SEMINARS

BASIC SCIENCE Recent Advances in Specific Immunotherapy of Cancer Hawaii: March 21 -22, 2001
	Organizers:	Dr. Douglas SCHWARTZENTRUBER Dr. Kyogo ITOH
	Participants:	US: 10 Japan: 10
CLINICAL SCIENCE Development of Tailor-Made Cancer Therapy Hawaii: February 17- 18, 2001
	Organizers:	Dr. Francesco M. MARINCOLA Dr. Tetsuya TOGE
	Participants:	US: 9 Japan: 9
EPIDEMIOLOGY/BEHAVIORAL SCIENCE Molecular Epidemiology of Nicotine Addiction and Lung Cancer Hawaii: February 9- 11, 2001
	Organizers:	Dr. Peter SHIELDS Dr. Haruhiko SUGIMURA
	Participants:	US: 10 Japan: 10

SEMINAR REPORT
UNDER THE U.S.-JAPAN COOPERATIVE CANCER RESEARCH PROGRAM

1. Recent Advances in Specific Immunotherapy of Cancer
Period: from March 21^st to March 22^nd, 2001 for 2 days
Site: Maui Prince Hotel, Maui, Hawaii

Japanese Organizer:
Itoh Kyogo
Affiliation Kurume University School of Medicine
Department of lmmunology
Professor and Chairman

US Organizer:
Schwartzentruber Douglas
Affiliation National Cancer Institute
Surgery Branch
Senior staff

List of Japanese-side Participants (except for Organizer):
Ryuzo Ueda
Professor
Nagoya City University Medical School

Hiroshi Shiku
Professor
Mie University School of Medicine

Keigo Endo
Professor
Gunma University School of Medicine

Yutaka Kawakami
Professor
Keio University School of Medicine

Nobuo Hanai
Vice-Chief
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co. Ltd.,

Yuichi Obata
Vice-Chief
Aichi Cancer Center Research Institute

Keiko Udaka Associate Professor
Kyoto University

Yuji Noguchi
Associate Professor
Okayama Medical School

Mamoru Harada
Lecturer
Kurume University School of Medicine

List of US-side Participants (except for Organizer) :

David Avigan
Director
Beth Israel Deaconness Medical center

Olivera J. Finn
Professor
University of Pittsburgh Cancer Institute

Eli Gilboa
Senior Researcher
Duke University Medical Center

Philip Greenberg
Professor
University of Washington and Fred Hutchinson Cancer Research Center

Ralph Reisfeld
Professor
The Scripps Research Institute

Craig Slingluff
Professor
University of Virginia

Louis Weiner
Chairman and Senior Member
Fox Chase Cancer Center

James Yang
Senior Staff
National Cancer Institute

Sacha Gnjatic
Assistant Member
Memorial Sloan-Ketterin Cancer Center

Itinerary and Topics

Tuesday, March 20th
	Check in/Arrival
Wednesday, March 21st
7:40 am	Introductions/Plan for Meeting
Session I Peptide-Based Immunotherapy of Cancer (Chair - Kyogo Itoh)
7:50 am	Identification and clinical application of peptides of epithelial cell-derived cancer	Kyogo Itoh
8:30 am	Humoral and cellular immune responses to cancer-testis antigens in cancer patients	Yuji Noguchi
9:10 am	Identification of new melanoma antigens using an in vitro immunoselected melanoma cell line	Mamoru Harada
9:50 am	Coffee Break
10:10 am	WT tumor antigens and tumor immunity from a view of self-nonself discrimination	Keiko Udaka
10:50 am	Role of helper T cells in activation of CTL	Hiroshi Shiku
11:30 am	Lunch
Session 11 Peptide-Based Immunotherapy of Cancer (Chair - Douglas J. Schwartzentruber)
1:00 pm	New and old epithelial tumor antigens as cancer vaccine candidates	Olivera J. Finn
1:40 pm	Cellular and molecular engineering of T cell responses to overcome obstacles to effective T cell therapy of human tumors	Phil Greenberg
2:20 pm	Immunity to NY-ES0-1: Monitoring T cell responses and clonical trials	Sacha Gnjatic
3:00 pm	Coffee Break
3:20 pm	A step by step approach to development of peptide vaccines for melanoma	Craig Slingluff
4:00 pm	Efficacy of peptide vaccines in the treatment of metastatic melanoma	Douglas J. Schwartzentruber
6:00 pm	Banquet,
Thursday, March 22nd Session III Antibody-Based Immunotherapy of Cancer (Chair - Kyogo Itoh)
7:50 am	Human tumor antigens in various cancers, detected by SEREX	Yutaka Kawakami
8:30 am	Significance and clinical application of cancer-associated antigens identified by SEREX	Yuichi Obata
9:10 am	Development of humanized antibodies against gangliosides and VEGF receptors	Nubuo Hanai
9:50 am	Coffee Break
10:10 am	Radioimmunotherapy of cancer in Japan	Keigo Endo
10:50 am	Recent clinical trials for cancer therapy with monoclonal antibodies in Japan	Ryuzo Ueda
Session IV Antibody-Based Immunotherapy of Cancer (Chair - Douglas J. Schwartzentruber)
1:00 pm	Clinical and Laboratory Developments in the Immunotherapy of Renal Cancer	James Yang
1:40 pm	Engineered antibodies for cancer therapy	Louis Weiner
2:20 pm	Dual function DNA vaccine for cancer immunotherapy	Ralph Reisfeld
3:00 pm	Coffee Break
Session V Dendritic Cell-Based Immunotherapy of Cancer (Chair - Douglas J. Schwartzentruber)
3:20 pm	Tumor RNA transfected dendritic cells: From vaccination to antigen discovery	Eli Gilbo
4:00 pm	Use of dendritic cell/tumor fusions as a novel cancer	David Avigan
4:40 pm	Discussion/Conclusions
6:00 pm	Farewell Dinner - Hakone's Restaurant at the hotel
Friday, March 23rd
	Check out

