Drug Metab. Pharmacokinet. 28 (3): 274–277 (2013).

Copyright © 2013 by the Japanese Society for the Study of Xenobiotics (JSSX)

Short Communication
Influence of SLCO1B3 Genetic Variations on Tacrolimus
Pharmacokinetics in Renal Transplant Recipients
Andrée-Anne B OIVIN 1 , Héloïse C ARDINAL 2, Azemi B ARAMA 3 , Judith N AUD 4 ,
Vincent P ICHETTE 4 , Marie-Josée H ÉBERT 2 and Michel R OGER 1, *
1
Laboratoire d’Immunogénétique, Centre de Recherche du Centre Hospitalier de l’Université de Montréal
et Département de Microbiologie et Immunologie de l’Université de Montréal, Montréal, Canada
2
Département de Médecine, Service de Néphrologie, Centre Hospitalier de l’Université de Montréal,
Montréal, Canada
3
Département de Chirurgie, Service de Chirurgie Greffe Rénale et Pancréatique,
Centre Hospitalier de l’Université de Montréal, Montréal, Canada
4
Département de Médecine, Service de Néphrologie, Hôpital Maisonneuve-Rosemont, Montréal, Canada

Full text of this paper is available at http://www.jstage.jst.go.jp/browse/dmpk

Summary: The immunosuppressive drug tacrolimus requires strict therapeutic monitoring due to its
narrow therapeutic index and high interindividual variability. Organic anion transporting polypeptide 1B3
(OATP1B3) is a human hepatocyte transporter involved in the hepatobiliary elimination of diverse
endogenous and exogenous substances. Genetic variations within the solute carrier (SLCO) 1B3 gene that
encodes OATP1B3 may contribute to interindividual differences in tacrolimus disposition. The purpose
of the present study is to investigate the association between SLCO1B3 polymorphisms and tacrolimus
pharmacokinetics in renal transplant recipients. We found significant correlations between two linked
coding nonsynomymous polymorphisms, T334G and G699A, and mean dose-adjusted tacrolimus trough
blood concentrations during the first week post-transplantation (p = 0.04) and when the target dose (10­
12 ng/ml) was obtained (p = 0.01). Patients carrying the homozygous mutant haplotype had 14.3-fold
higher risk (95% confidence interval: 1.43­100; p = 0.02) of having blood tacrolimus concentrations
above the median level, and thus being classified as poor OATP1B3 transporters, than carriers of one or two
copies of the wild-type haplotype. This study shows, for the first time, that SLCO1B3 polymorphism is
associated with tacrolimus exposure in the early post-transplant period.

Keywords: tacrolimus; drug level; renal transplants; SLCO1B3; genetic polymorphisms

rejection.1–3) Therefore, the identification of parameters predictive

Introduction
Tacrolimus, a member of the calcineurin inhibitor family, is
widely used in solid organ transplantation to prevent allograft
rejection. It is a critical dose-drug with a narrow therapeutic index.
Moreover, its pharmacokinetic characteristics may vary greatly
among individuals, and daily doses must be adjusted according
to its whole blood trough concentrations. Achieving therapeutic
trough levels is of paramount importance during the period
immediately after transplantation to prevent subsequent graft
of the optimal tacrolimus dosage would be a great clinical asset in
the determination of adequate tacrolimus administration.
Genetically inherited differences in drug disposition and
metabolism have been shown to significantly influence an indi-
vidual response to therapy and have been estimated to account
for 20 to 95% of the variability of drug pharmacokinetics and
pharmacodynamics.4) As tacrolimus is metabolized by the mem-
bers of the cytochrome P450 (CYP) 3A family, we and others
have shown that the CYP3A5*3 genotype is associated with its

Received August 24, 2012; Accepted October 19, 2012

J-STAGE Advance Published Date: November 13, 2012, doi:10.2133/dmpk.DMPK-12-SH-093
*To whom correspondence should be addressed: Michel ROGER, M.D., Ph.D., Département de Microbiologie et Infectiologie, Hôpital Notre-Dame
du CHUM 1560 Sherbrooke Est, Montréal, Québec, H2L 4M1, Canada. Tel. ©1-514-890-8000 (ext. 25802), Fax. ©1-514-412-7512, E-mail:
michel.roger@ssss.gouv.qc.ca
This work was supported by Astellas Pharma Canada Inc. The opinions expressed in this paper are those of the authors and Astellas Pharma
Inc. had no role in the design of the study, data analysis or interpretation, or in the manuscript preparation. A. A. Boivin holds a Student Research
award from Fonds de la Recherche en Santé du Québec (FRSQ). V. Pichette, M. J. Hébert and M. Roger are supported by career awards from
FRSQ.

