Targeting the secretory pathway in cytotoxic T lymphocytes to modulate their cytolytic capacity in cancer immunotherapy

雌激素调节基因EBAG9抑制细胞毒性T淋巴细胞的杀伤活性，提示雌激素及其受体抑制物也可能与T细胞介导的肿瘤免疫监视密切相关。为了治愈癌症，过继的T细胞转移必须采取策略来克服耐受环境，要么通过耗尽耐受因子，要么通过改善效应T细胞的能力。虽然许多实验和一些临床试验都集中在变量上，如a)抗原类型和呈递方式，b)转移的T细胞的数量和激活状态，c)肿瘤细胞的来源，d)患者的条件，但到目前为止，T细胞反应的细胞生物学方面的研究还很少。我们将在手术中将T细胞亲和力定义为一个受细胞溶解效应分子的合成和储存、细胞内小泡运输和细胞器成熟影响的变量。这一建议的具体目标是：1)通过药物雌激素调节增强过继转移的T细胞的杀伤能力；2)通过shRNA介导的EBAG9下调产生高度活跃的抗肿瘤T细胞；3)评估增强的细胞溶解能力与CTL记忆形成的关系；4)量化T细胞受体谱系多样性与调节溶解颗粒成熟的关系；5)研究过度的细胞毒性功能与免疫稳态之间的联系。

The estrogen-tunable gene EBAG9 tempers the killing activity of cytotoxic T ymphocytes, implicating that estrogen and its receptor inhibitors could also be intimately linked with T-cell mediated cancer immunosurveillance. Adoptive T cell transfer for the cure of cancer has to implicate strategies to overcome a tolerizing milieu, either by depleting tolerogenic factors, or by ameliorating the capacity of effector T cells. While many experimental and some clinical trials have focused on variables such as a) antigen type and mode of presentation, b) number and activation status of transferred T cells, c) origin of tumor cells, and d) conditioning of the patient, cell biological aspects of the T cell response have received little attention thus far. We will operationally define T cell avidity as a variable that is influenced by synthesis and storage of cytolytic effector molecules, intracellular vesicle transport, and organelle maturation. Specific aims within this proposal are: 1) Enhancement of cytolytic capacity of adoptively transferred T cells, as mediated by pharmacologic estrogen modulation; 2) Generation of highly avid anti tumor T cells through shRNAmediated EBAG9-downregulation; 3) Assessment of the relationship between enhanced cytolytic capacity and CTL memory formation; 4) Quantification of T cell receptor repertoire diversity in relationship to the modulation of lytic granule maturation; 5) Investigations on the link between excessive cytotoxic function and immune homeostasis.