(1) UEDA, Ryozo
Nagoya City University Medical School
SPONSOR AND HOST INSTITUTION :
Professor Riccardo Dall-Favera
College of Physician & Surgeons of Columbia University
DATE OF VISIT : from October 3, 1998 to October 14,1998
SUMMARY OF ACTIVITY
The purpose of my visit to the following four researchers in USA was to discuss and learn the present status of "Molecular targets of cancer diagnosis and therapy " in USA. The following information was obtained through this program.
Dr. Riccardo Dalla-Favera (URIS Professor of Pathology and Genetics & Development, Director, Division of Oncology, College of Physicians & Surgeons of Columbia University) has been studying on molecular mechanism of B cell lymphoma, and has previously found the translocation (8;14) in Burkitt's lymphoma, and has shown that this alteration lead to juxtaposition of MYC oncogene located on chromosome 8 with heavy chain gene of immunoglobulin (Ig) located on chromosome 14. Recently, he has been focusing on the analysis of chromosome 13q (13q14) deletion, which is observed around 60 % of chronic lymphocytic leukemia and, 70 % of mantle cell lymphoma. His group has sequenced genomic DNAs corresponding to minimal deleted lesion spanning more than 300 Kilo-bases, and found three novel transcripts, which are candidates for suppressor oncogenes located on 13q14. In addition, he has found that the incidence of mutation at the promoter region of the BCL-6 and that of hyper-mutation of Ig heavy-chain gene correlated well. Moreover, we discussed the on going joint project as for MUM1 gene whose importance has been found in oncogenesis of multiple myeloma.
Dr. Gary A Silverman (Joint Program in Neonatology, Harvard University, Department of Pediatrics) has discussed the recent molecular progress of 18q- syndrome analyzed in his group. He succeeded in isolation of a two gene encoding squamous cell carcinoma antigen 2 (SCCA2), which is a novel serpin that inhibits the chymotrypsin-like proteinases Cathepsin G and mast cell chymase. SCCA serve as a serological marker for more advanced squamous cell tumors of the cervix, lung, and oropharynx. His data showed that SCCA2 is a novel inhibitor of two physiologically important chymotrypsin-like serine proteases, so it might be closely related to cancer invasion and metastasrs.
Dr. Sydney Welt (Member Sloan-Kettering Cancer Center) has been engaged in utilization of humanized monoclonal antibody A33 against colon cancer. He organized the clinical phase I and II trials according to their clinical protocol. The results obtained showed no severe toxicity, although induction of human anti-humanized A33 (HAHA) reactivity was unexpectedly observed in almost all cases. No clinical response has been obtained in phase II trials so far.
Dr. Robert C Bast Jr., M.D. (Head, Division of Medicine, MD Anderson Cancer Center, The Univ. of Texas) has been promoted the clinical study of monoclonal antibodies CA125 for ovarian cancer and HER-2/neu for breast cancer patients supported by FDA and companies of the venture business strongly. The system of clinical phase trials in USA is totally different from that Japan in terms of financial supports by both government and venture business and of the evaluation system organized of by FDA.
(2) UMEZAWA, Kazuo
Keio University
SPONSOR AND HOST INSTITUTION:
Massachussetts Institute of Technology
DATES OF VISIT: Feb-Mar 1999
SUMMARY OF ACTIVITIES
1 . Schedule
I stayed in USA between Feb 15 and Mar 6, 1 999. I stayed in Cambridge at MIT for collaboration with Dr. Jun Liu from Feb 15 to Feb 21 , in Bethesda attending to the Angiogenesis Conference from Feb 21 to Feb 23, in New York at Columbia University for collaboration with Dr. I. Bernard Weinstein from Feb 23 to Mar 6. During the stay in New York I also visited University of Texas for giving the lecture.
2. Visit to MIT
In Dr. Jun Liu's lab at MIT I learned the technique of conjugation of our compound conophylline to biotin. Conophylline is a new vinca alkaloid that induces G1 block, inhibits TGF-beta action and inhibits TNF-alpha action. But its molecular target is not elucidated yet. Jun's methodology to determine the target was successfully used for the anti-angiogenesis drug TNP-470. I did several experiments trying to open the epoxide moiety of conophylline. My former graduate student Dr. Kakeya who is a post -doc in Jun's lab helped me. I will continue this project in Japan. In Boston I met Dr. T. Asahara at St. Elizabeth Medical Center, Tufts University. He recently found that endothelial cells are derived from the bone marrow. For our future prospects on angiogenesis research discussion with him was extremely fruitful.
