Mouse models of anemia of cancer

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Abstract

Anemia of cancer (AC) may contribute to cancer-related fatigue and impair quality of life. Improved understanding of the pathogenesis of AC could facilitate better treatment, but animal models to study AC are lacking. We characterized four syngeneic C57BL/6 mouse cancers that cause AC. Mice with two different rapidly-growing metastatic lung cancers developed the characteristic findings of anemia of inflammation (AI), with dramatically different degrees of anemia. Mice with rapidly-growing metastatic melanoma also developed a severe anemia by 14 days, with hematologic and inflammatory parameters similar to AI. Mice with a slow-growing peritoneal ovarian cancer developed an iron-deficiency anemia, likely secondary to chronically impaired nutrition and bleeding into the peritoneal cavity. Of the four models, hepcidin mRNA levels were increased only in the milder lung cancer model. Unlike in our model of systemic inflammation induced by heat-killed Brucella abortus, ablation of hepcidin in the ovarian cancer and the milder lung cancer mouse models did not affect the severity of anemia. Hepcidin-independent mechanisms play an important role in these murine models of AC. © 2014 Kim et al.

Figures

  • Figure 1. Mice with metastatic lung cancer develop anemia with reticulocytosis. C57BL/6 mice were injected intraperitoneally with 0.16106–0.56106 murine TC-1 or Lewis lung carcinoma (LLC) cells and euthanized after 14 days. Compared to controls, the tumor-bearing mice have: (A) decreased hemoglobin; (B) increased reticulocytosis. N = 8–19 mice per treatment and control group. *P,0.05, **P,0.001; P by Mann-Whitney rank sum test. Bars and error bars are median 675th/25th percentile. doi:10.1371/journal.pone.0093283.g001
  • Figure 2. Mice with metastatic lung cancer have iron-restricted erythropoiesis with hypoferremia and increased tissue iron stores.
  • Figure 3. TC-1 and LLC tumor-bearing mice show evidence of systemic inflammation but only TC-1 has increased hepcidin mRNA.
  • Figure 4. The melanoma-bearing mice develop anemia with iron-restricted erythropoiesis and inflammation. C57BL/6 mice were injected intraperitoneally with 0.16106–0.36106 B16-F10 cells, and euthanized after 14 days. The melanoma-bearing mice develop: (A) a significant anemia; (B) increased reticulocytosis compared to controls; (C) elevated zinc protoporphyrin (ZPP) levels compared to controls, indicating iron-restricted heme synthesis; (D) increased levels of liver SAA-1 mRNA, a marker of inflammation and IL-6 activity; (E) lower levels of hepcidin mRNA. N = 8–11 mice per treatment and control group. *P,0.05, **P,0.001; P by Mann-Whitney rank sum test. Bars and error bars are median 675th/25th percentile. doi:10.1371/journal.pone.0093283.g004
  • Figure 5. ID8-bearing mice develop iron deficiency anemia. C57BL/6 mice were injected intraperitoneally with 16106 ID8 cells and euthanized after 18.5 weeks. Compared to controls, ID8-bearing mice: (A) are more anemic; (B) have elevated ZPP levels; (C,D) have lower liver and spleen iron levels. N = 10–16 mice per treatment and control group. *P,0.05, **P,0.001; P by Mann-Whitney rank-sum test (A,B) or student t-test (C,D). Bars and error bars are median 6 median 675th/25th percentile (A,B) or mean 6 SD (C,D). doi:10.1371/journal.pone.0093283.g005
  • Figure 6. ID8-bearing mice have decreased liver hepcidin mRNA responsive to decreased iron stores. At 18.5 weeks, ID8-bearing mice compared to controls have (A) suppressed hepcidin mRNA; (B) significant positive correlation between liver iron stores and hepcidin mRNA expression. N = 11–16 mice per treatment and control group. *P,0.05; P Mann-Whitney rank sum test in (A), and by Pearson Correlation in (B). Bars and error bars are median 675th/25th percentile. doi:10.1371/journal.pone.0093283.g006
  • Figure 7. Peripheral blood smears from wild-type tumor and control mice show no significant schistocytosis. (A) PBS control (B) TC-1 (C) LLC (D) ID8. doi:10.1371/journal.pone.0093283.g007
  • Figure 8. Tumor-induced hemoglobin decrease in TC-1 or ID8 -bearing mice is not affected by hepcidin ablation. (A) By day 14, there is no significant difference between the hemoglobin changes in TC-1-bearing wild-type and hepcidin knockout mice (P = 0.908). (B) Absolute hemoglobin levels of mice shown in A. (C) By 18.5 weeks, there is no significant difference between the hemoglobin changes of ID8-bearing wildtype and hepcidin knockout mice (P = 0.739). (D) Absolute hemoglobin levels of mice shown in C. N = 4–19 mice per treatment and control group. P by t-test. Bars and error bars are mean 6 SD. doi:10.1371/journal.pone.0093283.g008

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APA

Kim, A., Rivera, S., Shprung, D., Limbrick, D., Gabayan, V., Nemeth, E., & Ganz, T. (2014). Mouse models of anemia of cancer. PLoS ONE, 9(3). https://doi.org/10.1371/journal.pone.0093283

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