology as well as to the development of new therapies for a range of age-related pathologies.