3. Attending to Angiogenesis Conference in Bethesda
I attended to the Angiogenesis Conference in Bethesda on Feb 22 and 23 with my student. We present an article in the poster session on inhibition of in vitro angiogenesis by a proteasome inhibitor. Each session was stimulatiing. But for me the discussion with Dr. Asahara was more valuable.
4. Visit to Columbia University
I worked as a post doc in 1979 for 4 months in Professor Bernard Weinstein's lab. I worked on the new tumor promoters and published two papers one in Nature and another in Biochem. Biophys. Res. Commun. The stay of 20 years ago was thus very productive. This time the stay was only for two weeks and short. However, it was also a very valuable experience. First I learned the technique of cyclin D1 promoter assay. A post doc Jae taught me the use of reporter plasmid. Then, I tested our sample conophylline in this assay system for the effect on serum-induced activation of cyclin D1 promoter. I also proposed to collaborate with Bernie to screen inhibitors of cyclin D1 promoter from microbial culture filtrates. Bernie agreed. Also it was nice to talk with another post doc Haim from Israel on the effect of Helicobacter pylori on the cell cycle of gastric cells. I gave a seminar on tyrosine phosphatase inhibitors. It was also nice to see Dr. Raymon Persons in Columbia University. He characterized a new tumor suppressor gene Pten, - which product should be important to regulate various signal tranduction pathways. We discussed to screen inhibitors of Pten product from natural sources.
5.Visit to University of Texas
I was invited to give a lecture in University of Texas, Austin by Professor Laurence H. Hurley. I introduced our signal transduction inhibitors and discussed the possibility of development of tyrosine phosphatase inhibitors to antidiabetic drugs. The discussions after lecture was pleasant one. In addition to the lecture, Laurence, myself and Hal Korn from Houston discussed having the 2nd US-Japan Chemistry-Biology Interface Symposium. It will be in Hawaii on Dec 12-13, 2000. I was a chairman of the 1st Symposium in Tokyo in 1 997.
6. Summary
This trip to USA was accomplished exactly as planned. It was really a wonderful and fruitful trip.
(3) MASANORI HATAKEYAMA
DEPARTMENT OF VIRAL ONCOLOGY CANCER INSTITUTE
JAPANESE FOUNDATION FOR CANCER RESEARCH
SPONSOR AND HOST INSTITUTION :
DR. ROBERT A. WEINBERG
MEMBER
WHITEHEAD INSTITUTE OF BIOMEDICAL RESEARCH
PROFESSOR
DEPARTMENT OF BIOLOGY
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
DATES OF VISIT : FROM AUGUST I TO AUGUST 25, 1998
SUMMARY OF ACTIVITIES :
l . RESEARCH TITLE
The role of RB tumor suppressor gene family in the proliferation and differentiation of hematopoietic cells
2. PURPOSE
The purpose of the work is to study physiological role of the RB family proteins (pRB, p107 and p130) on the growth and differentiation of hematopoietic cells with the use of in vitro cellular differentiation system and cells derived from genetically engineered mice.
3 . BACKGROUNDS
It is well recognized that highly regulated cell cycle control is essential for both growth and differentiation of normal cells. In general, arrest of cells in G0/G1 phase of the cell cycle is an essential prerequisite for initiating cell differentiation. The retinoblastoma gene product (pRB) and its related proteins, p107 and p130, are strong inhibitors of cell growth and are suspected to play crucial roles in the arrest of cell cycle in G0/G1 phases. Importance of the pRB family proteins in normal cell growth is also supported from the observation that functional inactivation of pRB is associated with a wide variety of human cancers. In addition to their roles in cell cycle, evidence accumulates that the pRB family proteins are critically involved in the differentiation of certain cell types such as myocytes, adipocytes and osteocytes. However, little is known as for the role of pRB family proteins in the growth and differentiation of hematopoietic cells.
