DOCSLIB.ORG

The Pharmacogenomics of Vincristine-Induced Neurotoxicity

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

THE PHARMACOGENOMICS OF VINCRISTINE-INDUCED NEUROTOXICITY
IN PAEDIATRIC CANCER PATIENTS WITH WILMS TUMOR OR
RHABDOMYOSARCOMA

by
Tenneille Loo

A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF
THE REQUIREMENTS FOR THE DEGREE OF

MASTER OF SCIENCE in
THE FACULTY OF GRADUATE STUDIES
(Experimental Medicine)

THE UNIVERSITY OF BRITISH COLUMBIA
(Vancouver)

July 2011
© Tenneille Loo, 2011

Abstract

Vincristine is one of the most effective and widely utilized antineoplastic agents.
However, the clinical utility of this drug is limited by severely debilitating vincristineinduced neurotoxicities (VIN). Previous studies have associated VIN with genetic polymorphisms in genes involved in the metabolism and transportation of vincristine, including CYP3A4, CYP3A5, and ABCB1. However, the findings of such studies have not been consistently reproduced. This study hypothesizes that there are specific variants in genes involved in general drug absorption, metabolism, distribution, excretion, and toxicity (ADME-Tox) that affect the individual susceptibility to VIN in patients with Wilms tumor and rhabdomyosarcoma.
Detailed clinical data was collected from 140 patients with Wilms tumor and rhabdomyosarcoma by retrospective chart review. VIN cases were characterized by type of neurotoxicity, and severity was evaluated using a validated clinical grading system for adverse events (NCI-CTCAE v4.03). A customized Illumina GoldenGate Panel was used to genotype 4,536 single nucleotide polymorphisms (SNPs) in candidate genes involved in the metabolism and transportation pathway of vincristine, as well as in genes broadly involved in ADME-Tox.
None of the SNPs that were previously reported to be associated with VIN were found to be significantly associated (p-value < 0.05). With similar effect sizes, six novel

genetic variants in five genes (PON1, ABCA4, ABCG1, CY51A1, SLCO1C1) were

significantly associated with VIN in both tumor types. Whereas none of these genes have been previously associated with VIN or the biotransformation of vincristine, interestingly,

ii the biological functions of the encoded proteins have been indirectly linked to nerve function, neuropathy, or neurodegenerative diseases. Therefore, I hypothesize that the genetic basis of VIN is likely polygenic and that the six genes influence individual susceptibility to VIN by affecting: nerve regeneration (PON1, PPP1R9A), and cholesterol homeostasis and remyelination (ABCA4, ABCG1, CYP51A1), as well as the metabolism of vincristine (PON1, CYP51A1) and the transportation of lipids, vincristine, metabolites, or neuroprotectants

(SLCO1C1, ABCA4, ABCG1).

This study adds to the literature by identifying new potential biomarkers for VIN, providing novel hypotheses for the mechanisms underlying VIN susceptibility, and is a point of origin for replication studies.

iii

Preface

The work contained in this thesis has been approved by the University of British
Columbia and the Children’s & Women’s Health Centre of BC Research Ethics Board, the Ethics Certificate Numbers for this study are H10-01568 and H04-70358.

iv

Table of Contents

Abstract ................................................................................................................................... ii Preface .....................................................................................................................................iv Table of Contents.....................................................................................................................v List of Tables............................................................................................................................x List of Figures .........................................................................................................................xi List of Symbols and Abbreviations..................................................................................... xii Acknowledgements............................................................................................................. xvii Chapter 1: Introduction.........................................................................................................1

1.1 Adverse Drug Reactions...............................................................................................1
1.1.1 Factors that Contribute to Adverse Drug Reactions...............................................2 1.1.2 Adverse Drug Reactions in Children......................................................................2 1.1.3 Pharmacogenomics.................................................................................................4
1.2 Vincristine ....................................................................................................................5
1.2.1 The Antineoplastic Mechanism of Action..............................................................6 1.2.2 Metabolism and Transportation..............................................................................6 1.2.3 Clinical Use of Vincristine in Children ..................................................................9
1.3 Vincristine-Induced Neurotoxicity.............................................................................10
1.3.1 Signs and Symptoms ............................................................................................10 1.3.2 Clinical Diagnosis of Vincristine-Induced Neurotoxicity....................................11 1.3.3 Management .........................................................................................................13 1.3.4 Future Management: Neuroprotectants ................................................................14

v
1.3.5 Pathophysiology ...................................................................................................15
1.4 Factors Affecting the Exposure to Vincristine ...........................................................17
1.4.1 Dose Intensity, Frequency, and Cumulative Dose................................................17 1.4.2 CYP3A Inhibitors.................................................................................................19 1.4.3 Genetic Variation in CYP3A4 and CYP3A5, and Ancestry ................................20 1.4.4 Vincristine Transporters .......................................................................................21 1.4.5 Previous Pharmacogenomic Studies.....................................................................22
1.5 Patient Cohort Selection: Wilms Tumor and Rhabdomyosarcoma............................24

Chapter 2: Hypothesis, Project Goal and Objectives........................................................28

