Exposure to High-Altitude Environment is Associated with Drug Transporters Change: miR-873-5p-Mediated Alteration of Function and Expression Levels of Drug Transporters under Hypoxia

Abstract Abstract 972 Successful treatment of diffuse large B-cell lymphoma (DLBCL) is frequently hindered by development of resistance to conventional chemotherapy resulting in disease relapse and high mortality. High expression of anti-apoptotic and/or drug transporter proteins induced by oncogenic signaling pathways has been implicated in the development of chemoresistance in cancer. However, the cellular mechanisms responsible for drug resistance and treatment failure in DLBCL are poorly understood and their elucidation could lead to development of new therapeutic approaches. Activated Hedgehog (HH) signaling was identified in our laboratory to contribute to DLBCL cell survival and proliferation. We also found that inhibiting HH signaling resulted in decrease of BCL-2 protein and mRNA levels in ABC DLBCL cells but not in GC DLBCL cells, and that the resistance to apoptosis to HH inhibition observed in GC DLBCL cells can be overcome using the BCL2 inhibitor YC137. (Leukemia 2010;24:1025). We also found that the drug transporter protein ABCG2 is a potential prognostic factor in DLBCL given that its expression (as detected by immunohistochemistry) inversely correlated with overall survival and failure-free survival (Mod Pathol 2009;22:1312). Here, we investigate potential mechanistic connections between HH signaling and chemoresistance in DLBCL. Using chromatin immunoprecipitation (ChIP), site directed mutagenesis and luciferase reporter based assays, we confirmed the presence of a GLI-1 transcription factor-binding site in the ABCG2 gene promoter and we established ABCG2 gene as a direct downstream target of HH signaling. We also characterized the baseline expression and functionality of ABCG2 in DLBCL cell lines and explored the role of stroma in modulating its expression levels as well as the expression of other drug transporters (MDR1 and ABCC1) and the levels of the anti-apoptotic proteins, BCL-2, BCL2A1 and BCL-xL. Co-culturing DLBCL cell lines (BJAB and DOHH2) with human bone marrow stromal cells (HS5) resulted in decreased chemosensitivity to doxorubicin and methotrexate that was associated to marked increased expression levels of the drug transporters ABCG2, MDR1 and ABCC1 as well as of the expression levels of BCL2, BCL2A1 and BCL-xL. Our results also showed that pharmacologic inhibition of the activity of ABCG2, using fumitremorgin C and glefanine, significantly increased the chemosensitivity of BJAB and DOHH2 cells in the presence of stromal cells. However, this effect was not seen in the absence of stromal cells. Collectively, our data confirm that the stromal cells plays a role in inducing chemoresistance in DLBCL and that this effect is in part mediated by inducing high expression of drug protein transporters and antiapoptotic proteins. Inhibition of HH signaling as well as inhibition of drug transporters abrogates the stroma-induced chemoresistance suggesting that targeting these molecules may have a therapeutic value for the treatment of DLBCL. Disclosures: No relevant conflicts of interest to declare.

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Hepatitis C Virus-related Cirrhosis is a Major Determinant of the Expression Levels of Hepatic Drug Transporters

Drug Metabolism and Pharmacokinetics ◽

10.2133/dmpk.25.190 ◽

2010 ◽

Vol 25 (2) ◽

pp. 190-199 ◽

Cited By ~ 49

Author(s):

Ken Ogasawara ◽

Tomohiro Terada ◽

Toshiya Katsura ◽

Etsuro Hatano ◽

Iwao Ikai ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Drug Transporters ◽

Major Determinant ◽

Hepatic Drug ◽

Expression Levels

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O30 A comprehensive analysis of ontogeny of renal drug transporters: mRNA analyses, quantitative proteomics and localization

Archives of Disease in Childhood ◽

10.1136/archdischild-2019-esdppp.30 ◽

2019 ◽

Vol 104 (6) ◽

pp. e13.2-e13

Author(s):

KWK Cheung ◽

BD van Groen ◽

E Spaans ◽

MD van Borselen ◽

ACJM de Bruijn ◽

...

