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

项目概要/摘要 我们已经确定Ly 6 K表达增加与三阴性患者预后不良相关， 乳腺癌（TNBC）。从机制上讲，Ly 6 K是激活TGFβ信号传导所必需的，并且在TGF β 1/2细胞中增加。 体内肿瘤发生。我们认为，生物标志物Ly 6 K是一个理想的治疗靶点，用于治疗 TNBC，因为这种蛋白质在正常细胞中不表达，除了睾丸。Ly 6 K也不需要用于重要的 器官功能，除了精子生成。因此，靶向这种蛋白质用于治疗TNBC， 主要影响女性，是合适和理想的。正因为如此，Ly 6 K是一个最佳的治疗靶点， 使用CAR T细胞的TNBC免疫疗法。CAR T细胞是表达嵌合T细胞的遗传修饰的T细胞。 抗原受体融合蛋白（CAR），其衍生自识别靶蛋白的单克隆抗体（mAb）， 癌细胞的表面。CAR T细胞可以识别并杀死表达靶蛋白的靶细胞，而不需要免疫细胞。 需要HLA配型。我们已经产生了三种针对人Ly 6 K的高亲和力mAb，其识别 癌细胞表面的Ly 6 K。来自Ly 6 K mAb的序列用于在第三实施方案中产生CAR。 在一个实施方案中，所述方法用于产生慢病毒CAR载体以产生Ly 6 K特异性CAR T细胞。我们将测试Ly 6 K特异性CAR T细胞将识别表达细胞表面Ly 6 K的癌细胞，并在原发性和转移性肿瘤中消除它们。 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.