Influence of serum inflammatory cytokines on cytochrome P450 drug metabolising activity during breast cancer chemotherapy: a patient feasibility study

Abstract

Individual response to chemotherapy in patients with breast cancer is variable. Obesity and exercise are associated with better and worse outcomes, respectively, and it is known that both impact the systemic cytokine milieu. Cytochrome P450 (CYP) enzymes are responsible for the metabolism of many chemotherapy agents, and CYP enzyme activity has been shown to be modified by inflammatory cytokines in vitro and in vivo. Cytokine-associated changes in CYP metabolism may alter chemotherapy exposure, potentially affecting treatment response and patient survival. Therefore, better understanding of these biological relationships is required. This exploratory single arm open label trial investigated changes in in vivo CYP activity in twelve women treated for stage II or III breast cancer, and demonstrated for the first time the feasibility and safety of utilising the Inje phenotyping cocktail to measure CYP activity in cancer patients receiving chemotherapy. Relative CYP activity varied between participants, particularly for CYP2C9 and CYP2D6, and changes in serum concentrations of the inflammatory cytokine monocyte chemoattractant protein 1 inversely correlated to CYP3A4 activity during chemotherapy. Future use of phenotyping cocktails in a clinical oncology setting may help guide drug dosing and improve chemotherapy outcomes.Clinical Trial Registration: Trial was retrospectively registered to the Australia New Zealand Clinical Trial Registry (ANZCTR). ACTRN12620000832976, 21 Aug 2020, https://www.anzctr.org.au/ACTRN12620000832976.aspx .

Figure 1

Changes in CYP metabolising ratios during chemotherapy for breast cancer. The change in CYP2C9 (n = 8), CYP2C19 (n = 9), CYP2D6 (n = 8) and CYP3A4 (n = 9) metabolising activity was determined by comparing the probe drug to metabolite ratios from after chemotherapy to before chemotherapy (log10). Horizontal black solid lines represent median values. Horizontal black dotted line represents no change in CYP metabolising activity from baseline to paclitaxel dose six, and points above or below the red dotted horizontal lines represent a decrease or increase, respectively, in CYP metabolising activity of 1.25-fold or greater (log10 of 0.80–1.25 =  ± 0.097) from before chemotherapy to after chemotherapy. Statistical analysis was performed using Wilcoxon matched-pairs signed rank testing, and significance was determined as p < 0.05.

Figure 2

Change in circulating inflammatory cytokines measured during chemotherapy. Inflammatory cytokines ANG2, BAFF, CRP, GDF-15, IL-10, and MCP-1 were measured in participant serum using enzyme-linked immunosorbent assays before chemotherapy (n = 10), and after chemotherapy (after paclitaxel dose six; n = 10). Black horizontal solid lines represent median values. The black horizontal dotted line represents no difference in cytokine concentration from baseline to paclitaxel dose six, and points above or below the dotted line represent an increase or decrease in cytokine concentration during chemotherapy, respectively. Statistical analysis was performed using the Wilcoxon matched-pairs signed rank test. *p < 0.05; **p < 0.01.

Figure 3

Correlation between changes in serum MCP-1 and changes in CYP3A4 metabolising activity during chemotherapy. The change in MCP-1 cytokine concentrations from before to after chemotherapy (log10), was correlated with the change in CYP3A4 metabolising activity from before to after chemotherapy (log10; n = 9). Black solid lines represent linear regression line of best fit. Horizontal black dotted lines represent no change in CYP3A4 metabolising activity from baseline to paclitaxel dose six, and points above or below this represent a decrease or increase in CYP3A4 metabolising activity, respectively. Statistical analysis was performed using Spearman correlation analysis, and significance was determined as p ≤ 0.05.

Figure 4

Monocyte chemoattractant protein 1 (MCP-1) mediated decrease in CYP3A4 activity in the liver of human breast cancer patients; proposed hypothesis. (A) The liver of breast cancer patients is exposed to increased circulating levels of MCP-1 during chemotherapy. Liver kupffer cells (hepatic macrophages) express the MCP-1 cell surface receptor C–C chemokine receptor type 2 (CCR2), and thus, MCP-1 can bind and induce an increase in the production of other inflammatory cytokines, such as interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-α), and further increase levels of MCP-1 molecules. IL-6 and TNF-α bind their membrane receptors, interleukin 6 receptor (IL-6R) and tumour necrosis factor receptor 1 (TNFR1), on the surface of nearby hepatocytes, inducing inflammatory signalling cascades that regulate CYP3A4 transcription. (B) Schematic of one of the mechanisms by which inflammatory cytokines inhibit CYP3A4 transcription; as reported by Jover et al., Intracellular signalling, following IL-6 binding, induces translation of CCAAT-enhancer-binding protein beta isoform LIP (C/EBPβ-LIP), an antagonist of CCAAT-enhancer-binding protein alpha (C/EBPα); C/EBPα is a known transcription factor that constitutively promotes CYP3A4 expression in hepatocytes.