Epidermal growth factor receptor tyrosine kinase inhibitor (EGFR–TKI), including osimertinib, have been reported to cause nephrotoxicity. However, distinguishing true renal impairment from pseudo-renal dysfunction due to transporter inhibition (pseudo-AKI) remains challenging, particularly in patients with chronic kidney disease (CKD). We report a case of a 76-year-old man with pre-existing CKD stage G3bA3 who developed a rapid increase in serum creatinine levels 8 days after initiating osimertinib for EGFR L858R mutation-positive non-small cell lung cancer (NSCLC). Anticancer therapy was subsequently modified by switching to dose-adjusted afatinib, allowing for continued EGFR–TKIs treatment without further renal deterioration. Renal function remained stable for more than 2 years under joint oncological and nephrological management. This case highlights the clinical utility of an integrated renal assessment using cystatin C and tubular injury markers in differentiating true renal injury from pseudo-AKI. This approach supports the clinical decision to switch to an alternative EGFR–TKI in patients with CKD, thereby enabling the safe continuation of targeted cancer therapy.

Abstract Lorlatinib, a third-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor, is widely used to treat ALK-positive nonsmall cell lung cancer. However, lorlatinib has occasionally been associated with severe liver injury. Immune-mediated adverse events are frequently provoked by certain pharmacological agents or their reactive metabolites. However, the specific contribution of immune pathways to hepatotoxicity caused by lorlatinib is yet to be fully elucidated. In the present study, the potential for direct inflammasome stimulation by lorlatinib was examined in differentiated THP-1 cells. This was accompanied by an evaluation of inflammasome-activating factors present in the culture supernatants of FLC-4 cells treated with lorlatinib. Our results showed that lorlatinib did not directly activate inflammasomes in differentiated THP-1 cells; however, the supernatants from FLC-4 cells activated inflammasomes in differentiated THP-1 cells following treatment with lorlatinib. Detection of pyrido-pyrazole-containing metabolites in FLC-4 cells suggests that these reactive metabolites activate inflammasomes. Exposure of FLC-4 cells to lorlatinib resulted in elevated extracellular concentrations of heat shock protein 90 (HSP90), a damage-associated molecular pattern (DAMP). These findings suggest that the reactive metabolites of lorlatinib promote hepatocellular release of damage-associated molecular patterns, which in turn activate inflammasomes. Thus, inflammasome activation may be an important step in the activation of the immune system by lorlatinib and may cause immune-related adverse events in some patients.

Vildagliptin has been reported to cause liver injury, with clinical findings suggesting the involvement of the immune response. However, the underlying mechanism remains unclear. Vildagliptin possesses a covalent-binding group that may induce an immune response via inflammasome activation. In this study, we examined whether covalent binding of vildagliptin to proteins in differentiated THP-1 or FLC-4 cells leads to inflammasome activation either directly or via damage-associated molecular patterns (DAMPs). We also performed biochemical and histopathological assessments of liver injury in PD-1-/- mice treated with anti-CTLA-4 antibody and vildagliptin (0.13%). Vildagliptin didn't directly induce IL-1β production and Caspase-1 activity in differentiated THP-1 cells. In contrast, the culture medium of FLC-4 cells incubated with vildagliptin exhibited increased levels. The levels of heat shock protein 40 (HSP40), a DAMP that triggers inflammasome activation, were significantly increased in the culture supernatant. In addition, adducts with a trapping agent were detected in FLC-4 cells, suggesting that covalent binding of vildagliptin induces the release of DAMPs. In mice with impaired immune tolerance due to immune checkpoint blockade, serum AST and ALT levels were significantly elevated 4 weeks after treatment with vildagliptin, and marked granulomatous inflammation was observed in the liver tissues. These results indicate that vildagliptin-induced liver injury occurs via a mechanism whereby the covalent binding of vildagliptin induces the release of HSP40 from hepatocytes, in turn activating inflammasomes. We further demonstrated that the risk of vildagliptin-induced liver injury may be increased by impaired immune tolerance, such as that caused by the co-administration of immune checkpoint inhibitors.

Anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitors (TKI), including brigatinib, are widely used to treat ALK-positive non-small cell lung cancer. However, severe adverse effects associated with brigatinib, such as interstitial pneumonia and liver dysfunction, may involve immune system activation. The precise mechanisms underlying these immune-related adverse effects remain unclear. In this study, we evaluated the direct activation of inflammasomes by brigatinib and other ALK TKI (crizotinib, alectinib, ceritinib) in differentiated THP-1 cells. Additionally, we analyzed the inflammasome-activating potential of supernatants from functional liver cell (FLC)-4 cells treated with these drugs. Our results demonstrate that brigatinib directly activates inflammasomes in THP-1 cells, inducing the production of interleukin-1β and the activation of caspase-1. In contrast, no inflammasome activation was observed with the other ALK TKIs. Furthermore, supernatants from FLC-4 cells, characterized by high drug-metabolizing activity, were shown to activate inflammasomes in differentiated THP-1 cells following treatment with brigatinib. Brigatinib treatment significantly increased the levels of damage-associated molecular patterns (DAMPs), including heat shock protein 90 and S100A6, in the supernatants of FLC-4 cells. These findings suggest that brigatinib induces the release of DAMPs from hepatocytes, which subsequently activate inflammasomes. This mechanism may be essential for brigatinib-induced immune system activation and the development of immune-related adverse events.

Pulmonary arterial hypertension (PAH) is a progressive condition that frequently leads to right ventricular (RV) remodeling. Aldosterone promotes vascular and RV remodeling. The upregulation of steroidogenic acute regulatory protein (StAR) stimulates aldosterone synthesis. However, the expression of StAR in the myocardium under PAH conditions remains unknown. To investigate the expression of StAR in the myocardium and its association with RV remodeling in PAH, utilizing spironolactone as a treatment. A PAH model was created using male Sprague-Dawley rats, which received a subcutaneous injection of Sugen5416 (20 mg/kg) and were exposed to hypoxia (10% O2) for 2 weeks, followed by 2 weeks of normoxia. The animals were then divided into two groups, with one group receiving spironolactone (25 mg/kg/day) for an additional 4 weeks, while the other group did not. H9c2 cells were cultured under hypoxic conditions (37 °C, 1% O2, 5% CO2) with or without spironolactone treatment. In the model rats, RV systolic pressure and the Fulton index, both of which increased upon exposure to Sugen5416 and hypoxia, significantly decreased with spironolactone treatment. In H9c2 cells, hypoxic exposure elevated aldosterone levels, while spironolactone treatment significantly suppressed aldosterone production. Suppression of StAR expression in the myocardium via spironolactone contributes to the improvement of RV remodeling in PAH. Spironolactone may offer a valuable therapeutic strategy for RV remodeling in patients with PAH.

Trovafloxacin is a quinolone antibiotic drug with broad-spectrum activity, which was withdrawn from a global market relatively soon after approval because of serious liver injury. The characteristics of trovafloxacin-induced liver injury are consistent with an idiosyncratic reaction; however, the details of the mechanism have not been elucidated. We examined whether trovafloxacin induces the release of damage-associated molecular patterns (DAMPs) that activate inflammasomes. We also tested ciprofloxacin, levofloxacin, gatifloxacin, and grepafloxacin for their ability to activate inflammasomes. Drug bioactivation was performed with human hepatocarcinoma functional liver cell-4 (FLC-4) cells, and THP-1 cells (human monocyte cell line) were used for the detection of inflammasome activation. The supernatant from the incubation of trovafloxacin with FLC-4 cells for 7 days increased caspase-1 activity and production of IL-1ß by THP-1 cells. In the supernatant of FLC-4 cells that had been incubated with trovafloxacin, heat shock protein (HSP) 40 was significantly increased. Addition of a cytochrome P450 inhibitor to the FLC-4 cells prevented the release of HSP40 from the FLC-4 cells and inflammasome activation in THP-1 cells by the FLC-4 supernatant. These results suggest that reactive metabolites of trovafloxacin can cause the release of DAMPs from hepatocytes that can activate inflammasomes. Inflammasome activation may be an important step in the activation of the immune system by trovafloxacin, which, in some patients, can cause immune-related liver injury.

