Use of Ly6K specific CAR T-cells to treat primary and metastatic triple negative breast cancer

项目摘要/摘要 我们已经发现，Ly6K表达增加与三重阴性的预后不良相关 乳腺癌(TNBC)。从机制上讲，Ly6K是转化生长因子β信号通路激活所必需的，并且在 体内的肿瘤发生。我们认为生物标记物Ly6K是治疗骨肉瘤的理想靶点。 TNBC，因为这种蛋白在正常细胞中不表达，除了在睾丸中。Ly6K也不是至关重要的 器官功能，但精子发生除外。因此，靶向该蛋白用于治疗TNBC这一疾病 主要影响女性，这是适当和理想的。正因为如此，Ly6K是治疗的最佳靶点 使用CAR T细胞进行TNBC免疫治疗。CAR T细胞是一种表达嵌合体的转基因T细胞 抗原受体融合蛋白(CAR)来源于识别靶蛋白的单抗 癌细胞的表面。CAR T细胞可以识别和杀伤表达靶蛋白的靶细胞，而不需要 人类白细胞抗原配型的必要性。我们已经制备了三株抗人Ly6K的高亲和力单抗，它们可以识别 癌细胞表面的Ly6K。来自Ly6K单抗的序列被用来在三分之一中产生一辆汽车 产生慢病毒CAR载体以产生Ly6K特异性CAR T细胞。我们将测试特定于Ly6K的汽车 T细胞将识别表达细胞表面Ly6K的癌细胞，并在原发和转移中消除它们 TNBC小鼠模型。

Project Summary/abstract We have identified that increased Ly6K expression is associated with poor outcome in triple negative breast cancer (TNBC). Mechanistically, Ly6K is required for the activation of TGFβ signaling and is increased in tumorigenesis in vivo. We propose that the biomarker Ly6K is an ideal therapeutic target for the treatment of TNBC because this protein is not expressed in normal cells, except in testis. Ly6K is also not required for vital organ function, except for spermatogenesis. Thus, targeting this protein for the treatment of TNBC, a disease affecting mostly females, is appropriate and ideal. Because of this, Ly6K is an optimal therapeutic target for TNBC immunotherapy using CAR T-cells. CAR T-cells are genetically modified T cells which express a chimeric antigen receptor fusion protein (CAR) derived from a monoclonal antibody (mAb) recognizing a target protein on the surface of cancer cells. CAR T-cells can recognize and kill target cells expressing the target protein without the need for HLA matching. We have generated three high affinity mAbs against human Ly6K, which recognize the cell surface Ly6K on cancer cells. The sequences from Ly6K mAbs were used to generate a CAR in a third generation lentiviral CAR vector to generate Ly6K-specific CAR T-cells. We will test whether Ly6K-specific CAR T-cells will recognize cancer cells expressing cell surface Ly6K and eliminate them in primary and metastatic TNBC mouse models.