ToxSci Advance Access originally published online on September 1, 2004
Toxicological Sciences 2005 83(2):282-292; doi:10.1093/toxsci/kfh264
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Toxicological Sciences vol. 83 no. 2 © Society of Toxicology 2005; all rights reserved.
HIGHLIGHTED ARTICLE |
A Toxicogenomic Approach to Drug-Induced Phospholipidosis: Analysis of Its Induction Mechanism and Establishment of a Novel in Vitro Screening System
* Biomedical Research Laboratories, Discovery Research Center, and Development Research Center, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Osaka 532-8686, Japan
Received August 2, 2004; accepted August 8, 2004
 |
ABSTRACT |
|---|
 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Phospholipidosis is a lipid storage disorder in which excess phospholipids accumulate within cells. Some cationic amphiphilic compounds are known to have the potential to induce phospholipidosis. This study was undertaken to examine the molecular mechanisms that contribute to the development of phospholipidosis and to identify specific markers that might form the basis of an in vitro screening test. Specifically, we performed a large-scale gene expression analysis using DNA microarrays on human hepatoma HepG2 cells after they were treated with each of 12 compounds known to induce phospholipidosis. In electron microscopy, HepG2 cells developed lamellar myelin-like bodies in their lysosomes, the characteristic change of phospholipidosis, after treatment with these compounds for 72 h. DNA microarray analysis performed 6 and 24 h after treatment showed alterations in gene expression reflecting the inhibition of lysosomal phospholipase activity and lysosomal enzyme transport, and the induction of phospholipid and cholesterol biosynthesis. Seventeen genes that showed a similar expression profile following treatment were selected as candidate markers. Real-time PCR analysis confirmed that 12 gene markers showed significant concordance with lamellar myelin-like body formation. Furthermore, the average fold change values of these markers correlated well with the magnitude of this pathological change. In conclusion, microarray analysis revealed that factors such as alterations in lysosomal function and cholesterol metabolism were involved in the induction of phospholipidosis. Furthermore, comprehensive gene expression analysis enabled us to identify biomarkers of this condition that we then used to develop a rapid and sensitive in vitro screening test for drug-induced phospholipidosis.
Key Words: drug-induced phospholipidosis; DNA microarray; real-time PCR; HepG2 cells; toxicogenomics; in vitro screening test.
|
INTRODUCTION |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Phospholipidosis is a lipid storage disorder in which phospholipids accumulate in lysosomes. More than 50 cationic amphiphilic drugs (CADs), including antidepressants as well as antianginal, antimalarial, and cholesterol-lowering agents, have been reported to induce phospholipidosis (Lullmann et al., 1978
; Halliwell, 1997, Reasor, 1989). While CADs are thought to induce phospholipidosis by inhibiting lysosomal phospholipase activity, the mechanism by which this occurs has not been extensively studied and is not well understood (Hostetler and Matsuzawa, 1981; Joshi et al., 1988; Reasor and Kacew, 2001; Xia et al., 2000). Electron microscopy has been the most reliable method for identifying phospholipidotic cell damage (Drenckhahn et al., 1976), but because it is a time-consuming and expensive procedure, its use is impractical as a rapid screening tool. It was recently reported that phospholipidotic cell damage could be rapidly assessed in a human monocyte-derived U937 cell line using a Nile red fluorescent stain with a high affinity for lipids (Casartelli et al., 2003).
DNA microarray technology enables investigators to monitor and quantify the expression of thousands of genes simultaneously. Use of this technology in combination with conventional tools is rapidly contributing to our understanding of the mechanisms underlying cellular toxicity, and has emerged as the field of "toxicogenomics" (Aardema and MacGregor, 2002). DNA microarray technology has the potential to more comprehensively contribute to our understanding of toxicity than any available traditional approach, since toxic changes in cells generally result from alterations not just in a single or few molecules, but in many molecular cascades. It may also help to identify early, sensitive biomarkers of toxicity, since alterations in cellular molecules are thought to precede the toxic outcome. These markers could then be used to develop screening tests to predict the toxicity of particular compounds. In pharmaceutical research, such tests are an invaluable tool for cost-effectively selecting candidate drugs, prioritization and compound modification, especially in the early stages of their development.
