scholarly journals Evaluation of biomarkers for in vitro prediction of drug-induced nephrotoxicity in RPTEC/TERT1 cells

2020 ◽  
Vol 9 (2) ◽  
pp. 91-100 ◽  
Author(s):  
Xuan Qiu ◽  
Yufa Miao ◽  
Xingchao Geng ◽  
Xiaobing Zhou ◽  
Bo Li
Keyword(s):  
Transcriptional Activation ◽  
Messenger Rna ◽  
Aristolochic Acid ◽  
Kidney Injury ◽  
Drug Induced ◽  
Vitro Assay ◽  
Protein Levels ◽  
Aristolochic Acid I

Abstract There have been intensive efforts to identify in vivo biomarkers that can be used to monitor drug-induced kidney damage before significant impairment occurs. Kidney injury molecule-1, neutrophil gelatinase-associated lipocalin, clusterin, β2-microglobulin and cystatin C (CysC) have been validated as clinical or preclinical biomarkers in urinary and plasma predictive of acute and chronic kidney injuries and diseases. A high-throughput in vitro assay predictive of nephrotoxicity could potentially be implemented in early drug discovery stage to reduce attrition at later stages of drug development. To assess the potential of these known in vivo biomarkers for in vitro evaluation of drug-induced nephrotoxicity, we selected four nephrotoxic agents (cisplatin, cyclosporin, aristolochic acid I and gentamicin) and detected their effects on the protein levels of nephrotoxic biomarkers in RPTEC/TERT1 cells. The protein levels of clusterin, CysC, GSTπ and TIMP-1 significantly increased in the conditioned media of RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin. The messenger RNA levels of clusterin, CysC, GSTπ and TIMP-1 also increased in RPTEC/TERT1 cells treated with cisplatin, cyclosporin, aristolochic acid I and gentamicin, indicating that drug-induced upregulation involves transcriptional activation. Taken together, the results clearly demonstrate that among the known in vivo nephrotoxic biomarkers, clusterin, CysC, GSTπ and TIMP-1 can be effectively used as in vitro biomarkers for drug-induced nephrotoxicity in RPTEC/TERT1 cells.

scholarly journals CHIP promotes Runx2 degradation and negatively regulates osteoblast differentiation

2008 ◽  
Vol 181 (6) ◽  
pp. 959-972 ◽  
Author(s):  
Xueni Li ◽  
Mei Huang ◽  
Huiling Zheng ◽  
Yinyin Wang ◽  
Fangli Ren ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Osteoblast Differentiation ◽  
Degradation Mechanism ◽  
Interacting Protein ◽  
E3 Ligases ◽  
Protein Levels ◽  
C Terminus ◽  
Osteoblast Lineage

Runx2, an essential transactivator for osteoblast differentiation, is tightly regulated at both the transcriptional and posttranslational levels. In this paper, we report that CHIP (C terminus of Hsc70-interacting protein)/STUB1 regulates Runx2 protein stability via a ubiquitination-degradation mechanism. CHIP interacts with Runx2 in vitro and in vivo. In the presence of increased Runx2 protein levels, CHIP expression decreases, whereas the expression of other E3 ligases involved in Runx2 degradation, such as Smurf1 or WWP1, remains constant or increases during osteoblast differentiation. Depletion of CHIP results in the stabilization of Runx2, enhances Runx2-mediated transcriptional activation, and promotes osteoblast differentiation in primary calvarial cells. In contrast, CHIP overexpression in preosteoblasts causes Runx2 degradation, inhibits osteoblast differentiation, and instead enhances adipogenesis. Our data suggest that negative regulation of the Runx2 protein by CHIP is critical in the commitment of precursor cells to differentiate into the osteoblast lineage.

