Annual Report 2017

Akinobu Hamada, Mituhiro Hayashi, Shigehiro Yagishita, Tomomi Yoshino, Shoraku Ryu, Mayu Ohuchi, Ryosuke Matsukane, Yuki Murai

Introduction

 The Division of Molecular Pharmacology and the Department of Pharmacology and Therapeutics are focused on the development of pharmacokinetics/ pharmacodynamics/ pharmacogenetics (PK/PD/PGx) analyzing systems to evaluate the efficacy and toxicity in patients and animal models treated with chemotherapy. The system provides drug exposure in blood and tissues by using high-sensitivity liquid chromatography tandem mass spectrometry (LC-MS/ MS), a spatial drug distribution on tissue by using mass spectrometry imaging without labeling reagents, and next generation sequencer systems. Recently, we, the National Cancer Center (NCC), National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN), and LSI Medience Corporation (LSIM) started a Japanese Cancer Patient-derived Xenograft (J-PDX) Repository Establishment Program supported by the Japan Agency for Medical Research and Development (AMED) on March 7, 2018.

Our team and what we do

 Our laboratory aims for implementing anticancer drug discovery and development in Japan. In this regard, we are working on "Clinical pharmacological research at a molecular level" by using next generation PK/PD/PGx analyses, a novel Molecular Drug Imaging system, and J-PDX library.

Research activities

1. PK/PD/PGx Project

 In recent years, anticancer drug development shifts from conventional cytotoxic agents to molecular target drugs, antibody-drug conjugates (ADCs), and immune checkpoint inhibitors. Our laboratory established next generation PK/PD/PGx analyses to achieve Precision Medicine of antibody drugs. Our novel analyses include precise PK/PD analyses of antibody drugs, an immune monitoring system, a molecular drug imaging system, and an originally developed NGS PGx panel. Moreover, in collaboration with clinicians and clinical pharmacologists, we are working on translational research of drug discovery and development.

2. Molecular Drug Imaging Project

 Molecular imaging is the medical practice and laboratory science of visualizing molecular process in living body. Our laboratory focused on molecular process of anticancer drugs and corresponding various factors. We already developed Liquid Chromatography/ Mass Spectrometry (LC-MS/MS) imaging system (Multispectrum Imaging: MSI). By using MSI, we published many reports about spatial drug distribution, intra-

tumoral drug concentration, and intra-tumoral drug heterogeneity. Moreover, we are also struggling with drug concentration analysis at a cell level. In cooperation with KONIKA-MINOLTA, we employed newly developed fluorescent nano-

particles (Phosphor Integrated Dot nanoparticles: PID). Using PID method, we expect to clarify the precise mode of action in antibody drugs.

3. J-PDX Project

 In the course of drug development, a pre-clinical screening model is a key to success in development.

 The objective of this program is to create an international-level research repository of J-PDX for use in the pharmaceutical industry in addition to nurturing expert personnel to promote academia-industry collaboration in their relevant fields.

 Following the requests of the Pharmaceuticals and Medical Devices Agency (PMDA), we are planning to: 1) refine bio-ethical rules for the industrial use of J-PDXs, 2) establish a world-class J-PDX repository that includes major cancers (lung, colon, breast, stomach, uterus) in addition to prostate, pancreatic and rare cancers, along with the associated donors' clinical information, 3) set criteria for the establishment and use of the J-PDX repository, 4) create standard operating procedures for the storage and use of J-PDXs in GLP-compliant, non-clinical studies, and 5) establish a research platform for fundamental and applied PDX science.

Education

 Dr. Akinobu Hamada is a visiting professor at the Department of Medical Oncology and Translational Research, Graduate School of Medical Sciences, Kumamoto University.

Future prospects

 The combination of PK/PD/PGx analyses, molecular drug imaging technique, and establishment of J-PDX library will provide us more accurate information about anticancer drugs. These systems must enable us to establish exceptional drug discovery infrastructure of Japan and establish personalized medicine in the future.