Scientific Achievements
1. Summary
The meeting was held for approximately 1000 minutes, that was 16 hours, and each of twenty attendants made a presentation for 40 -50 min including discussion. Throughout the meeting, almost attendants had joined the meeting and discussed about recent progress in specific anti-tumor immunotherapy.
1-1. Basic Research
The main subject of the first day was specific anti-tumor immunotherapy using peptides, and it was revealed that many genes, which encode tumor antigens and their peptides, have been identified not only in melanoma but also in epithelial cancers. Namely, approximately 200 tumor antigens, which have been identified using tumor-reactive cytotoxic T lymphocytes (CTLs) using the cDNA library expression cloning method, and approximately 300 peptides have also been identified. On the other hand, more than 1500 molecules, which can be recognized by IgG antibody in serum of cancer patients, have been identified by the SEREX method. These progresses suggest that further new tumor antigens will be identified within a couple of years, and that we will be able to get hold of the whole immune system participating in tumor-specific immune responses (T cells/ B cells). Specific immunotherapy using antibodies was the main subject of the second day. It was revealed that antibodies, which have the potential to show strong antitumor effects, have been developed based on methodological progresses in the preparation of humanized antibodies and molecular targeting techniques. In addition, several clinical trials revealed that antibodies targeting signal transduction molecules could show an efficient clinical effect and that this field of research is expected to make a rapid progress. Furthermore, anti-tumor vaccine using tumor-derived mRNA or dendritic cell (DC)/tumor cell fusion was suggested to be a potent immunotherapeutical protocol.
1-2. Clinical Research
It was suggested that efficient vaccine protocols using tumor antigen-derived peptides have not been established yet. Therefore, the following points were addressed in the meeting:
A) Which peptides should be administered into patients with different types of cancers?
B) Should either a single peptide or combined peptides be administered?
C) Should several peptides be administered at the same site or at different sites?
D) What volume, intervals and the number of times should be administered?
E) Does it need to measure CTL precursors before vaccination?
F) Should peptides be administered either subcutaneously, intradermally, or both?
G) What kinds of adjuvant should be used?
H) Are cytokines needed?
I ) Which cytokines including IL-2 and GM-CSF should be administered?
J) How should cytokines be administered?
K) Which systems are useful for monitering?
L) Is it needed to moniter humoral responses?
Because information using murine models has been not always applicable for clinical trials, questions described above can be resolved only by clinical trials. Therefore, it was concluded that further performance of clinical trials is needed to establish peptide-based anti-tumor immunotherapy. On the other hand, it was accepted that a major progress has been made in the filed of antibody-based anti-tumor immunotherapy, including anti-CD20 and anti-Her2/neu antibodies. In this meeting, the development of new antibodies and basic techniques to develop such antibodies were further discussed. The underlying mechanism of antibody-induced anti-tumor effect was further discussed. Although both antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) had been thought to be responsible for anti-tumor effect of antibodies, direct effect of antibodies was suggested in this meeting because antibodies against proteins participating in signal transduction could elicit anti-tumor effect clinically.
2. Individual Presentation
2-1. Dr. Kyogo Itoh: He reported that his group has identified more than 100 genes encoding human tumor rejection antigens and more than 200 tumor antigen-derived peptides. He also reported that a phase I trial using 13 peptides among them has been carrying out against advanced cancer patients (lung cancer and colon cancer). Based on this clinical trial, he asserted safety of vaccine with cyclophilin B-derived peptides and the induction of CTLS and antiboides against peptides administered. 2-2. Dr. Yuji Noguchi: He reported that the expression of NY-ESO-1 antigen, which was identified in the US by the SEREX method, was positively correlated with clinical stages, based on an analysis of NY-ESO-1 expression in pancreas and prostate cancers. He also reported that his group investigated the expression of an OY-TES-1 antigen, which was identified at the Okayama University, in colon cancers and cancers in the field of Urology, and identified several OY-TES-1 peptides, which could be recognized by CTLs.
2-3, Dr. Mamoru Harada: He reported 2 melanoma antigens, AIM-1 and AIM-2, which were identified at the Surgery Branch of NCI, USA, and have antigenic peptides recognized by HLA-A2-restricted and HLA-A1-restricted CTLs, respectively. These tumor antigens were newly 1identified using a cDNA library of melanoma cell line which had lost immunodominant melanoma antigens.
2-4. Dr. Keiko Udaka: She reported that WT-1 gene produce, which had identified as an oncogene, could be recognized by tumor-reactive CTLs. She also reported that, using murine CTL clones, whether self antigen-derived peptides could be recognized by CTLS or not was dependent on their affinity to HLA-class I molecules. She also reported that fyn, a member of src family, could participate in immunological tolerance of CTLs.
2-5. Dr. Hiroshi Shiku: He reported that mutated MAP kinase could be a tumor antigen on a murine fibrosarcoma line, CMS5a, and that his group identified its CTL epitopes. He also revealed the presence of tumor antigen on CMS5a by the cDNA array method. In addition, he reported that more than 20 tumor antigens were identified by the SEREX method, and that they contained antigenic epitopes recognized by CTLs, and that CD4+ T cells could participate in the induction of CTLs.
2-6. Dr. Olivera J. Finn: She overviewed MUC-1 antigen, which she has been working on for many years. She reported that CTL responses to this antigens was weak and humoral responses are dependent on IgM, but not on IgG. She also reported that a clinical trial using MUC- 1 peptides has been ongoing. She finally reported that her group identified cyclin B1-derived epitopes recognized by CTLs. She played an important role in this meeting as a leader.
2-7. Dr. Philip D. Greenberg: He reported a result of clinical trial of adoptive immunotherapy using tumor-reactive CD8+ CTL clones. It revealed that such CD8+ T cells could infiltrate into the tumor sites, but their retention was too short, approximately 7 days, to show clinical responses. He also reported that a clinical response was observed in the part of patients who received a low dose of IL-2, but that such patients developed recurrence shortly. Based on these findings, his group has been trying to transduce genes including CD28 and IL-2 receptor into CD8+ T cell clones using the retrovirus system. He also played a propulsive role in the meeting.