Prediction of in vivo nephrotoxicity using an in vitro-in silico approach: The case of aristolochic acid I

2015 ◽  
Vol 238 (2) ◽  
pp. S94
Author(s):  
R. Abdullah ◽  
W. Alhusainy ◽  
I.M.C. Rietjens ◽  
A. Punt
Keyword(s):  
In Silico ◽  
Aristolochic Acid ◽  
Aristolochic Acid I

scholarly journals Macrophages-Derived, LRG1-Enriched Extracellular Vesicles Exacerbate Aristolochic Acid Nephropathy Via A TGFβR1-Dependent Manner

2021 ◽  
Author(s):  
Wenjuan Jiang ◽  
Jiahui Dong ◽  
Changlin Du ◽  
Chuanting Xu ◽  
Songbing Xu ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aristolochic Acid ◽  
Herbal Medicines ◽  
Kidney Injury ◽  
Dependent Manner ◽  
Aristolochic Acid Nephropathy ◽  
Injury Model ◽  
Culture Model

Abstract Aristolochic acid nephropathy (AAN) is a progressive kidney disease caused by some herbal medicines, but treatment remains ineffective. We previously found NADPH oxidases 4 (NOX4), which regulates oxidative stress, play an important role in kidney injury model. However, its regulatory mechanism of action in AAN is still obscure. In this study, we established AAN model in vivo, a co-culture system of macrophage and TEC, and macrophage/TEC conditioned media culture model in vitro respectively. We found macrophages infiltration promoted injury，oxidative stress and apoptosis of TEC. Furthermore, the role of macrophage in AAN was dependent on macrophages-derived EV. Importantly, we found that macrophages-derived, Leucine-rich α-2-glycoprotein 1(LRG1)-enriched EV induced TEC injury and apoptosis of via a TGFβR1-dependent process. Mechanistically, macrophages-derived, LRG1-enriched EV mediating TECs injury by upregulating NOX4 in AAN model. This study may help design a better therapeutic strategy to treat AAN patients.

scholarly journals Targeting the Tumour-Specific Spliceosome Through In Silico Virtual Screening for Discovery of New SF3B1 Small Molecule Inhibitors

2018 ◽  
Vol 4 (Supplement 2) ◽  
pp. 201s-201s
Author(s):  
L.Z. Wong
Keyword(s):  
Virtual Screening ◽  
Vitamin D3 ◽  
Transcriptional Activation ◽  
Messenger Rna ◽  
Binding Free Energy ◽  
Specific Cell ◽  
Breast Cancers ◽  
Eukaryotic Genes

Background: The coding (exon) and noncoding (intron) eukaryotic genes are expressed as precursor messenger RNA (premRNA). mRNA splicing defines the process by which introns are excised from premRNA and flanking exons are ligated together. This process is catalyzed by the spliceosome in which combination of small nuclear ribonucleoproteins (U1, U2, U4, U5, U6) forms a spliceosome. The SF3B1 protein is a core component of the U2 snRNP that binds to the branch site and facilitate RNA splicing. Recent studies have identified mutations and dysregulation of in SF3B1 activities in subsets of human cancers including chronic lymphocytic leukemias (CLLs), uveal melanomas, pancreatic cancers and breast cancers. Despite promising in vivo results indicating the potential of spliceosome modulators in targeting the refractory breast cancers, the preclinical and clinical development of such modulators will take several years to complete. Aim: In the current study, we sought to identify new SF3B1 modulators using massive virtual screening of FDA-approved drugs or novel agents for drug repurposing. Methods: A total of 3000 compounds were screened and the hits were identified based on the binding free energy (kcal/mol) of the molecules to the predicted binding sites. Of the 90 hits, vitamin D3 and its analogs (calcipotriol and calcitriol) were identified as a putative SF3B1 modulators. Results: Further in vitro testing revealed that vitamin D3 and its analogs induced significant mRNA misplicing and tumor-specific cell death in MCF7 and MDA-MB-468. Further analyses revealed that vitamin D3 and its analogs significantly reduce SF3B1 protein expression with no changes in its mRNA expression. Conclusion: These results suggest that vitamin D3 and its analogs might interact with SF3B1 to induce protein degradation rather than transcriptional activation.