List of papers published in January 2017 - March 2018

Journal

 1. Iwama E, Goto Y, Murakami H, Harada T, Tsumura S, Sakashita H, Mori Y, Nakagaki N, Fujita Y, Seike M, Bessho A, Ono M, Okazaki A, Akamatsu H, Morinaga R, Ushijima S, Shimose T, Tokunaga S, Hamada A, Yamamoto N, Nakanishi Y, Sugio K, Okamoto I. Alectinib for Patients with ALK Rearrangement-Positive Non-Small Cell Lung Cancer and a Poor Performance Status (Lung Oncology Group in Kyushu 1401). J Thorac Oncol, 12:1161-1166, 2017

 2. Iwamoto N, Shimomura A, Tamura K, Hamada A, Shimada T. LC-MS bioanalysis of Trastuzumab and released emtansine using nano-surface and molecular-orientation limited (nSMOL) proteolysis and liquid-liquid partition in plasma of Trastuzumab emtansine-treated breast cancer patients. J Pharm Biomed Anal, 145:33-39, 2017

 3. Nishidate M, Yamamoto K, Masuda C, Aikawa H, Hayashi M, Kawanishi T, Hamada A. MALDI mass spectrometry imaging of erlotinib administered in combination with bevacizumab in xenograft mice bearing B901L, EGFR-mutated NSCLC cells. Sci Rep, 7:16763, 2017

 4. Sasada S, Kurihara H, Kinoshita T, Yoshida M, Honda N, Shimoi T, Shimomura A, Yonemori K, Shimizu C, Hamada A, Kanayama Y, Watanabe Y, Fujiwara Y, Tamura K. Visualization of HER2-specific breast cancer intratumoral heterogeneity using 64Cu-DOTA-trastuzumab PET. Eur J Nucl Med Mol Imaging, 44:2146-2147, 2017

 5. Tsuchiya K, Hayashida T, Hamada A, Oki S, Oka S, Gatanaga H. High plasma concentrations of dolutegravir in patients with ABCG2 genetic variants. Pharmacogenet Genomics, 27:416-419, 2017

 6. Tanabe Y, Shimizu C, Hamada A, Hashimoto K, Ikeda K, Nishizawa D, Hasegawa J, Shimomura A, Ozaki Y, Tamura N, Yamamoto H, Yunokawa M, Yonemori K, Takano T, Kawabata H, Tamura K, Fujiwara Y. Paclitaxel-induced sensory peripheral neuropathy is associated with an ABCB1 single nucleotide polymorphism and older age in Japanese. Cancer Chemother Pharmacol, 79:1179-1186, 2017

 7. Tsubata Y, Hayashi M, Tanino R, Aikawa H, Ohuchi M, Tamura K, Fujiwara Y, Isobe T, Hamada A. Evaluation of the heterogeneous tissue distribution of erlotinib in lung cancer using matrix-assisted laser desorption ionization mass spectrometry imaging. Sci Rep, 7:12622, 2017

 8. Iwamoto N, Hamada A, Shimada T. Antibody drug quantitation in coexistence with anti-drug antibodies on nSMOL bioanalysis. Anal Biochem, 540-541:30-37, 2018

 9. Nishimura M, Hayashi M, Mizutani Y, Takenaka K, Imamura Y, Chayahara N, Toyoda M, Kiyota N, Mukohara T, Aikawa H, Fujiwara Y, Hamada A, Minami H. Distribution of erlotinib in rash and normal skin in cancer patients receiving erlotinib visualized by matrix assisted laser desorption/ionization mass spectrometry imaging. Oncotarget, 9:18540-18547, 2018

10. Tsuchiya K, Ohuchi M, Yamane N, Aikawa H, Gatanaga H, Oka S, Hamada A. High-performance liquid chromatography-tandem mass spectrometry for simultaneous determination of raltegravir, dolutegravir and elvitegravir concentrations in human plasma and cerebrospinal fluid samples. Biomed Chromatogr, 32:2018

11. Ogata-Aoki H, Higashi-Kuwata N, Hattori SI, Hayashi H, Danish M, Aoki M, Shiotsu C, Hashiguchi Y, Hamada A, Kobayashi H, Ihn H, Okada S, Mitsuya H. Raltegravir blocks the infectivity of red-fluorescent-protein (mCherry)-labeled HIV-1JR-FL in the setting of post-exposure prophylaxis in NOD/SCID/Jak3-/- mice transplanted with human PBMCs. Antiviral Res, 149:78-88, 2018

12. Ryu S, Hayashi M, Aikawa H, Okamoto I, Fujiwara Y, Hamada A. Heterogeneous distribution of alectinib in neuroblastoma xenografts revealed by matrix-assisted laser desorption ionization mass spectrometry imaging: a pilot study. Br J Pharmacol, 175:29-37, 2018