外科的同所移植マウスを用いる抗がん剤の薬効評価 受託サービス. Services of Efficacy Testing of AntiCancer Drug by SOI mouse model 1/21

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1 1/21 外科的同所移植マウスを用いる抗がん剤の薬効評価 受託サービス Services of Efficacy Testing of AntiCancer Drug by SOI mouse model AntiCancer Japan Inc. Tatushi Goto May 8 th, 2013 Biotech2013

2 AntiCancer Japan Inc. Business Product sales GFP/RFP-expressing cell lines(wako Pure Chemical Industries Ltd.) GFP-expressing nude mouse(japan SLC Inc.) Contract research using in vivo fluorescent imaging technology Tumor imaging (SOI model, SC model, TVI model) Drug distribution Demonstration Location Saito Bioincubator 106, Saitoasagi, Ibaraki,Osaka URL: 2/21

3 GFP/RFP-expressing cell lines 3/21 PC3-GFP PC3-RFP HCT116-GFP HT1080-Dual color Now on sale!! Wako Pure Chemical Industries AntiCancer Japan

4 GFP-expressing nude mouse RFP nude mouse 4/21 CFP nude mouse Japan SLC AntiCancer Japan

5 Implantation of human tumor tissue into the corresponding organ of mouse SOI model 5/21 SOI = Surgical Orthotopic Implantation Prostate cancer Cell line:pc3-gfp 1 Cut subcutaneous tumor into small pieces. 2 Expose prostate. 3 Implant on prostate using nylon suture. 4 Close a wound.

6 Implantation of human tumor tissue into the corresponding organ of mouse SOI model Prostate cancer Cell line:pc3-gfp 6/21 Day 0 Day 7 Day 14 Day 24 SC Day 0 Day 7 Day 14 Day 25 SOI SOI model mouse shows tumor metastasis.

7 Implantation of human tumor tissue into the corresponding organ of mouse 7/21 SOI model Breast cancer No. of mice No. of mice with Primary tumor No. of mice with lung metastasis SOI SC (Fu et al, Anticancer Res., 1993) Colon cancer Local tumor growth Regional lymph node Metastasis Peritoneum Liver Lung SOI 12 / 12 6 / / 12 9 / 12 6 / 12 Cell suspension 3 / 12 0 / 12 0 / 12 0 / 12 0 / 12 (Furukawa et al, Jpn J. Surgery., 1993) SOI model enhances metastatic ability of tumor.

8 Implantation of human tumor tissue into the corresponding organ of mouse SOI model 8/21 Ovary Stomach Colon Various SOI models are available.

9 Implantation of human tumor tissue into the corresponding organ of mouse SOI model 9/21 Ovarian cancer Cell line:skov3-gfp Day 14 Day 28 Day 42 Day 56 Day 56 GFP Various SOI models are available.

10 Implantation of human tumor tissue into the corresponding organ of mouse SOI model 10/21 Stomach cancer Cell line:nugc4-gfp Day 13 Day 28 Day 42 Day 56 Day 60 GFP Various SOI models are available.

11 Implantation of human tumor tissue into the corresponding organ of mouse SOI model 11/21 Colon cancer Cell line:hct-116-gfp Day 14 Day 22 Day 28 Day 41 Various SOI models are available.

12 Evaluation of anticancer drug efficacy in SOI model Colon cancer Cell line:ht29-gfp 12/21 Treatment Inhibition rate (%) A B C D E F G H 1760 mg/kg, single 880 mg/kg, q7d x 2 MEN4901/T mg/kg, single 440 mg/kg, q7d x mg/kg, q4d x 4 80 mg/kg, q4d x 4 Irinotecan 40 mg/kg, q4d x 4 Untreated (Ma et al, J. Surgical Res., 2011)

13 Leukemia TVI model 13/21 Leukemia Cell line:u937-gfp Dorsal view Day 10 Day 21 Day 29 Ventral view Bone metastases of leukemia were observed.

14 Cancer studies by fluorescent imaging 14/21 Correlation between fluorescent area and tumor volume Regression 95% confidence r=0.89, p < 0.05 (Katz et al, J. Surgical Res., 2003) Detection of micro-metastasis Liver Lung

15 Cancer studies by fluorescent imaging 15/21 Time course of angiogenesis Vascular migration of cancer cell Skin flap Migration (Yang et al, PNAS., 2001) Extravasation * Movie * Movie

16 AntiCancer s SOI mouse model is. 16/21 Growth and metastasis model of human cancer Available model of various human cancers - Colon, Stomach, Pancreas, Breast, Ovary, Bladder, Prostate - (TVI model are used in case of Lung cancer, Leukemia, Melanoma ) Less invasive method

17 Fluorescent imaging systems 17/21 NIPPON ROPER Lumazone FL UVP ibox INDEC Biosystems FluorVivo100 OLYMPUS OV110

18 Our challenges for advanced imaging research 1. Near infrared (NIR) probes for cancer BTP derivatives (Zhang et al, Cancer Res., 2010) 18/21 CHO HeLa Pre-inj. 60 min 20% O 2 5% O 2 Excitation: nm Emission : > 645 nm

19 Our challenges for advanced imaging research 19/21 2. Sensitivity improvement of fluorescent signal Usage of low fluorescent diet Normal diet Day 0 Day 2 Day 4 Day 7 NIR NIR NIR NIR Low-fluorescent diet * CE-2 (without alfalfa) (CLEA Japan Inc.) Day 0 Day 2 Day 4 Day 7 NIR NIR NIR NIR Intrinsic NIR fluorescence decreased.

20 Flow of contract research 20/21 Contact Anticancer Japan or Wako Pure Chemical Industries Discussion Experiment Cell line : GFP/RFP cells (See our catalog), Custom-made GFP/RFP cells Mouse : Nude mouse, Fluorescent nude mouse (GFP, RFP, CFP) Model : SOI model, SC model, TVI model Dosing : Intraperitoneal administration, Tail vein injection, Oral administration Imager : OV110(OLYMPUS), Lumazone FL(NIPPON ROPER) ibox (UVP), FluorVivo 100 (INDEC Biosystems) Photo : Whole body, Necroscopy, cell observation Report Within one month from end of the experiments

21 Please visit to Wako pavilion in BIOtech 2013 and Anticancer Japan s Saito lab. 21/21 Contact to AntiCancer Japan Tel: / Fax: / info@anticancerjapan.com Contact to Wako Pure Chemical Industries Tel: / Fax: labchem-tec@wako-chem.co.jp

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