The present study was undertaken to examine the molecular mechanisms that contribute to the development of phospholipidosis and to identify specific markers that might form the basis of an in vitro screening test. Specifically, we performed a large-scale gene expression analysis using DNA microarrays on human hepatoma HepG2 cells after they were treated with one of the following 12 compounds known to induce phospholipidosis: amiodarone (Lewis et al., 1990; Martin and Standing, 1988; Reasor, 1989), amitriptyline (Drenckhahn et al., 1976), AY-9944 (Sakuragawa et al., 1977), chlorcyclizine (Lullmann-Rauch and Stoermer, 1982), chlorpromazine (Drenckhahn et al., 1976; Lullmann-Rauch, 1974), clomipramine (Lullmann-Rauch, 1974; Lullmann-Rauch and Scheid, 1975; Xia et al., 2000), fluoxetine (Gonzalez-Rothi et al., 1995), imipramine (Lullmann-Rauch, 1974; Lullmann-Rauch and Scheid, 1975), perhexiline (Hauw et al., 1980; Pessayre et al., 1979), tamoxifen (Drenckhahn et al., 1983; Lullmann and Lullmann-Rauch, 1981), thioridazine (Lullmann-Rauch, 1974), and zimelidine (Bockhardt and Lullmann-Rauch, 1980). HepG2 cells were used because they are widely accepted as a good model for in vitro toxicology studies. In vitro phospholipidosis was confirmed by electron microscopic observation of lamellar myelin-like bodies in the lysosomes of these cells.
|
MATERIALS AND METHODS |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Materials. The following compounds were purchased from the following vendors: amiodarone and clozapine from ICN Biomedicals (Irvine, CA); imipramine, clarithromycin, disopyramide, erythromycin, haloperidol, ketoconazole, quinidine, sertraline, and sulfamethoxazole from Wako Pure Chemicals (Osaka, Japan); amitriptyline, AY-9944, chlorcyclizine, chlorpromazine, clomipramine, fluoxetine, perhexiline, tamoxifen, thioridazine, zimelidine, acetaminophen, flecainide, ofloxacin, and sotalol from Sigma (St. Louis, MO); levofloxacin from Apin Chemicals (Abingdon, U.K.); loratadine and sumatriptan from KEMPROTEC (Middlesbrough, U.K.); pentamidine from Toronto Research Chemicals (North York, Canada), and procainamide from Aldrich Chemical (Milwaukee, WI). All other chemicals and solvents were of the highest grade commercially available.
Cell culture and drug treatment. The human hepatocellular carcinoma cell line (HepG2) was purchased from the American Type Culture Collection (ATCC, Manassas, VA). Cells in their log growth-phase were seeded (2 x 105 cells) in 24-well plates and incubated with 8.3 or 25 µmol/l of a particular compound in 0.25% dimethylsulfoxide (vehicle) in Dulbecco's modified Eagle's medium (DMEM; Gibco BRL, Grand Island, NY) supplemented with 5% heat-inactivated fetal calf serum (FCS; Bio Whittaker, Walkersville, MD), 100 U/ml penicillin, and 100 g/ml streptomycin (Gibco BRL). Cells were incubated in a 37°C incubator in an atmosphere of 5% CO2 and 95% air, and were cultured for 72 h, 6 and 24 h, or 24 h, depending on whether they were being used for electron microscopy, DNA microarray analysis, or real-time PCR, respectively.
Transmission electron microscopy. Following incubation, cells were fixed in 1% glutaraldehyde for 2 h after which they were washed with sodium phosphate buffer and post-fixed with 2% osmium tetroxide for 2 h. They were then dehydrated in increasing concentrations of ethanol and embedded in epoxy resin (Quetol 812). Ultrathin sections (80 nm) were cut using an ultramicrotome (LKB-8800 Ultrotome), double stained with uranyl acetate and lead acetate, and observed in an electron microscope (H-300; Hitachi, Tokyo, Japan). The pathological changes indicative of phospholipidosis (formation of lamellar myelin-like bodies in lysosomes) were scored on a scale of 0 to 3 (– = none, + = slight, ++ = moderate, and +++ = severe) in a blinded fashion.