Keyword(s):

Drug Transporters ◽

Mrna Levels ◽

Potential Conflict ◽

Renal Handling ◽

Expression Levels ◽

Protein Levels ◽

Renal Transporters ◽

Age Related ◽

Exogenous Compounds ◽

Endogenous Substrates

BackgroundPostnatal developmental changes of human renal membrane transporters, which are key players of disposition of renally cleared drugs and endogenous substrates, are largely unknown. This study aimed to characterize the ontogeny of 11 human renal transporters to understand changes in the renal clearance of substrate drugs in children.Methods mRNA levels of known renal transporters: BCRP, MATE1, MATE2-K, MDR1, MRP2, MRP4, URAT1, GLUT2, OAT1, OAT3 and OCT2, and the transcription factor PXR were measured with RT-qPCR in 184 human postmortem frozen renal cortical tissues (preterm newborns - adults; 1 day-75 yrs old) from individuals of European and African descent. Protein expression of all but MRP2, MRP4 and PXR was quantified with LC-MS/MS SRM in 62 of those samples (term newborns - adults; 1 day-29 yrs old). Localization of MRP4 was tested with immunohistochemistry.ResultsExpression levels of MDR1, URAT1, OAT1, OAT3, and OCT2 increased with age, but levels of MATE1 and GLUT2 were stable from birth. Protein levels of MATE2-K and BCRP showed no difference from newborns to adults despite age-related changes in mRNA expression. MRP2, MRP4 and PXR expression levels were stable. MRP4 localization in pediatric samples was similar to that in adult samples.ConclusionRenal drug transporters exhibited different rates and patterns of maturation, suggesting that renal handling of both endogenous and exogenous compounds may change with age. It is important to consider ontogeny of renal transporters during pediatric drug development.Disclosure(s)The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Views expressed in this paper are those of authors and do not necessarily reflect the official views or policies of the FDA; nor does any mention of trade names, commercial practices, or organization imply endorsement by the U.S. Government.*Contributed equally,**Contributed equally

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Comparative analysis of skeleton muscle proteome profile between yak and cattle provides insight into high-altitude adaptation

Current Proteomics ◽

10.2174/1570164617666200127151931 ◽

2020 ◽

Vol 17 ◽

Author(s):

Jin-Wei Xin ◽

Zhi-Xin Chai ◽

Cheng-Fu Zhang ◽

Yu-Mei Yang ◽

Qiang Zhang ◽

...

Keyword(s):

High Altitude ◽

Pyruvate Dehydrogenase ◽

Molecular Mechanisms ◽

Expression Profiles ◽

Pyruvate Dehydrogenase Complex ◽

Pulmonary Arteries ◽

Adenosine Monophosphate ◽

Oxygen Utilization ◽

Antibody Microarray ◽

Expression Levels

Background:: Mechanisms underlying yak adaptation to high-altitude environments have been investigated at levels of morphology, anatomy, physiology, genome and transcriptome, but not at the proteome level. Objective: To explore for the first time the protein expression profiles in yak to reveal molecular mechanisms underlying adaptation to high altitude, up to now investigated by genome sequencing. Methods: In the present study, an antibody microarray chip was developed, which included 6,500 mouse monoclonal antibodies. Immunoprecipitation and mass spectrometry on 12 selected antibodies showed that the chip was highly specific. Using this chip, muscle tissue proteome was compared between yak and cattle, and 12 significantly differentially expressed proteins (DEPs) were identified and their expression levels were validated by Western blot. Results: Compared with cattle, higher levels of Rieske iron-sulfur protein (RISP), Cytochrome C oxidase subunit 4 isoform 1, mitochondrial (COX4I1), ATP synthase F1 subunit beta (ATP5F1B), Sarcoplasmic/endoplasmic reticulum calcium ATPase1 (SERCA1) and Adenosine monophosphate deaminase1 (AMPD1) in yak might increase oxygen utilization and energy metabolism. Pyruvate dehydrogenase protein X component (PDHX) and Acetyltransferase component of pyruvate dehydrogenase complex (DLAT) showed higher expression levels and L-lactate dehydrogenase A chain (LDHA) showed lower expression level, which might help yak reduce accumulation of lactic acid. In addition, higher expression levels of Filamin C (FLNC) and low levels of AHNAK and Four and a half LIM domains 1(FHL1) in yak might contribute to inhibition of pulmonary arteries vasoconstriction, remodeling and hypertension. Conclusion: Overall, the present study reported new data at protein level in comparison between yak and cattle, which might be helpful to further understand molecular mechanisms underlying yak adaptation to high altitude environments.