2.1 Hypothesis ..................................................................................................................28 2.2 Project Goal and Objectives .......................................................................................28

Chapter 3: Methodology ......................................................................................................29

3.1 Recruitment, Consent, and Establishing Causality.....................................................29 3.2 Defining Cases and Controls......................................................................................31 3.3 Clinical Characterization of Vincristine-induced Neurotoxicity Cases .....................33
3.3.1 Classifying Each Vincristine-Induced Neurotoxicity Event by Type of
Neuropathy ...........................................................................................................33
3.3.2 Evaluating the Severity of Each Vincristine-Induced Neurotoxicity Event.........33
3.4 Study Inclusion and Exclusion Criteria......................................................................40 3.5 Calculating Dose and Time to First Adverse Drug Reaction.....................................41 3.6 Association Studies.....................................................................................................42
3.6.1 General Drug Biotransformation Genotyping ......................................................42 3.6.2 Candidate Gene Analysis......................................................................................43

vi
3.7 Genotyping: Illumina GoldenGate Assay...................................................................43 3.8 Quality Control and Assurance in Genotypic Data ....................................................44
3.8.1 Quality of Patient Samples and SNPs...................................................................44 3.8.2 Data Cleaning .......................................................................................................45
3.9 Statistical Analyses: Logistic Regression Models and Manhattan Plot .....................46 3.10 Prediction Model ........................................................................................................48 3.11 Linkage Disequilibrium Studies.................................................................................49 3.12 Imputation Analyses...................................................................................................49

Chapter 4: Results ................................................................................................................51

4.1 Patient Cohort.............................................................................................................51 4.2 Patient Demographics and Clinical Factors................................................................52 4.3 Assessing the Severity and Types of Vincristine-Induced Neurotoxicity and the
Interventions Provided ...............................................................................................55
4.4 Variation by Ancestry.................................................................................................60 4.5 Genetic Analyses........................................................................................................62
4.5.1 Screening for Genes Involved in the General Drug Biotransformation Pathways
.............................................................................................................................63
4.5.2 Candidate Study....................................................................................................78

Chapter 5: Discussion...........................................................................................................84

5.1 The Incidence of Vincristine-Induced Neurotoxicity.................................................84 5.2 Clinical Factors...........................................................................................................85
5.2.1 Age........................................................................................................................85 5.2.2 Tumor Type ..........................................................................................................87

vii
5.2.3 Ancestry................................................................................................................88 5.2.4 Concomitant Medications: Steroids and CYP3A4 Inhibitors...............................88
5.3 Vincristine-Induced Neurotoxicity: Types of Neurotoxicity and Severity ................89 5.4 Novel Genes and Genetic Variants Potentially Associated with Vincristine-Induced
Neurotoxicity..............................................................................................................90
5.4.1 PON1 (Paraoxonase 1) and PPP1R9A (Neurabin 1)............................................92 5.4.2 ABCA4, ABCG1, and CYP51A1 .........................................................................102 5.4.3 SLCO1C1............................................................................................................108
5.5 Previously Identified Polymorphisms and Candidate Genes Involved in the
Biotransformation of Vincristine .............................................................................108
5.5.1 CYP3A5 and CYP3A 4.........................................................................................110 5.5.2 ABCB1 ................................................................................................................111 5.5.3 Other Candidate Genes.......................................................................................112
5.6 Inter-Ethnic Differences ...........................................................................................113 5.7 Limitations................................................................................................................114 5.8 Future Directions......................................................................................................116
5.8.1 Expansion, Replication, and Validation of Novel SNPs and Genes...................116 5.8.2 Detailed Genetic Investigation of Novel Candidate Genes................................117 5.8.3 Pharmacokinetic and Functional Validation Studies..........................................118
5.8.3.1 PON1 (Paraoxonase 1) and PPP1R9A (Neurabin 1)...................................119 5.8.3.2 ABCA4, ABCG1, and CYP51A1: Cholesterol Biosynthesis and Intracellular
Transport......................................................................................................120
5.8.3.3 Metabolizers and Transporters ....................................................................121

viii
5.8.4 Investigation of Additional Genes Potentially Involved in VIN ........................121 5.8.5 Observational Trials............................................................................................123
5.9 Future Implications...................................................................................................123

Chapter 6: Conclusion........................................................................................................126 References.............................................................................................................................128

Appendix A........................................................................................................................139
A.1 Excluded SNPs: Less than 98% Completion Rate Across Patients.....................139 A.2 Excluded SNPs: Hardy-Weinberg Disequilibrium..............................................140 A.3 Significant SNPs in the ADME-Tox Panel of General Drug Biotransformation141

