TY - JOUR
T1 - NADPH Oxidase 1 in Liver Macrophages Promotes Inflammation and Tumor Development in Mice
AU - Liang, Shuang
AU - Ma, Hsiao Yen
AU - Zhong, Zhenyu
AU - Dhar, Debanjan
AU - Liu, Xiao
AU - Xu, Jun
AU - Koyama, Yukinori
AU - Nishio, Takahiro
AU - Karin, Daniel
AU - Karin, Gabriel
AU - Mccubbin, Ryan
AU - Zhang, Cuili
AU - Hu, Ronglin
AU - Yang, Guizhi
AU - Chen, Li
AU - Ganguly, Souradipta
AU - Lan, Tian
AU - Karin, Michael
AU - Kisseleva, Tatiana
AU - Brenner, David A.
N1 - Funding Information:
We thank Dennis R. Petersen and Kevin J. Tracey for providing us with the antibodies against 4-hydroxynonenal and HMGB1, respectively. Zhenyu Zhong was supported by a Prevent Cancer Foundation Board of Directors Award and an American Association for the Study of Liver Diseases Pinnacle Research Award. Debanjan Dhar was supported by the American Liver Foundation Liver Scholar Award. Research was supported by grants from the National Institutes of Health (R01AI043477, R01CA211794, and R01CA198103) to Michael Karin.
Funding Information:
Funding Supported by the National Institutes of Health ( R01 DK101737-01A1 , U01 AA022614-01A1 , R01 DK099205-01A1 , and P50 AA011999 to Tatiana Kisseleva and David A. Brenner).
Publisher Copyright:
© 2019 AGA Institute
PY - 2019/3
Y1 - 2019/3
N2 - Background & Aims: Although there are associations among oxidative stress, reduced nicotinamide adenine dinucleotide phosphate oxidase (NOX) activation, and hepatocellular carcinoma (HCC) development, it is not clear how NOX contributes to hepatocarcinogenesis. We studied the functions of different NOX proteins in mice after administration of a liver carcinogen. Methods: Fourteen-day-old Nox1 –/– mice, Nox4 –/– mice, Nox1 –/– Nox4 –/– (double-knockout) mice, and wild-type (WT) C57BL/6 mice were given a single intraperitoneal injection of diethylnitrosamine (DEN) and liver tumors were examined at 9 months. We also studied the effects of DEN in mice with disruption of Nox1 specifically in hepatocytes (Nox1 ΔHep ), hepatic stellate cells (Nox1 ΔHep ), or macrophages (Nox1 ΔMac ). Some mice were also given injections of the NOX1-specific inhibitor ML171. To study the acute effects of DEN, 8–12-week-old mice were given a single intraperitoneal injection, and liver and serum were collected at 72 hours. Liver tissues were analyzed by histologic examination, quantitative polymerase chain reaction, and immunoblots. Hepatocytes and macrophages were isolated from WT and knockout mice and analyzed by immunoblots. Results: Nox4 –/– mice and WT mice developed liver tumors within 9 months after administration of DEN, whereas Nox1 –/– mice developed 80% fewer tumors, which were 50% smaller than those of WT mice. Nox1 ΔHep and Nox1 ΔHSC mice developed liver tumors of the same number and size as WT mice, whereas Nox1 ΔMac developed fewer and smaller tumors, similar to Nox1 –/– mice. After DEN injection, levels of tumor necrosis factor, interleukin 6 (IL6), and phosphorylated signal transducer and activator of transcription 3 were increased in livers from WT, but not Nox1 –/– or Nox1 ΔMac , mice. Conditioned medium from necrotic hepatocytes induced expression of NOX1 in cultured macrophages, followed by expression of tumor necrosis factor, IL6, and other inflammatory cytokines; this medium did not induce expression of IL6 or cytokines in Nox1 ΔMac macrophages. WT mice given DEN followed by ML171 developed fewer and smaller liver tumors than mice given DEN followed by vehicle. Conclusions: In mice given injections of a liver carcinogen (DEN), expression of NOX1 by macrophages promotes hepatic tumorigenesis by inducing the production of inflammatory cytokines. We propose that upon liver injury, damage-associated molecular patterns released from dying hepatocytes activate liver macrophages to produce cytokines that promote tumor development. Strategies to block NOX1 or these cytokines might be developed to slow hepatocellular carcinoma progression.
AB - Background & Aims: Although there are associations among oxidative stress, reduced nicotinamide adenine dinucleotide phosphate oxidase (NOX) activation, and hepatocellular carcinoma (HCC) development, it is not clear how NOX contributes to hepatocarcinogenesis. We studied the functions of different NOX proteins in mice after administration of a liver carcinogen. Methods: Fourteen-day-old Nox1 –/– mice, Nox4 –/– mice, Nox1 –/– Nox4 –/– (double-knockout) mice, and wild-type (WT) C57BL/6 mice were given a single intraperitoneal injection of diethylnitrosamine (DEN) and liver tumors were examined at 9 months. We also studied the effects of DEN in mice with disruption of Nox1 specifically in hepatocytes (Nox1 ΔHep ), hepatic stellate cells (Nox1 ΔHep ), or macrophages (Nox1 ΔMac ). Some mice were also given injections of the NOX1-specific inhibitor ML171. To study the acute effects of DEN, 8–12-week-old mice were given a single intraperitoneal injection, and liver and serum were collected at 72 hours. Liver tissues were analyzed by histologic examination, quantitative polymerase chain reaction, and immunoblots. Hepatocytes and macrophages were isolated from WT and knockout mice and analyzed by immunoblots. Results: Nox4 –/– mice and WT mice developed liver tumors within 9 months after administration of DEN, whereas Nox1 –/– mice developed 80% fewer tumors, which were 50% smaller than those of WT mice. Nox1 ΔHep and Nox1 ΔHSC mice developed liver tumors of the same number and size as WT mice, whereas Nox1 ΔMac developed fewer and smaller tumors, similar to Nox1 –/– mice. After DEN injection, levels of tumor necrosis factor, interleukin 6 (IL6), and phosphorylated signal transducer and activator of transcription 3 were increased in livers from WT, but not Nox1 –/– or Nox1 ΔMac , mice. Conditioned medium from necrotic hepatocytes induced expression of NOX1 in cultured macrophages, followed by expression of tumor necrosis factor, IL6, and other inflammatory cytokines; this medium did not induce expression of IL6 or cytokines in Nox1 ΔMac macrophages. WT mice given DEN followed by ML171 developed fewer and smaller liver tumors than mice given DEN followed by vehicle. Conclusions: In mice given injections of a liver carcinogen (DEN), expression of NOX1 by macrophages promotes hepatic tumorigenesis by inducing the production of inflammatory cytokines. We propose that upon liver injury, damage-associated molecular patterns released from dying hepatocytes activate liver macrophages to produce cytokines that promote tumor development. Strategies to block NOX1 or these cytokines might be developed to slow hepatocellular carcinoma progression.
KW - Hepatocellular carcinoma
KW - Inflammation
KW - Macrophage
KW - Reactive oxygen species
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U2 - 10.1053/j.gastro.2018.11.019
DO - 10.1053/j.gastro.2018.11.019
M3 - Article
C2 - 30445007
AN - SCOPUS:85062339861
SN - 0016-5085
VL - 156
SP - 1156-1172.e6
JO - Gastroenterology
JF - Gastroenterology
IS - 4
ER -