Michael Hirth1, Jagadeesh Gandla2, Christiane Höper2, Matthias M Gaida3, Nitin Agarwal2, Manuela Simonetti2, Aykut Demir2, Yong Xie2, Cleo Weiss4, Christoph W Michalski5, Thilo Hackert6, Matthias P Ebert7, Rohini Kuner8. 1. Institute of Pharmacology, Medical Faculty Heidelberg, Heidelberg University, Heidelberg, Germany; Department of Medicine II, University Medical Center Mannheim, Medical Faculty at Mannheim, University of Heidelberg, Mannheim, Germany. 2. Institute of Pharmacology, Medical Faculty Heidelberg, Heidelberg University, Heidelberg, Germany. 3. Institute of Pathology, University Hospital Heidelberg, Heidelberg, Germany; Institute of Pathology, University Hospital Johannes Gutenberg-University Mainz, Mainz, Germany. 4. Institute of Pathology, University Medical Center Mannheim, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany. 5. Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, Heidelberg, Germany; Department of Surgery, Halle University Hospital, Halle, Germany. 6. Department of General, Visceral and Transplantation Surgery, Heidelberg University Hospital, Heidelberg, Germany. 7. Department of Medicine II, University Medical Center Mannheim, Medical Faculty at Mannheim, University of Heidelberg, Mannheim, Germany. 8. Institute of Pharmacology, Medical Faculty Heidelberg, Heidelberg University, Heidelberg, Germany. Electronic address: rohini.kuner@pharma.uni-heidelberg.de.
Abstract
BACKGROUND & AIMS: Pancreatic ductal adenocarcinoma (PDAC) is frequently accompanied by excruciating pain, which has been associated with attraction of cancer cells and their invasion of intrapancreatic sensory nerves. Neutralization of the chemokine CCL2 reduced cancer-associated pain in a clinical trial, but there have been no systematic analyses of the highly diverse chemokine families and their receptors in PDAC. METHODS: We performed an open, unbiased RNA-interference screen of mammalian chemokines in co-cultures of mouse PDAC cells (K8484) and mouse peripheral sensory neurons, and confirmed findings in studies of DT8082 PDAC cells. We studied the effects of chemokines on migration of PDAC cell lines. Orthotopic tumors were grown from K8484 cells in mice, and mice were given injections of neutralizing antibodies against chemokines, antagonists, or control antibodies. We analyzed abdominal mechanical hypersensitivity and collected tumors and analyzed them by histology and immunohistochemistry to assess neural remodeling. We collected PDAC samples and information on pain levels from 74 patients undergoing resection and measured levels of CXCR3 and CCR7 by immunohistochemistry and immunoblotting. RESULTS: Knockdown of 9 chemokines in DRG neurons significantly reduced migration of PDAC cells towards sensory neurons. Sensory neuron-derived CCL21 and CXCL10 promoted migration of PDAC cells via their receptors CCR7 and CXCR3, respectively, which were expressed by cells in orthotopic tumors and PDAC specimens from patients. Neutralization of CCL21 or CXCL10, or their receptors, in mice with orthotopic tumors significantly reduced nociceptive hypersensitivity and nerve fiber hypertrophy and improved behavioral parameters without affecting tumor infiltration by T cells or neutrophils. Increased levels of CXCR3 and CCR7 in human PDAC specimens were associated with increased frequency of cancer-associated pain, determined from patient questionnaires. CONCLUSIONS: In an unbiased screen of chemokines, we identified CCL21 and CXCL10 as proteins that promote migration of pancreatic cancer cells toward sensory neurons. Inhibition of these chemokines or their receptors reduce hypersensitivity in mice with orthotopic tumors, and patients with PDACs with high levels of the chemokine receptors of CXCR3 and CCR7 had increased frequency of cancer-associated pain.