RNA preparation. Following incubation with a test compound, cells were harvested and stored at –80°C until their RNA was extracted using an RNeasy Mini Kit (QIAGEN, Hilden, Germany). For the microarray analysis, RNA was extracted from similarly treated cells that were harvested from six wells and pooled. The concentration and purity of their total RNA were determined by measuring absorbance at 260 and 280 nm with an Ultrospec 2000 spectrophotometer (Amersham Biosciences, Piscataway, NJ). The integrity of the purified total RNA was confirmed using an RNA 6000 Nano Assay kit and Agilent 2100 Bioanalyzer (Agilent Technologies, Berlin, Germany). RNA samples were stored at –80°C until assayed.
Microarray experiments. Microarray analysis was carried out using the GeneChip System (Affymetrix, Santa Clara, CA). The targets of GeneChip analysis were prepared as described in the Expression Analysis Technical Manual. Briefly, total RNA (5 µg) was converted into double-stranded cDNA using a Super Script Choice cDNA synthesis kit (Invitrogen, Carlsbad, CA) with an oligo(dT)24 primer containing a T7 polymerase promoter site at its 3' end (Amersham Biosciences). Biotin-labeled cRNA was generated from double-stranded cDNA using a BioArray HighYield RNA transcript labeling kit (Enzo Biochem, Farmingdale, NY) and was purified using a GeneChip Sample Cleanup Module (QIAGEN). Each cRNA sample (20 µg) was fragmented and hybridized with the Human Genome Array containing 22283 human specific probe sets (HG-U133A; Affymetrix, Santa Clara, CA) for 16 h at 45°C with rotation at 60 rpm. Each array was then washed and detected by consecutive exposure to phycoerythrin-streptavidin (Molecular Probes, Eugene, OR), biotinylated antibodies to streptavidin (Vector Laboratories, Burlingame, CA), and phycoerythrin-streptavidin, after which each array was washed again with a nonstringent wash buffer. All washing and staining procedures were performed with a Fluidics Station 400 (Affymetrix). The array was scanned using a confocal microscope scanner (Hewlett-Packard). To achieve wider signal dynamic range, each chip was scanned three times: before, after the first, and after the second signal amplification using an anti-streptavidin antibody.
The output fluorescence was captured using Affymetrix Microarray Analysis Suite 5.0 software (Affymetrix). This software qualitatively rates the abundance of mRNA of genes (detection call) as present, marginal, or absent, and can also perform comparative analyses between vehicle and drug treated samples at each time point to provide a change call (increased, decreased, marginally increased, marginally decreased, no change) and to generate a signal log ratio (SLR), i.e., the change in expression level for a transcript. This SLR was expressed as the log 2 ratio. A SLR of 1 is the same as a fold change of 2.
Probe sets that satisfied all of the following three conditions in two or three of the three scan data were extracted as up-regulated probe sets: (1) more than 0.6 in their SLR; (2) assigned as "increased"; and (3) determined as "present in the drug-treated data." Similarly, probe sets that satisfied the following three conditions in two or three of the three scan data were extracted as down-regulated genes: (1) less than –0.6 in their SLR; (2) assigned as "decreased"; and (3) determined as "present in vehicle-treated data."
All probe sets representing genes of interest for this study were functionally annotated by the NetAffx database (Affymetrix) and HumanPSD (Incyte, Beverly, MA).
Reverse transcription and real-time PCR analysis. Reverse transcription (RT) was performed using total RNA (1 µg) and oligo-dT oligonucleotide primer in 100 µl volumes using standard methods with MultiScribe Transcriptase in order to synthesize cDNA (TaqMan Reverse Transcription Reagent; Applied Biosystems).