2-8. Dr. Sacha Gnjatic: He reported basic and clinical research on the NY-ESO-1 antigen. The percentages of detecting anti-NYESO-1 antibody were 50, 25 and less than 25% of patients with melanoma, lung cancers, and colon cancers, respectively He also reported the result of a phase I trial of peptide vaccination (100 microgram) with NY-ESO-1-derived peptides against HLA-A2+ melanoma patients. Vaccine effects such as the induction of CTLS and enhanced humoral responses were observed in patients having antibodies against the antigen prior to vaccination. He also reported a new NY-ESO-1 epitopes which could be presented by HLA-CW3 molecules using the transduction of this gene into EBVB cell line.
2-9. Dr. Craig Slingluff: He reported a comparative trial in which one group was administered DC pulsed with 4 kinds of melanoma antigenic peptides and the other group was subcutaneously administered with these peptides with GM-CSF/IFA. The result was that the latter group (6/12) was superior to induce CTLS than the former group (1/9). He also reported that IL-2 administration after peptide vaccination resulted in one partial response among 13 melanoma patients. He also reported that his group has been comparing antitumor effects between the group administered with mixed peptides and the group administered 4 peptides separately.
2-10. Dr. Douglas J. Schwartzentruber: He introduced both clinical trials of immunotherapy done at the Surgery Branch/ NCI in recent 20 years and ongoing clinical trials. He reported that 33% of melanoma patients who received vaccination with a gp100-2M peptide in combination with a high dose of IL-2 showed a partial response (PR). He also reported that 2 out of 16 melanoma patients showed clinical responses after vaccination with 4 different kinds of peptides. In addition, he reported that no clear response was observed by vaccination of virus vectors inserted with either MART-1 or gp100 gene. He pointed out that determination of optimal interval of peptide vaccination is an important problem. He was an organizer of the US side and took the chair throughout the meeting.
2-11. Dr. Yutaka Kawakami: He suggested that CD4+ T cells recognizing melanosomal proteins were suggested to participate in VKH disease, of which intraocular melanocytes are destroyed as the result of autoimmune responses. He reported newly identified melanoma antigens, including KU-MEL-1 and MART-3. In addition, he reported many melanoma antigens which were identified by newly-developed methods such as SAGE, SEREX, and gene-chip.
2-12. Dr. Yuichi Obata: He reported that more than 400 antigens were identified by the SEREX method using serum from cancer patients. He also reported that more than 1500 genes were registered in the database of SEREX home-page in cooperation with colleagues in the Memorial Sloan-Kettering Cancer Center. Aggressive discussion was done concerning clinical implication of these many tumor antigens identified by the SEREX method.
2-13. Dr. Nobuo Hanai: He reported basic and clinical research on humanized anti-Flt-1 antibody, He reported that this antibody could selectively block migration of cells and that another antibody, anti-KDR antibody, could selectively block cell proliferation. He also reported promising results such as no induction of HAMA, no adverse effect, and selective accumulation of antibody administered. Two out of 15 patients showed a partial response. He also addressed that his group has been preparing a new clinical trial using IL-2-fusion antibody and adriamycin/antibody conjugate.
2-14. Dr. Keigo Endo: He introduced the present status of radioimmunotherapy in Japan. He pointed out that facilities for radioimmunotherapy were behind compared with those in the US/Europe. He reported that a few percentage of patients with Graves' disease had been received radioimmunotherapy in Japan, but this therapy has been the first choice in the US. He also reported that this was the case with patients with gastric cancer and B cell lymphoma. He reported that his group has been planning to prepare 90Y-CD20 antibody and clinical trial using the antibody.
2-15. Dr. Ryzo Ueda: He overviewed 30 year's trials of cancer treatment with antibody, and suggested the possibility that new effective antibodies would be developed in the following 10 years. He pointed out that there were few original studies done by Japanese in this field. He also pointed out that new drugs, such as anti-CD20 and anti-Her2/neu antibodies, took a long time to obtain the governmental permission. He reported that such time-consuming procedure has been gradually improving after the establishment of PMDFC. He also reported basic research on anti-GD3 and anti-CD2 antibodies. He took a leadership for preparation of this meeting.
2-16. Dr. James Yang: He reported that immunotherapy could be effective in treating patients with melanoma or renal carcinoma, although the frequency was relatively low, based on his experience for 20 years at the Surgery Branch/NCI. He reported that more than 20 patients in both cancer patients were completely cured by administration of IL-2 alone. He also reported that administration of a high dose of IL-2 induced the most long effective period in spite of adverse effects. He also reported a clinical trial of humanized anti-VEGF antibody against renal cancer. The result was that although the rate of partial response was less than 10%, there was no severe adverse effect He also pointed out that time to progression (TTP) should be considered as a new estimation factor. He also reported a basic research on renal tumor antigen, such as FGF5, which could be recognized by HLA-A3-restricted CTLs.
2-17. Dr. Louis M. Weiner: He pointed out that 3 antibodies (anti-Her/2neu, anti-CD20, and anti-VEGF antibodies), which were recently revealed to be effective, were against molecules of signal transduction, and that other many antibodies, which are irrelevant to signal transduction, were ineffective. He showed his idea that ADCC seemed not to be responsible of antitumor effects by antibodies. He also referred to his new idea using bispecific (anti-Her2/neu + anti-Fc!!!RIII) antibody, Fab-SEA fusion protein, and C5a molecules.
2-18. Dr. Ralph A Reisfeld: He reported that, in a murine model targeting CEA antigen, anti-tumor effect of CFA/DNA vaccine could be enhanced by the cooperation with CD40 molecules and IL-2-fusion antibody. He has been a leader of cancer immunotherapy in the US, and played an advisory role in the meeting.
2-19. Dr. Elo Gilboa: He reported that tumor-specific CTLS could be generated by in vitro stimulation of DC pulsed with tumor mRNA, which could be obtained from a tiny biopsy. He reported that anti-tumor CTLS were able to generate in all cases tested. He reported that adoptive transfer of DC pulsed with tumor mRNA could generate CTLS in the periphery in 8 out of 9 cases, but no clinical responses was observed. He also reported that DC, transduced with telomerase gene could induce telomerase-derived antigen-specific CTLs.
2-20. Dr. David Avigan: He reported a basic and clinical trial in which tumor/DC fusion was used as anti-cancer vaccine. He reported that the induction of anti-tumor CTLS could be induced by vaccine with tumor/DC fusion, and that the rate of fusion was about 25% when DC and tumor cells were mixed at a ratio of 10:1. He reported that vaccine with tumor/DC fusion was applicable in 10 out of 19 patients with breast cancer, and that one patient who had received no chemotherapy showed a complete response. He also reported that this therapy was applicable in all 8 patients with renal cancer or melanoma.
3. Conclusion
Although the meeting was held only for 2 days, aggressive discussion about recent progress in specific antitumor immunotherapy was done. This meeting will contribute to establish the treatment of cancer patients with immunotherapy