A.4 PON1, ABCA4, SLCO1C1, CYP51A1, and ABCG1 Risk Alleles Increase the

Chances of Developing Vincristine-Induced Neurotoxicity...............................146
A.5 Imputed Regions with Base Pair Limits Used.....................................................147 A.6 Manhattan Plot for Imputed SNPs.......................................................................148

ix

List of Tables

Table 1 Grading Vincristine-Induced Neurotoxicity with the CTCAE Criteria (v4.03)....34 Table 2 Grading Vincristine-Induced Neurotoxicity by Intervention ................................38 Table 3 Patient Demographics and Clinical Factors Between Cases and Controls............53 Table 4 Vincristine-Induced Neurotoxicity and the Interventions Provided to Wilms
Tumor and Rhabdomyosarcoma Cases ................................................................56
Table 5 The Frequency of Each Type of Vincristine-Induced Neurotoxicity Sign or
Symptom...............................................................................................................58
Table 6 Genetic Variants that are Significantly Associated with Vincristine-Induced
Neurotoxicity........................................................................................................65
Table 7 Subgroup Analysis of Genetic Variants that are Significantly Associated with
Vincristine-Induced Neurotoxicity: Stratification of Patients by European Ancestry................................................................................................................69
Table 8 Linkage Disequilibrium of SNP Pairs: PON1, PPP1R9A, PON1, and ABCA 4....71 Table 9 PON1 rs854549, PPP1R9A rs705377, and PON1 rs854560 and their Association with Vincristine-Induced Neurotoxicity...............................................................76
Table 10 The Effects of SNPs Previously Identified to be Associated with Vincristine-
Induced Neurotoxicity ..........................................................................................79
Table 11 The Effects of Previously Identified Genes Involved in the Metabolism and
Transportation Pathway of Vincristine.................................................................82

x

List of Figures

Figure 1 The Pharmacokinetics and Pharmacodynamics of Vincristine...............................7 Figure 2 Ethnic Distribution of the Patients in the Cohort..................................................60 Figure 3 Manhattan Plot ......................................................................................................63

  • Figure 4
  • SNP Clusters of PON1 rs854549, ABCA4 rs3789433, SLCO1C1 rs10770704,

ABCG1 rs221948, CYP51A1 rs7797834, and ABCA4 rs549848..........................66

Figure 5 PON1, ABCA4, SLCO1C1, CYP51A1, and ABCG1 Risk Alleles Increase the

Chances of Developing Vincristine-Induced Neurotoxicity.................................69
Figure 6 Linkage Disequilibrium Plot of PON1, PPP1R9A, and the Intergenic Region in the European (CEU) Hapmap Populations...........................................................71
Figure 7 Linkage Disequilibrium Plot of PON1, PPP1R9A, and the Intergenic Region in the African (YRI) Hapmap Populations ...............................................................72
Figure 8 Linkage Disequilibrium Plot of PON1, PPP1R9A, and the Intergenic Region in the Asian (CHB, JPT) Hapmap Populations.........................................................73
Figure 9 SNP Clusters of PPP1R9A rs705377 and PON1 rs854560 ..................................76 Figure 10 Clustering of Candidate SNPs: CYP3A5 and CYP3A4........................................79 Figure 11 Clustering of Candidate SNPs: ABCB 1...............................................................80 Figure 12 The Theorized Role of the Novel Genes in the Metabolism and
Biotransformation of Vincristine..........................................................................90
Figure 13 Chemical Structures of Methyl Phenylacetate, Vincristine, and Clopidogrel......96 Figure 14 Cholesterol Biosynthesis ....................................................................................103

xi

List of Symbols and Abbreviations

ABCA4 ABCB1
ATP-binding cassette, sub-family B, member 4; multidrug resistance protein 3 ATP-binding cassette, sub-family B, member 1; multidrug resistance protein 1; P-glycoprotein
ABCB4 ABCC1
ATP-binding cassette, sub-family B, member 4; multidrug resistance protein 4 ATP-binding cassette, sub-family C, member 1; multiple drug resistance protein 1
ABCC2

ABCC3 ABCC4 ABCC5 ABCC6 ABCC7 ABCC8 ABCC10
ATP-binding cassette, sub-family C, member 2; multiple drug resistance protein 2 ATP-binding cassette, sub-family C, member 3; multiple drug resistance protein 3 ATP-binding cassette, sub-family C, member 4; multiple drug resistance protein 4 ATP-binding cassette, sub-family C, member 5; multiple drug resistance protein 6 ATP-binding cassette, sub-family C, member 6; multiple drug resistance protein 6 ATP-binding cassette, sub-family C, member 7; multiple drug resistance protein 7 ATP-binding cassette, sub-family C, member 8; multiple drug resistance protein 8 ATP-binding cassette, sub-family C, member 10; multiple drug resistance protein 10

xii
ABCG1 ADL
ATP-binding cassette, subfamily G, member 1 Activities of Daily Living

  • ADME
  • Absorption, Distribution, Metabolism, and Excretion

ADME-Tox Absorption, Distribution, Metabolism, Excretion, and Toxicity ADR ALL
Adverse Drug Reaction Acute Lymphoblastic Leukemia

  • Amyotrophic Lateral Sclerosis
  • ALS

  • AN
  • Autonomic Neuropathy

  • ANS
  • Autonomic Nervous System

AUC BSA
Area-Under-the-Concentration Body Surface Area

  • CEU
  • Northern and Western European Ancestry (Utah, United States)

