Imaging CXCL12-CXCR4 signaling in ovarian cancer therapy.
Chemokine CXCL12 and receptor CXCR4 have emerged as promising therapeutic targets for ovarian cancer, a disease that continues to have a dismal prognosis. CXCL12-CXCR4 signaling drives proliferation, survival, and invasion of ovarian cancer cells, leading to tumor growth and metastasis. Pleiotropic effects of CXCR4 in multiple key steps in ovarian cancer suggest that blocking this pathway will improve outcomes for patients with this disease. To quantify CXCL12-CXCR4 signaling in cell-based assays and living mouse models of ovarian cancer, we developed a click beetle red luciferase complementation reporter that detects activation of CXCR4 based on recruitment of the cytosolic adapter protein β-arrestin 2. Both in two-dimensional and three-dimensional cell cultures, we established that bioluminescence from this reporter measures CXCL12-dependent activation of CXCR4 and inhibition of this pathway with AMD3100, a clinically-approved small molecule that blocks CXCL12-CXCR4 binding. We used this imaging system to quantify CXCL12-CXCR4 signaling in a mouse model of metastatic ovarian cancer and showed that treatment with AMD3100 interrupted this pathway in vivo. Combination therapy with AMD3100 and cisplatin significantly decreased tumor burden in mice, although differences in overall survival were not significantly greater than treatment with either agent as monotherapy. These studies establish a molecular imaging reporter system for analyzing CXCL12-CXCR4 signaling in ovarian cancer, which can be used to investigate biology and therapeutic targeting of this pathway in cell-based assays and living mice.
Center for Molecular Imaging, Department of Radiology, University of Michigan Medical School, Ann Arbor, Michigan, United States of America.
Emma
E
Salomonnson
Amanda C
AC
Stacer
Anna
A
Ehrlich
Kathryn E
KE
Luker
Gary D
GD
Luker
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D019390:Arrestins / Q000235:genetics / Q000378:metabolism; D001596:Benzylamines; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D054377:Chemokine CXCL12 / Q000235:genetics* / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology; D000080027:Cyclams; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D017930:Genes, Reporter; D006571:Heterocyclic Compounds / Q000494:pharmacology*; D006801:Humans; D008156:Luciferases; D008163:Luminescent Measurements; D051379:Mice; D016513:Mice, SCID; D057054:Molecular Imaging / Q000379:methods*; D009368:Neoplasm Transplantation; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D019718:Receptors, CXCR4 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
C000605254:ARRB2 protein, human; D000970:Antineoplastic Agents; C000605258:Arrb2 protein, mouse; D019390:Arrestins; D001596:Benzylamines; D054377:Chemokine CXCL12; D000080027:Cyclams; D006571:Heterocyclic Compounds; D019718:Receptors, CXCR4; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D008156:Luciferases; D002945:Cisplatin; C088327:plerixafor