Genetic Engineering of T Cells to Target HERV-K, an Ancient Retrovirus on Melanoma.
Genetic Engineering of T Cells to Target HERV-K, an Ancient Retrovirus on Melanoma.
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DOI：
10.1158/1078-0432.ccr-14-3197
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发表时间：
2015-07-15
期刊：
影响因子：
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通讯作者：
Cooper LJ
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文献类型：
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作者：
Krishnamurthy J;Rabinovich BA;Mi T;Switzer KC;Olivares S;Maiti SN;Plummer JB;Singh H;Kumaresan PR;Huls HM;Wang-Johanning F;Cooper LJ
人内源性逆转录病毒（HERV-K）包膜（env）蛋白是在黑色素瘤上表达的肿瘤相关抗原，而不是在正常细胞上表达。这项研究旨在设计T细胞表面的嵌合抗原受体（CAR），使其靶向黑色素瘤晚期的肿瘤。使用免疫组织化学（IHC）分析在220个黑素瘤样本（具有不同疾病阶段）和139个正常器官供体组织中分析HERV-K蛋白的表达。使用基于转座子的睡美人（SB）系统将源自小鼠单克隆抗体的HERV-K env特异性CAR引入T细胞中。在活化和增殖细胞（AaPC）上离体扩增HERV-K env特异性CAR+ T细胞，并表征CAR表达和特异性。这包括评估HERV-K特异性CAR+ T细胞在小鼠异种移植模型中杀死A375-SM转移肿瘤的能力。我们在黑色素瘤中检测到HERV-K env蛋白，但在正常组织中没有检测到。在电穿孔T细胞并在HERV-K+ AaPC上选择后，超过95%的遗传修饰的T细胞表达具有效应记忆表型的CAR，并以抗原特异性方式裂解HERV-K env+肿瘤靶标。尽管这种TAA从肿瘤细胞中明显脱落，可以被HERV-K env特异性CAR+ T细胞识别，但我们观察到了显著的抗肿瘤作用。使用HERV-K env特异性CAR+ T细胞的连续性细胞免疫疗法代表了晚期黑色素瘤的临床上有吸引力的治疗策略，并提供了将这种TAA靶向其他实体瘤的方法。
The human endogenous retrovirus (HERV-K) envelope (env) protein is a tumor- associated antigen expressed on melanoma, but not normal cells. This study was designed to engineer a chimeric antigen receptor (CAR) on T cell surface, such that they target tumors in advanced stages of melanoma. Expression of HERV-K protein was analyzed in 220 melanoma samples (with various stages of disease) and 139 normal organ donor tissues using immuno-histochemical (IHC) analysis. HERV-K env-specific CAR derived from mouse monoclonal antibody was introduced into T cells using the transposon-based Sleeping Beauty (SB) system. HERV-K env-specific CAR+ T cells were expanded ex vivo on activating and propagating cells (AaPC), and characterized for CAR expression and specificity. This includes evaluating the HERV-K-specific CAR+ T cells for their ability to kill A375-SM metastasized tumors in a mouse xenograft model. We detected HERV-K env protein on melanoma, but not in normal tissues. After electroporation of T cells and selection on HERV-K+ AaPC, over 95% of genetically-modified T cells expressed the CAR with an effector memory phenotype and lysed HERV-K env+ tumor targets in an antigen specific manner. Even though there is apparent shedding of this TAA from tumor cells which can be recognized by HERV-K env-specific CAR+ T cells, we observed a significant anti-tumor effect. Adoptive cellular immunotherapy with HERV-K env-specific CAR+ T cells represents a clinically-appealing treatment strategy for advanced-stage melanoma and provides an approach for targeting this TAA on other solid tumors.