Endocytosis of Calcium Oxalate Crystals and Proliferation of Renal Tubular Epithelial Cells in a Patient with Type 1 Primary Hyperoxaluria

Lieske, John C.; Spargo, Benjamin H.; Toback, F. Gary

doi:10.1016/s0022-5347(17)36954-9

Cited by 88 publications

(52 citation statements)

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“…Some are later internalized (9,25) and/or transported to the interstitium (9 -19, 25, 53). Given the presence of tubular intracellular crystals in hyperoxaluric and idiopathic stone formers (13,35) as well as in the renal interstitium of nonstone formers (8) and in patients with primary hyperoxalosis (10,35), it is clear that any potential role of intracrystalline proteins in CaOx crystals is not confined to tubular epithelial cells. It is well documented that once crystals have been transported to the interstitium they dissolve or are destroyed (10 -11, 23), probably by lysosomal enzymes in the macrophages and multinucleated giant cells that engulf them (9, 11-12, 23, 53), since CaOx crystals have been observed inside the phagolysosomes of renal tubular cells of rats with chronic hyperoxaluria (25).…”

Section: Discussionmentioning

confidence: 99%

“…of studies performed in the early 1990s, which demonstrated for the first time that CaOx crystals irreversibly adhere to and are phagocytosed by cultured renal epithelial cells (35,39). These findings introduced a new paradigm to urolithiasis research because they validated the credibility of using this experimental approach to examine and manipulate factors affecting the regulation of crystal retention, which had not previously been possible.…”

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confidence: 85%

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Intracrystalline urinary proteins facilitate degradation and dissolution of calcium oxalate crystals in cultured renal cells

Grover¹,

Thurgood²,

Fleming³

et al. 2008

American Journal of Physiology-Renal Physiology

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134: 5-14, 2001. The aim of this investigation was to determine the effect of increasing concentrations of intracrystalline protein on the rate of CaOx crystal dissolution in Madin-Darby canine kidney (MDCKII) cells. Crystal matrix extract (CME) was isolated from urinary CaOx monohydrate (COM) crystals. Cold and [ 14 C]oxalate-labeled COM crystals were precipitated from ultrafiltered urine containing 0 -5 mg/l CME. Crystal surface area was estimated from scanning electron micrographs, and synchrotron X-ray diffraction was used to determine nonuniform strain and crystallite size. Radiolabeled crystals were added to MDCKII cells and crystal dissolution, expressed as radioactive label released into the medium, was measured. Increasing CME content did not significantly alter crystal surface area. However, nonuniform strain increased and crystallite size decreased in a dose-response manner, both reaching saturation at a CME concentration of 3 mg/ and demonstrating unequivocally the inclusion of increasing quantities of proteins in the crystals. This was confirmed by Western blotting. Crystal dissolution also followed saturation kinetics, increasing proportionally with final CME concentration and reaching a plateau at a concentration of ϳ2 mg/l. These findings were complemented by field emission scanning electron microscopy, which showed that crystal degradation also increased relative to CME concentration. Intracrystalline proteins enhance degradation and dissolution of CaOx crystals and thus may constitute a natural defense against urolithiasis. The findings have significant ramifications in biomineral metabolism and pathogenesis of renal stones. nephrocalcinosis; urolithiasis THE PATHOGENESIS OF RENAL calculi requires the sequential combination of two processes, namely, the nucleation of crystals and their retention within the kidney. While crystal nucleation requires supersaturation of urine with calcium oxalate (CaOx), renal crystal trapping has been explained by either the "free particle" or "fixed particle" theory. Although both these theories appear equally plausible (26), current consensus favors a fixed particle mechanism. This view is supported by the results of studies performed in the early 1990s, which demonstrated for the first time that CaOx crystals irreversibly adhere to and are phagocytosed by cultured renal epithelial cells (35,39). These findings introduced a new paradigm to urolithiasis research because they validated the credibility of using this experimental approach to examine and manipulate factors affecting the regulation of crystal retention, which had not previously been possible. Consequently, there followed a series of reports investigating factors affecting interactions between CaOx monohydrate (COM) crystals and renal cells (reviewed in Refs. 30 and 38). Those studies showed that COM crystals, which are the predominant form of CaOx occurring in human kidney stones (41), are highly membranolytic (55), and also that their adherence to renal epithelial cells is very rapid, concentration ...

