TY - JOUR
T1 - Transport mechanisms of hepatic uptake and bile excretion in clinical hepatobiliary scintigraphy with 99mTc-N-pyridoxyl-5-methyltryptophan
AU - Kobayashi, Masato
AU - Nakanishi, Takeo
AU - Nishi, Kodai
AU - Higaki, Yusuke
AU - Okudaira, Hiroyuki
AU - Ono, Masahiro
AU - Tsujiuchi, Takafumi
AU - Mizutani, Asuka
AU - Nishii, Ryuichi
AU - Tamai, Ikumi
AU - Arano, Yasushi
AU - Kawai, Keiichi
N1 - Funding Information:
The author would like to thank the laboratory staff of Kanazawa and Chiba University. This study was partly funded by a Grant-in-Aid for Scientific Research from Japan Society for the Promotion of Science ( 23590176, 24601008, 24659558 and 25293260 ) and the Japanese Society of Nuclear Medicine Technology .
PY - 2014/4
Y1 - 2014/4
N2 - Introduction: In clinical hepatobiliary scintigraphy, 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) is an effective radiotracer among the 99mTc-pyridoxylaminates. However, the mechanisms of human hepatic uptake and bile excretion transport of 99mTc-PMT have not been determined. We thus investigated the transport mechanisms of human hepatic uptake and bile excretion in hepatobiliary scintigraphy with 99mTc-PMT. Methods: Four solute carrier (SLC) transporters involved in hepatic uptake were evaluated using human embryonic kidney (HEK) and HeLa cells with high expression of SLC transporters (organic anion transporting polypeptide (OATP)1B1, OATP1B3, OATP2B1, organic anion transporters (OAT)2 and organic cation transporters (OCT)1) after 5min of 99mTc-PMT incubation. Metabolic analysis of 99mTc-PMT was performed using pooled human liver S9. Adenosine triphosphate (ATP)-binding cassette (ABC) transporters for bile excretion were examined using hepatic ABC transporter vesicles human expressing multiple drug resistance 1 (MDR1), multidrug resistance-associated protein 2 (MRP2), breast cancer resistance protein or bile salt export pump. 99mTc-PMT was incubated for 1, 3 and 5min with ATP or adenosine monophosphate and these vesicles. SPECT scans were performed in normal and Eisai hyperbilirubinemic (EHBR) model rats, deficient in Mrp2 transporters, without and with verapamil (rat Mdr1 and human MDR1 inhibitor) after intravenous injection of 99mTc-PMT. Results: Uptake of 99mTc-PMT in HEK293/OATP1B1 and HeLa/OATP1B3 was significantly higher than that in HEK293- and HeLa-mock cells. 99mTc-PMT was not metabolized in the human liver S9. In vesicles with high expression of ABC transporters, uptake of MDR1 or MRP2 was significantly higher at all incubation times. Bile excretion of 99mTc-PMT was also identified by comparison between normal and EHBR rats with and without verapamil on in-vivo imaging. Conclusions: Human hepatic uptake of 99mTc-PMT was transferred by OATP1B1 and OATP1B3, and excretion into bile canaliculi via MDR1 and MRP2. 99mTc-PMT hepatobiliary scintigraphy may be a useful ligand as a noninvasive method of visualizing and quantifying hepatobiliary transporter functionality, which could predict drug pharmacokinetics.
AB - Introduction: In clinical hepatobiliary scintigraphy, 99mTc-N-pyridoxyl-5-methyltryptophan (99mTc-PMT) is an effective radiotracer among the 99mTc-pyridoxylaminates. However, the mechanisms of human hepatic uptake and bile excretion transport of 99mTc-PMT have not been determined. We thus investigated the transport mechanisms of human hepatic uptake and bile excretion in hepatobiliary scintigraphy with 99mTc-PMT. Methods: Four solute carrier (SLC) transporters involved in hepatic uptake were evaluated using human embryonic kidney (HEK) and HeLa cells with high expression of SLC transporters (organic anion transporting polypeptide (OATP)1B1, OATP1B3, OATP2B1, organic anion transporters (OAT)2 and organic cation transporters (OCT)1) after 5min of 99mTc-PMT incubation. Metabolic analysis of 99mTc-PMT was performed using pooled human liver S9. Adenosine triphosphate (ATP)-binding cassette (ABC) transporters for bile excretion were examined using hepatic ABC transporter vesicles human expressing multiple drug resistance 1 (MDR1), multidrug resistance-associated protein 2 (MRP2), breast cancer resistance protein or bile salt export pump. 99mTc-PMT was incubated for 1, 3 and 5min with ATP or adenosine monophosphate and these vesicles. SPECT scans were performed in normal and Eisai hyperbilirubinemic (EHBR) model rats, deficient in Mrp2 transporters, without and with verapamil (rat Mdr1 and human MDR1 inhibitor) after intravenous injection of 99mTc-PMT. Results: Uptake of 99mTc-PMT in HEK293/OATP1B1 and HeLa/OATP1B3 was significantly higher than that in HEK293- and HeLa-mock cells. 99mTc-PMT was not metabolized in the human liver S9. In vesicles with high expression of ABC transporters, uptake of MDR1 or MRP2 was significantly higher at all incubation times. Bile excretion of 99mTc-PMT was also identified by comparison between normal and EHBR rats with and without verapamil on in-vivo imaging. Conclusions: Human hepatic uptake of 99mTc-PMT was transferred by OATP1B1 and OATP1B3, and excretion into bile canaliculi via MDR1 and MRP2. 99mTc-PMT hepatobiliary scintigraphy may be a useful ligand as a noninvasive method of visualizing and quantifying hepatobiliary transporter functionality, which could predict drug pharmacokinetics.
KW - ABC transporter
KW - Hepatobiliary scintigraphy
KW - SLC transporter
KW - Transport mechanism
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U2 - 10.1016/j.nucmedbio.2014.01.004
DO - 10.1016/j.nucmedbio.2014.01.004
M3 - Article
C2 - 24607436
AN - SCOPUS:84895500566
VL - 41
SP - 338
EP - 342
JO - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology
JF - International journal of radiation applications and instrumentation. Part B, Nuclear medicine and biology
SN - 0969-8051
IS - 4
ER -