Other Seminar Activities (Reception, Excursion, other meetings):
March 21, 2001
6:00 pm Banquet (sponsored by the US side)
March 22, 2001
6:00 pm Farewell Dinner (sponsored by the Japanese side)



2. Development of Tailor-Made Cancer Therapy
Period: 17-18 February 2001
Site: Maui

Japanese Organizer:
Toge Tetsuya
Research Institute for Radiation Biology and Medicine, Hiroshima University
Department of Surgical Oncology
Position: Professor and Chairman

US Organizer
Marincola M Francesco
National Cancer Institute, National Institutes of Health
Surgery Branch
Senior Investigator Bethesda, MD 20892

List of Japanese-side Participants (except for Organizer):
Masaki MORI
Professor
Department of Surgical Oncology, Medical Institute of Bioregulation, Kyushu University

Takashi NISHIMURA
Professor
Division of Immunoregulation, Institute for Genetic Medicine, Hokkaido University

Shinzaburo NOGLTCHI
Professor
Molecular Oncology, Department of Surgical Oncology, Faculty of Medicine, Osaka University

Masahiko NISHIYAMA
Professor
Department of Biochemistry and Biophysics, Research Institute for Radiation Biology and Medicine, Hiroshima University

Hideaki TAHARA
Professor
Department of Surgery, Institute of Medical Science, The University of Tokyo

Takao YAMORI
Senior Investigator
Cancer Chemotherapy Center, Japanese Foundation for Cancer Research

Yoshiyuki YAMAGUCHI
Assistant Professor
Department of Surgical Oncology, Research Institute for Radiation Biology and Medicine, Hiroshima University

Yuko KITAGAWA
Assistant Professor
Department of Surgery, School of Medicine, Keio University

List of US-side Participants (except for Organizer):
Douglas Hanahan
Professor
Dept. Biochem. Biophys., U Cal-San Francisco

David Botstein
Chairman
Dept. Genetics-, L329, Stanford University School of Medicine

David L. Bartlett
Surgery Branch, NCI, NIH

Ena Wang
Surgery Branch, NCI, NIH

Thomas Look
Vice Chair
Dana-Farber Cancer Institute

Richard O. Hynes
Department of Biology, Massachusetts In -stitute of Technology

John N. Weinstein
Laboratory of Molecular Pharmacology, NCI

Dean H. Kedes
Departments of Microbiology and Medicine, Thaler Center

Itinerary and Topics

Saturday, 17 February 2001 (The 1st day)
09:00	Opening remarks	Masahiko NISHIYAMA
Session I: Genetic diagnosis of cancer and tailor-made cancer therapy Chair persons: Thomas Look, Masaki MORI
09:10	Angiogenesis switch during tumorgenesis	Douglas Hanahan
09:40	Molecular Oncology	Masaki MORI
10:10	Apoptosis dys-regulation in cancer	John C. Reed
10:40	Mechanisms of apoptosis avoidance in cancer	Thomas Look
11:10	Viral pathogenesis in Kaposi's sarcoma	Dean H. Kedes
11:40	Discussion
Session IV: DNA technology and bioengineering for tailor-made cancer therapy Chairpersons: Francesco M Marincola, Hideaki TAHARA
13:30	Augmentation of DC-based tumor vaccination therapy by Th1 cells	Takashi NISHIMURA
14:00	TCR analysis for cancer therapy	Yoshiyuki YAMAGUCHI
14:30	Bioengineering therapy of cancer	Hideaki TAHARA
15:00	Cancer vaccines at the NCI	Francesco M Marincola
15:30	Discussion
Session II: Chemotherapy and tailor-made cancer therapy Chairpersons: John N. Weinstein, Masahiko NISHIYAMA
16:10	Functional genomics and cancer	David Botstein
16:40	Chemotherapy of cancer	Takao YAMORI
17:10	Gene expression database for the molecular pharmacology of cancer	John N. Weinstein
17:40	Tailor-made chemotherapy of cancer	Masahiko NISHIYAMA
18:10	Discussion
Sunday, 18 February 2001 (The last day) Session III: Surgical treatment and tailor-made cancer therapy Chairperson: Edison Liu, Shinzaburo NOGUCHI
09:00	Molecular portraits of metastatic melanoma in vivo	Ena Wang
09:30	Metastatic progression in cancer	Richard O. Hynes
10:00	Molecular Oncology	Shinzaburo NOGUCHI
10:50	Sentinel node-navigation surgery	Yuko KITAGAWA
11:20	Prospective for cancer research	Edison Liu
11:50	Discussion
12:10-12:30	Summary and Discussion
	Chairpersons: Francesco M Marincola, Masahiko NISHIYAMA
12:30	Closing remarks	Francesco M Marincola