Although the mechanism underlying flutamide- or bicalutamide-induced liver injury may be immune related, the details remain unclear. If this mechanism is immune related, steroid use may be considered as a treatment option. Disproportionality analysis was conducted to evaluate the effect of concomitant steroid use on flutamide- and bicalutamide-induced liver injury. Male patients aged 20 years or older who were receiving nonsteroidal anti-androgens from April 2004 to October 2023 were screened from the Japanese Adverse Drug Event Report database. Data on liver injury, age, weight, height, steroid use, obesity, hepatic stenosis, alcohol-related hepatic disorders, hepatitis B and C, and common drugs known to cause drug-induced liver injury were analyzed. Liver injury was defined by the Standardized Medical Dictionary for Regulatory Activities query index (code 20000006, version 27.0). Among 142,430 patients, 2,316 were administered nonsteroidal anti-androgens. Reports of liver injury were disproportionate depending on the agents used (reporting odds ratio [ROR], 1.29; 95

INTRODUCTION:Kamebakaurin is an active constituent of both Rabdosia japonica and Rabdosia excisa, which are utilized in Chinese traditional medicine for improving symptoms in patients with allergies. We investigated the molecular mechanisms of the anti-allergic effects of kamebakaurin using BMMCs. METHODS:The degranulation ratio, histamine release, and the interleukin (IL)-4, leukotriene B4 (LTB4), and cysteinyl leukotriene productions on antigen-triggered BMMC were investigated. Additionally, the effects of kamebakaurin on signal transduction proteins were examined by Western blot and binding to the Syk and Lyn kinase domain was calculated. The effects of kamebakaurin on antigen-induced hyperpermeability were investigated using mouse model. RESULTS:At 10 μm, kamebakaurin partially inhibited degranulation, histamine release, and IL-4 production. At 30 μm, kamebakaurin partially reduced LTB4 and cysteinyl leukotriene productions and suppressed degranulation, histamine release, and IL-4 production. Phosphorylation of both Syk Y519/520 and its downstream protein, Gab2, was reduced by kamebakaurin, and complete inhibition was observed with 30 μm kamebakaurin. In contrast, phosphorylation of Erk was only partially inhibited, even in the presence of 30 μm kamebakaurin. Syk Y519/520 is known to be auto-phosphorylated via intramolecular ATP present in its own ATP-binding site, and this auto-phosphorylation triggers degranulation, histamine release, and IL-4 production. Docking simulation study indicated kamebakaurin blocked ATP binding to the ATP-binding site in Syk. Therefore, inhibition of Syk auto-phosphorylation by kamebakaurin binding to the Syk ATP-binding site appeared to cause a reduction of histamine release and IL-4 production. Kamebakaurin inhibited antigen-induced vascular hyperpermeability in a dose-dependent fashion but did not reduce histamine-induced vascular hyperpermeability. CONCLUSION:Kamebakaurin ameliorates allergic symptoms via inhibition of Syk phosphorylation; thus, kamebakaurin could be a lead compound for the new anti-allergic drug.

