Ferrari Francesca, Bellone Stefania, Black Jonathan, Schwab Carlton L, Lopez Salvatore, Cocco Emiliano, Bonazzoli Elena, Predolini Federica, Menderes Gulden, Litkouhi Babak, Ratner Elena, Silasi Dan-Arin, Azodi Masoud, Schwartz Peter E, Santin Alessandro D
Department of Obstetrics, Gynecology & Reproductive Sciences, Yale University School of Medicine, New Haven, CT, USA.
, 333 Cedar Street, LSOG 305, PO Box 208063, New Haven, CT, 06520-8063, USA.
J Exp Clin Cancer Res. 2015 Oct 17;34:123. doi: 10.1186/s13046-015-0241-7.
Uterine and ovarian carcinosarcomas (CS) are rare but highly aggressive gynecologic tumors which carry an extremely poor prognosis. We evaluated the expression levels of EpCAM and the in vitro activity of solitomab, a bispecific single-chain antibody construct which targets epithelial-cell-adhesion-molecule (EpCAM) on tumor cells and also contains a CD3 binding region, against primary uterine and ovarian CS cell lines.
EpCAM expression was evaluated by flow cytometry in a total of 5 primary CS cell lines. Sensitivity to solitomab-dependent-cellular-cytotoxicity (ADCC) was tested against the panel of primary CS cell lines expressing different levels of EpCAM in standard 4 h (51)Cr release-assays. The proliferative activity, activation, cytokine secretion (i.e., Type I vs Type II) and cytotoxicity of solitomab in autologous tumor-associated-T cells (TAL) in the pleural fluid of a CS patient were also evaluated by CFSE and flow-cytometry assays.
Surface expression of EpCAM was found in 80.0 % (4 out of 5) of the CS cell lines tested by flow cytometry. EpCAM positive cell lines were found resistant to NK or T-cell-mediated killing after exposure to peripheral blood lymphocytes (PBL) in 4-h chromium-release assays (mean killing ± SEM = 1.1 ± 1.6 %, range 0-5.3 % after incubation of EpCAM positive cell lines with control BiTE®). In contrast, after incubation with solitomab, EpCAM positive CS cells became highly sensitive to T-cell-cytotoxicity (mean killing ± SEM of 19.7 ± 6.3 %; range 10.0-32.0 %; P < 0.0001). Ex vivo incubation of autologous TAL with EpCAM expressing malignant cells in pleural effusion with solitomab, resulted in a significant increase in T-cell proliferation in both CD4+ and CD8+ T cells, increase in T-cell activation markers (i.e., CD25 and HLA-DR), and a reduction in number of viable CS cells in the exudate (P < 0.001).
Solitomab may represent an effective treatment for patients with recurrent/metastatic and/or chemo-resistant CS overexpressing EpCAM.
子宫和卵巢癌肉瘤（CS）是罕见但侵袭性很强的妇科肿瘤，预后极差。我们评估了上皮细胞黏附分子（EpCAM）的表达水平以及索利妥单抗的体外活性，索利妥单抗是一种双特异性单链抗体构建体，可靶向肿瘤细胞上的上皮细胞黏附分子（EpCAM），并且还含有一个CD3结合区域，用于检测原发性子宫和卵巢CS细胞系。
通过流式细胞术评估了总共5种原发性CS细胞系中EpCAM的表达。在标准4小时（51）铬释放试验中，针对表达不同水平EpCAM的原发性CS细胞系面板测试了对索利妥单抗依赖性细胞毒性（ADCC）的敏感性。还通过CFSE和流式细胞术试验评估了索利妥单抗在CS患者胸腔积液中自体肿瘤相关T细胞（TAL）中的增殖活性、激活、细胞因子分泌（即I型与II型）和细胞毒性。
通过流式细胞术检测的CS细胞系中，80.0%（5个中的4个）发现有EpCAM的表面表达。在4小时铬释放试验中，EpCAM阳性细胞系在暴露于外周血淋巴细胞（PBL）后对NK或T细胞介导的杀伤具有抗性（EpCAM阳性细胞系与对照双特异性T细胞衔接器（BiTE®）孵育后，平均杀伤率±标准误=1.1±1.6%，范围为0-5.3%）。相比之下，与索利妥单抗孵育后，EpCAM阳性CS细胞对T细胞细胞毒性变得高度敏感（平均杀伤率±标准误为19.7±6.3%；范围为10.0-32.0%；P<0.0001）。用索利妥单抗将自体TAL与胸腔积液中表达EpCAM的恶性细胞进行体外孵育，导致CD4+和CD8+T细胞中T细胞增殖显著增加，T细胞激活标志物（即CD25和HLA-DR）增加，渗出液中存活的CS细胞数量减少（P<0.001）。
索利妥单抗可能是治疗复发/转移和/或化疗耐药的过表达EpCAM的CS患者的有效疗法。
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