SUMMARY OF ACTIVITIES
a. MD Anderson Cancer Research Institute
1) I discussed the results and future directions of our collaraborative work to elucidate the roles of chemokines in tumor angiogenesis, with Dr. Fidler. I gave a seminar on the roles of monocyte chemoattractant protein- ÇP in interleukin-4-mediated tumor rejection mechanisms in Dr. Fidler’s laboratory.
2) I visited Dr. Roth's laboratory and discussed the present status and the basic aspects of p53 gene therapy in his lab with Dr. Roth's colleagues.
b. Department of Pathology, University of Michigan, School of Medicine
1) I discussed the roles of chemokines in various types of tissue injuries with Dr. Kunkel and his colleagues
2) I discussed the biological functions and pathophysiological roles of nod 2, which Dr. Nunez reported as the causative gene for familial Crohn's disease, with Dr. Nunez and his colleagues. We agreed to do collaborative work to elucidate the biological functions of nod 2.
c. University of Pennsylvania, School of Medicine
1) I discussed the roles of chemokines in GVHD reactions with Dr. Zhang and gave a seminar on the roles of monocyte chemoattractant protein-1 in interleukin-4-mediated tumor rejection mechanisms in Dr. Zhang's laboratory.
2) I discussed their clinical trials on brain tumors using interferon gene with Dr. Eck, the director of Cancer Gene Therapy Program in University of Pennsylvania.
3) I discussed the present status of adeno-associated virus vector system in the United States with Dr. Xiao, who is in charge to prepare adeno-associated vectors in University of Pennsylvania.
d. National Institutes ofÅ@Health (Bethesda)
1) I discussed the present status of the gene therapy against adenosine deaminase deficiency with Dr. Candotti, the director of gene therapy in NIH.
2) I discussed the future directions on our collaborative work on the pathophysiological roles of chemokines with Dr. Murphy. Dr. Murphy is going to give our group the mouse deficient in chemokine receptors such as CCR1 and CX3CR1, so that we will be able to elucidate the roles of these chemokines in the tumor progression and the establishment of tumor immunity.
e. NIH (Frederick)
I discussed our collaborative work to elucidate the pathophyiological roles of chemokines with Dr. Oppenheim and his colleagues and gave a seminar on the roles of monocyte chemoattractant protein-1 in interleukin-4?mediated tumor rejection mechanisms in Dr. Oppenheim’s laboratory.
(5) Takaaki Akaike
Kumamoto University School of Medicine
SPONSOR AND HOST INSTITUTION:
Victor Darley-Usmar (The University of Alabama at Birmingham, UAB) Bruce Freeman (UAB)
DATES OF VISIT: 29th July - 29th September, 2001
SUMMARY OF ACTIVITIES:
1) Collaboration with Prof. Darley-Usmar:
The main purpose of my visit to UAB was to perform joint studies on the pathophysiology of peroxynitrite focussing on its chemical modifications of sulfhydryls and nucleic acids. By the present collaboration, we made two major achievements, which revealed novel reaction products of peroxynitrite with biological molecules. One is formation of glutathione nitroso sulfoxide from the reaction of peroxynitrite and glutathione. This unique compound is also found to have a profound biological relevance in terms of modification of various sulflrydryl-containing proteins and enzymes. For example, we reported recently that a series of matrix metalloproteinases (MMPs) are activated by glutathione nitroso sulfoxide through a unique mixed disulfide formation between glutathione and a sulfhydryl moiety located in the autoinhibitory domain of the protein (J. Biol. Chem, 276, 29596, 2001). Because MMPS are critically involved in the solid tumor growth and cancer metastasis, peroxynitrite formation may have a tumorigenic potential and contributing to tumor progression. The other breakthrough we made in the present study is successful verification of 8-nitroguanine formation in the biological systems. In a separate experiment, we developed a specific antibody for 8-nitroguanine. By using this antibody, it is now clarified that appreciable amount of 8-nitroguanine was generated possibly by peroxynitrite produced in excess in the lung tissue sections of virus-induced pneumonia in mice. This result strongly supports the notion we currently proposed that free radicals such as NO and oxygen radicals are highly mutagenic through formation of reactive nitrogen oxides, so that free radicals are the important pathogenic molecules in inflammation- and infection-induced carcinogenesis.