Aside from the COVID-19 pandemic, the obesity and diabetes pandemics have threatened global health. Patients with diabetes are more likely to experience serious complications from COVID-19; thus, preventing obesity-associated diabetes is of paramount important. Furthermore, the development of a method to prevent diabetes and elucidation of its pathology is a currently urgent issue. We previously reported that thiamine plays a key role in suppressing abnormal glycolipid metabolism in Otsuka Long-Evans Tokushima fatty (OLETF) rats, an animal model of obesity-associated diabetes. However, whether thiamine affects only OLETF rats or other animal models including a type 2 diabetes model with a different pathology requires elucidation. In this study, leptin-receptor deficiency mice were used as a model of type 2 diabetes with a different pathology to evaluate the efficacy of thiamine. The mice had free access to water containing 0.2% thiamine for 9 weeks, and the results showed that food and water consumption decreased in db/db-homo mice. Urine output, body weight gains and blood glucose levels decreased in mice that received thiamine. There were 5 mice and 1 mouse with a fasting glucose level of ≥ 300 mg/dL in the db/db-homo control group (n = 10) and db/db-homo thiamine group (n = 10), respectively, suggesting that thiamine intake may suppress an increase in blood glucose levels. The results of the present study suggest that demand for thiamine may exceed the normal range in in vivo mouse models of diabetes and continuous thiamine intake affects diabetes onset and progression.

Flutamide is a non-steroidal anti-androgen agent, which is mainly used for the treatment of prostate cancer. Flutamide is known to cause severe adverse events, which includes idiosyncratic liver injury. However, details of the mechanism of these adverse reactions have not been elucidated. We investigated whether flutamide induces the release of damage-associated molecular patterns (DAMPs) that activate inflammasomes. We also tested bicalutamide, enzalutamide, apalutamide, and darolutamide for their ability to activate inflammasomes in differentiated THP-1 cells. The supernatant from the incubation of flutamide and bicalutamide with human hepatocarcinoma functional liver cell-4 (FLC-4) cells increased caspase-1 activity and production of IL-1ß by differentiated THP-1 cells. In the supernatant of FLC-4 cells with flutamide and bicalutamide, the heat shock protein (HSP) 40 or 60 was significantly increased. Addition of a carboxylesterase or a CYP inhibitor to the FLC-4 cells prevented release of HSPs from the FLC-4 cells. These results suggested that the reactive metabolites of flutamide and bicalutamide can cause the release of DAMPs from hepatocytes and activate inflammasomes. Inflammasome activation may be an important step in the activation of the immune system by flutamide or bicalutamide, which in some patients, can cause immune-related adverse events.

Diabetes mellitus and brain toxicity are closely linked. Oxidative stress, obesity, insulin resistance, and glucose toxicity can affect the brain. Orexin-A, also known as hypocretin-1, participates in many physiological processes through its activated receptor. Orexin-A has been associated with feeding behavior, obesity, and pathogenesis of Alzheimer's disease. We reported that high-dose thiamine in obese diabetic Otsuka Long–Evans Tokushima Fatty (OLETF) rats leads to reduced obesity and metabolic disorders. In addition, we found that plasma orexin-A levels in OLETF rats can be modulated by thiamine supplementation under conditions of oxidative stress. Herein, we focused on orexin-A in obese diabetic OLETF rats. At 58 weeks of age, the rats showed an increase in body weight and blood glucose levels. Plasma orexin-A was measured by ELISA and tended to be higher in obese diabetic OLETF rats than in non-obese diabetic control rats. We evaluated hypocretin receptor 1 (Hcrtr1, also orexin-A receptor) gene expression in the brain of diabetic OLETF rats by reverse transcription-polymerase chain reaction and found that diabetic OLETF rats exhibited higher orexin-A receptor gene expression in the brain than controls. The results presented here are expected to provide a better understanding of the role of orexin-A and its contribution to diabetic brain.