Han Chinese Ancestry (Beijing, China) Chinese Ancestry (Denver, Colorado) Confidence Interval
CHB CHD CI CMT CNS
Charcot Marie Tooth Central Nervous System
COG CPNDS CTEP CTCAE CYP450 CYP3A4
Children’s Oncology Group Canadian Pharmacogenomics Network for Drug Safety Cancer Therapy Evaluation Program Common Terminology Criteria for Adverse Events Cytochromes P450 Cytochrome P450, family 3, subfamily A, polypeptide 4

xiii

  • CYP3A5
  • Cytochrome P450, family 3, subfamily A, polypeptide 5

Cytochrome P450, family 51, subfamily A, polypeptide 1; Lanosterol; 14-α- Demethylase
CYP51A1 DNA DTR
Deoxyribonucleic Acid Deep Tendon Reflex
GWAS HDL HWE IADL IBS
Genome-Wide Association Studies High-density Lipoprotein Hardy-Weinberg Equilibrium Instrumental Activities of Daily Living Identity-by-State

  • IBD
  • Identity-by-Descent

IGF-1 JPT
Insulin-like Growth Factor 1 Japanese Ancestry (Tokyo, Japan)

  • Linkage Disequilibrium
  • LD

m-ABC MAF MAPT MAP2 Nb1
Motor Assessment Battery for Children Minor Allele Frequency Microtubule-Associated Protein Tau Microtubule-Associated Protein 2 Neurabin 1 ncRNA NCS
Non-Protein-Coding RNA Nerve Conduction Studies
NCI-CTCAE National Cancer Institute – Common Terminology Criteria for Adverse Events NGF Nerve Growth Factor

Recommended publications
  • ABCG1 (ABC8), the Human Homolog of the Drosophila White Gene, Is a Regulator of Macrophage Cholesterol and Phospholipid Transport

    ABCG1 (ABC8), the Human Homolog of the Drosophila White Gene, Is a Regulator of Macrophage Cholesterol and Phospholipid Transport

    ABCG1 (ABC8), the human homolog of the Drosophila white gene, is a regulator of macrophage cholesterol and phospholipid transport Jochen Klucken*, Christa Bu¨ chler*, Evelyn Orso´ *, Wolfgang E. Kaminski*, Mustafa Porsch-Ozcu¨ ¨ ru¨ mez*, Gerhard Liebisch*, Michael Kapinsky*, Wendy Diederich*, Wolfgang Drobnik*, Michael Dean†, Rando Allikmets‡, and Gerd Schmitz*§ *Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, 93042 Regensburg, Germany; †National Cancer Institute, Laboratory of Genomic Diversity, Frederick, MD 21702-1201; and ‡Departments of Ophthalmology and Pathology, Columbia University, Eye Research Addition, New York, NY 10032 Edited by Jan L. Breslow, The Rockefeller University, New York, NY, and approved November 3, 1999 (received for review June 14, 1999) Excessive uptake of atherogenic lipoproteins such as modified low- lesterol transport. Although several effector molecules have been density lipoprotein complexes by vascular macrophages leads to proposed to participate in macrophage cholesterol efflux (6, 9), foam cell formation, a critical step in atherogenesis. Cholesterol efflux including endogenous apolipoprotein E (10) and the cholesteryl mediated by high-density lipoproteins (HDL) constitutes a protective ester transfer protein (11), the detailed molecular mechanisms mechanism against macrophage lipid overloading. The molecular underlying cholesterol export in these cells have not yet been mechanisms underlying this reverse cholesterol transport process are characterized. currently not fully understood. To identify effector proteins that are Recently, mutations of the ATP-binding cassette (ABC) trans- involved in macrophage lipid uptake and release, we searched for porter ABCA1 gene have been causatively linked to familial HDL genes that are regulated during lipid influx and efflux in human deficiency and Tangier disease (12–14).
  • Ligand, Receptor, and Cell Type–Dependent Regulation of ABCA1 and ABCG1 Mrna in Prostate Cancer Epithelial Cells

    Ligand, Receptor, and Cell Type–Dependent Regulation of ABCA1 and ABCG1 Mrna in Prostate Cancer Epithelial Cells

    Published OnlineFirst June 16, 2009; DOI: 10.1158/1535-7163.MCT-09-0020 Published Online First on June 16, 2009 as 10.1158/1535-7163.MCT-09-0020 OF1 Ligand, receptor, and cell type–dependent regulation of ABCA1 and ABCG1 mRNA in prostate cancer epithelial cells Steven E. Trasino,1 Young S. Kim,2 that ABCA1 gene expression is differentially regulated by and Thomas T.Y. Wang1 synthetic and natural LXR ligands, possibly involving kinase mediated signal transduction. [Mol Cancer Ther 2009;8(7): 1 Diet, Genomics, and Immunology Laboratory, Beltsville Human OF1–12] Nutrition Research Center, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, Maryland and 2Nutritional Sciences Research Group, Division of Cancer Introduction Prevention, National Cancer Institute, NIH, Bethesda, Maryland Advanced and metastatic prostate cancer (PCa) remains the most prominent cancer type and the second leading cause of Abstract all cancer deaths among males in the United States (1). Recent evidence suggests that the liver X receptor (LXR) Treatment of recurrent or advanced PCa with hormone ab- is a potential anticancer target in prostate carcinoma. lation therapy typically results in disease regression with There is little characterization, however, of which of the eventual recurrence of PCa with a more aggressive and two LXR isoforms, LXRα or LXRβ, regulates the LXR- untreatable phenotype (2). In the absence of any effective responsive genes ATP-binding cassette subfamily members long-term therapy and with such an overwhelming social A1 (ABCA1)andG1(ABCG1) in transformed prostatic ep- burden, characterization of novel targets for effective che- ithelial cells. In this study, small interfering RNA (siRNA) moprevention or treatment of PCa has been a priority for α β cancer researchers.
  • Overall and Sex-Specific Associations Between Methylation of the ABCG1