4. METHODS
The role of the pRB family proteins in the growth and / or differentiation of hematopoietic cells was examined either by overexpression of the protein in in vitro cell lines or by analyzing primary hematopoietic cells derived from genetically engineered mice that lack pRB family proteins.
5. Results
Each of pRB family proteins was conditionally overexpressed with the use of the modified tetracycline-inducible promoter in interleukin-3 (IL-3)-dependent 32D myeloid cells that differentiate into granulocytes in the presence of granulocyte-differentiation factor (G-CSF). Overexpression of wild-type pRB or a phosphorylation-resistant pRB mutant did not have any effect on the growth or differentiation of 32D cells. In striking contrast, conditional overexpression of p130 in the same cells induced cell growth suppression that was associated with granulocyte differentiation despite the absence of G-CSF. This indicates that p130 has a critical role in the myeloid differentiation. The role of pRB family proteins in hematopoietic cell growth and/or differentiation was also examined with the use of lymphoid cells prepared from mice lacking one of the three members of pRB family proteins by using the gene targeting technique. As far as examined to date, lack of one of any pRB family proteins does not appear to affect cellular responsiveness to cytokines or mitogens significantly. This may indicate the existence of functional redundancy among the pRB family proteins. To address this issue, we have started to generate lymphoid cells that lack all of the three pRB family proteins. Such mutant cells are generated by injecting ES cells that are nullizygous for RB, p107 and p130 genes into blastocysts derived from RAG2-knock out mice. Since the RAG2 knock-out mice cannot generate mature lymphoid cells of their own because of the lack of VDJ recombinase, chimera mice made from the blastocysts should possess mature T and B cells that are exclusively derived from ES cells that lack all pRB family proteins. Generation of such chimera mice is now underway.
6. Future plan
Through this collaboration work, we will establish a panel of hematopoietic cell lines derived from genetically engineered mice that lack one or various combinations of pRB family proteins. By comparing their growth and differentiation properties with normal control cells that possess all three pRB farnily proteins, we believe that we can understand physiological roles of these proteins in the hematopoietic cell growth and differentiation.
(4) Takaaki Akaike
Kumamoto University School of Medicine
SPONSOR AND HOST INSTITUTION:
Louis Ignarro (University of California, Los Angeles)
Hilary Koprowski (Thomas Jefferson University)
Bruce Freeman (The University of Alabama at Birmingham)
DATES OF VISIT:
10th July - 15th July: Professor Louis Ignarro (UCLA)
16th July - 21st July: Professor Hilary Koprowski (Thomas Jefterson University)
21st July - 26th July: Professor Bruce Freeman (UAB)
SUMMARY OF ACTIVITIES:
1) 3rd International Conference on NO at UCLA:
I attended the NO Conference at UCLA, and four poster papers and an oral talk were presented by our group. One of the poster papers was my own presentation, which reported acceleration of genetic mutation of virus caused by peroxynitrite. In the poster session, there were a couple of paper presented showing NO-induced mutation of some eukariyotic cells. I was impressed that increasing attention is now paid to an important role of oxidative stress in the carcinogenesis mechanism induced by inflammation and infection. Other achievement reported in this recent conference was a proposal for a novel cytotoxic mechanism due to enhanced formation of nitrotyrosine in biological systems. Specifically, Eiserich et al. from Alabama clearly showed that a selective incorporation of nitrotyrosine into!!
!-tubulin in the cells, resulting in delayed apoptotic cell death. Overall, I could exchange uptodate information on NO research with many scientists at the meeting. 2) Visit to Thomas Jefferson Univ.:
I was invited by Prof. H. Koprowski at Thomas Jefferson Univ., where I gave a talk entitled "Viral mutation caused by peroxynitrite". My presentation was very well appreciated by the audience, who are the members of Center for Neurovirology at the University. Many questions asked by the audience were those regarding the possible implication of oxidative stress-induced mutation to other viruses such as HIV and hepatitis C virus, which are well known to be frequently mutated, and thus not only vaccination but also anti-viral chemotherapy are not effective as people expect. In addition, Koprowski's group is now testing therapeutic effect of uric acid as a peroxynitrite scavenger in multiple sclerosis, in which peroxynitrite is shown to be critically involved by their recent study. Thus, the therapeutic approach with use of peroxynitrite scavenger appears to be useful for not only antiinflammatory treatment but also cancer prevention, because peroxynitrite is recently suggested to contribute the carcinogenesis.