Diabetes mellitus and brain toxicity are closely linked, and oxidative stress, obesity, insulin resistance, and glucose toxicity can affect the brain. Orexin-A, also known as hypocretin-1, through its activated receptor, participates in many physiological processes. Orexin-A has been associated with feeding behavior, obesity, and pathogenesis of Alzheimer's disease. We have recently established that high-dose thiamine in obese diabetic Otsuka Long–Evans Tokushima Fatty (OLETF) rats leads to reduced obesity and metabolic disorders. Additionally, we found that plasma orexin-A levels in OLETF rats can be modulated by thiamine supplementation under conditions of oxidative stress. Here, we focused on orexin-A in obese diabetic OLETF rats, which at 58 weeks of age and as expected, showed an increase in body weight and blood glucose levels. Plasma orexin-A was measured by ELISA and tended to be higher in obese diabetic OLETF rats than in non-obese diabetic control rats. We evaluated hypocretin receptor 1 (Hcrtr1, also orexin-A receptor) gene expression in the brain of diabetic OLETF rats by reverse transcription (RT)- polymerase chain reaction (PCR) and show that, compared to controls, diabetic OLETF rats exhibited greater orexin-A receptor gene expression in the brain. The results presented here are expected to provide a better understanding of the role of orexin-A and its contribution to brain toxicity in obese diabetic rats.

Obesity and type 2 diabetes mellitus have become worldwide epidemics. Evidence indicates that glucose-dependent insulinotropic polypeptide (GIP) secreted by the intestines may partially underlie these conditions, considering that GIP levels are associated with lipid deposition and fat mass expansion. However, recent studies have found that GIP is also present in other tissues, such as the liver. Notably, one study discovered through microarray analyses of livers from obese diabetic rats that the transcriptional modulation of GIP also occurred in the liver. Otsuka Long-Evans Tokushima Fatty (OLETF) rats were chosen for this experiment because previous studies have shown that thiamine (vitamin B1) could successfully decrease the tendency of the animal toward obesity and mitigate the complications of diabetes. Here, the rats were randomly assigned to either the control (non-supplemented) or thiamine-supplemented (2 g thiamine/L in drinking water) groups. For this investigation, unlike that for young rats, OLETF rats were chosen for the experimental period at 93 weeks of age. Ageing is also a risk factor for diabetes and its complications. In this study, hepatic GIP expression was analysed using western blotting, suggesting that GIP was present in the livers of both obese diabetic OLETF rats and obese diabetic rats that received ongoing thiamine supplementation. Results showed that hepatic GIP expression had occurred and that liver-derived GIP may exist. Moreover, results showed that ongoing thiamine supplementation modified the hepatic GIP expression and prevented additional weight gain and complications arising from obesity and diabetes.

Orexin-A has been suggested to control hypertension, feeding behavior, and obesity. We recently established that long-term consumption of thiamine water by obese diabetic rats leads to reduced obesity and metabolic disorders. In addition, we found that drinking thiamine water daily may modulate oxidative stress-related diseases, such as diabetes and its complications. In the present study, we focused on obesity-related hypertension and plasma orexin-A levels in Otsuka Long–Evans Tokushima Fatty (OLETF) rats under oxidative stress conditions and assessed their cerebral ADP-ribosylated protein expression after drinking thiamine water. The thiamine water-drinking group was administered 2 g thiamine/L in drinking water. Plasma orexin-A content was measured by ELISA testing. ADP-ribosylated protein expression was analyzed in the brain of OLETF rats using Western blotting. Primary experimental characteristics, body weight, and caudal blood pressure were similar among the groups. However, at 28 weeks of thiamine water-drinking, significant decreases in body weight and systolic blood pressure were observed in the diabetic-thiamine group compared to those in the diabetic-control group. Moreover, obese diabetic rats exhibited increased plasma orexin-A levels and poly-ADP-ribosylated protein levels in the brain. Notably, the enhanced plasma orexin-A level and cerebral oxidative stress conditions of the obese diabetic rats were attenuated by drinking thiamine water. The relationship between consumption of thiamine in drinking water and obesity-related hypertension and cerebral oxidative stress status via modulation of plasma orexin-A levels requires further investigation. It is noteworthy that the upregulation of orexin signaling may not only cause hypertension, but also maintain obesity in polyphagia-induced OLETF rats.

