Pain is the cancer-related event that is most disruptive to a cancer patient's quality of life. Although bone cancer pain is one of the most severe and common sources of chronic cancer pain, relatively little is known about the mechanisms that generate and maintain this pain. The proposed experiments are designed to study the role of two chemokines, CCL2 and CCL3 in the genesis and maintenance of bone cancer pain using an established mouse bone tumor model and a novel ex vivo model of cancer pain.
Four Specific Aims will be pursued: 1) Measure the spontaneous release of CCL2 and CCL3 in vivo and in vitro, from three different tumor types (osteosarcoma, fibrosarcoma and nonpainful melanoma). Based on preliminary data we hypothesize that CCL2 is released only by the fibrosarcoma and osteosarcoma, while CCL3 is released only by the osteosarcoma. 2) Identify the cell types that are producing CCL2 and CCL3 by using immunocytochemistry to localize each mediator to cells at the tumor site. We hypothesize that CCL2 will be localized to fibrosarcoma and osteosarcoma cells and to osteoclasts and macrophages at the tumor site, while CCL3 will be found predominately in the osteosarcoma cells. 3) Investigate the effect of CCL2 and CCL3 on the functional responses of DRG neurons using a novel in vitro tumor/bone-explant/DRG culture system. We hypothesize that CCL2 and CCL3 released from tumor cells or bone produce acute and long-term changes in stimulus- evoked Ca2+ transients that reflect changes in voltage dependent Ca2+ channels and TRPV1 receptors. We further hypothesize that the knockdown of these chemokines will prevent the effect of tumor cells on Ca2+ transients. 4) Determine whether manipulation of expression of CCL2 and CCL3 in tumors cells alters the pain producing phenotype of the cells. We hypothesize that knock-in of CCL2 and CCL3 in nonpainful melanoma cells will result in pain producing tumors, while knock-down of CCL2 and CCL3 in fibrosarcoma or osteosarcoma cells will reduce bone tumor-associated nociceptive responsiveness in behavioral pain tests. The proposed experiments are unique in that they will utilize the technique of microperfusion to directly measure algogens produced in vivo by different tumor types, while an innovative ex vivo tumor/bone-explant/DRG culture system will allow us to measure these substances in the culture media and to analyze their effects on DRG neurons. In addition the use of a new sleeping beauty transposon-shRNA procedure for long-term knockdown or production of these mediators will provide an excellent opportunity to investigate their role in nociception both in vitro and in vivo. Results will extend our knowledge of the mechanisms underlying tumor nociception and will provide a template for developing novel, life-enhancing treatments of cancer pain. For cancer patients, pain relief is central to a high quality of life while they battle the disease, yet current therapies are often distressingly ineffective for some types of cancer pain. To address this critical need and develop new therapies that successfully diminish cancer pain, we need to understand at the cellular level how cancer pain is generated and why it persists. This research focuses on two proteins, CCL2 and CCL3, that are secreted by tumors [and other tissues], and whose absence appears to be linked to decreased cancer pain. Use of an animal model of bone cancer pain and a new ex vivo cancer pain model to test the effects of CCL2 and CCL3 knockdown or antagonism will provide important data that may lead to the development of novel therapies for this dreaded condition.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA084233-10
Application #
8111113
Study Section
Somatosensory and Chemosensory Systems Study Section (SCS)
Program Officer
O'Mara, Ann M
Project Start
1999-12-01
Project End
2013-07-31
Budget Start
2011-08-01
Budget End
2013-07-31
Support Year
10
Fiscal Year
2011
Total Cost
$330,342
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Smeester, Branden A; Lee, Jang-Hern; Beitz, Alvin J (2017) Influence of social interaction on nociceptive-induced changes in locomotor activity in a mouse model of acute inflammatory pain: Use of novel thermal assays. Brain Res Bull 134:47-54
Smeester, B A; O'Brien, E E; Michlitsch, K S et al. (2016) The relationship of bone-tumor-induced spinal cord astrocyte activation and aromatase expression to mechanical hyperalgesia and cold hypersensitivity in intact female and ovariectomized mice. Neuroscience 324:344-54
O'Brien, E E; Smeester, B A; Michlitsch, K S et al. (2015) Colocalization of aromatase in spinal cord astrocytes: differences in expression and relationship to mechanical and thermal hyperalgesia in murine models of a painful and a non-painful bone tumor. Neuroscience 301:235-45
Seo, Hyoung-Sig; Roh, Dae-Hyun; Kwon, Soon-Gu et al. (2011) Acidic pH facilitates peripheral **meATP-mediated nociception in rats: differential roles of P2X, P2Y, ASIC and TRPV1 receptors in ATP-induced mechanical allodynia and thermal hyperalgesia. Neuropharmacology 60:580-6
Schmidt, Brian L; Hamamoto, Darryl T; Simone, Donald A et al. (2010) Mechanism of cancer pain. Mol Interv 10:164-78
Pacharinsak, Cholawat; Beitz, Alvin (2008) Animal models of cancer pain. Comp Med 58:220-33
Schreiber, Kristin L; Beitz, Alvin J; Wilcox, George L (2008) Activation of spinal microglia in a murine model of peripheral inflammation-induced, long-lasting contralateral allodynia. Neurosci Lett 440:63-7
Lynch, Jessica L; Alley, Jeremy F; Wellman, Lori et al. (2008) Decreased spinal cord opioid receptor mRNA expression and antinociception in a Theiler's murine encephalomyelitis virus model of multiple sclerosis. Brain Res 1191:180-91
Lynch, Jessica L; Gallus, Nathan J; Ericson, Marna E et al. (2008) Analysis of nociception, sex and peripheral nerve innervation in the TMEV animal model of multiple sclerosis. Pain 136:293-304
Khasabova, Iryna A; Stucky, Cheryl L; Harding-Rose, Catherine et al. (2007) Chemical interactions between fibrosarcoma cancer cells and sensory neurons contribute to cancer pain. J Neurosci 27:10289-98

Showing the most recent 10 out of 14 publications