Antagonism of EG-VEGF Receptors as Targeted Therapy for Choriocarcinoma Progression In Vitro and In Vivo

Abstract

Choriocarcinoma (CC) is the most malignant gestational trophoblastic disease that often develops from complete hydatidiform moles (CHM). Neither the mechanism of CC development nor its progression is yet characterized. We recently identified endocrine gland-derived vascular endothelial growth factor (EG-VEGF) as a novel key placental growth factor that controls trophoblast proliferation and invasion. EG-VEGF acts via two receptors, PROKR1 and PROKR2. Here, we demonstrate that EG-VEGF receptors can be targeted for CC therapy. Three approaches were used: (i) a clinical investigation comparing circulating EG-VEGF in control ( = 20) and in distinctive CHM ( = 38) and CC ( = 9) cohorts, (ii) an study investigating EG-VEGF effects on the CC cell line JEG3, and (iii) an study including the development of a novel CC mouse model, through a direct injection of JEG3-luciferase into the placenta of gravid SCID-mice. Both placental and circulating EG-VEGF levels were increased in CHM and CC (×5) patients. EG-VEGF increased JEG3 proliferation, migration, and invasion in two-dimensional (2D) and three-dimensional (3D) culture systems. JEG3 injection in the placenta caused CC development with large metastases compared with their injection into the uterine horn. Treatment of the animal model with EG-VEGF receptor's antagonists significantly reduced tumor development and progression and preserved pregnancy. Antibody-array and immunohistological analyses further deciphered the mechanism of the antagonist's actions. Our work describes a novel preclinical animal model of CC and presents evidence that EG-VEGF receptors can be targeted for CC therapy. This may provide safe and less toxic therapeutic options compared with the currently used multi-agent chemotherapies. .

The Emerging Role of the Prokineticins and Homeobox Genes in the Vascularization of the Placenta: Physiological and Pathological Aspects

“…PROKs regulate a stunning array of biological functions such as gastrointestinal motility (LeCouter et al, 2001;Lin et al, 2002), circadian rhythm regulation, neurogenesis, angiogenesis, pain perception, mood regulation, and reproduction (Brouillet et al, 2010(Brouillet et al, , 2012bAlfaidy et al, 2016;Zhao et al, 2019). Dysregulation of PROKs/PROKRs signaling pathways have been reported in a variety of diseases, such as cancer, abnormal angiogenesis and pregnancy pathologies (Brouillet et al, 2010(Brouillet et al, , 2012bAlfaidy et al, 2016;Traboulsi et al, 2017;Zhao et al, 2019).…”

“…Ongoing validations of the newly discovered EG-VEGF-regulated homeobox genes should bring more insights into the role of these homeobox genes in the control of angiogenic processes at both intra-villi and extra-villi sites, throughout human pregnancy, Figure 1. Hoffmann et al, 2006Hoffmann et al, , 2009Brouillet et al, 2012aBrouillet et al, ,b, 2013bHolloway et al, 2014;Garnier et al, 2015;Murthi et al, 2015;Traboulsi et al, 2015Traboulsi et al, , 2017Sergent et al, 2016 HLX, DLX3, DLX4, MSX2, GAX,TLX1,TLX2 Murthi et al, 2007Murthi et al, , 2008Rajaraman et al, 2008Rajaraman et al, , 2010Chui et al, 2013;Liang et al, 2016;Novakovic et al, 2017;Harris et al,…”

“…These finding strongly suggest that antagonisation of EG-VEGF signaling might contribute to the attenuation of vascular-pregnancy pathologies. Importantly, we have recently demonstrated that treatment of animal model of choriocarcinoma with PROKR2 antagonist significantly reduced tumor growth, vascularization and metastasis (Traboulsi et al, 2017). Hence, one can speculate that the benefit upon the antagonisation of the prokineticin signaling might well trigger an upstream regulation of the EG-VEGF-dependent homeobox genes including NKX3.1, HOXA9, HOXC8, HOXC10, HOXD1, HOXD8, HOXD9, and HOXD11 as potential target genes for aberrant angiogenesis associated with the pathogenesis of PE and FGR.…”

The Angiogenic Paracrine Potential of Mesenchymal Stem Cells

“…It seems that [11] Angiopoietin-1 +/+ Activates TEK/TIE2 receptor; promotes angiogenic processes, endothelial cell survival, migration, proliferation, and stabilization; and during embryogenesis has a role in heart development [12] Angiopoietin-2 +/− Binds to TEK/TIE2, in the presence of VEGF and Ang-2 and promotes neovascularization [13,14] Angiopoietin-4 +/+ Binds to TEK/TIE2, modulating ANGPT1 signaling, can induce tyrosine phosphorylation of TEK/TIE2, and promotes endothelial cell survival, migration, and angiogenesis [15] Amphiregulin +/− An EGF-like ligand that binds to the EGFR, enhanced lymphangiogenesis, and stimulates the growth of normal epithelial cells [16] Artemin +/− Binds for the GFR-alpha-3-RET and GFR-alpha-1-RET receptor and promotes angiogenesis [17] Tissue factor +/− Stimulates PDGF receptor signaling pathway, angiogenesis, endothelial cell migration, chemotaxis and proliferation, and coagulation factor III/CD142; improves transcription of VEGF; and reduces transcription of the thrombospondins [18] CXCL16 +/+ Encourages a chemotactic response, pro-angiogenic [19] DPPIV +/− A membrane-bound oligopeptidase acting on and modulating the pro-angiogenic chemokine CXCL12 [20] Epidermal growth factor +/− Encourages the growth of epithelial tissues, is anti-apoptotic, induces lymphangiogenesis, and improves MSC survival [21] EG-VEGF +/− Also called Prokineticin 1. Binds to PROKR1 and PROKR2, pro-angiogenic [22] Endothelin-1 +/+ Derived from the endothelium with vasoconstrictor and angiogenic effects, prolymphoangiogenic [23] Endoglin +/− Also called CD 105. Modulates TGF-β1 and β3 responses, vascular development, and angiogenic effects [24] FGF-7 +/+ Has positive effects on cell proliferation, migration and division, chemotaxis, and arteriogenesis [25] Acidic FGF/FGF-1 +/− Binds to for FGFR1 and integrins and induces angiogenesis [26] Basic FGF/FGF-2 +/− Ligand for FGFR1, FGFR2, FGFR3, and FGFR4, Vascular regeneration; role in cell migration and proliferation involved in angiogenesis, stimulates arteriogenesis [27] FGF-4 +/− Has positive effects in MSC proliferation, pro-angiogenic [28] GDNF +/− Has positive effects in angiogenesis [29] GM-CSF +/− Has positive effects in angiogenesis [30] Heparanase +/+ Has positive effects in angiogenesis [31] Heparin binding-EGF +/+ Has positive effects in angiogenesis [32] Hepatocyte growth factor +/− Has positive effects in angiogenesis …”

Involvement of the Prion Protein in the Protection of the Human Bronchial Epithelial Barrier Against Oxidative Stress

“…Prion mRNA expression was quantified by real-time RT-qPCR using CFX96 TM Real-Time detection system (Biorad) as previously described(66).Downloaded by YORK UNIVERSITY LIBRARIES from www.liebertpub.com at 12/21/18. For personal use only.…”