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Section: Discussionmentioning

confidence: 99%

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confidence: 85%

Intracrystalline urinary proteins facilitate degradation and dissolution of calcium oxalate crystals in cultured renal cells

Grover¹,

Thurgood²,

Fleming³

et al. 2008

American Journal of Physiology-Renal Physiology

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“…We have observed renal deposits of calcium oxalate crystals in a patient with primary hyperoxaluria after kidney-liver transplantation when the high body oxalate load was being excreted by the transplanted kidney (2). Crystals were observed within tubular epithelial cells and were associated with proliferation and formation of multinucleated giant cells.…”

mentioning

confidence: 99%

Renal epithelial cells rapidly bind and internalize calcium oxalate monohydrate crystals.

Lieske

Swift

Martin

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

136

114

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Renal tubular fluid is supersaturated with calcium and oxalate ions, which can nucleate to form crystals of calcium oxalate monohydrate (COM), the most abundant constituent of kidney stones. However, the mechanisms by which nascent crystals are retained in the nephron and then grow into kidney stones are unclear. An interaction of COM crystals with the surface of renal epithelial cells could be a critical initiating event in nephrolithiasis. To investigate this possibility we used cultures of monkey kidney epithelial cells (BSC-1 line) as a model system and found that [14CJCOM crystals bound to the cell surface within seconds. Scanning electron microscopy revealed that crystals bind first to apical microvilli, which subsequently migrate over the crystalline surface. When visualized by transmission electron microscopy, intracellular crystals were located within vesicles. Cytoskeletal responses to crystal uptake were sought by immunofuorescence microscopy, which revealed concentration of (3). Before use, crystals were sterilized by heating to 180TC overnight and then suspended in distilled water to form a slurry from which they were added to the culture medium (3). X-ray crystallography, performed by S. Deganello (University of Chicago), demonstrated that heating did not alter the structure of COM crystals.[14C]COM crystals (10-300 Mig/ml; 2.4-70.8 pg/cm2 cell surface) were added to high-density, quiescent cultures in 60-mm dishes (Nunc) to define the kinetics of association between a cell and crystal. After a specified period, the medium was aspirated, and the monolayer was washed three times with phosphate-buffered saline (PBS; 5 ml). Each culture was inspected under a microscope and the number of cells with adherent crystals was counted in five separate fields. Subsequently, the cell monolayer with adherent crystals was scraped directly into a scintillation vial containing 6 M HC (0.5 ml), 4.5 ml of Ecoscint (National Diagnostics) was added, and the amount of radioactivity was measured. To investigate the effect of crystal exposure for up to 15 days on renal epithelial cells, COM crystals (50 pg/ml) were added to near-confluent cultures of BSC-1 cells (106 cells per 60-mm dish) containing 0.5% calf serum. Every 4 days thereafter, the medium was aspirated and replaced with fresh medium containing 0.5% calf serum with no additional crystals. At 1, 8, and 15 days after addition of crystals, the medium was aspirated, and a solution of crystalline trypsin was used to detach the cells, which were then inspected under a microscope as described (4). The total number of cells in each culture was counted with a hemocytometer, and 100 cells from each culture were scored for the presence of internalized crystals.Abbreviations: COM, calcium oxalate monohydrate; TEM, transmission electron microscopy; SEM, scanning electron microscopy. tTo whom reprint requests should be addressed. 6987The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "adv...