2) Seminar at the UAB Center for Free Radical Biology:
I was also invited to give a talk at the UAB Center for Free Radical Biology. My seminar was kindly organized by Prof. Darley-Usmar and. also by Prof. Bruce Freeman, director of the Center for Free Radical Biology at UAB. The audience are mostly students and scientists from the Free Radical Center, where many groups including Drs. Freeman's and Darley-Usmar's groups are working enthusiastically on pathophysiology of NO and oxygen radicals. The title of my talk was: "Activation of Matrix Metalloproteinases in Oxidative Stress and Inflammation", in which I reported the activation mechanisms of MMPs by peroxynitrite and oxidative stress. There were many questions and comments at the seminar. The most important question was as to the formation of glutathione nitroso sulfoxide which is thought to be most responsible for the MMP activation during peroxynitrite-induced oxidative tissue damage, and many audience are interested in its very unique chemical reactivity. In general, our hypothesis regarding a novel reaction of peroxynitrite was well appreciated by the scientists at UAB.
It was a great opportunity for me to obtain most up-to-date information on NO/peroxynitrite from all groups at the free radical center. I would sincerely thank for the generous and courteous financial support for my trip to and stay in the US. I hope this kind of exchange program should be extended and organized successfully in the future, and also I believe this program will contribute greatly to the promotion of scientific activity in Japan as well as US.
(6)Motoshi Suzuki
Nagoya graduate School of Medicine
SPONSOR AND HOST INSTITUTION:
Universities of Utah. Washington, and Medicine and Dentistry of New Jersey
DATES OF VISIT:
SUMMARY OF ACTIVITIES:
1 . Dr. Bradley Preston at University of Utah and I started a new collaboration; yeast DNA polymerase , delta and alpha in the maintenance of genome stability.
2. Dr. Lawrence Loeb at University of Washington and I have collaborated for years. We discussed further collaboration. In this year, part of the results has been published in J Mol Biol 3O8. p823-837. Our patent application has been accepted by US patent and trademark office.
3. Dr. Mukund Modak and I started a new collaboration; structure and functions of DNA polymerases I and alpha Part of the collaboration will be published as a review article in SEIKAGAKU.
(7) Yoshiko AKAMATSU
SPONSOR AND HOST INSTITUTION:
Marjorie A. Oettinger, PhD. Massachusetts General Hospital Harvard Medical School
DATES OF VISIT:
SUMMARY OF ACTIVITIES:
I had two major objectives to my visit of Marjorie A. Oettinger laboratory at Massachusetts General Hospital (Boston, MA). First, to meet reviewers request of my once- submitted paper, I need to show the size difference in gel shift assay between the truncated RAG2 and full-length or truncation with MBP tag. However it was difficult to purify the active RAG2 protein for the new post-doc of Dr. Oettinger to repeat my results. Thus. I showed the tip to successfully express it in vaccinia virus expression system, and did total of 160 plates (in 15 cm dish) myself to save the pellet. I did not have to purify them all, but I have completed the infection itself which is the most critical step to obtain active protein. Second, I planned to quantify the reduction level in V-to-DJ rearrangement in the knock-in mouse with mutant RAG2. Since I could not determine the actual data in my university mainly due to the equipment, I tried to complete it by using theirs. As a results, I estimated the eduction level of V-to-DJ rearrangement was approx. 10 fold,
In addition I have visited Frederick W. Alt laboratory at Children's Hospital (Boston, MA) to discuss our recent progress in the study of the regulation V(D)J rearrangement. We shared our new data each other and got the idea what to be asked next. Dr. Alt’s suggestions were very helpful for me to continue the research in my group.
(8) Tetsuyuki KOBAYASHI
Ochanomizu University
SPONSOR AND HOST INSTITUTION:
Prof. Gabor Tigyi, The University of Tennessee Memphis
DATES OF VISIT: from August 20, 2001 to September 18, 2001
SUMMARY OF ACTIVITIES :
The main objective of study during my stay in the U.S. was to determine a biosynthetic enzyme activity for lysophosphatidic acid (LPA) and cyclic phosphatidic acid (cPA) in serum from ovarian cancer patients. As a result we found that the cPA-synthetic activity in serum from patients with malignant ovarian tumor was significantly higher than that from patients with benign tumor. In addition, it was demonstrated that a ratio of LPA and cPA synthesis in serum was pathologically correlated with its malignancy.