3) Visit to UAB:
I visited Prof. Bruce Freeman, director of the Center for Free Radical Biology at UAB. Many groups including Freeman's and Beckman's group are working at the center, and they are enthusiastically studying pathophysiology of NO and oxygen radicals. There are a number of recent great achievement in the NO research done by the groups at UAB. I gave two lectures for the groups in the center. One was a talk entitled "Oxidative stress-induced viral mutation", in which I demonstrated very nicely acceleration of RNA virus mutation by peroxynitrite. A lot of questions and comments were given at the seminar. One of the most important question was as to the formation of nitroguanine in viral RNA, and many audience are interested in the tremendous stability of nitroguanine in the viral genome. In general, I could have very helpful discussion with regard to the mutagenesis potential of peroxynitrite.
Also, it was a great chance for me to obtain most uptodate information on NO/peroxynitrite from all groups at the free radical center. Moreover, I could learn details of newly developed nitrotyrosine assay based on HPLC/EC detector analysis, which have been established lately over there. In future, I believe that the analysis for endogenous nitrated substances in biological system will become the most important approach to verify the involvement of oxidative stress in carcinogenesis.
It was fruitful opportunity with great intellectual profit to visit this time many eminent scientists of NO. I appreciate gratefully of the generous and courteous financial support for my trip to and stay in the US. I hope this kind of exchange program should be executed 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.
(5) Shoichiro Tsugane
National Cancer Center Research Institute East
SPONSOR AND HOST INSTITUTION:
National Cancer Institute (NCI)
Division of Cancer Epidemiology and Genetics(DCEG)
Nutritional Epidemiology Branch
Dr. Rashmi Shinha
Harvard School ofPublic Health (HSPH)
Department of Nutrition
Prof. Walter Willett
DATE OF VISIT: February 3, 1999 - February 13, 1999
SUMMARY OF ACTIVITIES :
Objectives:
To make information exchange in the area of Nutritional Epidemiology on diet and cancer by visiting Nutritional Epidemiology Branch, National Cancer Institute and Department of Nutrition, Harvard School of Public Health.
Achievement :
National Cancer Institute, Rockville, MD
I visited Drs. Fraumeni (Director ofDivision of Cancer Epidemiology and Genetics (DCEG)), Sinha, Ziegler, Schatzkin, Neely (Nutritional Epidemiology Branch, DCEG), Rothman (Occupational Epidemiology Branch, DCEG), Manns (Viral Epidemiology Branch, DCEG), Ballard-Barbash Subar Thompson (Applied Research Branch, Division of Cancer Control and Population Science) and obtained much information on diet and cancer research in NCI. I also provided a seminar "A population-based cohort on diet and chronic diseases in Japan" and about 30 epidemiologists in NCI attended. We made a fruitful discussion after my seminar.
Harvard School of Public Health, Boston, MA
I visited Drs. Willett, Rimm, Giovanucci (Department of Nutrition) and obtained much information on three ongoing cohort study (Health Professional Follow-up Study, Nurses' Health Study I and II) and recent outcomes regarding diet and cancer from these cohorts. We also made information exchange on dietary assessment method in epidemiological study and validation study for developed food frequency questionnaire. I conducted a research on how we calculate nutrient intake from simple food frequency questionnaire which include 38 food group items and used in the baseline survey in our cohort study in Japan. Some researchers in the department made significant contribution to my research in Harvard.
Implication:
I believe that this experience will give a great contribution in my future research on diet and cancer through our ongoing prospective study on diet and cancer.
ACKNOWLEDGMENTS :
I would like to express my appreciation to JSPS for giving me this important visit to US institutions.
(6) Hirokazu Nagawa
Department of Surgical Oncology, Graduate School of Medicine, The University of Tokyo
Title and degree: Associate Professor, M.D., Ph.D.