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“…Several groups of investigators have suggested that an important early event in the formation of kidney stones is the attachment of calcium oxalate (CaOx) crystals to the apical surface of renal tubular epithelium (1)(2)(3). Well-polarized, undisturbed tubular epithelial cells in culture demonstrate little capacity for crystal attachment (4 -6).…”

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confidence: 99%

An Acidic Peptide Sequence of Nucleolin-Related Protein Can Mediate the Attachment of Calcium Oxalate to Renal Tubule Cells

Сорокина¹,

Wesson²,

Kleinman³

2004

Journal of the American Society of Nephrology

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Abstract. Crystals that form in tubular fluid must be retained in the kidney to become stones. Nucleolin-related protein (NRP) is found on the surface of inner medullary collecting duct (IMCD) cells in culture (cIMCD) and selectively adsorbs to calcium oxalate (CaOx). We proposed that NRP mediates attachment to the renal tubular epithelium of Ca stone crystals through an electrostatic interaction with a highly acidic region (acidic fragment [AF]) similar to those of other proteins that have been reported to affect urinary crystal formation. The current studies demonstrate that nucleolin is expressed on both apical and basolateral cell surfaces of cIMCD, reaching a peak in the late stages of mitosis and gradually declining to undetectable levels with maturation of the polarized epithelium. Scraping areas of mature monolayers stimulated the cells surrounding the defects to migrate and proliferate so as to repair them, and these areas demonstrate surface NRP expression and enhanced attachment of CaOx monohydrate crystals. Surface expression of the NRP AF was produced by cloning the NRP AF into a display vector. Transfected cIMCD demonstrating copious surface expression of AF enhanced CaOx attachment 6.7-fold compared with control cIMCD, whereas cells transfected with a vector without the AF did not differ from control. AF was also cloned into a replication-deficient adenovirus and expressed in 293 cells, resulting in AF secretion into the nutrient medium. This medium inhibited CaOx attachment to cIMCD, compared with conditioned medium from cells infected with wild-type virus. These results demonstrate that surface-bound AF can mediate CaOx attachment and that secreted AF can inhibit attachment. These results support the notion that surface-associated NRP could mediate attachment of CaOx to the renal tubule epithelium, thereby causing retention of crystals that might eventually become kidney stones.Several groups of investigators have suggested that an important early event in the formation of kidney stones is the attachment of calcium oxalate (CaOx) crystals to the apical surface of renal tubular epithelium (1-3). Well-polarized, undisturbed tubular epithelial cells in culture demonstrate little capacity for crystal attachment (4 -6). This resistance to attachment may be due to the confinement of molecules with crystal-binding properties to domains other than the apical membrane. When tight junctions between cells are disrupted in the course of cellular proliferation or migration or when cells are exposed to a number of different agents, apical membranes seem to express constituents with affinity for CaOx.Nucleolin-related protein (NRP) is a surface-associated protein of cultured inner medullary collecting duct (IMCD) cells that selectively adsorbs to CaOx and binds to membrane skeletal elements in a Ca-dependent manner (7). NRP has a highly acidic region composed of 62% Asp and Glu residues (135KNGKNAKKEDSDEEDDDDSEEEDEEDEDEEDEFE PTVMKAAAAAPAS-DEEEDDEEDDDEDDDEEDEDED-EDDSEEEAMETTPAKGKKAPVKAVPVKAKSTAEE-DEEEDDEDEEDDEE...

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Endocytosis of Calcium Oxalate Crystals and Proliferation of Renal Tubular Epithelial Cells in a Patient with Type 1 Primary Hyperoxaluria

Cited by 88 publications

References 8 publications

Intracrystalline urinary proteins facilitate degradation and dissolution of calcium oxalate crystals in cultured renal cells

Intracrystalline urinary proteins facilitate degradation and dissolution of calcium oxalate crystals in cultured renal cells

Renal epithelial cells rapidly bind and internalize calcium oxalate monohydrate crystals.

An Acidic Peptide Sequence of Nucleolin-Related Protein Can Mediate the Attachment of Calcium Oxalate to Renal Tubule Cells

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