During the study on LPA/cPA synthetic enzyme activities, we also found that there was an inhibitor for these reactions in human serum. Since the inhibitor may play a significant role in the regulation of serum level of LPA and/or cPA, we started studying on identification of the inhibitor.
During my stay in Memphis I could meet with many scientists including Prof. Serhan from Harvard Medical School, Prof. Spiegel from Georgetown University Medical Center, and 12 Professors in University of Tennessee Memphis to make a very useful discussion about our researches. I also made a seminar presentation for the Department of Physiology on September 6, 2001 (cf. the attached copy of letter).
From an educational point of view I had a chance to talk with many graduate students in the Department of Physiology, University of Tennessee Memphis and was able to discuss about the difference in research and educational systems in a doctor course between Japan and the U.S. It should be noticed here that one of the graduate students in Ochanomizu University is now studying in Prof. Tigyi’s laboratory as a research trainee to continue our collaborative work.
I make a grateful acknowledgment for your financial support of my visit.
(9)Suong- Hyu Hyon
Institute for Frontier Medical Sciences, Kyoto University
SPONSOR AND HOST INSTITUTION:
Japan Society for the Promotion of Science Center for Controlled Chemical Delivery, University of UTAH
DATE OF VISIT: from July 3, 2001 to September 30, 2001 for 90 days.
SUMMARY OF ACTIVITIES :
The objective of cytokine gene-mediated immunotherapy of cancer lies in effective retardation of established tumor metastasis, confinement of tumor and its elimination, and prevention of reoccurrence of tumor. Cancer gene therapy has been attempted with virtually every cytokine belonging to the family of interleukins, interferons, tumor necrosis factors, and colony stimulating factors. Among them IL- 12 has proven to be one of the most effective in the induction of potent antitumor immunity.
Them, we tried the complex formation of plasmid DNA encoding murine interleukin 12 (mIL-12) by epigallo-catechin-gallate (EGCg) extracted from green tea for cytokine gene delivery.
As the result, mIL-12 complex was able to formed by EGCg, and it was proven that the size of the complex was able to adjusted by changing the concentration of mIL-12 and EGCg.
In the future, we will examine in vitro cytotoxicity and in vivo effectiveness of mIL- 12 complexs.
(10) Jiyang O-Wang
Chiba Cancer Center Research Institute
SPONSOR AND HOST INSTITUTION:
Dr.Martin Dolle; South Texas Centers for Biology in Medicine
DATES OF VISIT: July 16 Å` Aug. 16
SUMMARY OF ACTIVITIES:
To investigate the role of the translesional DNA polymerases in mutagenesis, we have generated transgenic mice that express pol and pol , and crossed with the transgenic mice harboring the lacZ gene that can be retrieved from chromosomal DNA to determine mutation rates. During this short visit to Dr. Dolle’s laboratory, I isolated genomic DNA from 10 week old double transgenic mice, rescued the PUR288 plasmid containing the lacZ gene and determined the mutation frequency as shown in the initial proposal.
The results showed that the mutation frequency was not increased in these young (10 week old) transgenic mice. In the next few months, we will examine the mutation frequency in mutagen-treated as well as in older, untreated mice. We have also transfected the PUR288 DNA into several cell lines, from which we will try to rescue the PUR288 DNA and establish a quick, easy system to determine mutation frequency in eukaryotic cells. The short visit to Dr. Dolle’s laboratory has made all these experiments possible and should greatly help clarify the function of the translesional DNA polymerases.
(11) Isao Oishi
Kobe University Graduate School of Medicine
SPONSOR AND HOST INSTITUTION:
National Institute of Arthritis and Musculoskeletal and skin Diseases, National Institute of Health
DATES OF VISIT: 03/11/02 to 03/17/02
SUMMARY OF ACTIVITIES:
The purpose of the visit was:
To discuss about the results of our research concerning molecular mechanisms of Chk2 signaling and Ror signaling, and to obtain feedback on our work.