Primers and TaqMan probes (SigmaGenosys, Hokkaido, Japan) used in this study were designed with Primer Express version 1.5 software (PE Applied Biosystems, Foster City, CA). The mRNA sequences used to design the primer and probe sets were obtained from the Affymetrix database (NetAffx). The TaqMan probes had 6-carboxyfluorescein (6-FAM) as the reporter dye and 6-carboxytetramethylrhodamine (6-TAMRA) as the quencher dye at their 5' and 3' ends, respectively. The primer and TaqMan probe sequences used in this assay are listed in Table 1.
|
Quantitative real-time PCR was performed using 5 µl of the cDNA solution, 1x TaqMan Universal PCR Master Mix, and 200 nM of primer/probe set or 1x TaqMan GAPDH control reagents (PE Applied Biosystems) in an ABI PRISM 7000 Sequence Detection System with the following schedule: 2 min at 50°C and 10 min at 95°C, followed by 40 cycles of denaturation for 15 s at 95°C, and annealing and elongation for 1 min at 60°C in a final volume of 50 µl. Relative gene expression levels were normalized to the expression of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and calculated using the comparative Ct method, described in User Bulletin #2 that was provided with the ABI PRISM 7700 Sequence Detection System. The expression data represent the average values from three replicates in a given experiment.
|
RESULTS |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Electron Microscopy
The ability of thirty compounds to induce phospholipidosis in HepG2 cells was evaluated by electron microscopy. Ultrastructural analysis of cells treated with vehicle revealed an extensive endoplasmic reticulum, numerous mitochondria and glycogen granules, and a few small lysosomes with an internal structure that consisted of thin, parallel, concentric lamellae. No lysosomal abnormalities were found in, and only a few lipid droplets were seen in the cytoplasm of, cells that were treated with vehicle or each of 13 of our test compounds (Fig. 1A). However, treatment with 17 compounds, including all of the 12 compounds used in the DNA microarray analysis portion of the study, resulted in an increased number and size of abnormal lysosomes that contained electron-dense deposits and membraneous structures arranged in whorled arrays (myelin figures) suggestive of lysosomal phospholipidosis (Figs. 1B and 1C). The phospholipidotic pathology scores for these compounds are listed in Table 2.
|
|
Large-Scale DNA Microarray Gene Expression Analysis and Nomination of Candidate Markers for Phospholipidosis
Probe sets that were up- or down-regulated by the 12 test compounds that induced phospholipidosis in HepG2 cells were determined based on the criteria described in Materials and Methods. The number of probe sets that were up- or down-regulated is listed in Table 3.
|
Probe sets that were up- or down-regulated by more than 6 out of 12 compounds at each time point were identified as phospholipidosis-related genes; these probe sets are listed and categorized in Tables 4 and 5. Major functional categories included lipid metabolism, cell cycle/proliferation/death, transport, proteolysis and peptidolysis, and endopeptidase inhibition. Genes involved in lipid metabolism were particularly numerous and included those that mediated phospholipid degradation, and cholesterol and fatty acid biosynthesis.
|
|
The number of phospholipidosis-related genes was greater in cells that were treated for 24 h, compared to those that were treated for only 6 h. Accordingly, 17 genes from various functional categories were nominated as candidate markers for phospholipidosis from the group of phospholipidosis-related genes that were identifiable after 24 h of treatment (Table 1).
Real-Time PCR Analysis of Candidate Markers for Phospholipidosis and Selection of Phospholipidosis Markers
The relative expression levels of the candidate markers for phospholipidosis are shown in Table 6. The expression of five genes (PHYH; phytanoyl-CoA hydroxylase [Refsum disease], INHBE; activin beta E, P8; p8 protein [candidate of metastasis 1], ASNS; asparagine synthetase, AP1S1; adaptor-related protein complex 1, sigma 1 subunit) did not correspond well to the pathology score of the HepG2 cells. On the other hand, the following 12 genes were selected as phospholipidosis markers: ASAH1—N-acylsphingosine amidohydrolase (acid ceramidase) 1; MGC4171—hypothetical protein MGC4171; LSS—lanosterol synthase (2,3-oxidosqualene-lanosterol cyclase); NR0B2—nuclear receptor subfamily 0, group B, member 2; FABP1—fatty acid binding protein 1, liver; HPN—hepsin (transmembrane protease, serine 1); SERPINA3—serine (or cysteine) proteinase inhibitor; clade A (alpha-1 antiproteinase, antitrypsin), member 3—C10orf10—chromosome 10 open reading frame 10; FLJ10055mdash;hypothetical protein FLJ10055 FRCP1—likely ortholog of mouse fibronectin type III repeat containing protein 1; SLC2A3—solute carrier family 2 (facilitated glucose transporter) member 3; and TAGLN—transgelin.