    Overall and Sex-Specific Associations Between Methylation of the ABCG1

    Qin et al. Clinical Epigenetics (2019) 11:189 https://doi.org/10.1186/s13148-019-0784-0 RESEARCH Open Access Overall and sex-specific associations between methylation of the ABCG1 and APOE genes and ischemic stroke or other atherosclerosis-related traits in a sibling study of Chinese population Xueying Qin1*† , Jin Li1, Tao Wu1, Yiqun Wu1, Xun Tang1, Pei Gao1, Lin Li2, Mengying Wang1, Yao Wu1, Xiaowen Wang1, Dafang Chen1 and Yonghua Hu1*† Abstract Background: Identifying subjects with a high risk of ischemic stroke is fundamental for prevention of the disease. Both genetic and environmental risk factors contribute to ischemic stroke, but the underlying epigenetic mechanisms which mediate genetic and environmental risk effects are not fully understood. The aim of this study was to explore whether DNA methylation loci located in the ATP-binding cassette G1 (ABCG1) and apolipoprotein E (APOE)genes, both involved in the metabolism of lipids in the body, are related to ischemic stroke, using the Fangshan/Family-based Ischemic Stroke Study in China. We also tested if these CpG sites were associated with early signs of cardiovascular atherosclerosis (carotid intima–media thickness (cIMT), ankle–brachial index (ABI), and brachial–ankle pulse wave velocity (baPWV)). Results: DNA methylation at the cg02494239 locus in ABCG1 was correlated with ischemic stroke after adjusting for gender, previous history of diabetes and hypertension, smoking, drinking, body mass index, and blood lipid levels (above vs below mean, OR = 2.416, 95% CI 1.024–5.700, P =0.044;75–100% percentile vs 0–25% percentile, OR = 4.461, 95% CI 1.226–16.225, P = 0.023).
  • Interindividual Differences in the Expression of ATP-Binding

    Interindividual Differences in the Expression of ATP-Binding

    Supplemental material to this article can be found at: http://dmd.aspetjournals.org/content/suppl/2018/02/02/dmd.117.079061.DC1 1521-009X/46/5/628–635$35.00 https://doi.org/10.1124/dmd.117.079061 DRUG METABOLISM AND DISPOSITION Drug Metab Dispos 46:628–635, May 2018 Copyright ª 2018 by The American Society for Pharmacology and Experimental Therapeutics Special Section on Transporters in Drug Disposition and Pharmacokinetic Prediction Interindividual Differences in the Expression of ATP-Binding Cassette and Solute Carrier Family Transporters in Human Skin: DNA Methylation Regulates Transcriptional Activity of the Human ABCC3 Gene s Tomoki Takechi, Takeshi Hirota, Tatsuya Sakai, Natsumi Maeda, Daisuke Kobayashi, and Ichiro Ieiri Downloaded from Department of Clinical Pharmacokinetics, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan (T.T., T.H., T.S., N.M., I.I.); Drug Development Research Laboratories, Kyoto R&D Center, Maruho Co., Ltd., Kyoto, Japan (T.T.); and Department of Clinical Pharmacy and Pharmaceutical Care, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan (D.K.) Received October 19, 2017; accepted January 30, 2018 dmd.aspetjournals.org ABSTRACT The identification of drug transporters expressed in human skin and levels. ABCC3 expression levels negatively correlated with the methylation interindividual differences in gene expression is important for understanding status of the CpG island (CGI) located approximately 10 kilobase pairs the role of drug transporters in human skin. In the present study, we upstream of ABCC3 (Rs: 20.323, P < 0.05). The reporter gene assay revealed evaluated the expression of ATP-binding cassette (ABC) and solute carrier a significant increase in transcriptional activity in the presence of CGI.
  • ABCA7 and Pathogenic Pathways of Alzheimer's Disease