scholarly journals Deficiency of Mettl3 in Bladder Cancer Stem Cells Inhibits Bladder Cancer Progression and Angiogenesis

2021 ◽  
Vol 9 ◽  
Author(s):  
Ganping Wang ◽  
Yarong Dai ◽  
Kang Li ◽  
Maosheng Cheng ◽  
Gan Xiong ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Progression ◽  
Messenger Rna ◽  
Conditional Knockout ◽  
Stem Cell Population ◽  
Protein Levels ◽  
Rna Immunoprecipitation ◽  
Endothelial Growth Factor

RNA N6-methyladenosine is a key step of posttranscriptional modulation that is involved in governing gene expression. The m6A modification catalyzed by Mettl3 has been widely recognized as a critical epigenetic regulation process for tumorigenic properties in various cancer cell lines, including bladder cancer. However, the in vivo function of Mettl3 in bladder cancer remains largely unknown. In our study, we found that ablation of Mettl3 in bladder urothelial attenuates the oncogenesis and tumor angiogenesis of bladder cancer using transgenic mouse model. In addition, conditional knockout of Mettl3 in K14+ bladder cancer stem cell population leads to inhibition of bladder cancer progression. Coupled with the global transcriptome sequencing and methylated RNA immunoprecipitation sequencing results, we showed that deletion of Mettl3 leads to the suppression of tyrosine kinase endothelial (TEK) and vascular endothelial growth factor A (VEGF-A) through reduced abundance of m6A peaks on a specific region. In addition, the depletion of Mettl3 results in the decrease in both messenger RNA (mRNA) and protein levels of TEK and VEGF-A in vitro. Taken together, Mettl3-mediated m6A modification is required for the activation of TEK–VEGF-A-mediated tumor progression and angiogenesis. Our findings may provide theoretical basis for bladder cancer treatment targeting Mettl3.

scholarly journals Nephrotoxicity Biomarkers: Role and Significance in the Diagnosis of Drug-Induced Kidney Injury

2021 ◽  
Vol 9 (4) ◽  
pp. 173-184
Author(s):  
O. V. Muslimova ◽  
V. A. Evteev ◽  
I. A. Mazerkina ◽  
E. A. Sokova ◽  
A. B. Prokofiev ◽  
...  
Keyword(s):  
Kidney Injury ◽  
Experimental Models ◽  
Injury Incidence ◽  
Drug Induced ◽  
Blood Concentrations ◽  
Incidence And Mortality ◽  
Hospital Patients

Drug-induced kidney injury (DIKI) accounts for 8 to 60% of episodes of acute kidney injury (AKI) among hospital patients. Early DIKI detection and timely adjustment of therapy will help reduce the kidney injury incidence and mortality. The aim of the study was to analyse scientific literature on the biomarkers used in DIKI diagnosis. The study revealed that the use of such kidney damage markers as serum creatinine, urinary output, urea nitrogen, sodium excretion, urinary sediment microscopy is limited because they do not give a full picture of the kidney injury degree and progression and do not allow for early AKI diagnosis. It was demonstrated that some of the most promising biomarkers are KIM-1, L-FABP, NAG, NGAL, cystatin C, clusterin, β2-microglobulin, МСР-1, IGFBP7, and TIMP-2. However, recommendations for determination of these biomarkers’ urine or blood concentrations for AKI diagnosis are somewhat preliminary, because there have been insufficient clinical and preclinical studies to establish validity of such tests. No precise algorithms based on determination of the biomarkers levels in urea and/or blood serum have been developed for AKI risk assessment, diagnosis, monitoring, and treatment. Thus, further research is necessary to investigate different AKI biomarkers and improve experimental models (both in vivo and in vitro), which will support assessment of potential nephrotoxic properties of existing and new medicinal products.

scholarly journals Drug-induced thrombocytopenia is associated with increased binding of IgG to platelets both in vivo and in vitro