|
Establishment and Validation of an in Vitro Screening System
In order to establish a representative value for phospholipidosis markers, we calculated average fold change values and referred to them as the phospholipidosis mRNA scores. There was a significant correlation between these mRNA scores and the pathological scores obtained from electron microscopic analysis of the HepG2 cells (Fig. 2).
|
In order to validate these mRNA scores, we examined the mRNA and pathology scores twice more on another set of 14 compounds. Three of these compounds that had an mRNA score of <1.5 failed to induce pathology in the HepG2 cells, while the 11 compounds that had a score above 1.5 induced phospholipidotic changes in these cells. These results were highly reproducible (Fig. 3).
|
|
DISCUSSION |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Ultrastructural analysis revealed lamellar myelin-like bodies in the lysosomes of HepG2 cells after they were treated with 12 compounds that were reported to cause phospholipidosis (Bockhardt and Lullmann-Rauch, 1980
; Drenckhahn et al., 1976, 1983; Gonzalez-Rothi et al., 1995; Hauw et al., 1980; Lewis et al., 1990; Lullmann and Lullmann-Rauch, 1981; Lullmann-Rauch, 1974; Lullmann-Rauch and Scheid, 1975; Lullmann-Rauch and Stoermer, 1982; Martin and Standing, 1988; Pessayre et al., 1979; Reasor, 1989; Sakuragawa et al., 1977; Xia et al., 2000). These findings validated the use of these cells as an in vitro study model for the assessment of the phospholipidosis-inducing potential of various compounds.
In this study, we examined the molecular mechanisms that contribute to the development of phospholipidosis and identified specific markers that might form the basis of an in vitro screening test. Specifically, we performed a large-scale gene expression analysis using DNA microarrays on human hepatoma HepG2 cells after they were treated with each of 12 compounds known to induce phospholipidosis. Probe sets that were up- or down-regulated by at least six compounds were identified as phospholipidosis-related genes, a large number of which were genes that played a role in lipid metabolism (Tables 4 and 5). Functional annotation and categorization of these genes suggested that the following four processes were involved in the induction of phospholipidosis (Fig. 4): (1) Inhibition of lysosomal phospholipase activity—this is generally regarded as the primary mechanism of induction, as confirmed by the up-regulation of phospholipid degradation-related genes such as N-acylsphingosine amidohydrolase 1 (ASAH1), sphingomyelin phosphodiesterase (SMPDL3A), and hypothetical protein MGC4171 (MGC4171). (2) Inhibition of lysosomal enzyme transport, as demonstrated by the down-regulation of genes involved in lysosomal enzyme transport such as adaptor-related protein complex 1 sigma 1 subunit (AP1S1). AP1S1 is responsible for the transport of newly synthesized lysosomal enzymes between the trans-golgi network and lysosomes (Zhu et al., 1999). (3) Enhanced phospholipid biosynthesis, which is supported by the up-regulation of fatty acid biosynthesis-related genes such as ELOVL family member 6 (ELOVL6) and stearoyl-CoA desaturase (SCD). (4) Enhanced cholesterol biosynthesis, as shown by the up-regulation of cholesterol biosynthesis-related genes such as 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 (HMGCS1), 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGCR), squalene epoxidase (SQLE), lanosterol synthase (LSS), and 7-dehydrocholesterol reductase (DHCR7).