274
 Tacrolimus Pharmacogenetics
pharmacokinetics.5,6) Organic anion transporting polypeptides
(OATPs) are transporter proteins that mediate intracellular uptake
of many molecules in various tissues7,8) contributing to the
overall drug absorption, distribution, elimination, and conse-
quently, influencing the systemic levels of many drugs.9) OATP1B3
is mainly expressed on the hepatocyte sinusoidal (basolateral)
membrane and transports diverse endogenous and exogenous
compounds from portal blood into hepatocytes.10,11) OATP1B3 has
also been identified on human tumor cells derived from brain, lung,
colon, gastric, pancreatic, gallbladder and prostate.12,13) OATP1B3
is encoded by the solute carrier (SLCO) 1B3 gene. Several
SLCO1B3 nucleotide sequence variations have been identified with
frequencies varying between different ethnic groups.14–16) Among
them, the T334G and G699A polymorphisms were recently
associated with an improved survival rate in patients with pros-
tatic cancer due to impaired testosterone transport in cancer cells
featuring these mutations13) and with mycophenolate mofetil
pharmacokinetics in renal transplant patients co-treated with
tacrolimus or sirolimus.17) Tacrolimus has been identified as an
inhibitor of OATP1B1.18) Two mutations in SLCO1B1 gene that
encode OATP1B1, A388G and T521C, have been shown to
influence tacrolimus trough blood concentrations after liver
transplantation.19) Since OATP1B3 exhibits overlapping substrate
specificities with OATP1B1,20) it is reasonable to assume that
tacrolimus might equally be transported by OATP1B3. The
purpose of the present study is to investigate whether SLCO1B3
polymorphism is associated with tacrolimus pharmacokinetics in
renal transplant recipients.
Materials and Methods
Thirty-eight white Canadian renal transplant recipients were
enrolled in this prospective and observational clinical study. The
study group was composed of 18 males and 20 females with a
mean age at the time of transplantation of 49.5 « 13.9 years. All
patients were given an immunosuppressive therapy consisting
of tacrolimus, mycophenolate mofetil, and steroids. Our standard
steroid regimen was characterized by 125 mg of methylpredniso-
lone given intravenously on the day of surgery, followed by
100 mg of oral prednisolone for day 1 post-operation, reducing by
20 mg every day from day 2 to 5 to a maintenance dose of 5 mg
daily thereafter. Plasma creatinine concentrations were measured
3 times a week to quantify creatinine clearance. Acute graft rejec-
tion was confirmed by kidney biopsy according to Banff criteria.21)
Post-transplantation diabetes was defined as a new need for
oral hypoglycemic agents or insulin after transplantation. Daily
tacrolimus trough blood concentration (C0) (ng/ml) and tacrolimus
dosage (mg/kg) were noted for the first 3 months post-trans-
plantation. Blood concentrations of tacrolimus were determined
using a validated Enzyme Multiplied Immunoassay Technique
(EMIT) (DADE Behring, Missisauga, Ontario, Canada). Daily
tacrolimus dosage was adjusted to achieve the target blood trough
levels of 10–12 ng/ml during the first month post-transplantation
and 8–10 ng/ml thereafter. This investigation has been conducted
in accordance with the Declaration of Helsinki and approved by
the ethics committee of the Centre Hospitalier de l’Université
de Montréal (CHUM). Written informed consent was obtained
from all patients. SLCO1B3 polymorphisms were determined using
direct DNA sequencing procedures as described previously.16)
The statistical analysis was performed using SPSS software.
Differences were considered statistically significant for a p-value
Copyright © 2013 by the Japanese Society for the Study of Xenobiotics (JSSX)
275
<0.05. Continuous variables were compared using unpaired the
Student’s t-test when continuous variables were normally dis-
tributed or with the Mann-Whitney U test otherwise. Categorical
values were compared using Fisher’s exact tests. Genotypic
frequencies were compared by Hardy-Weinberg expectations using
the »2 test.
Results and Discussion
The full extent of SLCO1B3 nucleotide sequence diversity in
the white Canadian population was reported elsewhere.16) Specific
SLCO1B3 polymorphisms with frequencies above 5% and with the
potential to affect the expression or protein structure of OATP1B3
were selected to determine their contributions to tacrolimus
pharmacokinetics (Table 1). The genotypic distribution of the
selected SLCO1B3 variants in the study population was in Hardy-
Weinberg equilibrium.
Mean dose-adjusted tacrolimus trough blood concentrations
(C0/dose weight-adjusted ratio) were determined for all patients
at two time points; during the first week (between day 3 and 7)
and at 3 months after transplantation. We established the dis-
tinction since tacrolimus blood levels can be influenced by co-
administration of additional medication, and during the first
week after transplantation no other drug known to interact with
tacrolimus was administered to the patients. Furthermore, because
gastro-intestinal motility is seriously disturbed the first 48 h
after transplantation in most patients, we determined tacrolimus
trough blood concentrations between day 3 and 7 post-op to
avoid this potential confounding effect. We found no correla-
tion between clinical outcomes (creatinine clearance, acute graft
rejection and post-transplant diabetes) during the first 3 months
post-transplantation and the selected key SLCO1B3 polymor-
phisms (Table 2). Although none of the SLCO1B3 promoter
or 5AUTR variants tested correlated with tacrolimus pharmaco-
kinetics, we found significant correlations between two frequent
coding nonsynomymous polymorphisms, T334G and G699A,
and tacrolimus exposure in the early post-transplant period.
This association remained significant after adjusting for previ-
ously reported CYP3A5 polymorphisms associated with blood
tacrolimus levels in these patients.5) These two variants are in
complete linkage disequilibrium in the white Canadian popula-
tion16) and in other populations.13–15) Therefore all individuals
carry the same genotype for both variants that are transmitted
together on the same chromosome (haplotype). The homozygous
mutant haplotype (334GG and 699AA) was significantly associ-
Table 1. Selected SLCO1B3 polymorphisms
Amino acid Allele
Location Nucleotide substitution
variation frequency (%)
Promoter ¹5838 CATCACA ¹5831 > del 19.5
Promoter ¹5828A > C 25.6
Promoter ¹5538T > C 9.8
5AUTR ¹28 ATATTCACTTGGTATCTG 24.4
¹11 > del
5AUTR ¹7 TTTA ¹4 > del 24.4
Exon 4 334T > G Ser112Ala 86.6
Exon 7 699G > A Met233Ile 86.6
Exon 8 767G > C Gly256Ala 12.2
UTR: untranslated region, del: deletion polymorphism.
Polymorphism positions are given from the initiation translation site (ATG) or
the nearest exon according the reference SLCO1B3 sequence (GenBank
NC_000012 region 20853800 to 20961400).
276 Andrée-Anne BOIVIN, et al.