To make new contacts and get various information about cell proliferation and differentiation. To discuss about the future orientation of our research and a possible collaboration. I visited Dr. O’Shes (NIAMS), and give a talk at his laboratory. We discussed about our up-to-date results and future orientation of our study. We also discussed about the roles of signaling molecules in lymphocyte proliferation and differentiation. Dr. Samelson (NCI) introduced me his elegant imaging studies on molecular dynamics of the immunoreceptor signaling. During my stay in US, I also visited Dr. Yamashita (NIAMS), Dr. Shepard (NIAMS), Dr. Schwartzberg (NHGRI), Dr. Nuckolls (NIAMS), Dr. Shum (NIAMS), Dr. Saitoh (NCI), Dr. Rubin (NCI), Dr. Sommers (NCI), Dr. Tuan (NIAMS), and Dr. Hall (NIAMS). We discussed about our studies on Chk2 and/or Ror-mediated signaling pathways. We also discussed about the roles of signaling molecules in the proliferation and differentiation of various cells including lymphocyte, chondrocyte, osteoblast, osteoclast, and mesenchimal stem cells. From this visit, I gained many knowledge and benefits for my future work. The visit to US was extremely worthwhile.
(12)Makoto Emoto, MD, PhD
Fukuoka University, School of Medicine
SPONSOR AND HOST INSTITUTION:
Harvard University Medical School
DATES OF VISIT: From 4th November, 2001 to 29th March, 2002
SUMMARY OF ACTIVITIES:
I firstly visited the Department of Ceil Biology, Harvard University Medical School in Boston. I studied a few . experimental works in angiogenesis. including an advanced organ culture system using a fetal mouse immature scalp tissue. The researchers in this laboratory tried to search some new angiogenic markers that might be expressed in such immature tissue. I also attended this unique work so that I could get a new idea as to angiogenesis in gynecology. The Endostatin. one of the new anti-angiogenic agents, was firstly made in this laboratory as the recombinant one. I discussed with Prof. Olsen and Dr. Fukai for my following research in Japan that would use this agent, and then a co-laboratory work in anti-angiogenic therapy has been planned. In the meeting of this laboratory. I had a lecture as to my previous work in angiogenesis in 13th November 2001. Secondary, I visited the Department of Pathology in Massachusetts General Hospital (Harvard Medical School). I reviewed a lot of important cases in gynecologic cancers including some high angiogenic tumors everyday. I also attended a teaching conference for gynecologic pathologists in this hospital twice a week. Conclusively, I could get new excellent ideas in my following works in angiogenesis that would be re-started in Japan from this April.
(13) Saburo Sone
The University of Tokushima School of Medicine
DATE OF VISIT: August 18/01 - September 3/01
SUMMARY OF ACTIVITIES:
The objectives of this study was to investigate recent progress of development of emerging molecular target-based therapeutics against cancer metastasis in the U.S.A. For this purpose I visited one venture business company (EntreMed), and several famous cancer centers in the U.S.A. where clinical trials with new therapeutic agents targeting the molecules involved in in autocrine/paracrine cell proliferation, adhesion, invasion, angiogenesis and signal transduction pathways. Particularly in MD Adnerson Cancer Center, Prof. IJ Fidler as main host arranged very well so that I could meet many basic scientists and clinical oncologists to learn how cancer clinical trials under collaboration with the related companies were performed, and I also obtained useful informations of the infrastructure to support high quality of cancer clinical trials with new drugs. I also exchanged valuable informations regarding recent progress of developing molecular targeted drugs against cancer metastasis through a lecture with discussion. In NCI I learned present status of the translational research in the U.S.A. Particulary Dr. L. Kwak. demonstrated that his original preparation of new tumor-specific antigen peptide could be found to be clinically an effective vaccine to control lymphoma in the translational research project. I have this time realized that in the U.S.A. the translational research from bench to bedside in clinics have developed very well with financial support by the government funds. In Japan priority and originality of the cancer research seem to be comparable to that in the U.S.A. Nevertheless, the infrastructure of translational research system seems to be very poor as compared to the U.S.A. Based on these experiences I as clinical oncologist would like to contribute a lot to establish the infrastructure for achievement of cancer clinical trials with high quality through a translational research. This time Cooperative Cancer Research Program was very helpful to me for learning and recognizing what was mostly important in term of the patient-oriented cancer research.
