Melanoma-associated chondroitin sulfate proteoglycan (MCSP) as target antigen for specific immunotherapy; from the bench to the bed-side.

针对黑色素瘤相关硫酸软骨素蛋白多糖（MCSP）的特异性免疫治疗MCSP是一种很有前途的黑色素瘤和肿瘤基质相关表面抗原。在之前的资助期内，我们参与了mcsp特异性T细胞反应的诱导，并建立了对芯片预测的HLA-A*02限制性表位特异性的T细胞。这些T细胞不能识别内源性MCSP，肽洗脱方法显示MCSP显然没有被加工和正常呈现。因此，我们寻求使用MCSP特异性嵌合抗原受体（CARs）来靶向这种抗原，因为它们以不依赖hla的方式识别MCSP。此外，MCSP被确定为肿瘤特异性T细胞的巨噬细胞获得性标志物，T细胞刺激因子SLAMF6被表征。巴勒斯坦伙伴建立了细胞离体和体外工作所需的基础设施。在目前的申请中，我们希望通过进行I期临床试验将这种方法应用于临床，我们打算用自体T细胞治疗皮肤黑色素瘤和葡萄膜黑色素瘤患者，这些T细胞被编码mcsp特异性CAR的mRNA电穿孔。纳入10例患者，主要终点为安全性和可行性。临床疗效将作为次要终点。以色列合作伙伴将继续他们对SLAMF6的研究，以确定它是否可以用于扩增更好的T细胞进行过继转移，并将对这些T细胞进行首次体内实验，在小鼠中转染mcsp特异性CAR。巴勒斯坦小组将检查非皮肤黑色素瘤的突变和抗原表达，非皮肤黑色素瘤占巴勒斯坦人口黑色素瘤的很大一部分。在来自Erlangen的荧光显微镜专家的帮助下，他将在巴勒斯坦呆一段时间，巴勒斯坦首席研究员是一名癌症病理学家，他将对来自巴勒斯坦、以色列和德国的肿瘤样本进行免疫病理学调查，包括试验患者，以及来自以色列组car治疗小鼠的肿瘤样本，以确定炎症环境、T细胞和其他免疫细胞的浸润以及mcsp的表达。

Targeting Melanoma-associated chondroitin sulfate proteoglycan (MCSP) for specific immunotherapy; from bench to bedside MCSP is a promising melanoma- and tumor-stroma-associated surface antigen. Within the previous funding period, we engaged in the induction of MCSP-specific T-cell responses, and established T cells specific for in silico-predicted HLA-A*02-restricted epitopes. These T cells failed to recognize endogenous MCSP and a peptide elution approach revealed that MCSP is apparently not processed and presented normally. Therefore we pursued the use of MCSP-specific Chimeric Antigen Receptors (CARs) to target this antigen, because they recognize MCSP in an HLA-independent fashion. In addition, MCSP was established as trogocytosis-acquired marker of tumor-specific T cells and the T-cell-stimulatory factor SLAMF6 was characterized. The Palestinian partner established the infrastructure required for cellular ex vivo and in vitro work. In the current application, we wish to take this approach to the clinic by performing a phase I clinical trial in which we intend to treat cutaneous melanoma and uveal melanoma patients with autologous T cells, which are electroporated with mRNA encoding an MCSP-specific CAR. Ten patients shall be included with the primary end-points safety and feasibility. Clinical efficacy will be addressed as secondary end-point. The Israeli partner will continue their investigation on SLAMF6 to determine whether it can be used to expand better T cells for adoptive transfer and will perform first in vivo experiments with these T cells, transfected with the MCSP-specific CAR in mice. The Palestinian group will examine the mutations and antigen-expression of non-cutaneous melanomas, which represent a large part of melanomas in the Palestinian population. With the help of a specialist for fluorescence microscopy from Erlangen, who will spend some time in Palestine, the Palestinian principal investigator, who is a cancer pathologist, will perform immunopathological investigations of tumor samples from Palestine, Israel, and Germany, including the trial patients, as well as from the CAR-treated mice of the Israeli group, to determine the inflammatory milieu, infiltration of T cells and other immune cells, and the MCSP-expression.