Blood ◽  
1981 ◽  
Vol 58 (3) ◽  
pp. 524-529 ◽  
Author(s):  
JG Kelton ◽  
D Meltzer ◽  
J Moore ◽  
AR Giles ◽  
WE Wilson ◽  
...  
Keyword(s):  
Diagnostic Techniques ◽  
In Vitro Assay ◽  
Suspected Drug ◽  
Drug Induced ◽  
Vitro Assay ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
In Vitro Diagnostic

Abstract Thrombocytopenia is a common serious adverse effect of drug treatment. A variety of in vitro diagnostic techniques to confirm the diagnosis are available, but the majority lack sufficient sensitivity to detect all cases of drug-induced thrombocytopenia. We studied 19 patients with suspected drug-induced thrombocytopenia and demonstrated that platelet- associated IgG (PAIgG) was elevated in all at the time of thrombocytopenia, and PAIgG returned to normal levels as the thrombocytopenia resolved. In the majority of patients, the platelet count rapidly returned to normal after the drug was discontinued; however, in six patients, the thrombocytopenia persisted well beyond the period of time that the offending drug would be expected to be cleared from the blood. In 13 patients, serum obtained after recovery was used to identify the drug responsible for the thrombocytopenia in an in vitro assay. In all cases, the addition of the drug historically associated with the thrombocytopenic episode was associated with an increased binding of IgG to control platelets. For uncertain reasons, the concentration of drug required to increase the in vitro binding of IgG to test platelets was often more than the concentration usually achieved in vivo. Wider application of these techniques may provide better understanding of the clinical characteristics and mechanisms responsible for drug-induce thrombocytopenia.

DNA adduct formation of aristolochic acid I and II in vitro and in vivo

1988 ◽  
Vol 9 (2) ◽  
pp. 297-303 ◽  
Author(s):  
H.H. Schmeiser ◽  
K.-B. Schoepe ◽  
M. Wiessler
Keyword(s):  
Aristolochic Acid ◽  
Adduct Formation ◽  
Dna Adduct ◽  
Aristolochic Acid I

scholarly journals Drug-induced thrombocytopenia is associated with increased binding of IgG to platelets both in vivo and in vitro

Blood ◽  
1981 ◽  
Vol 58 (3) ◽  
pp. 524-529 ◽  
Author(s):  
JG Kelton ◽  
D Meltzer ◽  
J Moore ◽  
AR Giles ◽  
WE Wilson ◽  
...  
Keyword(s):  
Diagnostic Techniques ◽  
In Vitro Assay ◽  
Suspected Drug ◽  
Drug Induced ◽  
Vitro Assay ◽  
In Vitro Binding ◽  
Vitro Binding ◽  
In Vitro Diagnostic

Thrombocytopenia is a common serious adverse effect of drug treatment. A variety of in vitro diagnostic techniques to confirm the diagnosis are available, but the majority lack sufficient sensitivity to detect all cases of drug-induced thrombocytopenia. We studied 19 patients with suspected drug-induced thrombocytopenia and demonstrated that platelet- associated IgG (PAIgG) was elevated in all at the time of thrombocytopenia, and PAIgG returned to normal levels as the thrombocytopenia resolved. In the majority of patients, the platelet count rapidly returned to normal after the drug was discontinued; however, in six patients, the thrombocytopenia persisted well beyond the period of time that the offending drug would be expected to be cleared from the blood. In 13 patients, serum obtained after recovery was used to identify the drug responsible for the thrombocytopenia in an in vitro assay. In all cases, the addition of the drug historically associated with the thrombocytopenic episode was associated with an increased binding of IgG to control platelets. For uncertain reasons, the concentration of drug required to increase the in vitro binding of IgG to test platelets was often more than the concentration usually achieved in vivo. Wider application of these techniques may provide better understanding of the clinical characteristics and mechanisms responsible for drug-induce thrombocytopenia.

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