Unravelling Pathophysiology of Crystalline Nephropathy in Ceftriaxone-Associated Acute Kidney Injury: A Cellular Proteomic Approach

Abstract

Background: Previous studies showed that ceftriaxone can cause acute kidney injury (AKI) in the pediatric population. This study proposed a cellular model of crystalline nephropathy in ceftriaxone-associated AKI and explored the related pathophysiology by using a proteomic approach. Methods: Ceftriaxone was crystallized with calcium in artificial urine. Madin-Darby Canine Kidney (MDCK) cells, a model of distal renal tubular cell, were cultured in the absence (untreated control) or presence of ceftriaxone crystals for 48-h (n = 5 each). MDCK cells were harvested and subsequently analyzed by proteomic analysis. Protein bioinformatics (i.e., STRING and Reactome) was used to predict functional alterations, and subsequently validated by Western blotting and cellular studies. p < 0.05 was considered statistically significant. Results: Phase-contrast microscopy showed increased intracellular vesiculation and cell enlargement as a result of ceftriaxone crystal exposure. Proteome analysis revealed a total of 20 altered proteins (14 increased, 5 decreased and 1 absent) in ceftriaxone crystal-treated MDCK cells as compared to untreated cells (p < 0.05). Protein bioinformatics and validation studies supported heat stress response mediated by heat shock protein 70 (Hsp70) and downregulation of annexin A1 as the proposed pathophysiology of crystalline nephropathy in ceftriaxone-associated AKI, in which impaired proliferation and wound healing of crystal-induced distal tubular cells were outcomes. Conclusions: This study, for the first time, used the in vitro model of crystalline nephropathy to investigate the underlying pathophysiology of ceftriaxone-associated AKI, which should be investigated in vivo for potential clinical benefits in the future.

Meeting report of the “Symposium on kidney stones and mineral metabolism: calcium kidney stones in 2017”

“…The dysproteinemia-related CN includes light chain cast nephropathy, also known as myeloma cast nephropathy, which is the most common dysproteinemia-related renal disease [53,54]. Crystalline nephropathy may develop during the use of drugs (ceftriaxone [55]), sulfadiazine [56], acyclovir [57], triamterene [58], atanazavir [59], and others that are excreted by the kidney (see details in recent review [60]). Calcium crystals CN is characterized by abundant tubular and interstitial deposits of calcium phosphate or CaOx crystals, easily distinguished by their tinctorial properties.…”

Cefazolin sodium pentahydrate cause urolithiasis: a case report and literature review

“…Chatchen et al . [ 10 ] studied the underlying pathophysiology of ceftriaxone-associated acute kidney injury via cellular proteomic in vitro cell model approach, whereby cell survival and adaptation were mediated by the concurrent upregulation of Hsp70, whereas downregulation of annexin A1 was proposed as the negative-feedback mechanism to mitigate the stressful effects of ceftriaxone crystallization, cell growth arrest and delayed wound healing of distal tubular cells after ceftriaxone crystal exposure, shedding light on collateral damage in combination therapy involving ceftriaxone and other stressors. Kimata et al .…”

Drug-Induced Urolithiasis in Pediatric Patients

“…Ceftriaxone is a third-generation cephalosporin and is characterized by its broad spectrum, high safety profile and long plasma half-life, rendering it among the commonly used antibacterial drugs during the childhood period for the treatment of different bacterial infections [24]. Ceftriaxone has been known for its biliary side effects, including biliary sludge and pseudolithiasis since the late 1980s [25].…”

“…This may be the underlying mechanism for its biliary side effects [29]; however, the mechanism of ceftriaxone-induced nephrolithiasis is not yet completely understood [31]. In this setting, several authors have tried to investigate the pathophysiological mechanism for ceftriaxone-associated renal side effects [24,46]. Zhang et al [46] demonstrated in their animal study that the rats that were administered ceftriaxone and calcium showed a significantly lower volume of 24-h urine, with a significant increase in serum creatinine and blood urea nitrogen levels, suggesting that calcium chloride is important for stone formation.…”

“…This is in contrast to Avci et al [33], who reported unchanged calcium excretion in children treated with ceftriaxone. Moreover, Chatchen et al [24] microscopically examined a Madin-Darby canine model (a model of distal renal tubular cells) in the presence and absence of ceftriaxone crystals for 48 h, demonstrating that heat stress response and heat shock protein 70 play an important role in the process of crystalline nephropathy and post-ceftriaxone acute renal injury.…”