GASTRIC CANCER

|

RESEARCH PAPER

HLA-B54 is a candidate of response to Fluoropyrimidine plus PSK therapy in gastric cancer 2 Kyoji Ogoshi1 and Kaichi Isono Department of Medicine, Massachusetts General Hospital, Harvard Medical School 1 Department of Gastroenterological Surgery, Tokai University 2 Department of Surgery, Chiba University (Retired)

Recieved: 11/9/2009, Reviewed: 17/9/2009, Accepted: 1/10/2009 Keywords: HLA-B54, FPSK, gastric cancer DOI:10.5083/apjom.20424965.09

ABSTRACT

CORRESPONDENCE

It is not well known that the correlation between HLA antigens and response to cancer therapy. To evaluate the correlation among patients’ outcome, HLA antigens and therapies, we used the hierarchical clustering analysis (Ward method). We examined HLA antigens as biomarkers as well as models of gene polymorphism in 1753 gastric cancer patients followed over 25 years after treatment. Among these patients, we identified that HLA haplotypes; HLA-B54-Cw1-DQ4-DR4, might be restricting to Fluoropyrimidine drugs plus PSK therapy in gastric cancer. HLA haplotypes including HLA-B54 were candidate predictors for response to FPSK therapy in gastric cancer. HLAs will be useful biomarkers for response to therapy in Japanese patients.

Kyoji Ogoshi, Department of Gastroenterological Surgery, Tokai University, Shimokasuya 143, Isehara, Kanagawa, 259-1193, Japan.

INTRODUCTION Human diseases are different, complex, and multifactorial, and even with the same disease, patients can exhibit different disease patterns. With the individual variation among patients in the manifestations of cancer, there is an acknowledged heterogeneity of disease and differences in individual response to drugs; thus, some patients respond to specific therapeutic approaches, and some do not. One possible source of this individual variation may be single nucleotide polymorphisms (SNPs) or mutations. SNPs are abundant in the human genome, and their ultimate effects may be to alter the expected or predicted pharmacokinetics and pharmacodynamics of a drug. Human leukocyte antigens (HLAs) are glycoproteins present on the surface membranes of nearly every cell in the body. They occur in high concentrations on the surface of white blood cells (i.e., leukocytes) and are the major histocompatibility antigens for tissue recognition. In people, the complex is also called the HLA system and serves as a model of gene polymorphism. MHC molecules are also important components of the immune response.1,2 The phenomenon of MHC restriction was first recognized by Doherty and Zinkernagel,3 who hypothesized that the great diversity in HLA class I and class II was maintained through exposure to infectious diseases.

We also reported this overdominant selection, that is, the incidences of cancers in heterozygotes were lower than those in homozygotes in human malignant diseases.4,5 We hypothesized that HLAs might correlate with response to therapy in patients with gastric cancer. To investigate this idea, we selected HLAs as biomarkers for correlation to specific therapies and began analyzing them in cancer patients from January 1977. After decades of data collection, we assessed the correlation between HLA antigens and response to therapies from the perspective of patient outcomes.

Tel: +81-0-463-96-6163, Fax: +81-0-463-96-4120, E-mail: [email protected] Competing interests The authors declare that they have no competing interests.

Previously, we found correlations between specific patterns of HLA antigens and metastasis and secondary cancer, in addition to the response to therapies after gastrectomies.6-9 However, it remains difficult to predict the response to therapy during the decision making process about treatment. By applying a multivariate analysis method that we developed, we classified Japanese patients into 4 groups10 and followed their outcomes. Based on these outcomes for each group, we recommended specific therapies that had proved suitable for a given HLA group.11,12 In this study, under our results, we focused on confirmation whether the specific HLA antigen or haplotype relates to the response to specific therapy, or not. ISSN: 2042-4965