Table 2. Pharmacokinetic parameters of tacrolimus according to SLCO1B3 T334G-G699A genotypes

T334G-G699A genotypes
Homozygotes 334T-699G p-valuea
Homozygotes 334G-699A
or heterozygotes
(n = 29)
(n = 9)
log tacrolimus C0/Dose at target dose (SD)b 4.28 (0.47) 4.94 (0.63) 0.010
log tacrolimus C0/Dose 3–7 days (SD)c 4.33 (0.47) 4.84 (0.66) 0.040
log tacrolimus C0/Dose 3 months (SD)d 4.69 (0.36) 4.95 (0.57) 0.120
GFR at 3 months (SD)e 53 (15) 61 (14) 0.200
Graft rejection (%)f 3 (33) 6 (21) 0.430
Post-transplantation diabetes (%)g 3 (37) 9 (36) 0.930
a
p-value as determined by the Mann-Whitney U test for GFR; Student’s t-test for log C0/Dose when target dose was obtained, log C0/Dose 3–7 days and 3 months;
Fisher’s test for graft rejection and post-transplantation diabetes.
b
Average of log tacrolimus C0/dose (ng/mL)/(mg/kg) when target dose (10–12 ng/mL) was obtained.
c
Average of log tacrolimus C0/dose (ng/mL)/(mg/kg) between day 3 and 7 post-transplantation.
d
Average of log tacrolimus C0/dose (ng/mL)/(mg/kg) for 3 months post-transplantation.
e
Average glomerular filtration rate in mL/min/1.73 m2 for 3 months post-transplantation.
f
Graft rejection episode up to 1 year post-transplantation.
g
Defined as a new need for oral hypoglycemic agents or insulin up to 1 year post-transplantation.
C0: Trough concentration of tacrolimus, GFR: glomerular filtration rate, SD: standard deviation.