Diabetic hyperglycemia is typically accompanied by various protein modifications, indicating hyperglycemic glucotoxicity. Overactivation of poly [adenosine diphosphate (ADP)-ribose] polymerase 1 (PARP-1) has been implicated in the pathogenesis of oxidative stress-related diseases including diabetes and its complications. Furthermore, obesity and diabetes are known to be associated with a substantial risk of chronic liver disease. We have previously reported that thiamine supplementation prevented obesity and diabetes-related liver disease. As a step forward, in the present study, we focus on hepatic ADP-ribosylation that reflects PARP-1 activation and an increased oxidative stress condition. Otsuka Long-Evans Tokushima Fatty (OLETF) rats were randomly divided into the following groups: thiamine-supplemented and unsupplemented control groups. The thiamine-supplemented group received 2 g of thiamine/L of drinking water for 33 weeks. ADP-ribosylated protein expression was analyzed in the livers of OLETF rats using Western blotting. Moreover, the fasting blood glucose level was measured in these rats. The obese diabetic OLETF rats exhibited high ADP-ribosylated protein expression in the liver. Interestingly, hepatic ADP-ribosylated protein expression and fasting blood glucose levels were lower in the thiamine-supplemented OLETF group than in the control OLETF group. These results suggest that thiamine supplementation attenuates oxidative stress by inhibiting hepatic ADP-ribosylation in OLETF rats. The beneficial effect of high-dose thiamine on oxidative stress-related diseases could be attributed to its inhibitory effect on PARP-1 activation, in addition to its role as a coenzyme. Furthermore, we found that thiamine supplementation prevented fasting hyperglycemia, suggesting that high-dose thiamine modifies the hepatic glucose metabolism and obesity-induced hepatic insulin resistance.

Obesity is linked with type 2 diabetes in terms of increasing the risk of developing type 2 diabetes and that of its associated morbidity. We previously reported that thiamine supplementation decreases body weight and visceral fat mass in rats with obesity-related diabetes. Glucose-dependent insulinotropic polypeptide (GIP) acts on pancreatic β cells to promote insulin secretion. According to established theory, GIP is derived from the gastrointestinal tract. We previously discovered increased expression of the incretin GIP gene (Gip) in the livers of obese rats with diabetes receiving high-dose thiamine. We focused on liver-derived GIP to demonstrate GIP protein expression in the liver and visually present localization of GIP in the liver.

Glucose toxicity and lipotoxicity are important states in obesity and diabetes. We previously reported that thiamine supplementation decreases body weight and visceral fat mass in rats with obesity-related diabetes. Glucose-dependent insulinotropic polypeptide (GIP) acts on pancreatic β cells to promote insulin secretion. According to established theory, GIP is derived from the gastrointestinal tract. We previously discovered increased expression of the GIP gene in the livers of obese rats with diabetes receiving high-dose thiamine. We referred to our previous dataset of gene expression analysis using a microarray for livers, which led to the new idea for the present study. We focused on “liver-derived GIP” to demonstrate GIP protein expression in the liver and visually present localization of GIP in the liver. Four-week-old male Otsuka Long-Evans Tokushima Fatty (OLETF) rats were randomly divided into two groups: an unsupplemented control group and a thiamine-supplemented group receiving 2 g of thiamine/L in drinking water for 51 weeks. GIP protein expression in the livers of OLETF rats at 55 weeks of age were determined by western blotting and immunohistochemical analysis. GIP protein expression in the liver was increased in thiamine-supplemented rats compared with that in controls, suggesting that it is involved in preventing and controlling obesity-related diabetic complications. The novel findings of this study that GIP is expressed in the liver, is likely to be added to the story regarding GIP modification of the obese diabetic state.