Wakileh Gamal A, Bierholz Philipp, Kotthoff Mara, Skowron Margaretha A, Bremmer Felix, Stephan Alexa, Anbuhl Stephanie M, Heukers Raimond, Smit Martine J, Ströbel Philipp, Nettersheim Daniel
Department of Urology, Urological Research Laboratory, Translational UroOncology, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, Moorenstraße 5, 40225, Düsseldorf, Germany.
Department of Urology, University Hospital Ulm, Ulm, Germany.
Exp Hematol Oncol. 2023 Nov 23;12(1):96. doi: 10.1186/s40164-023-00460-9.
Being stimulated by the chemokine CXCL12, the CXCR4 / CXCR7 cascade is involved in tumor proliferation, migration, and metastasis. The interaction between CXCL12, secreted by cells from the microenvironment, and its receptors is complex and has been ascribed to promote chemotherapy resistance. However, the role of this signaling axis and its targetability in germ cell tumors (GCT) is not fully understood. Thus, this study investigated the therapeutic efficacy of a nanobody-drug-conjugate targeting CXCR4 (CXCR4-NDC) and functionally characterized this signaling pathway in GCT using small molecule inhibitors and nanobodies. As shown by diminished cell viability, enhanced apoptosis induction, and detection of mitotic catastrophes, we confirmed the cytotoxic efficacy of the CXCR4-NDC in CXCR4-GCT cells (i.e. seminoma and yolk-sac tumor), while non-malignant CXCR4-fibroblasts, remained largely unaffected. Stimulation of CXCR4 / CXCR7-GCT cells with CXCL12 resulted in an enhanced proliferative and migratory capacity, while this effect could be reverted using CXCR4 inhibitors or a CXCR7-nanobody. Molecularly, the CXCR4 / CXCR7-signaling cascade could be activated independently of MAPK (ERK1 / 2)-phosphorylation. Although, in CXCR4 / CXCR7-embryonal carcinoma cells, CXCR7-expression was re-induced upon inhibition of ERK1 / 2-signaling. This study identified a nanobody-drug-conjugate targeting CXCR4 as a putative therapeutic option for GCT, i.e. seminoma and yolk-sac tumors. Furthermore, this study shed light on the functional role of the CXCR4 / CXCR7 / CXCL12-signaling cascade in GCT, demonstrating an important influence on proliferation and migration.
趋化因子CXCL12刺激后，CXCR4 / CXCR7级联反应参与肿瘤增殖、迁移和转移。微环境细胞分泌的CXCL12与其受体之间的相互作用复杂，且被认为可促进化疗耐药。然而，该信号轴在生殖细胞肿瘤（GCT）中的作用及其靶向性尚未完全明确。因此，本研究调查了靶向CXCR4的纳米抗体 - 药物偶联物（CXCR4 - NDC）的治疗效果，并使用小分子抑制剂和纳米抗体对GCT中的该信号通路进行功能表征。通过细胞活力降低、凋亡诱导增强以及有丝分裂灾难的检测表明，我们证实了CXCR4 - NDC对CXCR4阳性GCT细胞（即精原细胞瘤和卵黄囊瘤）具有细胞毒性作用，而对非恶性的CXCR4阳性成纤维细胞基本无影响。用CXCL12刺激CXCR4 / CXCR7阳性GCT细胞会导致增殖和迁移能力增强，而使用CXCR4抑制剂或CXCR7纳米抗体可逆转这种作用。在分子水平上，CXCR4 / CXCR7信号级联反应可独立于MAPK（ERK1 / 2）磷酸化而被激活。尽管在CXCR4 / CXCR7阳性胚胎癌细胞中，抑制ERK1 / 2信号后CXCR7表达会再次诱导。本研究确定靶向CXCR4的纳米抗体 - 药物偶联物是GCT（即精原细胞瘤和卵黄囊瘤）的一种潜在治疗选择。此外，本研究揭示了CXCR4 / CXCR7 / CXCL12信号级联反应在GCT中的功能作用，证明其对增殖和迁移有重要影响。