44

THE ASIA-PACIFIC JOURNAL OF ONCOLOGICAL MEDICINE

VOL I ISSUE I

HLA-B54 IS A CANDIDATE OF RESPONSE TO FLUOROPYRIMIDINE PLUS PSK THERAPY IN GASTRIC CANCER

PATIENTS AND METHODS The patients all had been diagnosed with gastric cancer, and the group had a median age of 60 y (range: HLA-B54 is a candidate of response to FPSK therapy 41 22–93 y) (1245 men, median age 61 y, range: 22–92 y and 508 women, median age 58 y, range: 23–93 y). All patients gave informed consent. A total of 1719 of the 1753 patients had undergone some type of resection. For peri- and postoperative adjuvant therapy, mitomycin C (MMC; Kyowa Hakko Co., Ltd., Tokyo, Japan) at a dose of 0.4 mg/kg was given intravenously during the operation and at a dose of 0.2 mg/kg on postoperative day 1. In addition, PSK (Krestin, Kureha Chemical Industry Co., Ltd., Tokyo, Japan) at a dose of 3.0 g/day, a nonspecific immunomodulator, and fluoropyrimidine drugs (5-fluorouracil (5FU; Kyowa Hakko Kogyo Co., Ltd, Tokyo, Japan) at a dose of 150 mg/ day, Futrafur at a dose of 600 mg/day, UFT (Taiho Pharmaceutical Co., Ltd., Tokyo, Japan) at a dose of 300 mg/day), were given orally from within 30 days after treatment and continued until tumor regression. PSK is virtually unrecognized in the US. PSK is a polysaccharide (large molecules made up of chains of linked sugar molecules) complex with immune stimulating effects, and is derived from the edible mushroom Coriolus Versicolor (Fr.) Quel, a member of the Basisiomycetes. It is composed of proteins and polysaccharides and has a molecular mass approximately 100kDa. All patients without tumor regressions received drug therapy for one year or longer and continued until a satisfactory outcome was achieved. Blood samples were collected at the Department of Surgery, Tokai University, beginning in Jan 1977, and HLA antigens were examined in the Department of Transplantation Immunology, Tokai University, by using the NIH standard microlymphocytotoxicity method for HLA-A, B, C, DR, and DQ antigens, and then throughout Japan at hospitals belonging to the Japanese Society of Strategies for Cancer Research and Therapy from 1987 to Sep 2005. HLA antigens were serologically tested at the Mitsubishi Chemical Medicine Corporation (Tokyo) . We examined HLA antigens as follows; HLA-A; A1, A2, A11, A24, A26, A30, A31, A33, HLA-B; B7, B13, B17, B27, B35, B37, B38, B39, B44,B46, B48, B51, B52, B54, B55, B56, B59, B60, B61, B62, B67, B70, B75, HLA-C; Cw1,Cw3, Cw4, Cw5, Cw6, Cw7, Cw8, HLADR; DR1, DR3, DR4, DR7, DR8, DR9, DR10, DR11, DR12, DR13, DR14, DR15, DR16, and HLA-DQ; DQ2, DQ4, DQ5, DQ6, and DQ7. The therapies were classified into groups, as follows: without postoperative adjuvant therapy (No adjuvant group, n=607); with PSK (PSK therapy group, n=123); F (F therapy group, n=136); F plus PSK (FPSK therapy group, n=168); MMC (MMC therapy group, n=93); MMC plus F (MF therapy group, n=354); and MMC plus F plus PSK (MFPSK therapy group, n=272). All statistical analyses were carried out using SPSS software, version 15 (SPSS Inc., Chicago, IL, USA.). Mean values were compared by Student’s-test. The chisquare test was used to compare the prevalence of incidence of HLAs and the prognoses of patients. For evaluate the correlation between HLA haplotype and right patients with possible right therapy, we performed the hierarchical cluster analysis by using Ward method. Results were considered significant when the P value was less than 0.05.

THE ASIA-PACIFIC JOURNAL OF ONCOLOGICAL MEDICINE

VOL I ISSUE I

One thousand fifty/1753 (65.6%) patients were followed over 10 years. The survival period of cancer patients was defined as the interval from operation to death (overall survival rate included death within 30 days after treatment), with data regarding survivors censored at the last follow-up visit (August 2007). Survival curves were calculated using the Kaplan-Meier product-limit estimate, and differences in survival were assessed by the log-rank test.