(14) Riichi Tawa
Kyoto Pharmaceutical University, Faculty of Pharmaceutical Science
SPONSOR AND HOST INSTITUTION:
Dr. Peter C. Dedon, Massachusetts Institute of Technology
DATES OF VISIT: July 28, 2001 - September 19, 2001
SUMMARY OF ACTIVITIES:
[Objection]
Specific modulation of DNA damage in a reconstituted nucleosome: Anticancer drug-mediated gene recognition
Under the guidances of Dr. Dedon, the following experiments were carried out.
(1) Isolation of the nucleosomes from chicken erythrocytes and purification of the histone core.
(2) Isolation of 5S rDNA fragments (X. borealis) from the plasmid pXP-10 and 5'-end labeling the fragments with!!
!32P-ATP. (3) Reconstitution of the labeled 5S rDNA fragments with the histone core.
(4) Analysis of enediyne-mediated DNA damages in the reconstituted nucleosome by radicalfootprinting assay. The experiment "Reconstitution of nucleosomes containing the specific transcriptionally active genes" could not carried out during the stay in Dedon’s laboratory, but Dr. Dedon promised me to support further experimental advices and to send the plasmid DNA for in vitro reconstitution of nucleosomes in my laboratory. In near future we have a schedule applying these techniques to the investigations of
(1)Site-specific DNA damages in specific genes reconstituted in nucleosome by anticancer agents
(2)Conformational changes of the reconstituted nucleosome by base modification and its interaction with anticancer agents.
(15) Yasushi Nagata
Kyoto University, Graduate School of Medicine,
SPONSOR AND HOST INSTITUTION:
Ritsuko Komaki MD Anderson Cancer Center
DATES OF VISIT: June 1-June 30, 2001
SUMMARY OF ACTIVITIES:
Objectives: To survey on the current status of radiotherapy for lung cancer, and multi-institute clinical studies on lung cancer. Achievements :
1 . The current ongoing protocols of radiotherapy for lung cancer at the MD Anderson Cancer Center were surveyed.
2. The previous and current protocols of RTOG study for lung cancer were surveyed, and the procedures to manage the RTOG studies were also surveyed.
3. The most advanced radiotherapy techniques for lung cancer including IMRT and respiratorygating irradiation were surveyed. For future: More sophiscated multi-institutional protocols of radiotherapy for lung cancer should be established in Japan in cooperation with radiation oncologists, surgeons and medical oncologists. On the other hand, a more advanced technology should be utilized for the treatment of lung cancer.
(16) Uhi Toh
Kurume University, School of Medicine
SPONSOR AND HOST INSTITUTION:
Thomas Sayers, PhD; Laboratory of Expremental Immunology, National Cancer Institute-Frederick
DATES OF VISIT: 6/30/01~9/27/01
SUMMARY OF ACTIVITIES:
Purpose: To. investigate the synerginetic interaction in inducing tumor cell apoptosis between the proteasome inhibitor . (PS341) and TRAIL (TNF-related apoptosis inducing ligand).
Objectives:
1) Screening the sensitivity of the human tumor cell lines (breast cancer: 2, prostate cancer: 1 and melanoma: 9)to the combination of PS341 and TRAIL by MTT assay and Crystal violet assay.
2) Investigating molecular basis of the synergy between the PS341 and TRAIL by examining levels of pro- and anti-apoptotic proteins(cFlip, IAPs, bcl-2 etc. ) in the cells using Western blotting
Results:
1) All of 12 tumor cell lines showed high sensitivity to the combination of PS341 and TRAIL, but the dose response curve of the positive interaction between PS341 and TRAIL was very tight, With regards to the levels of PS341. Therefore high dose of PS341 alone (50-100um) is toxic to the most cells.
2) After pretreating the tumor cell lines by PS341 , the expression of cFlip, cIAP and XIAP demonstrated significant decreasing in the levels of the proteins detected by immunological stain (Western Blotting). The level of DR5 receptor protein also showed a decrease after the pretreatment of PS341. The expression of Caspase 8 was enhanced by a short treatment time(2-4hrs), but was decreased by a long treatment time (overnight). These results suggested TRAIL activity against multiple tumor cells could be enhanced or recovered by proteasome inhibitor (PS341) depend on blocking pro- and anti-apoptotic proteins (cFlip, IAPs etc.). The combination of TRAIL and PS341 showed unique cytotoxic effects on the various tumor cell types in vitro. The response of the antitumor activity of this combination should be investigated in vivo in future study.