Scientific Achievements
On February 17th through 18th 2001 the U.S.-Japan Cooperative Cancer Research Program sponsored a workshop to update scientists from both Countries about each other's contributions to the "Development of Tailor Made Cancer Therapies". The assumption of the Workshop was that human cancers may represent at the extreme, a mosaic of unique entities whose heterogeneity explains the unpredictability of treatment outcome. This assumption is supported by two observations. First the obvious disparity of clinical responsiveness to identical treatments of patients* affected by homonymous diseases. The second, more recent observation that sub-sets of disease can be identified through molecular depictions of their global transcript profiles. Present at the Workshop were leaders in this field. For instance, David Botstein, from Stanford University, CA, who together with Pat Brown developed the concept of high-throughput functional genomics, demonstrated the usefulness of this approach for the depiction of basic biological processes pertinent to cancer such as cell cycle regulation. In addition, he demonstrated the applicability of this strategy in the clinical context as his group has recently identified among morphologically similar breast cancer lesions subsets with markedly different transcriptional programs. Such sub-classification may have important clinical significance has one of the subsets expressed marker genes associated with specific prognostic behavior. Because of the recognized importance of the need for a novel strategy to address clinical oncology the Workshop was heavily weighted toward the promising field of high-throughput (micro-array) technology with the hope of revitalizing old concepts with fresh air from this unprejudiced discovery driven approach. Dr Weinstein, for instance, gave an in depth overview of the statistical challenges the field of bio-informatics is confronting. As observed by Dr. Look, a participant to the workshop, "after performing a couple of micro-arrays, we spent the following eight months trying to interpret the data". Analysis of large data sets is a result of using these powerful tools and can some times discouraging. However, the approach cannot be blamed for the complexity of biology as it only provides a realistic glance of the intricacy of events regulating biological phenomena. Although the data generated may seem overwhelming, it would not help to ostrich this complexity by following the old dogma that scientific knowledge can be achieved through the study of "Individual hypothesis / one variable at the time". As Dr. Weinstein and Dr Botstein pointed out there are ways to overcome the load of information using mathematicastatistical tools such as the hierarchical clustering model developed by Michael Eisen. In addition, the large throughput of information generated by array technology can be filtered through "common sense" or, more specifically, "biological sense". An important point brought up by Dr. Botstein was the ongoing controversy between "discovery" vs "hypothesis" driven research. In the 21st century many scientists feel uncomfortable to look at the big picture and they would rather study one hypothesis at the time by knocking out or putting new genes in mice. These scientists consider high-throughput efforts purely descriptive and not sufficiently hypothesis driven. As pointed out by Dr Botstein this criticism may or may not be valid depending upon the definition of hypothesis. Indeed, it is questionable what hypothesis has driven crystallography and or gene mapping beyond the reasoning that "most proteins might have a structure" and "most genes are located somewhere". Yet nobody can deny the scientific achievement derived from these discovery driven approaches. Thus, it seems that discovery vs hypothesis driven research is more a matter of semantics not worthy of further scientific discussion. Other participants were able to describe applications of this strategy for the study of clinical questions. For instance, Dr. Masaki Mori summarized is experience using cDNA arrays to link molecular portraits of gastric cancer to clinical parameters. His approach appeared particularly interesting since he identified specific genes that correlated with invasiveness of tumor and prognosis. Similarly, Dr. Ena Wang by prospectively studying gene expression patterns of melanoma metastases in patients undergoing immunotherapy identified few genes possibly associated with immune responsiveness. This was achieved through the adoption of a RNA amplification method she recently described that maintains proportional the abundance of different transcripts. Although conceptually interesting both investigators* results were derived from relatively few clinical samples and it was felt that the results needed confirmation through a larger, prospective series. A very useful application of microarray technology was discussed by Dr Weinstein, Dr. Takao Yamori and Dr. Masahiko Nishiyama who have separately pioneered the use of panels of human cancer cell lines as a data base for drug discovery and characterization.
Dr. Richard Hynes gave the best demonstration of how microarray analysis can lead to the discovery of biological mechanisms. By developing an elegant model whereby melanoma cell lines were selected according to metastatic potential, Dr. Hynes described how his group could segregate few genes that appear to govern invasiveness from a 60,000-gene array chip. By focusing on these few genes they could demonstrate how metastatic potential may be switched on and off. These models are not only of obvious biological interest but they may lead to direct therapeutic application by identifying drugs or biological agents inhibiting specifically those mechanisms.
The importance of discovery driven approaches was emphasized by several talks from investigators that have studied tumor behavior in the context of pre-clinical or clinical models. An example was Dr Hanahan discussion of putative factors within the tumor microenvironment that could lead to tumor rejection by the immune system. In particular, Dr Hanahan described mechanisms of tumor angiogenesis in well-characterized pre-clinical models developed by his laboratory. His pioneering work on tumor / host interactions has already identified various chemokines that might be responsible for the different biological and immune behavior of tumors. Interestingly, Dr. Hanahan suggested that part of its future work will rely on the adoption of array-based research to frame within the big picture his findings. A similar conclusion was suggested by Dr. Marincola’s presentation. This participant described his efforts to identify the algorithm governing tumor regression in response to treatment with biological response modifiers and, in particular, antigen-specific vaccination. Using various technologies developed by his lab, Dr. Marincola could monitor vaccine-specific responses at the systemic level and in the tumor microenvironment. In particular, he could follow the natural history of individual tumors through serial gene expression analysis of specimens obtained by fine needle aspirates. This strategy allows to directly related clinical outcome to laboratory findings since the lesion studied can be left in situ. Using this approach, he observed that vaccine-elicited T lymphocytes localize at tumor site, engage a dialogue with tumor cells but this is not sufficient to induce tumor regression. The conclusion was that factors other than the engagement of T cell receptor with target molecules on the cancer cell surface might govern tumor responsiveness. However, the number of possibility is so large that only a high throughput approach will efficiently lead to the discovery of likely pathway governing immunologically-mediated tumor regression in humans.
