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Strain- and sex-dependent circadian changes in abcc2 transporter expression: implications for irinotecan chronotolerance in mouse ileum.

Okyar A, Piccolo E, Ahowesso C, Filipski E, Hossard V, Guettier C, La Sorda R, Tinari N, Iacobelli S, Lévi F - PLoS ONE (2011)

Bottom Line: Irinotecan was also best tolerated in males (p = 0.05) and in B6CBAF(1) (p = 0.0006).Strain- and sex-dependent circadian patterns in abcc2 expressions displayed robust relations with the chronotolerance of ileum mucosa for irinotecan.This finding has strong potential implications for improving the intestinal tolerability of anticancer drugs through circadian delivery.

View Article: PubMed Central - PubMed

Affiliation: INSERM, U776 Rythmes Biologiques et Cancers, Hôpital Paul Brousse, Villejuif, France.

ABSTRACT

Background: ATP-binding cassette transporter abcc2 is involved in the cellular efflux of irinotecan. The drug is toxic for mouse ileum, where abcc2 is highly expressed. Here, we investigate whether circadian changes in local abcc2 expression participate in the circadian rhythm of irinotecan toxicity for ileum mucosa, and further assess whether genetic background or sex modify this relation.

Methodology/principal findings: Ileum mucosa was obtained every 3-4 h for 24 h in male and female B6D2F(1) and B6CBAF(1) mice synchronized with light from Zeitgeber Time (ZT)0 to ZT12 alternating with 12 h of darkness. Irinotecan (50 mg/kg i.v. daily for 4 days) was administered at the sex- and strain-specific times corresponding to least (ZT11-15) or largest drug-induced body weight loss (ZT23-03-07). Abcc2 expression was determined with qRT-PCR for mRNA and with immunohistochemistry and confocal microscopy for protein. Histopathologic lesions were graded in ileum tissues obtained 2, 4 or 6 days after treatment. Two- to six-fold circadian changes were demonstrated for mRNA and protein mean expressions of abcc2 in mouse ileum (p<0.05). ZT12 corresponded to high mRNA and protein expressions, with circadian waveforms differing according to genetic background and sex. The proportion of mice spared from ileum lesions varied three-fold according to irinotecan timing, with best tolerability at ZT11-15 (p = 0.00003). Irinotecan was also best tolerated in males (p = 0.05) and in B6CBAF(1) (p = 0.0006).

Conclusions/significance: Strain- and sex-dependent circadian patterns in abcc2 expressions displayed robust relations with the chronotolerance of ileum mucosa for irinotecan. This finding has strong potential implications for improving the intestinal tolerability of anticancer drugs through circadian delivery.

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Related in: MedlinePlus

Tolerance of mouse small intestine for irinotecan, as a function of circadian timing, genetic background and sex.According to previous studies, the circadian timing (as ZT) corresponding to best tolerability of irinotecan was ZT11 in male B6D2F1 and ZT15 in female B6D2F1 as well as in male or female B6CBAF1 mice. The worst tolerability resulted from irinotecan dosing at ZT23 in male B6D2F1, ZT3 in female B6D2F1, and ZT7 in male or female B6CBAF1. Transverse sections of ileum 2 days after the fourth daily dose of irinotecan (50 mg/kg/d×4 days). Hemalun-erythrosine-safran staining. A, control, Grade 0; B, Grade 1; C, Grade 2. Bar graph of the proportion (±SEM) of mice free of ileum lesions (Grade 0) according to circadian timing of irinotecan (χ2, p = 0.00003). Bar graph of the proportion (± SEM) of toxicity grade according to circadian timing of irinotecan (ANOVA, p = 0.016). (D) Histogram of mean (± SEM) grade of ileum toxicity according to circadian timing, genetic background.and sex. The incidence of mice without any ileum damage was significantly higher in male as compared to female mice, irrespective of strain (χ2, p = 0.05), and in B6CBAF1 as compared to B6D2F1 (χ2, p = 0.0006). Three-way ANOVA validated the relevance of “best” vs. “worst” circadian timing (p = 0.018).
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pone-0020393-g005: Tolerance of mouse small intestine for irinotecan, as a function of circadian timing, genetic background and sex.According to previous studies, the circadian timing (as ZT) corresponding to best tolerability of irinotecan was ZT11 in male B6D2F1 and ZT15 in female B6D2F1 as well as in male or female B6CBAF1 mice. The worst tolerability resulted from irinotecan dosing at ZT23 in male B6D2F1, ZT3 in female B6D2F1, and ZT7 in male or female B6CBAF1. Transverse sections of ileum 2 days after the fourth daily dose of irinotecan (50 mg/kg/d×4 days). Hemalun-erythrosine-safran staining. A, control, Grade 0; B, Grade 1; C, Grade 2. Bar graph of the proportion (±SEM) of mice free of ileum lesions (Grade 0) according to circadian timing of irinotecan (χ2, p = 0.00003). Bar graph of the proportion (± SEM) of toxicity grade according to circadian timing of irinotecan (ANOVA, p = 0.016). (D) Histogram of mean (± SEM) grade of ileum toxicity according to circadian timing, genetic background.and sex. The incidence of mice without any ileum damage was significantly higher in male as compared to female mice, irrespective of strain (χ2, p = 0.05), and in B6CBAF1 as compared to B6D2F1 (χ2, p = 0.0006). Three-way ANOVA validated the relevance of “best” vs. “worst” circadian timing (p = 0.018).

