Vonoprazan as an adjuvant to cisplatin: enhancing antitumor efficacy while mitigating nephrotoxicity and gastrointestinal adverse effects

Figures

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  FIGURE 1. Influences of vonoprazan on cisplatin-mediated alterations in the percent change in bodyweight, kidney/bodyweight ratio, and serum kidney function biomarkers and LDH. The data are presented as the mean ± standard deviation (SD) (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan was orally administered at two dose levels (10 or 20 mg/kg) for 10 consecutive days. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. BUN, blood urea nitrogen; LDH, lactate dehydrogenase.

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  FIGURE 2. Influences of vonoprazan on cisplatin-mediated histopathological alterations in the kidney, stomach, duodenal, and jejunal tissues based on hematoxylin and eosin staining. Top: Representative photomicrographs of tissues from different treatment groups: (A) control, (B) cisplatin, (C) vonoprazan-only, (D) cisplatin + vonoprazan (10 mg/kg), and (E) cisplatin + vonoprazan (20 mg/kg) groups. Bottom row: scoring for kidney tubular necrosis, stomach lesions, duodenal lesions, and jejunal lesions. In the stained images, thin arrows indicate fibrosis + inflammation, while thick arrows indicate tubular swelling and necrosis; the arrowheads indicate hyaline degeneration or intraluminal desquamated epithelium (cellular cast), while the circular structures indicate tubular dilation. V: villi, cr: crypt, thin arrow: inflammatory aggregation, line: proliferative crypt, thick arrow: glandular change as dilation, necrosis, or abscess. Image magnification = ×100, inset = ×400. Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan was administered at two dose levels (10 or 20 mg/kg) orally for 10 d. The data are expressed as the median ± interquartile…

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  FIGURE 3. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in the oxidant/antioxidant balance in the kidney, stomach, and intestinal tissues. The data are presented as the mean ± SD (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan (20 mg/kg) was given orally for 10 d. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. MDA, malondialdehyde; TAC, total antioxidant capacity.

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  FIGURE 4. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in the inflammasome component (NLRP3) and inflammatory signaling mediator (NF-κB). The data are presented as the mean ± SD (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan (20 mg/kg) was given orally for 10 d. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. NF-κB, nuclear factor kappa B subunit; NLRP3, NOD-, LRR-, and pyrin domain-containing protein 3.

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  FIGURE 5. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in kidney, stomach, and intestinal expression levels of IL-6 using immunohistochemical assay. Kidney sections: (A) control group showing negative IL-6 immunostaining of podocytes or renal tubules; (B) cisplatin group showing strong IL-6 immunostaining (thin arrows) of podocytes in the glomeruli and tubular cells (thick arrows); (C) cisplatin + vonoprazan (20 mg/kg) group showing weak IL-6 immunostaining (thick arrow). Image magnification = ×400. Stomach sections: (A) control group showing negative IL-6 immunostaining; (B) cisplatin group showing strong IL-6 immunostaining (thick arrows) in the fundic glands; (C) cisplatin + vonoprazan (20 mg/kg) group showing weak IL-6 immunostaining (thick arrow) in the fundic glands. Image magnification = ×400. Intestinal sections: (A) control group showing negative IL-6 immunostaining; (B) cisplatin group showing strong IL-6 immunostaining in the cells of lamina propria of the intestinal villi (thick arrows); (C) cisplatin + vonoprazan (20 mg/kg) group showing weak IL-6 immunostaining (thick arrow) in the cells of the lamina propria of the intestinal villi. Image…

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  FIGURE 6. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in the autophagy-lysosomal pathway. The data are presented as the mean ± SD (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan (20 mg/kg) was given orally for 10 d. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. LC3B, microtubule-associated protein 1A/1B-light chain 3 beta; Beclin-1, coiled-coil moesin-like BCL2-interacting protein.

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  FIGURE 7. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in the kidney, stomach, and intestinal expressions of SQSTM1 (p62) based on the immunohistochemical assay. Kidney sections: (A) control group showing negative p62 immunostaining in the tubular cells or podocytes; (B) cisplatin group showing strong p62 immunostaining (thin arrows) of the podocytes in the glomeruli and tubular cells (thick arrows); (C) cisplatin + vonoprazan (20 mg/kg) group showing weak p62 immunostaining (thick arrow). Image magnification = ×400. Stomach sections: (A) control group showing negative p62 immunostaining; (B) cisplatin group showing strong p62 immunostaining (thick arrows) in the fundic glands; (C) cisplatin + vonoprazan (20 mg/kg) group showing weak p62 immunostaining (thick arrow) in the fundic glands. Image magnification = ×400. Intestinal sections: (A) control group showing negative p62 immunostaining; (B) cisplatin group showing strong p62 immunostaining in the epithelial cells (thin arrows) and in cells of the lamina propria of the intestinal villi (thick arrows); (C) cisplatin + vonoprazan (20 mg/kg) group showing weak p62 immunostaining (thick arrow) in cells of the lamina…

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  FIGURE 8. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in the early biomarkers of acute kidney injury (KIM-1 and NGAL) in the kidney tissue. The data are presented as the mean ± SD (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan (20 mg/kg) was given orally for 10 d. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. KIM-1: kidney injury molecule-1; NGAL: neutrophil gelatinase-associated lipocalin.

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  FIGURE 9. Influences of vonoprazan (20 mg/kg) on cisplatin-mediated alterations in gastrointestinal protective mediators as well as intestinal epithelial and mucosal injury markers. The data are presented as the mean ± SD (n = 7). Cisplatin was administered as a single dose (8 mg/kg, i.p.) on the seventh day, while vonoprazan (20 mg/kg) was given orally for 10 d. Statistical evaluations were carried out using one-way ANOVA, followed by the Tukey–Kramer post hoc test; # denotes a significant difference over the control group, ## indicates a highly significant difference over the control group, * represents a significant difference over the cisplatin group, and ** indicates a highly significant difference over the cisplatin group. cGMP, cyclic guanosine monophosphate; PGI2, prostacyclin; 5-HT, serotonin; IFABP, intestinal fatty-acid-binding protein; TFF3, trefoil factor 3.

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  FIGURE 10. Influences of cisplatin and vonoprazan on human breast cancer (MCF-7) cells after 48 h of exposure; the cytotoxicity was evaluated using the MTT assay in terms of percentage of cell viability by administering increasing concentrations of cisplatin (1, 10, 15, 20, and 30 µM) and vonoprazan (1, 3, 10, 30, and 100 µM). The data are expressed as the mean ± SD. Statistical analyses were performed using one-way ANOVA, followed by Dunnett’s post hoc test; # indicates a significant difference over the control group. IC50, half of the maximal inhibitory concentration; cis, cisplatin; Vono, vonoprazan.

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  FIGURE 11. Consequences of combining vonoprazan and cisplatin on the apoptotic effects of cisplatin in MCF-7 cells. The MCF-7 cells were exposed to cisplatin (Cis; 11 µM) and vonoprazan (Vono; 16 µM) individually or in combination for 48 h. The data are expressed as mean ± SD. Statistical analyses were performed using one-way ANOVA followed by the Tukey–Kramer post hoc test; the symbols #, *, and $ indicate significant differences over the control (0 µM Cis + 0 µM Vono), vonoprazan-only treatment (0 µM Cis + 16 µM Vono), and cisplatin-only treatment (11 µM Cis + 0 µM Vono) groups, respectively. BCL2, B-cell lymphoma 2; p53, tumor protein p53; BAX, BCL2-associated X protein.