    ABCA7 and Pathogenic Pathways of Alzheimer's Disease

    brain sciences Review ABCA7 and Pathogenic Pathways of Alzheimer’s Disease Tomonori Aikawa, Marie-Louise Holm ID and Takahisa Kanekiyo * Department of Neuroscience, Mayo Clinic, 4500 San Pablo Road, Jacksonville, FL 32224, USA; [email protected] (T.A.); [email protected] (M.-L.H.) * Correspondence: [email protected]; Tel.: +1-904-953-2483 Received: 28 December 2017; Accepted: 3 February 2018; Published: 5 February 2018 Abstract: The ATP-binding cassette (ABC) reporter family functions to regulate the homeostasis of phospholipids and cholesterol in the central nervous system, as well as peripheral tissues. ABCA7 belongs to the A subfamily of ABC transporters, which shares 54% sequence identity with ABCA1. While ABCA7 is expressed in a variety of tissues/organs, including the brain, recent genome-wide association studies (GWAS) have identified ABCA7 gene variants as susceptibility loci for late-onset Alzheimer’s disease (AD). More important, subsequent genome sequencing analyses have revealed that premature termination codon mutations in ABCA7 are associated with the increased risk for AD. Alzheimer’s disease is a progressive neurodegenerative disease and the most common cause of dementia, where the accumulation and deposition of amyloid-β (Aβ) peptides cleaved from amyloid precursor protein (APP) in the brain trigger the pathogenic cascade of the disease. In consistence with human genetic studies, increasing evidence has demonstrated that ABCA7 deficiency exacerbates Aβ pathology using in vitro and in vivo models. While ABCA7 has been shown to mediate phagocytic activity in macrophages, ABCA7 is also involved in the microglial Aβ clearance pathway. Furthermore, ABCA7 deficiency results in accelerated Aβ production, likely by facilitating endocytosis and/or processing of APP.
  • Ohnologs in the Human Genome Are Dosage Balanced and Frequently Associated with Disease

    Ohnologs in the Human Genome Are Dosage Balanced and Frequently Associated with Disease

    Ohnologs in the human genome are dosage balanced and frequently associated with disease Takashi Makino1 and Aoife McLysaght2 Smurfit Institute of Genetics, University of Dublin, Trinity College, Dublin 2, Ireland Edited by Michael Freeling, University of California, Berkeley, CA, and approved April 9, 2010 (received for review December 21, 2009) About 30% of protein-coding genes in the human genome are been duplicated by WGD, subsequent loss of individual genes related through two whole genome duplication (WGD) events. would result in a dosage imbalance due to insufficient gene Although WGD is often credited with great evolutionary impor- product, thus leading to biased retention of dosage-balanced tance, the processes governing the retention of these genes and ohnologs. In fact, evidence for preferential retention of dosage- their biological significance remain unclear. One increasingly pop- balanced genes after WGD is accumulating (4, 7, 11–20). Copy ular hypothesis is that dosage balance constraints are a major number variation [copy number polymorphism (CNV)] describes determinant of duplicate gene retention. We test this hypothesis population level polymorphism of small segmental duplications and show that WGD-duplicated genes (ohnologs) have rarely and is known to directly correlate with gene expression levels (21– experienced subsequent small-scale duplication (SSD) and are also 24). Thus, CNV of dosage-balanced genes is also expected to be refractory to copy number variation (CNV) in human populations deleterious. This model predicts that retained ohnologs should be and are thus likely to be sensitive to relative quantities (i.e., they are enriched for dosage-balanced genes that are resistant to sub- dosage-balanced).
  • Hedgehog-GLI Signalling Promotes Chemoresistance Through The

    Hedgehog-GLI Signalling Promotes Chemoresistance Through The

    www.nature.com/scientificreports OPEN Hedgehog‑GLI signalling promotes chemoresistance through the regulation of ABC transporters in colorectal cancer cells Agnese Po1, Anna Citarella1, Giuseppina Catanzaro2, Zein Mersini Besharat2, Sofa Trocchianesi1, Francesca Gianno1, Claudia Sabato2, Marta Moretti2, Enrico De Smaele2, Alessandra Vacca2, Micol Eleonora Fiori3 & Elisabetta Ferretti2,4* Colorectal cancer (CRC) is a leading cause of cancer death. Chemoresistance is a pivotal feature of cancer cells leading to treatment failure and ATP-binding cassette (ABC) transporters are responsible for the efux of several molecules, including anticancer drugs. The Hedgehog-GLI (HH-GLI) pathway is a major signalling in CRC, however its role in chemoresistance has not been fully elucidated. Here we show that the HH-GLI pathway favours resistance to 5-fuorouracil and Oxaliplatin in CRC cells. We identifed potential GLI1 binding sites in the promoter region of six ABC transporters, namely ABCA2, ABCB1, ABCB4, ABCB7, ABCC2 and ABCG1. Next, we investigated the binding of GLI1 using chromatin immunoprecipitation experiments and we demonstrate that GLI1 transcriptionally regulates the identifed ABC transporters. We show that chemoresistant cells express high levels of GLI1 and of the ABC transporters and that GLI1 inhibition disrupts the transporters up-regulation. Moreover, we report that human CRC tumours express high levels of the ABCG1 transporter and that its expression correlates with worse patients’ prognosis. This study identifes a new mechanism where HH‑GLI signalling regulates CRC chemoresistance features. Our results indicate that the inhibition of Gli1 regulates the ABC transporters expression and therefore should be considered as a therapeutic option in chemoresistant patients. Colorectal cancer (CRC) is a leading cause of cancer-related death worldwide and is characterized by resistance mechanisms that lead to disease progression 1.
  • Rosmarinic Acid Increases Macrophage Cholesterol Efflux