RESULTS Figure 1 showed the results of the hierarchical cluster analysis by using Ward method in patients who received FPSK. Among them Figure 1a showed the patients who survived over 10 years, while, 1b showed those who were died within 10 years. In patients who survived over 10 years, HLA-B54, -Cw1, -DR4, and -DQ4 antigens were clustered, while in those who were died within 10 years HLA-A2, -Cw1, and-DR4 were clustered, and HAL-B54 and -DQ4 were separated. In other therapies analyzed, we could not find out the HLA antigens which correlated between death or alive and clustering or not.

Outcomes of the patients with HLA haplotypes including HLA-B54 HLA-B54, -Cw1, -DR4, and -DQ4 positive patients were 261/1753 (14.9%), 550/1753 (31.4%), 648/1753 (37.0%), and 396/1753 (22.6%), respectively. Figures 2 showed the survival curves of patients with HLA-B54 (Fig. 2a), HLA-B54-Cw1 (2b), HLAB54-Cw1-DR4 (2c), HLA-B54-Cw1-DQ4 (2d), HLAB54-Cw1-DR4-DQ4 (2e), and HLACw1-DR4-DQ4 (2f). Patients with B54 (7/134), those with B54-Cw1 (12/244), those with B54-Cw1-DR4 (8/172), those with B54-Cw1DQ4 (7/126), and B54-Cw1-DR4-DQ4 (7/125) who received FPSK therapy were all survived. Patients with HLA-B54 (log rank test, vs. MFPSK, compare to other therapies.

DISCUSSION We demonstrated the specific HLA antigen as well as haplotypes included HLA-B54 might be a predictor for response to FPSK therapy in gastric cancer. Unfortunately, at present, there was no data concerning between HLA and response to therapy clinically as well as basically. Unfortunately, these antigens are common in Japanese and Asian countries, not foreign countries. PSK is also familiar to Japanese clinicians and it’s clinical benefit was reported,13 not but foreign countries. As for HLA-B54, significantly increased frequency of HLA-B54 was reported in Japanese patients with diffuse panbronchiolitis, which is considered a Japanese specific disease. These are well coincided in Japan.14,15 We think that HLA system is the best example of polymorphisms for predicting the right patients with response to right therapy at the right time. The ability to assess an individual’s reaction to a drug before prescribing it will increase the physician’s confidence in prescribing and the patient’s confidence in taking the drug. In conclusion, HLA-B54 is a useful predictor for response to FPSK therapy before treatment, and we think that consideration of HLAs for predictor of response to therapy in the clinical setting is a useful tool.

45

HEALTHCARE BULLETIN

|

GASTRIC CANCER

Figure 1. Dendrograms by using hierarchical cluster analysis in patients who survived over 10 years (1a), while, those who were died within 10 years (1b).

46

THE ASIA-PACIFIC JOURNAL OF ONCOLOGICAL MEDICINE

VOL I ISSUE I

HLA-B54 IS A CANDIDATE OF RESPONSE TO FLUOROPYRIMIDINE PLUS PSK THERAPY IN GASTRIC CANCER

Figure 2. Survival curves of patients with HLA-B54 (2a), HLA-B54-Cw1 (2b), HLA-B54-Cw1-DR4 (2c), HLA-B54-Cw1-DQ4, HLAB54-Cw1-DR4-DQ4 (2d), and HLA-Cw1-DR4-DQ4 (2e) according to the actually administered therapy.

THE ASIA-PACIFIC JOURNAL OF ONCOLOGICAL MEDICINE

VOL I ISSUE I

47

HEALTHCARE BULLETIN

|

GASTRIC CANCER

ACKNOWLEDGEMENTS

8

We thank all participating patients, collaborators of The Japanese Society of Strategies for Cancer Research and Therapy, and Dr. K. Iwata for help with the analysis of the data. This study was supported by grants from The Japanese Society of Strategies for Cancer Research and Therapy.