(17) Hironobu Yanagie, M.D., Ph.D.
Department of Surgery Teikyo University School of Medicine Ichihara Hospital
SPONSOR AND HOST INSTITUTION:
Professor Leaf Huang, Ph.D. , University of Pittsburgh
DATES OF VISIT: March 17th, 2002 -- March 31st, 2002 ( 15 days )
SUMMARY OF ACTIVITIES:
The complex of cationic liposome and plasmid DNA, so called, " Lipoplex " are applied to gene therapy as non-viral vector,
Recently cationic liposome is used to clinical trials of gene therapy, so it is the most important issue that how the transfection efficiency using lipoplex is increased to the level of that of viral vector in non-viral vector gene therapy. In order to increase the transfection efficiency, the LPD is composed with cationic liposome and plasmid DNA which is condensed with protamine salfate in the Leaf Huang’s Lab. A novel fusogenic peptide, JTS-1, in Influenza virus are also conbined to plasmid DNA to increase the transfection efficiency in his Lab .
(1) Tumor growth suppression by NG-1 and p53 suppressor using LPD/JTS-1 gene delivery system
In this visiting period, I prepared the LPD conjugated with JTS~1 and evaluate the expression of NG-1 gene, a novel tumor suppressor, and p53 gene, well known as tumor suppressor, to pancreatic cancer cell line, AsPC-1. NG-1 and p53 induce apoptosis. We have reported that NG gene have cloned in the cytotoxicity inducing region of hepatitis C virus, and have homology from 1 to 5. The effect of apoptosis is strongest by NG-1 transfection. Every cancer cell line that was transfected with lipofection of NG genes showed the apoptosis in 72 hours in vitro.
The concentrations of cationic lipid and plasmid DNA in LPD were as below; DCchol : plasmid DNA : protamine : JTS-1 = 3nmol : 1µg : 2µg : 0.1µg The mean diameter of LPD is 50 nm and it is so compact because the diameter of lipoplex is between 300 nm and 1000nm.
I evaluate the expression of NG-1 / GFP protein and apoptosis in AsPC-1 cells by flow cytometer(FACS) and fluorescent microscopy. The induction of apoptosis by NG-1 was strongest according to the expression of NG-1 / GFP protein by the double staining of FACS with Annexin V and PI. The expression of NG / GFP protein was strongest in NG-1 gene transfectant cells by fluorescent microscopy. Almost cells were dead by induction of apoptosis that was exprained relative low numbers of PI staining of nucleus after 48 hour transfection of NG-1 gene.
The tumor growth suppression was shown by the induction of p53 and NG-1 protein with the transfection of tumor suppressor genes using LPD/JTS-1 systems.
(2) Enhancement of tumor growth suppression by using transferrin binding oxaliplatin entrapped stealth liposome
In order to examine the combination therapy with drug delivery system of anti-cancer agents, we prepared transferrin binding oxaliplatin entrapped stealth liposome(TF-PEG/oxali-lip). The oxaliplatin is a novel platimun anticancer agent, and used for gastric cancer , colon cancer or ovarian cancers in clinically. This compound has low toxicity for kidney or bone marrow, so that be hopeful for treatment of therapeutic-resistant cancers in future. The stealth liposome is coated with polyethylene-glycol, so the liposome is able to escape by phagocytosis from macrophages ( reticuloendotherial sysytem), and also increase the retension in the blood caused increasing the chance to reach the cancer tissues. The liposome is also conjugated with transferrin, so that the liposome can reach to cancer cells expressed the transferrin receptors on the surf aces .
Apoptosis was strongly induced by addition of TF-PEG/oxali-lip. The induction of apoptosis by the TF-PEG / oxali-lip was shown by the double staining of FACS with Annexin V and PI. The tumor growth suppression was shown by the reaction of oxaliplatin and induction of p53 and NG-1 protein using LPD / JTS-1 gene delivery systems.
These results suggest that the "LPD/JST-1" which prepared with cationic lipid and DNA plasmid which was condensed with protamine and fusogenic peptide has become an strong candidate for non viral vector for gene therapy The tumor suppressor gene delivery using "LPD " system has applied to nonviral vector targeting for cancer gene therapy. The combination therapy of anti-cancer agents and tumor suppressor gene delivery is also easily induced the apoptosis, so this combination will be recommended for treatment of therapeutic-resistant cancers in near future.