Mentions: Histologic lesions in ileum were found in mice from each strain and sex, with scores up to two (Figure 5A). Ileum mucosa was mostly damaged two days after treatment completion, with full recovery within four days in all groups. Irinotecan timing determined the incidence of mice free of ileum lesions (grade 0) as well as the average lesion score. Thus no ileum damage was observed in 46.4% of the mice receiving irinotecan at the ZT predicted to achieve best tolerability as compared to 11.5% of those treated at the ZT predicted to yield poorest tolerability (13/28 vs. 3/26; p fromχ2 = 0.00003) (Figure 5B). The respective mean scores were 0.64±0.11 and 1.00±0.01 for the respective “best” and “worst” times (p from ANOVA = 0.016) (Figure 5C).


Strain- and sex-dependent circadian changes in abcc2 transporter expression: implications for irinotecan chronotolerance in mouse ileum.

Okyar A, Piccolo E, Ahowesso C, Filipski E, Hossard V, Guettier C, La Sorda R, Tinari N, Iacobelli S, Lévi F - PLoS ONE (2011)

Tolerance of mouse small intestine for irinotecan, as a function of circadian timing, genetic background and sex.According to previous studies, the circadian timing (as ZT) corresponding to best tolerability of irinotecan was ZT11 in male B6D2F1 and ZT15 in female B6D2F1 as well as in male or female B6CBAF1 mice. The worst tolerability resulted from irinotecan dosing at ZT23 in male B6D2F1, ZT3 in female B6D2F1, and ZT7 in male or female B6CBAF1. Transverse sections of ileum 2 days after the fourth daily dose of irinotecan (50 mg/kg/d×4 days). Hemalun-erythrosine-safran staining. A, control, Grade 0; B, Grade 1; C, Grade 2. Bar graph of the proportion (±SEM) of mice free of ileum lesions (Grade 0) according to circadian timing of irinotecan (χ2, p = 0.00003). Bar graph of the proportion (± SEM) of toxicity grade according to circadian timing of irinotecan (ANOVA, p = 0.016). (D) Histogram of mean (± SEM) grade of ileum toxicity according to circadian timing, genetic background.and sex. The incidence of mice without any ileum damage was significantly higher in male as compared to female mice, irrespective of strain (χ2, p = 0.05), and in B6CBAF1 as compared to B6D2F1 (χ2, p = 0.0006). Three-way ANOVA validated the relevance of “best” vs. “worst” circadian timing (p = 0.018).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3108586&req=5