Ogoshi, K., Tajima, T., Mitomi, T., Makuuchi, H., Tsuji, K. (1997) HLA-A2 antigen Status predicts metastasis and response to immunotherapy in gastric cancer. Cancer Immunol Immunother 45: 53-59.

9

Ogoshi, K., Isono, K. (2004) Patients outcomes in a randomized trial according to the status of HLA antigenss in gastric anc colorectal cancers and the secondary malignancies. Ann Cancer Res Ther 12: 87-104.

REFERENCES

10

Hayashi, F., Hayashi, C., Ogoshi, K. (1994) Classification of gastric cancer patients based on HLA antigen expression using quantification method III. Ann Cancer Res Ther 3: 117-120.

11

Ogoshi, K., Iwata, K., Miyaji, M., Nakamura, K., Kajiura, Y., Nabeshima, K., Morita, M., Soeda, J., Kondoh, Y., Tajima, T., Makuuchi, H. (2000) A prospective study of HLA-oriented therapy: An attempt to predictive the response to anti-cancer therapy in gastric cancer. Ann Cancer Res Ther 8: 155-167.

1

Benacerraf, B., McDevitt, H.O. (1972) Histocompatibility-linked immune response genes. Science 175: 273-279.

2

Benacerraf, B. (1982) Role of MHC gene products in immune regulation. Science 212: 1229-1238.

3

Zinkernagel, R.M., Doherty, P.C. (1974) Restriction of in vitro T cell-mediated cytotoxicity in lymphocytic choriomeningitis within a syngeneic or semiallogeneic system. Nature 248: 701-702.

12

Ogoshi, K., Isono, K. (2001) Heterozygte advantage at major histocompatibility complex (MHC) molecules and cancer risk. Ann. Cancer Res Ther 9: 73-86.

Oogoshi, K., Koyanagi, Y., Tsuji, K., Isono, K. (2008) Outcome of HLA-oriented therapy for gastric cancer in retrospective and prospective study. Ann Cancer Res Ther 2: 36-43.

13

Ogoshi, K., Isono, K. (2001) Incidence of heterozygotes and homozygotes of major histocompatibility complex in Japanese compared to non-Japanese. Ann. Cancer Res Ther 9: 87-98.

Ogoshi , K., Kondoh, Y., Tajima , T., Mitomi , T. (1992) Glycosidically bound sialic acid levels as a predictive marker of postoperative adjuvant therapy in gastric cancer. Cancer Immunol Immunother 35: 175-180.

14

Ogoshi, K., Tajima, T., Mitomi, T., Tsuji, K. (1994) HLA-DR4 antigen and lymph node metastases in poorly differentiated adenocarcinoma of the stomach. Cancer 73: 2250-2252.

Sugiyama, Y., Kudoh, S., Maeda, H., Suzaki, H. and Takaku, F. (1990) Analysis of HLA antigens in patients with diffuse panbronchiolitis. Am. Rev. Respir. Dis. 141: 1459-1462.

15

Keicho, N., Tokunaga, K., Nakata, K., Taguchi, Y., Azuma, A., Bannai, M., Emi, M., Ohishi, N., Yazaki, Y. and Kudoh, S. (1998). Contribution of HLA genes to genetic predisposition in diffuse panbronchiolitis. Am. J. Respir. Crit. Care Med. 158: 846-850.

16

Originally published in Ann.Cancer.Res.Therap Vol 17 No2 40-44 2009

4

5

6

7

48

Ogoshi, K., Tajima, T., Mitomi, T., Tsuji, K. (1996) HLA antigens are candidate markers for prediction of lymph node metastasis in gastric cancer. Clin Exp Metastasis 14: 277-281.

THE ASIA-PACIFIC JOURNAL OF ONCOLOGICAL MEDICINE

VOL I ISSUE I