|
Inhibition of lysosomal phospholipase activity and lysosomal enzyme transport, coupled with enhanced phospholipid biosynthesis could directly trigger phospholipidosis. Increased cholesterol biosynthesis is considered to be an indirect trigger for the following two reasons: (1) The accumulation of sphingomyelin occurs concurrently with the increase in cholesterol in visceral tissues (e.g., spleen) in patients with Niemann-Pick type C disease (NPC), which is caused by a genetic defect in the cholesterol trafficking protein NPC1 or, in far fewer patients, the sterol regulating protein HE1 (Blanchette-Mackie, 2000
; Harzer et al., 2003; Vanier, 1983). (2) The induction of lamellar myelin-like bodies and the accumulation of free cholesterol occur in cultured mouse macrophages that have been incubated with acetylated low density lipoprotein or acyl-CoA:cholesterol acyltransferase inhibitor (McGookey and Anderson, 1983; Robenek and Schmitz, 1988). In addition, the up- or down-regulation of transporter genes (e.g., facilitated glucose transporter) and genes that control the cell cycle (e.g., cyclin G2) may also be involved, but modification of the expression of these genes likely reflects secondary changes that occur as a result of an increase in cellular phospholipids. An important, practical goal of our study was to identify phospholipidosis specific gene markers that could be used for the establishment of an in vitro screening test. Using DNA microarray and real-time PCR analyses, we identified 12 phospholipidosis marker genes. The average fold change values of these gene markers were calculated and termed their phospholipidosis mRNA scores, the latter of which correlated well with the cells' pathological scores. Since these phospholipidosis markers included genes whose functions included phospholipid degradation, cholesterol biosynthesis, fatty acid transport, proteolysis and peptidolysis, and endopeptidase inhibition, among others, by monitoring these 12 marker genes, we were able to also monitor multiple intracellular events that were involved in the induction of phospholipidosis. Our results showed that the mRNA score was a good index of phospholipidosis induction potential for a given compound and as such, we went ahead to establish a novel in vitro real-time PCR screening test.
The in vitro screening test required a far lower amount of the test compound and shorter periods than did conventional in vivo toxicity studies, and was readily able to detect the phospholipidosis induction potential of multiple compounds. The assay also provided a more detailed ranking score than electron microscopy that can be useful in structure-activity relationship studies by sorting compounds in the order of their phospholipidosis induction potential. This rapid and sensitive system should facilitate the efficient screening of new compounds for their potential for inducing phospholipidosis at an early developmental stage. Finally, transferring our PCR-based screening system into a 96- or 384-well microplate-based mRNA measuring format such as Array Plate (High Throughput Genomics, Tucson, AZ) and QuantiGene branched DNA (Bayer Diagnostics, Tarrytown, NY), should improve its throughput and cost-efficiency.
The toxicogenomics approach used in this study should be helpful in the examination of the mode of action and identification of gene markers for other toxic conditions as well. The hope is that in the near future new toxic gene markers will be identified that will allow for the testing of multiple drug toxicities simply by measuring gene expression.
In conclusion, microarray analysis revealed that factors such as alterations in lysosomal function and cholesterol metabolism were involved in the induction of phospholipidosis. Furthermore, comprehensive gene expression analysis enabled us to identify biomarkers of this condition that we then used to develop a rapid and sensitive in vitro screening test for drug-induced phospholipidosis.
|
ACKNOWLEDGMENTS |
|---|
We thank Dr. Yoshinobu Yoshimura, Dr. Kenji Okonogi, and Dr. Tetsuo Miwa for encouragement. We are grateful to our toxicogenomics and in vitro toxicology team members for their invaluable suggestions and helpful discussions. We would like to thank Mr. Kazuo Takabe for his excellent technical assistance.