pone-0020393-g005: Tolerance of mouse small intestine for irinotecan, as a function of circadian timing, genetic background and sex.According to previous studies, the circadian timing (as ZT) corresponding to best tolerability of irinotecan was ZT11 in male B6D2F1 and ZT15 in female B6D2F1 as well as in male or female B6CBAF1 mice. The worst tolerability resulted from irinotecan dosing at ZT23 in male B6D2F1, ZT3 in female B6D2F1, and ZT7 in male or female B6CBAF1. Transverse sections of ileum 2 days after the fourth daily dose of irinotecan (50 mg/kg/d×4 days). Hemalun-erythrosine-safran staining. A, control, Grade 0; B, Grade 1; C, Grade 2. Bar graph of the proportion (±SEM) of mice free of ileum lesions (Grade 0) according to circadian timing of irinotecan (χ2, p = 0.00003). Bar graph of the proportion (± SEM) of toxicity grade according to circadian timing of irinotecan (ANOVA, p = 0.016). (D) Histogram of mean (± SEM) grade of ileum toxicity according to circadian timing, genetic background.and sex. The incidence of mice without any ileum damage was significantly higher in male as compared to female mice, irrespective of strain (χ2, p = 0.05), and in B6CBAF1 as compared to B6D2F1 (χ2, p = 0.0006). Three-way ANOVA validated the relevance of “best” vs. “worst” circadian timing (p = 0.018).
Mentions: Histologic lesions in ileum were found in mice from each strain and sex, with scores up to two (Figure 5A). Ileum mucosa was mostly damaged two days after treatment completion, with full recovery within four days in all groups. Irinotecan timing determined the incidence of mice free of ileum lesions (grade 0) as well as the average lesion score. Thus no ileum damage was observed in 46.4% of the mice receiving irinotecan at the ZT predicted to achieve best tolerability as compared to 11.5% of those treated at the ZT predicted to yield poorest tolerability (13/28 vs. 3/26; p fromχ2 = 0.00003) (Figure 5B). The respective mean scores were 0.64±0.11 and 1.00±0.01 for the respective “best” and “worst” times (p from ANOVA = 0.016) (Figure 5C).

Bottom Line: Irinotecan was also best tolerated in males (p = 0.05) and in B6CBAF(1) (p = 0.0006).Strain- and sex-dependent circadian patterns in abcc2 expressions displayed robust relations with the chronotolerance of ileum mucosa for irinotecan.This finding has strong potential implications for improving the intestinal tolerability of anticancer drugs through circadian delivery.

View Article: PubMed Central - PubMed

Affiliation: INSERM, U776 Rythmes Biologiques et Cancers, Hôpital Paul Brousse, Villejuif, France.

ABSTRACT

Background: ATP-binding cassette transporter abcc2 is involved in the cellular efflux of irinotecan. The drug is toxic for mouse ileum, where abcc2 is highly expressed. Here, we investigate whether circadian changes in local abcc2 expression participate in the circadian rhythm of irinotecan toxicity for ileum mucosa, and further assess whether genetic background or sex modify this relation.

Methodology/principal findings: Ileum mucosa was obtained every 3-4 h for 24 h in male and female B6D2F(1) and B6CBAF(1) mice synchronized with light from Zeitgeber Time (ZT)0 to ZT12 alternating with 12 h of darkness. Irinotecan (50 mg/kg i.v. daily for 4 days) was administered at the sex- and strain-specific times corresponding to least (ZT11-15) or largest drug-induced body weight loss (ZT23-03-07). Abcc2 expression was determined with qRT-PCR for mRNA and with immunohistochemistry and confocal microscopy for protein. Histopathologic lesions were graded in ileum tissues obtained 2, 4 or 6 days after treatment. Two- to six-fold circadian changes were demonstrated for mRNA and protein mean expressions of abcc2 in mouse ileum (p<0.05). ZT12 corresponded to high mRNA and protein expressions, with circadian waveforms differing according to genetic background and sex. The proportion of mice spared from ileum lesions varied three-fold according to irinotecan timing, with best tolerability at ZT11-15 (p = 0.00003). Irinotecan was also best tolerated in males (p = 0.05) and in B6CBAF(1) (p = 0.0006).

Conclusions/significance: Strain- and sex-dependent circadian patterns in abcc2 expressions displayed robust relations with the chronotolerance of ileum mucosa for irinotecan. This finding has strong potential implications for improving the intestinal tolerability of anticancer drugs through circadian delivery.

Show MeSH
Related in: MedlinePlus