Tumor immunology was another component of the Workshop as evidence has emerged during the last decade that tumors are susceptible to immune mediated rejection. This phenomenon although amply documented in pre-clinical and clinical arenas is, however, capricious and sporadic. It is currently believed that most often natural or treatment induced anti-tumor immunity. might be insufficient and efforts aimed at enhancing its intensity may shift the balance in favor of the host. Dr. Hideaki Tahara gave a comprehensive overview of this approach by describing the relationship between immune effector cells, target cells and their intermediary: antigen-presenting cells. In particular is work focuses on the afferent loop of antigen presentation. He showed that various cytokines (and in particular IL-18) may have particularly strong effects in inducing effective immune responses. For instance, the intra-tumoral administration of IL-18 (quite toxic when administered systemically) can induce general anti-tumor responses effective also against tumors at distant sites. Thus, the possibility of extending this pre-clinical findings to the clinical settings appears intriguing. Dr. Takashi Nishimura again focusing on the afferent loop of the immune system suggested that T cell effector function is enhanced by improvement of antigen presentation. His work suggests that the maturation into a particular phenotype of antigen presenting cells can facilitate the expansion of effector anti-tumor T cells. A somewhat more complicated but highly tailored approach was suggested by Dr. Yoshiyuki Yamaguchi, who described the effort of his laboratory under Dr. Toge's direction to pre-select highly effective T cells against cancer based on their T cell receptor utilization. These cells could then be used for adoptive transfer after in vitro expansion or could be used to engineer highly potent killer cells.
Whether high-throughput analysis will eventually make a major contribution to the understanding of the neoplastic process in the context of heterogeneous human diseases remains to be seen. However, this approach is not exclusive of others and in fact various models could complement discovery science by validating its findings. Dr. Thomas Look provided a fascinating overview of his work focused on the discovery of relevant genetic modifiers such as tumor suppressor genes that might affect tumorigenesis. His laboratory has developed powerful tools for such analysis by modeling the effects of gene manipulation on lymphocyte maturation and in worms and fish. His works demonstrates the contribution of model organisms to the dissection of molecular mechanisms responsible for tumor formation and consequent targeting with candidate drugs or biological agents. Dr. Dean Kedes, demonstrated another approach to cancer therapy, whereby, oncogenic virus physiology is dissected to understand its pathophysiology. His work on the molecular characterization and structure of the capsid of the KSHV/HHV8, ethiologic agent of Kaposi's sarcoma, exemplifies how such information might provide new targets for anti-viral drugs leading to novel means to treat KS and other related malignancies.
Three more presentations concluded the Workshop each one addressing a unique and imaginative new form of tailored made anti-cancer therapy. Dr. Shinzaburo Noguchi discussed the frequency of BRCAI and BRCA2 mutations in Japanese breast cancer families. His work suggested that these mutations are only partially linked to those identified in the Caucasia population while new founder mutations could be identified among the Japanese. The clinical relevance of this information is presently under investigation. A very interesting and thorough presentation was given by Dr. Yuko Kitagawa, who presented is extensive experience in the Surgical arena. His work is focused on the identification of sentinel nodes draining metastatic cells away from visceral cancers, in particular gastric and esophageal. His work conclusively demonstrated that primary draining nodes in visceral organs cannot be easily predicted and follow a capricious pattern identifiable only be injection of radioactively marked colloids within the tumors. Interestingly, his elegant work demonstrated also that metastatic cells almost uniquely favor sentinel nodes while skip metastases (metastases in nodes down-stream or the primary draining node) occur extremely rarely. Thus, as a tailored made approach, his work suggests that individual patients should be subjected to sentinel node identification during resection of their cancer and subsequent nodal dissection should be driven by the physiological pattern of lymphatic drainage in each individual rather than by text book-based predictions.
The final presentation by Dr. David Bartlett was of particular interest since it suggested a totally novel approach to cancer therapy. Dr. Bartlett observed that certain viruses (in his case Pox-Viruses) can be utilized as vectors to deliver therapy directly within cancer cells. In particular, he identified a tumor selective Vaccinia virus mutant. This vector holds promise for the delivery of gene therapy directly into tumor cells and can in itself cause death of cancer cells without causing systemic toxicity in pre-clinical models. This approach has been recently investigated in primates and holds promise for future application in humans.
In summary, recent advances in the understanding of tumor biology have revealed that malignant cell transformation is the consequence of multiple genetic alterations. Furthermore, there is significant evidence suggesting the association between individual gene alteration and tumor behavior. This information suggests that optimal treatment for individual cancer patients could be achieved in the information on the tumor characteristics can be obtained and utilized. In this Workshop recent advancement toward the establishment of "Tailor-made cancer therapy" were extensively discussed in an informal atmosphere by experts taking different approaches including surgery, chemotherapy, radiotherapy, immunotherapy and gene therapy. It is hoped that this exchange among the scientists of the U.S. and Japan will avoid redundancy and will expedite the development of novel cancer therapies in the 2lst century.