BACKGROUND & AIMS:Pancreatic ductal adenocarcinoma (PDAC) is frequently accompanied by excruciating pain, which has been associated with attraction of cancer cells and their invasion of intrapancreatic sensory nerves. Neutralization of the chemokine CCL2 reduced cancer-associated pain in a clinical trial, but there have been no systematic analyses of the highly diverse chemokine families and their receptors in PDAC. METHODS: We performed an open, unbiased RNA-interference screen of mammalian chemokines in co-cultures of mouse PDAC cells (K8484) and mouse peripheral sensory neurons, and confirmed findings in studies of DT8082 PDAC cells. We studied the effects of chemokines on migration of PDAC cell lines. Orthotopic tumors were grown from K8484 cells in mice, and mice were given injections of neutralizing antibodies against chemokines, antagonists, or control antibodies. We analyzed abdominal mechanical hypersensitivity and collected tumors and analyzed them by histology and immunohistochemistry to assess neural remodeling. We collected PDAC samples and information on pain levels from 74 patients undergoing resection and measured levels of CXCR3 and CCR7 by immunohistochemistry and immunoblotting. RESULTS: Knockdown of 9 chemokines in DRG neurons significantly reduced migration of PDAC cells towards sensory neurons. Sensory neuron-derived CCL21 and CXCL10 promoted migration of PDAC cells via their receptors CCR7 and CXCR3, respectively, which were expressed by cells in orthotopic tumors and PDAC specimens from patients. Neutralization of CCL21 or CXCL10, or their receptors, in mice with orthotopic tumors significantly reduced nociceptive hypersensitivity and nerve fiber hypertrophy and improved behavioral parameters without affecting tumor infiltration by T cells or neutrophils. Increased levels of CXCR3 and CCR7 in human PDAC specimens were associated with increased frequency of cancer-associated pain, determined from patient questionnaires. CONCLUSIONS: In an unbiased screen of chemokines, we identified CCL21 and CXCL10 as proteins that promote migration of pancreatic cancer cells toward sensory neurons. Inhibition of these chemokines or their receptors reduce hypersensitivity in mice with orthotopic tumors, and patients with PDACs with high levels of the chemokine receptors of CXCR3 and CCR7 had increased frequency of cancer-associated pain.
Authors: William L Hwang; Karthik A Jagadeesh; Jimmy A Guo; Hannah I Hoffman; Payman Yadollahpour; Jason W Reeves; Rahul Mohan; Eugene Drokhlyansky; Nicholas Van Wittenberghe; Orr Ashenberg; Samouil L Farhi; Denis Schapiro; Prajan Divakar; Eric Miller; Daniel R Zollinger; George Eng; Jason M Schenkel; Jennifer Su; Carina Shiau; Patrick Yu; William A Freed-Pastor; Domenic Abbondanza; Arnav Mehta; Joshua Gould; Conner Lambden; Caroline B M Porter; Alexander Tsankov; Danielle Dionne; Julia Waldman; Michael S Cuoco; Lan Nguyen; Toni Delorey; Devan Phillips; Jaimie L Barth; Marina Kem; Clifton Rodrigues; Debora Ciprani; Jorge Roldan; Piotr Zelga; Vjola Jorgji; Jonathan H Chen; Zackery Ely; Daniel Zhao; Kit Fuhrman; Robin Fropf; Joseph M Beechem; Jay S Loeffler; David P Ryan; Colin D Weekes; Cristina R Ferrone; Motaz Qadan; Martin J Aryee; Rakesh K Jain; Donna S Neuberg; Jennifer Y Wo; Theodore S Hong; Ramnik Xavier; Andrew J Aguirre; Orit Rozenblatt-Rosen; Mari Mino-Kenudson; Carlos Fernandez-Del Castillo; Andrew S Liss; David T Ting; Tyler Jacks; Aviv Regev Journal: Nat Genet Date: 2022-07-28 Impact factor: 41.307
Authors: Alessandro Di Federico; Mirta Mosca; Rachele Pagani; Riccardo Carloni; Giorgio Frega; Andrea De Giglio; Alessandro Rizzo; Dalia Ricci; Simona Tavolari; Mariacristina Di Marco; Andrea Palloni; Giovanni Brandi Journal: Cancers (Basel) Date: 2022-05-14 Impact factor: 6.575