|
NOTES |
|---|
1 To whom correspondence should be addressed at Biomedical Research Laboratories, Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, 2-17-85 Juso-Honmachi Yodogawa-ku, Osaka 532-8686, Japan. Fax: +81-6-6300-6306. E-mail: Sawada_Hiroshi{at}takeda.co.jp
|
REFERENCES |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Aardema, M. J., and MacGregor, J. T. (2002). Toxicology and genetic toxicology in the new era of "toxicogenomics": impact of "-omics" technologies. Mutat. Res. 499, 13–25.[ISI][Medline]
Blanchette-Mackie, E. J. (2000). Intracellular cholesterol trafficking: Role of the NPC1 protein. Biochim. Biophys. Acta 1486, 171–183.[Medline]
Bockhardt, H., and Lullmann-Rauch, R. (1980). Zimelidine-induced lipidosis in rats. Acta Pharmacol. Toxicol. (Copenh). 47, 45–48.[Medline]
Casartelli, A., Bonato, M., Cristofori, P., Crivellente, F., Dal Negro, G., Masotto, I., Mutinelli, C., Valko, K., and Bonfante, V. (2003). A cell-based approach for the early assessment of the phospholipidogenic potential in pharmaceutical research and drug development. Cell Biol. Toxicol. 19, 161–176.[CrossRef][ISI][Medline]
Drenckhahn, D., Jacobi, B., and Lullmann-Rauch, R. (1983). Corneal lipidosis in rats treated with amphiphilic cationic drugs. Arzneimittelforschung 33, 827–831.[Medline]
Drenckhahn, D., Kleine, L., and Lullmann-Rauch, R. (1976). Lysosomal alterations in cultured macrophages exposed to anorexigenic and psychotropic drugs. Lab. Invest. 35, 116–123.[ISI][Medline]
Gonzalez-Rothi, R. J., Zander, D. S., and Ros, P. R. (1995). Fluoxetine hydrochloride (Prozac)-induced pulmonary disease. Chest 107, 1763–1765.[Medline]
Halliwell, W. H. (1997). Cationic amphiphilic drug-induced phospholipidosis. Toxicol. Pathol. 25, 53–60.[ISI][Medline]
Harzer, K., Massenkeil, G., and Frohlich, E. (2003). Concurrent increase of cholesterol, sphingomyelin and glucosylceramide in the spleen from non-neurologic Niemann-Pick type C patients but also patients possibly affected with other lipid trafficking disorders. FEBS Lett. 537, 177–181.[CrossRef][ISI][Medline]
Hauw, J. J., Boutry, J. M., Albouz, S., Harpin, M. L., Baudrimont, M., Escourolle, R., and Baumann, N. (1980). Perhexiline maleate-induced lipidosis in cultured human fibroblasts: cell kinetics, ultrastructural and biochemical studies. Virchows Arch. B. Cell Pathol. Incl. Mol. Pathol. 34, 239–249.[ISI][Medline]
Hostetler, K. Y., and Matsuzawa, Y. (1981). Studies on the mechanism of drug-induced lipidosis. Cationic amphiphilic drug inhibition of lysosomal phospholipases A and C. Biochem. Pharmacol. 30, 1121–1126.[CrossRef][ISI][Medline]
Joshi, U. M., Kodavanti, P. R., Coudert, B., Dwyer, T. M., and Mehendale, H. M. (1988). Types of interaction of amphiphilic drugs with phospholipid vesicles. J. Pharmacol. Exp. Ther. 246, 150–157.
Lewis, J. H., Mullick, F., Ishak, K. G., Ranard, R. C., Ragsdale, B., Perse, R. M., Rusnock, E. J., Wolke, A., Benjamin, S. B., Seeff, L. B., and et al. (1990). Histopathologic analysis of suspected amiodarone hepatotoxicity. Hum. Pathol. 21, 59–67.[CrossRef][ISI][Medline]
Lullmann, H., and Lullmann-Rauch, R. (1981). Tamoxifen-induced generalized lipidosis in rats subchronically treated with high doses. Toxicol. Appl. Pharmacol. 61, 138–146.[CrossRef][ISI][Medline]
Lullmann, H., Lullmann-Rauch, R., and Wassermann, O. (1978). Lipidosis induced by amphiphilic cationic drugs. Biochem. Pharmacol. 27, 1103–1108.[CrossRef][ISI][Medline]
Lullmann-Rauch, R. (1974). Lipidosis-like ultrastructural alterations in rat lymph nodes after treatment with tricyclic antidepressants or neuroleptics. Naunyn. Schmiedebergs Arch. Pharmacol. 286, 165–179.[CrossRef][ISI][Medline]
Lullmann-Rauch, R., and Scheid, D. (1975). Intraalveolar foam cells associated with lipidosis-like alterations in lung and liver of rats treated with tricyclic psychotropic drugs. Virchows Arch. B. Cell Pathol. 19, 255–268.[ISI][Medline]
Lullmann-Rauch, R., and Stoermer, B. (1982). Generalized lipidosis in newborn rats and Guinea pigs induced during prenatal development by administration of amphiphilic drugs to pregnant animals. Virchows Arch. B. Cell Pathol. Incl. Mol. Pathol. 39, 59–73.[ISI][Medline]
Martin, W. J., 2nd, and Standing, J. E. (1988). Amiodarone pulmonary toxicity: biochemical evidence for a cellular phospholipidosis in the bronchoalveolar lavage of human subjects. J. Pharmacol. Exp. Ther. 244, 774–779.