Other Seminar Activities (Reception, Excursion, other meetings):
Saturday, 17 February 2001 (The 1st day)
1830 Welcome Dinner

Comments and Opinions:
This Seminar was very exciting, stimulatory and informative for each participant. This type of joint meetings is highly appriciated to be continued.



3. Molecular Epidemiology of Nicotine Addiction and Lung Cancer
Period: from Feb. 9 to Feb.11, 2001, 3days including an arriving day
Site: Maui Prince Hotel, Maui, Hawaii, USA

Japanese Organizer:
Sugimura Haruhiko
Hamamatsu University School of Medicine
Department of Pathology
Professor

US Organizer:
Shields Peter
Georgetown University
Lombardi Cancer Center
Professor

List of Japanese-side Participants (except for Organizer):
Tetsuya KAMATAKI
Professor
Faculty of Pharmaceutical Science, Hokkaido University

Jun YOKOTA
Chief
Biology Division, National Cancer Center Research Institute

Takashi TAKAHASHI
Chief
Division of Molecular Oncology
Aichi Cancer Center Research Institute

Kei NAKACHI
Senior Investigator
Epidemiology Division
Saitama Cancer Center Research Institute

Eizaburo SUEOKA
Assistant Professor
Department of Internal Medicine
Saga Medical School of Medicine

Naoko SUEOKA
Assistant Professor
Department of Internal Medicine
Saga Medical School of Medicine

Shigeto YOSHII
Assistant Professor
Department of Medicine
Hamamatsu University School of Medicine

Kazuya SHINMURA
Research Scientist
Biology Division, National Cancer Center Research Institute

List of Japanese-side Participants (except for Organizer):

Peter SHIELDS
Professor
Georgetown University, Lombardi Cancer Center

Caryn LE RMAN
Professor
Georgetown University, Lombardi Cancer Center

Ross BROWNSON
Professor
Saint Louis University School of Public Health

Karl KELSEY
Professor
Harvard School of Public Health

Loic Le MARCHAND
Professor
University of Hawaii

David SIDRANSKY
Director Professor
Johns Hopkins Medical University

Adi GAZDAR
Professor
UT Southwestern Medical Center

Steve BELINSKY
Director
Lovelace Respiratory Cancer Program

Raymond Niaura
Professor
Brown Medical School

Stephen Lam
Professor
University of British Columbia


Itinerary and Time Table
Molecular Epidemiology of Nicotine Addiction and Lung Cancer
Tobacco and Lung Cancer
Maui Prince Hotel
Maui, Hawaii
Friday; February 9, 2001

February 9-11, 2001 Evening Dinner - 6pm Saturday, February 10, 2001
7:30am	Continental Breakfast
8:00am	Dosing oneself - Reducing exposures and risks	Peter Shields
8:40am	Smoking behavior and its consequences in the pathogenesis of lung cancer	Takashi Takahashi
9:20am	Genetic influences on smoking behavior	Caryn Lerman
10:00am	BREAK
10:20am	Candidate risk markers of lung cancer, case-control study in Okinawa	Haru Sugimura
11:00am	Genetic susceptibility to lung cancer in Hawaii	Loic LeMarchand
11:40am	Role of CYP2A6 in the metabolic activation of nitrosames and its effects on lung cancer risk in smokers	Tetsuya Kamatki
12:20am	LUNCH - (on own)
6:00pm	Dinner for participants
7:30pm	Defining and assessing tobacco use and dependence phenotypes: What is heritable?	Raymond Niaura
8:10pm	Environmental tobacco smoke and lung cancer: Epidemiologic findings and policies to control exposure.	Ross C. Brownson
8:50pm	Smoking dose-response and p16INK4a inactivation in lung cancer: does inverse dose-response identity susceptibles.	Karl T. Kelse
9:30pm	Molecular Epidemiology of Human Lung Cancer	Curt Harris
Sunday, February 11, 2000
8:00am	Continental Breakfast
8:30am	Molecular Genetics of Lung Cancer Progression	Jun Yokota
9:10am	Early detection of lung cancer: Strategies and obstacles.	Steve Belinsky
9:50am	Involvement of alpha PIX in the PAK1JNK1 activation and apoptosis induced by Benzo(a)pyrene	Shigeto Yoshii
10:30am	BREAK
10:50am	Promising Molecular Detection Approaches in Lung and Head and Neck Cancer	David Sidransky
11:30am	Significant effects of cigarette smoking on host biology, in association with lifestyle-related diseases	Kei Nakachi
12:10pm	LUNCH
7:30pm	Quantitative nuclear morphometry of bronchial intraeoithelial neoplasia detected by fluorescence bronchoscopy	Stephen Lam
8:10pm	Clinical significance of hnRNP B1 as a new molecular biomarker for lung cancer, and regulatory machinery of hnRNP A2/B1 gene expression	Eizaburo Sueoka
8:50pm	Smoking related damage in the bronchial epithelium	Adi Gazdar
9:30pm	IGFBP-6 induced by all-trans retinoic acid activates apoptpsis in lung cancer cells	Naoko Sueoka

