CXCL10 and CCL21 Promote Migration of Pancreatic Cancer Cells Toward Sensory Neurons and Neural Remodeling in Tumors in Mice, Associated With Pain in Patients
CXCL10 and CCL21 Promote Migration of Pancreatic Cancer Cells Toward Sensory Neurons and Neural Remodeling in Tumors in Mice, Associated With Pain in Patients
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DOI：
10.1053/j.gastro.2020.04.037
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发表时间：
2020-08-01
期刊：
影响因子：
29.4
通讯作者：
Kuner, Rohini
中科院分区：
文献类型：
--
作者：
Hirth, Michael;Gandla, Jagadeesh;Kuner, Rohini
背景与目的：胰腺导管腺癌（Pancreatic ductal adencarcinoma， PDAC）常伴有剧烈疼痛，这与癌细胞的吸引及其对胰腺内感觉神经的侵袭有关。在一项临床试验中，趋化因子CCL2的中和可以减少癌症相关的疼痛，但目前还没有对PDAC中高度多样化的趋化因子家族及其受体进行系统分析。方法：我们对小鼠PDAC细胞（K8484）和小鼠外周感觉神经元共培养的哺乳动物趋化因子进行了开放、无偏倚的RNA干扰筛选，并在DT8082 PDAC细胞的研究中证实了这些发现。我们研究了趋化因子对PDAC细胞系迁移的影响。用K8484细胞在小鼠体内培养原位肿瘤，并给小鼠注射针对趋化因子、拮抗剂或对照抗体的中和抗体。我们分析了腹部机械超敏反应，并收集肿瘤，通过组织学和免疫组织化学分析神经重塑。我们收集了74例切除患者的PDAC样本和疼痛水平信息，并通过免疫组织化学和免疫印迹法测量了CXCR3和CCR7的水平。结果：DRG神经元中9种趋化因子的下调显著降低了PDAC细胞向感觉神经元的迁移。感觉神经元来源的CCL21和CXCL10分别通过受体CCR7和CXCR3促进PDAC细胞的迁移，CCR7和CXCR3分别在原位肿瘤细胞和患者PDAC标本中表达。在原位肿瘤小鼠中，中和CCL21或CXCL10或其受体可显著降低伤害性超敏反应和神经纤维肥大，改善行为参数，而不影响T细胞或中性粒细胞的肿瘤浸润。从患者问卷中确定，人类PDAC标本中CXCR3和CCR7水平的增加与癌症相关疼痛频率的增加有关。结论：在一个无偏倚的切细胞因子筛选中，我们发现CCL21和CXCL10是促进胰腺癌细胞向感觉神经元迁移的蛋白。抑制这些趋化因子或其受体可减少原位肿瘤小鼠的超敏反应，并且具有高水平CXCR3和CCR7趋化因子受体的pdac患者癌症相关疼痛的频率增加。
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 anti-bodies. 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 immuno-blotting. RESULTS: Knockdown of 9 chemokines in DRG neurons significantly reduced migration of PDAC cells to-wards 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 fre-quency of cancer-associated pain, determined from patient questionnaires. CONCLUSIONS: In an unbiased screen of che-mokines, 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 re-ceptors of CXCR3 and CCR7 had increased frequency of cancer -associated pain.