    Rosmarinic Acid Increases Macrophage Cholesterol Efflux

    International Journal of Molecular Sciences Article Rosmarinic Acid Increases Macrophage Cholesterol Efflux through Regulation of ABCA1 and ABCG1 in Different Mechanisms Jean-Baptiste Nyandwi 1,2,3 , Young Shin Ko 1, Hana Jin 1, Seung Pil Yun 1, Sang Won Park 1,2 and Hye Jung Kim 1,2,* 1 Department of Pharmacology, Institute of Health Sciences, College of Medicine, Gyeongsang National University, Jinju 52727, Korea; [email protected] (J.-B.N.); [email protected] (Y.S.K.); [email protected] (H.J.); [email protected] (S.P.Y.); [email protected] (S.W.P.) 2 Department of Convergence Medical Science (BK21 Plus), Gyeongsang National University, Jinju 52727, Korea 3 Department of Pharmacy, School of Medicine and Pharmacy, College of Medicine and Health Sciences, University of Rwanda, Kigali 4285, Rwanda * Correspondence: [email protected] Abstract: Lipid dysregulation in diabetes mellitus escalates endothelial dysfunction, the initial event in the development and progression of diabetic atherosclerosis. In addition, lipid-laden macrophage accumulation in the arterial wall plays a significant role in the pathology of diabetes-associated atherosclerosis. Therefore, inhibition of endothelial dysfunction and enhancement of macrophage cholesterol efflux is the important antiatherogenic mechanism. Rosmarinic acid (RA) possesses benefi- cial properties, including its anti-inflammatory, antioxidant, antidiabetic and cardioprotective effects. We previously reported that RA effectively inhibits diabetic endothelial dysfunction by inhibiting inflammasome activation in endothelial cells. However, its effect on cholesterol efflux remains Citation: Nyandwi, J.-B.; Ko, Y.S.; unknown. Therefore, in this study, we aimed to assess the effect of RA on cholesterol efflux and its Jin, H.; Yun, S.P.; Park, S.W.; Kim, H.J.
  • ABC Transporter Genes and Risk of Type 2 Diabetes a Study of 40,000 Individuals from the General Population

    ABC Transporter Genes and Risk of Type 2 Diabetes a Study of 40,000 Individuals from the General Population

    Cardiovascular and Metabolic Risk ORIGINAL ARTICLE ABC Transporter Genes and Risk of Type 2 Diabetes A study of 40,000 individuals from the general population 1,2 2,3,4,6 JESPER SCHOU, MSC BØRGE G. NORDESTGAARD, MD, DMSC secretion in mice (2). In humans, family 1,2,3,4 1,2,4 ANNE TYBJÆRG-HANSEN, MD, DMSC RUTH FRIKKE-SCHMIDT, MD, DMSC 5 studies and small case-control studies HOLGER J. MØLLER, MD, PHD have suggested that loss-of-function muta- tions in ABCA1 associate with decreased in- sulin secretion (8) or type 2 diabetes (9). OBJECTIVEd b b Alterations of pancreatic -cell cholesterol content may contribute to -cell Further, macrophages from diabetic mice dysfunction. Two important determinants of intracellular cholesterol content are the ATP- (10) and humans (11) display decreased binding cassette (ABC) transporters A1 (ABCA1) and -G1 (ABCG1). Whether genetic variation ABCG1 expression. Finally, we recently re- in ABCA1 and ABCG1 predicts risk of type 2 diabetes in the general population is unknown. ported that two functional variants, ABCA1 RESEARCH DESIGN AND METHODSdWe tested whether genetic variation in the N1800H and ABCG1 g.-376C.T, were as- promoter and coding regions of ABCA1 and ABCG1 predicted risk of type 2 diabetes in the sociated with, respectively, substantial re- general population. Twenty-seven variants, identified by previous resequencing of both genes, ductions in levels of HDL cholesterol or were genotyped in the Copenhagen City Heart Study (CCHS) (n = 10,185). Two loss-of-function reduced ABCG1 mRNA expression levels mutations (ABCA1 N1800H and ABCG1 g.-376C.T) (n = 322) and a common variant (ABCG1 .
  • Characterizing the Interaction of the ATP Binding Cassette Transporters (G Subfamily) with the Intracellular Protein Lipid Environment

    Characterizing the Interaction of the ATP Binding Cassette Transporters (G Subfamily) with the Intracellular Protein Lipid Environment

    Characterizing the Interaction of the ATP Binding Cassette Transporters (G subfamily) with the Intracellular Protein Lipid Environment Sonia Gulati Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy under the Executive Committee of the Graduate School of Arts and Sciences Columbia University 2011 © 2011 Sonia Gulati All Rights Reserved Abstract Characterizing the Interaction of the ATP Binding Cassette Transporters (G subfamily) with the Intracellular Protein Lipid Environment Sonia Gulati Cholesterol is an essential molecule that mediates a myriad of critical cellular processes, such as signal transduction in eukaryotes, membrane fluidity, and steroidogenesis. As such it is not surprising that cholesterol homeostasis is tightly regulated, striking a precise balance between endogenous synthesis and regulated uptake/efflux to and from extracellular acceptors. In mammalian cells, sterol efflux is a key component of the homeostatic equation and is mediated by members of the ATP binding cassette (ABC) transporter superfamily. ATP-binding cassette (ABC) transporters represent a group of evolutionarily highly conserved cellular transmembrane proteins that mediate the ATP-dependent translocation of substrates across membranes. Members of this superfamily, ABCA1 and ABCG1, are key components of the reverse cholesterol transport pathway. ABCG1 acts in concert with ABCA1 to maximize the removal of excess cholesterol from cells by promoting cholesterol efflux onto mature and nascent HDL particles, respectively. To date, mammalian ABC transporters are exclusively associated with efflux of cholesterol. In Saccharomyces cerevisiae, we have demonstrated that the opposite (i.e inward) transport of sterol in yeast is also dependent on two ABC transporters (Aus1p and Pdr11p). This prompts the question what dictates directionality of sterol transport by ABC transporters.
  • Identification of Placental Nutrient Transporters Associated With