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Effects of Metabolic Acidosis on Expression Levels of Renal Drug Transporters

Pharmaceutical Research ◽

10.1007/s11095-010-0348-7 ◽

2010 ◽

Vol 28 (5) ◽

pp. 1023-1030 ◽

Cited By ~ 10

Author(s):

Arong Gaowa ◽

Hideyuki Motohashi ◽

Toshiya Katsura ◽

Ken-ichi Inui

Keyword(s):

Metabolic Acidosis ◽

Drug Transporters ◽

Expression Levels

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Regional Expression Levels of Drug Transporters and Metabolizing Enzymes along the Pig and Human Intestinal Tract and Comparison with Caco-2 Cells

Drug Metabolism and Disposition ◽

10.1124/dmd.116.072231 ◽

2017 ◽

Vol 45 (4) ◽

pp. 353-360 ◽

Cited By ~ 29

Author(s):

Stefan F.C. Vaessen ◽

Marola M.H. van Lipzig ◽

Raymond H.H. Pieters ◽

Cyrille A.M. Krul ◽

Heleen M. Wortelboer ◽

...

Keyword(s):

Intestinal Tract ◽

Drug Transporters ◽

Metabolizing Enzymes ◽

Expression Levels ◽

Human Intestinal Tract

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Expression profile of hepatic drug transporters in patients with hepatocellular carcinoma

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.e22026 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. e22026-e22026

Author(s):

K. Higuchi ◽

T. Terada ◽

K. Ogasawara ◽

M. Aoki ◽

M. Fukudo ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Dna Methylation ◽

Mrna Expression ◽

Expression Profile ◽

Promoter Region ◽

Drug Transporters ◽

Hepatic Drug ◽

Expression Levels ◽

Mrna Expression Levels ◽

Liver Tissues

e22026 Background: Hepatocellular carcinoma (HCC) does not respond sufficiently to chemotherapy. It has been demonstrated that several transporters can cause drug resistance in tumor cells. In human liver, various drug transporters are expressed and play pivotal roles in hepatic uptake and excretion of drugs. In HCC, the expression profile of hepatic drug transporters may be altered due to pathophysiological changes. However, limited data are available on their expression and regulation in HCC tissues. The aims of this study were to determine the expression levels of hepatic drug transporters in HCC patients and to identify transporter specifically regulated in HCC tissues. Methods: The mRNA expression levels of 17 drug transporters, including 9 solute carrier (SLC) transporters (OAT2, OAT7, OATP1B1, OATP1B3, OATP2B1, OCT1, OCTN2, MATE1 and PEPT1) and 8 ATP-binding cassette (ABC) transporters (R1, MRP1–6 and BCRP), were determined by real-time RT-PCR in cancerous and non-cancerous liver tissues from 57 patients with HCC. Protein expression level was evaluated by Western blot analysis. In addition, DNA methylation status in promoter region of transporter gene was analyzed using bisulfite sequencing. Results: The mRNA expression levels of 11 transporters (OAT7, OCTN2, MATE1, PEPT1, R1, MRP1–6) were significantly increased (1.45 to 4.74-fold, p<0.05) in HCC tissues compared with non-cancerous liver tissues, whereas only OCT1 mRNA was significantly decreased (0.61-fold, p=0.0004). Notably, MRP4 mRNA showed the highest increase, which was consistent with the marked augmentation in the protein level. The frequency of methylation in MRP4 promoter region was comparable between HCC and non-cancerous liver tissues, suggesting other mechanisms than DNA methylation for MRP4 up-regulation. Interestingly, the mRNA expression levels of MRP4 did not change with the differentiation of HCC. Conclusions: It was revealed that most of the hepatic drug transporters were elevated in HCC tissues. Furthermore, we first identified that MRP4 showed remarkable increase in protein as well as mRNA expression levels. These findings suggest that MRP4 is a novel candidate for diagnostic marker and a target molecule to reverse multidrug resistance in chemotherapy for HCC. No significant financial relationships to disclose.

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