McGookey, D. J., and Anderson, R. G. (1983). Morphological characterization of the cholesteryl ester cycle in cultured mouse macrophage foam cells. J. Cell Biol. 97, 1156–1168.
Pessayre, D., Bichara, M., Degott, C., Potet, F., Benhamou, J. P., and Feldmann, G. (1979). Perhexiline maleate-induced cirrhosis. Gastroenterology 76, 170–177.[ISI][Medline]
Reasor, M. J. (1989). A review of the biology and toxicologic implications of the induction of lysosomal lamellar bodies by drugs. Toxicol. Appl. Pharmacol. 97, 47–56.[CrossRef][ISI][Medline]
Reasor, M. J., and Kacew, S. (2001). Drug-induced phospholipidosis: Are there functional consequences? Exp. Biol. Med. 226, 825–830.
Robenek, H., and Schmitz, G. (1988). Ca++ antagonists and ACAT inhibitors promote cholesterol efflux from macrophages by different mechanisms. II. Characterization of intracellular morphologic changes. Arteriosclerosis 8, 57–67.[Medline]
Sakuragawa, N., Sakuragaw, M., Kuwabara, T., Pentchev, P. G., Barranger, J. A., and Brady, R. O. (1977). Niemann-Pick disease experimental model: Sphingomyelinase reduction induced by AY-9944. Science 196, 317–319.
Vanier, M. T. (1983). Biochemical studies in Niemann-Pick disease. I. Major sphingolipids of liver and spleen. Biochim. Biophys. Acta 750, 178–184.[Medline]
Xia, Z., Ying, G., Hansson, A. L., Karlsson, H., Xie, Y., Bergstrand, A., DePierre, J. W., and Nassberger, L. (2000). Antidepressant-induced lipidosis with special reference to tricyclic compounds. Prog. Neurobiol. 60, 501–512.[CrossRef][ISI][Medline]
Zhu, Y., Traub, L. M., and Kornfeld, S. (1999). High-affinity binding of the AP-1 adaptor complex to trans-golgi network membranes devoid of mannose 6-phosphate receptors. Mol. Biol. Cell 10, 537–549.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  B. A. Merrick The plasma proteome, adductome and idiosyncratic toxicity in toxicoproteomics research Brief Funct Genomic Proteomic, February 12, 2008; (2008) eln004v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Nioi, B. K. Perry, E.-J. Wang, Y.-Z. Gu, and R. D. Snyder In Vitro Detection of Drug-Induced Phospholipidosis Using Gene Expression and Fluorescent Phospholipid Based Methodologies Toxicol. Sci., September 1, 2007; 99(1): 162 - 173. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Atienzar, H. Gerets, S. Dufrane, K. Tilmant, M. Cornet, S. Dhalluin, B. Ruty, G. Rose, and M. Canning Determination of Phospholipidosis Potential Based on Gene Expression Analysis in HepG2 Cells Toxicol. Sci., March 1, 2007; 96(1): 101 - 114. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. P. Fanton, M. W. Rowe, E. J. Moler, M. Ison-Dugenny, S. K. De Long, K. Rendahl, Y. Shao, T. Slabiak, T. G. Gesner, and M. L. MacKichan Development of a screening assay for surrogate markers of chk1 inhibitor-induced cell cycle release. J Biomol Screen, October 1, 2006; 11(7): 792 - 806. [Abstract] [PDF]
|
|
|
|
|
|
T. Kasahara, K. Tomita, H. Murano, T. Harada, K. Tsubakimoto, T. Ogihara, S. Ohnishi, and C. Kakinuma Establishment of an In Vitro High-Throughput Screening Assay for Detecting Phospholipidosis-Inducing Potential Toxicol. Sci., March 1, 2006; 90(1): 133 - 141. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||