    Identification of Placental Nutrient Transporters Associated With

    Huang et al. BMC Genomics (2018) 19:173 https://doi.org/10.1186/s12864-018-4518-z RESEARCHARTICLE Open Access Identification of placental nutrient transporters associated with intrauterine growth restriction and pre-eclampsia Xiao Huang1,2, Pascale Anderle3,4, Lu Hostettler2, Marc U. Baumann1,5, Daniel V. Surbek1,5, Edgar C. Ontsouka1,2 and Christiane Albrecht1,2* Abstract Background: Gestational disorders such as intrauterine growth restriction (IUGR) and pre-eclampsia (PE) are main causes of poor perinatal outcomes worldwide. Both diseases are related with impaired materno-fetal nutrient transfer, but the crucial transport mechanisms underlying IUGR and PE are not fully elucidated. In this study, we aimed to identify membrane transporters highly associated with transplacental nutrient deficiencies in IUGR/PE. Results: In silico analyses on the identification of differentially expressed nutrient transporters were conducted using seven eligible microarray datasets (from Gene Expression Omnibus), encompassing control and IUGR/PE placental samples. Thereby 46 out of 434 genes were identified as potentially interesting targets. They are involved in the fetal provision with amino acids, carbohydrates, lipids, vitamins and microelements. Targets of interest were clustered into a substrate-specific interaction network by using Search Tool for the Retrieval of Interacting Genes. The subsequent wet-lab validation was performed using quantitative RT-PCR on placentas from clinically well- characterized IUGR/PE patients (IUGR, n =8;PE,n =5;PE+IUGR,n = 10) and controls (term, n = 13; preterm, n = 7), followed by 2D-hierarchical heatmap generation. Statistical evaluation using Kruskal-Wallis tests was then applied to detect significantly different expression patterns, while scatter plot analysis indicated which transporters were predominantly influenced by IUGR or PE, or equally affected by both diseases.
  • RAGE Mediates Cholesterol Efflux Impairment in Macrophages

    RAGE Mediates Cholesterol Efflux Impairment in Macrophages

    International Journal of Molecular Sciences Article RAGE Mediates Cholesterol Efflux Impairment in Macrophages Caused by Human Advanced Glycated Albumin Adriana Machado-Lima 1,2 , Raquel López-Díez 3 , Rodrigo Tallada Iborra 1,2 , Raphael de Souza Pinto 1,4 , Gurdip Daffu 3, Xiaoping Shen 3, Edna Regina Nakandakare 1, Ubiratan Fabres Machado 5 , Maria Lucia Cardillo Corrêa-Giannella 6,7 , Ann Marie Schmidt 3 1,7, , and Marisa Passarelli * y 1 Laboratório de Lípides (LIM 10), Hospital das Clínicas (HCFMUSP), Faculdade de Medicina da Universidade de São Paulo, São Paulo CEP 01246-000, Brazil; [email protected] (A.M.-L.); [email protected] (R.T.I.); [email protected] (R.d.S.P.); [email protected] (E.R.N.) 2 Programa de Pós-Graduação em Ciências do Envelhecimento, Universidade São Judas Tadeu, São Paulo CEP 03166-000, Brazil 3 Department of Medicine, Diabetes Research Program, New York University Langone Health, New York, NY 10016, USA; [email protected] (R.L.-D.); gdaff[email protected] (G.D.); [email protected] (X.S.); [email protected] (A.M.S.) 4 Curso de Biomedicina, Centro Universitário CESMAC, Maceió, Alagoas CEP 57051-160, Brazil 5 Laboratório de Metabolismo e Endocrinologia, Instituto de Ciências Biomédicas da Universidade de São Paulo, São Paulo CEP 05508-000, Brazil; [email protected] 6 Laboratório de Carboidratos e Radioimunoensaio (LIM 18), Hospital das Clínicas (HCFMUSP), Faculdade de Medicina da Universidade de São Paulo, São Paulo CEP 01246-000, Brazil; [email protected] 7 Programa de Pós-Graduação em Medicina, Universidade Nove de Julho, São Paulo CEP 01225-000, Brazil * Correspondence: [email protected] Present address: Marisa Passarelli, Laboratório de Lípides (LIM 10), Hospital das Clínicas (HCFMUSP), y Faculdade de Medicina da Universidade de São Paulo, Brazil.