Address: 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan
Subject of research:
Application of molecular biology to diagnosis and therapy in
gastrointestinal cancer
Period of stay: 13 days
from October 10, 1998 to October 22, 1998
Name of host scientists/institution
1) Isaiah J. Fidler/MD Anderson Cancer Center
2) Claudio Fiocchi/Case Western Reserve University
3) Victor W. Fazio/The Cleveland Clinic Foundation
Summary of research
Research into experimental equipments and system at three proposed laboratories was made focusing on cancer research for three days in each institute.
There was no difference between the three laboratories in U.S.A. and the University of Tokyo, Japan in terms of experimental equipments such as those used for cell biology; clean bench, incubator, temperature-adjusted centrifugal machine and fluorescent activated cell sorter (FACS), such as used for molecular biology; polymerase chain reaction machine, electrophoresis and gene-analyzer.
On the other hand, system of experimental research in U.S.A. was observed to be different from that in Japan. A labo-manager has responsible for administering an order, purchase, and maintenance of experimental tools and materials so that double order and/or purchase of materials by researchers can be avoided. It is reasonable and necessary for laboratories in Japan to introduce the system.
With respect to an exchange of opinion, the visitor made a presentation to several researchers at each visited institute concerning the results of biological markers to make a diagnosis of metastasis and those of genetic markers to select patients with cancer in terms of chemotherapy and radiotherapy. This presentation was highly evaluated by researches in U.S.A. And the investigators in U.S.A. made a presentation mainly concerning cancer metastasis entitled "Angiostatin-mediated suppression of cancer metastases by primary neoplasms engineered to produce granulocyte/macrophage colony-stimulating factor", "Interferon-!!
!prevents the upregulation of interleukin-8 expression in human melanoma cells" and "Prevention of intestinal toxic effects and intensification of irinotecan's therapeutic efficacy against murine colon cancer liver metastases by oral administration of the lipopeptide JBT 3002" followed by discussion between the visitor and the host scientists. Through the U.S.-Japan Cooperative Cancer Research Program, a productive visit and a fruitful experience were achieved in terms of obtaining U.S. opinions which have always a direction to application of basic science to clinical setting.
(7) TOSHIHARU YAMAGUCHI
DEPARTMENT OF SURGERY, KYOTO PREFECTURAL UNIVERSITY OF MEDICINE
SPONSOR AND HOST INSTITUTION :
Dr. Ira Pastan
Laboratory of Molecular Biology, National Cancer Institute, NlH
Building 37, Room 4E16, 37 CONVENT DR MSC 4255, BETHESDA MD 20892-4255, USA
DATES OF VISIT:
February 22, 1 999
I visited Dr. Ira Pastan at Laboratory of Molecular Biology of NCI and presented our experimental and clinical study on Missile cancer chemotherapy using monoclonal antibody A7 and anticancer drug Neocarzinostatin. We discussed about the problem and future of immunotargeting cancer chemotherapy.
February 23-24, 1999
I visited Dr.E.W.Martin at Division of Surgical Oncology of A.G.James Cancer Hospital and Research Institute and observed the imnrunoguided surgery at operating room. I presented our data and discussed on the problem and future of this new technique.
SUMMARY OF ACTIVITIES :
My main object to visit Dr. Pastan was to discuss about clinical application of immunoconjugates for cancer therapy. We discussed about advantages and disadvantages of three type of immunoconjugates. Dr. Pastan recommended the immunoconjugate of Fv fragment and the pseudomonas exotoxin. He was kind enough to show me latest data of clinical trial of his new immunotoxin LMB2. He pointed out that further characterization of epitopes defined by our monoclonal antibody A7 will be necessary to proceed our study for clinical trials.
My main object to visit Dr. Martin was to observe immuno-guided surgery and discuss about possibility of our monoclonal antibody A7 as a good tool to detect metastases using the technique of inrmuno-guided surgery. Through our discussion It was concluded that we should use 125-Iodine Instead of 131-Iodine for labeling of antibody. The dose of radiolabelled antibody and timing of counting were discussed. Such discussions were useful to improve our projects on immuno-guided surgery.
I could observe sentinel lymph node biopsy during operation of the breast cancer using handy gamma-detector. Also we made a round of visits to Dr. Martins patients and could have chance to realize clinics in USA. At the Kettering Medical Center, new 3D-Positron Emission Tomography was presented and the splendid images impressed us.