(18) Masuhiro Takahashi
Niigata University
SPONSOR AND HOST INSTITUTION:
Duke University
DATE OF VISIT: September 3-16, 2001
SUMMARY OF ACTIVITIES:
Objectives of study
1) To explore the efficient way of getting exosomes derived from human dendritic cells.
2) To exchange the information of establishing the dendritic cell and exosome therapy for the patients with malignancies .
Achievements
1) Tlnnor antigen RNA transfection into dendritic cells was thought to be one of the most efficient way of getting exosomes derived from human dendritic cells for anti-tumor immunotherapy. The procedure of RNA transfection was studied practically in Duke University.
2) Evaluation of the immune responses was shown to be one of the most important process for establishing the dendritic cell and exosome therapy for the patients with malignancies. Tetramer analysis, ELISPOT assay, proliferation assay and cytoplasmic IFN- expression were thought to be the most reliable ways for evaluating immune responses in the patients who have received the dendritic cell or exosome therapy for the patients with malignancies. Each procedures of these assays were studied in Duke University.
How study relates to future work
1) RNA transfection will be applied to the procedure of getting exosomes derived from human dendritic cells in Japan, Niigata.
2) Tetramer analysis, ELISPOT assay, proliferation assay and cytoplasmic IFN- expression were planned to be induced to the immune evaluation of the patients who have been and will be treated with dendritic cells in Japan, Niigata.
(19) Haruhiko Sugimura
Hamamatsu University School of Medicine
SPONSOR AND HOST INSTITUTION:
Laboratory of Human Carcinogenesis, NCI
DATES OF VISIT: Aug30, 2002 - Sepl9, 2002
SUMMARY OF ACTIVITIES:
Objectives:
To discuss about the possible application of human genome information to molecular epidemiology of cancer. Discussions covers
1. ulcerative colotis and colon cancer susceptibility interms of NO involvement
2. XPD polymorphism and lung cancer risk
3. Breast cancer case-control study
4. Usage of microtissue array to screen early detection of chromosomal change
5. Usage of pathology archives by innovate FISH technology
Achievements:
Introduction of microtissue array int our FISH study was achieved.
Updated discussion with specialists in this field provide an occasion to promote undergoing work related and brought upon several publications.
Publications resulting from this experience:
Takahisa Furuta, Naohito Shirai, Misako Takashima, Fang Xiao, Haruhiko Sugimura Effect of genotypic differences in interlueukin-1beta on gastric juice pH in Japanese patients infected with Helicobacter pylori. Am J of Med,Å@112:141-3.2002
Changming Gao, Toshiro Takezaki, Jianzhong Wu, Zhongyou Li, Jiandong Wang, Jianhua Ding, Yanting Liu, Xu Hu, Tianliang Xu, Kazuo Tajima, Haruhiko Sugimura Interaction between Cytochrome P450 2E1 Polymorphisms and Environmental Factors with Risk of Esophageal and Stomach Cancers in Chinese. Cancer Epidemiol Biomark & Prev, 1l:29-34, 2002
Takezaki T, Gao CM, Wu JZ, Li ZY, Wang JD, Ding JH, Liu YT, Hu X, Xu TL, Tajima K, Sugimura H. The hOGGI ser326cys polymorphism and modification by environmental factors of stomach cancer risk in Chinese. Int J Cancer, 99:624-7, 2002
Takahisa Furuta, Emad m El-Omar, Fang Xiao, Naohito Shirai, Misako Takashima, Haruhiko Sugimura Effect of genetic polymorphism in interleukin-1_ on gastritis, gastric juice pH, and recurrence of peptic ulcer disease in relation to H.pylori infection. Gastroenterol, 123:92-105, 2002.
Song JP, Kitayama Y, Igarashi H, Guo RJ, Wang YJ, Kobayashi T, Konno H, Kataoka H, Tanaka M, Sugimura H Centromere numerical abnormality in the papillary, papillotubular type of early gastri cancer, further characterization of a subset of gastric cancer.
Int J of Oncol. 21:, in_press, 2002