Scientific Achievements
Reflecting the broadness of the topic of the conferences, the distinguished scientists from the various original majors, from behavior science to chemoprevention, came together in a small room next to the lobby of the resort hotel in Maui (5400 Makena Alanui, Makena Resort, Kihei, Hawaii, 96753). The discussions were continuously activated by the questions, primitive, but pertaining to the essential matter by the people who have not been always involved with that particular issue which the speaker addressed. Each participant reverberated to any arguments casted upon this heterogeneous group.
Genetics of smoking behavior was reviewed by Dr.Shields and the data of the two country, presented by Dr. Lerman and Takahashi were far from consistent. In other way, the difference shown there was exciting. The discussion of this topic in the meeting impressed the participants the importance of this burgeoning field. Especially for Japanese participants, the genetic influence on human behavior is still a topic on the headline of tabloid newspaper, though we have a lot of basic scientists having neuropharmacological data using experimental animals. Description of smoking behavior, genotyping of several receptors in the brain, and basic information of population difference are urged to collect.
The second issue presented by Drs. Sugimura, LeMarchand, and Kamataki are lung cancer susceptibility due to CYP polymorphism. The first two speakers' conclusions overall consistent in terms of 2E1 and 1A1, and Dr. Sugimrua addressed Cyp1A2 data (the second assessment on Japanese lung cancer) in his data set. Dr. Kamataki strongly advocated the 2A6 usability in lung cancer. Dr. LeMarchant(only one local participant) extended his findings to Hawaians. These genetic markers all expect the tobacco related lung caner (small and squamous cell carcinoma)
Drs. Niaura and Brownson addressed exposure assessment of smoking from epidemiological standpoint. Dr. Niaura's presentation was about mental illness and smoking addiction and DRD4 VNTR polymorphism. The talk about why people require smoking was new to most of Japanese participants, in terms that craves for reward, relief, and obsession are related to dopamine, GABA, and serotonin receptor dysregulation respectively. This kind of study obviously requires recruitment of psychologists and other interdisciplinary team. Dr. Browonson further extend to the non-smokers' heart disease from the standpoint of environmental tobacco smoke (ETS). Investigation of ETS, also known as passive smoking, in Japan does not seem to draw much attention of experimental workers recently, and establishing a competing design of studies are necessary. Risk assessment should include dose-response relationship, exposure assessment, and risk characterization.
Dr. Kelsey in addition to Dr. Belinsky on the second day focused on methylation. Especially Dr. Kelsey took advantage of the exposure quantitation to delineate it to methylation status of pl6INK4a. Naturally, the status in premalignant or nonmalignant tissue of smokers was intensively argued. Furthermore these status may be related to individual difference of repair gene polymorphisms, (e.g. XRCC1).
Dr. Harris integratively closed the first day presenting his accumulating data.
The second day starts with molecular oncology by Drs. Yokota and Belinsky. Again early genetic changes were again of great interest of the participants. Drs. Yokota, Takahashi, Belinsky, Kesely, Sidransky and Gadzar were well-known players in this field, and exchanges of their comments reflected rivalry and emerging importance of these area.
Dr. Yoshii presented his meticulous work about benzopyrene signals (published in MCB in the fall of 2001, six months later), raising the issues to be resolved, such as BeP effect and mechanistic clarification of transcription level.
Dr. Sidransky presented the changes of Mitochondria DNA alteration in lung cancer and the possible analysis of it in plasma DNA. As already known, potential clinical or translational applications are extending rapidly.
Dr. Nakachi talked his long-term cohort study about NK activity and cancer risk.
In the last evening, Dr. Lum presented impressive follow-up study by bronchofluoroscopy to document early or small changes of broncial mucosa in large scale clinical study. The surveillance system like his will continuously produce useful and important findings in human. The numbers of the institutes involved including personnels and subjects were envious for the most clinical participants from Japan.
Dr. Sueoka, N presented her work done during her stay in united states and Dr. Sueoka, E showed us a promising markers of lung cancer detection.
Finally Dr. Gadzar presented his new data on lung cancer and addressed his idea about lung cancer carcinogenesis, early adenocarcinoma to invasive one. The figures of the two ends of this spectrum were strikingly different and his theory has been largely influenced by Dr. Shimosato's group in Japan in that putting adenomatous hyperplasia in the precancerous stage in the presumable multistage progression of lung cancer. We should evaluate effectiveness of the surveillance on the population to detect such small lesions, but if the samples were available during such performance, the research on pathogenesis of lung adenocarcinoma which are gradually increasing in many countries, will definitely progress.
As briefed above, two issues, genotype-behavior-phenotype and adenocarcinoma development were most actively discussed and these two fully covered the scope of the meeting. Addiction is still not so popular topic for Japanese cancer researchers, but obviously there seems to be a difference in two countries in distribution of genotypes or behavior pattern in a certain situation.

Other Seminar Activities (Reception, Excursion, other meetings):
A reception was held on Feb 9. Most of the Japanese participants and Dr. Harris took part in the JCA-AACR joint meeting held from Feb. 12 in the Mariott hotel in Maui.

Comments and Opinions:
Concentrated, short term, and closed meeting by a dozen of participants like this is the most effective way of exchange important information for both part. Comparison with the Joint meeting (10 and more so-called distinguished speakers, 100 posters, four days) clearly highlighted the characteristics of the two types (small, closed vs medium, open) of scientific meetings. Even if the topic looks like considerably wide, the scientists with wider perspective can cover all of them and integrate the discussions well in case of small meeting. During the small meeting, each participant can (must) be under good strenuous level of willingness to fight against the issues addressed by a speaker. On the other hand, young people would not object to the big speakers presenting data of 50 and more his lab members in an atmosphere of the concert event. I am not sure mixings in such environment (parties of 200 people) are easy for notoriously unsociable scientists.
Hawaii is a perfect resort and a jet lag is worse than a trip to the east coast of USA for Japanese participants. We can recruit many Japanese scientists to Bethesda if the participants are unique and in small numbers.
Genes and addiction is a currently burgeoning field especially in Japan, and scarcity of this topic presented by Japanese participants is definitely related to the state of Japanese society itself. Social problem of addiction (drug abuse, alcoholics, psychiatric problem) seems less serious at least comparably and apparently than US, and does not draw so much attention of the young lab investigators (still ordinary Japanese rarely visit psychiatrist). Intentional encouragement to Japanese scientists toward this filed may be necessary considering recent changes of life style in Japan. In such cases the participants can be recruited from the community other than that of cancer research, such as neuroscience, genetics, and psychology. Furthermore we cannot say that the differences grown in their own culture and history is not related to genetic difference. Genome diversity and its biological meaning may be a huge area to be studied.