ated with higher mean dose-adjusted tracrolimus trough blood
concentrations during the first week post-transplantation (p = 0.04)
and when the target dose (10–12 ng/ml) was obtained (p = 0.01)
(Table 2). Taken separately, the homozygote wild-type or hetero-
zygote genotypes were not associated with tacrolimus dosages,
suggesting that there is no dose-gene effect. These findings were
corroborated by a second statistical analysis where the subjects
were dichotomized into two subgroups; patients with good or
poor transporter activities, based on the median dose-adjusted
tacrolimus trough blood concentrations value of 124 (range 32–
417) ng/ml per mg/kg observed when target levels were achieved.
Patients carrying the homozygous mutant haplotype had 14.3-
fold higher risk (95% confidence interval: 1.43–100; p = 0.02)
of having blood tacrolimus concentrations above the median level,
and thus being classified as poor OATP1B3 transporters, than
carriers of one or two copies of the wild-type haplotype. We found
no significant correlation between the selected key polymorphisms
and blood tracrolimus concentrations at 3 months post-trans-
plantation. This suggests that the effect of SLCO1B3 polymor-
phisms on blood tacrolimus levels is limited to the early post-
transplant period. However, we cannot exclude the possibility that
some patients might have taken other drugs or suffered from mild
gastro-intestinal disorders during the first 3 months post-trans-
plantation that could have influenced blood tracrolimus levels and
confounded the analysis at the 3 months’ time point.
OATP1B3 is highly expressed in the human liver and is
involved in the uptake of endogenous substances and xenobiotics
into hepatocytes.10,11) Consequently, this protein is suspected to
play a central function in the disposition of drugs for both
metabolism and hepatobiliary elimination.22,23) Our study shows,
for the first time, that SLCO1B3 T334G and G699A variants are
associated with tacrolimus exposure in the early post-transplant
period. The T334G and G699A variants result in the amino-acid
substitutions Ser112Ala and Met233Ile located in the 3rd trans-
membrane-spanning domain and the 3rd extracellular loop of the
protein, respectively.24) The functional consequences of SLCO1B3
polymorphisms were investigated in stably transfected MDCKII
and HEK 293 cells and the uptake activities of the OATP1B3-
Ser112Ala and OATP1B3-Met233Ile proteins was similar to
those mediated by the OATP1B3-wild-type protein.25) However,
the latter study, did not investigate the consequence of the com-
bined effect of the two variants that are in complete linkage
disequilibrium. Hamada et al. found that COS-7 cells transfected
with the variant SLCO1B3 haplotype (334GG-699AA) showed
significant reduced uptake of testosterone as compared with cells
transfected with the SLCO1B3 wild-type sequence or a sequence
containing either the 334GG or the 699AA variants.13) Our results
are consistent with the decreased activity of the OATP1B3 variants
since subjects carrying the homozygous mutant haplotype (334GG
and 699AA) had higher tacrolimus blood levels than those found
in wild-type carriers. However, it should be noted that although
OATP1B3 exhibits overlapping substrate specificities with
OATP1B1,20) which is known to interact with tacrolimus,18) the
direct interaction between OATP1B3 and tacrolimus has not been
studied yet.
In conclusion, this study shows that SLCO1B3 T334G and
G699A variants are associated with tacrolimus exposure in the
early period following renal transplantation and warrants further
studies to demonstrate that the OATP1B3 variants exhibit reduced
tacrolimus hepatocellular uptake to support our clinical findings.
Acknowledgments: We thank Jo-Ann Fugère from the CHUM
Renal Transplant Unit for providing the clinical data.
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