小澤 敬也

Komatsu, N., Watanabe, T., Uchida, M., Mori, M., Kirito, K., Kikuchi, S., Liu, Q., Tauchi, T., Miyazawa, K., Endo, H., Nagai, T., and Ozawa, K.: A member of Forkhead transcription factor FKHRL1 is a downstream effector of STI571-induced cell cycle arrest in BCR-ABL expressing cells.<br />

Lu, Y.-Y, Wang, L.J., Muramatsu, S., Ikeguchi, K., Fujimoto, K., Okada, T., Mizukami, H., Matsushita, T., Hanazono, Y., Kume, A., Nagatsu, T., Ozawa, K., and Nakano, I.: Intramuscular injection of AAV-GDNF results in sustained expression of transgenic GDNF, and its delivery to spinal motoneurons by retrograde transport.

The application of adeno-associated virus (AAV) vectors to cancers is limited by their low transduction efficiency. Previously, we reported that gamma-ray enhanced the second-strand synthesis, leading to the improvement of the transgene expression, and cytocidal effect of the herpes simplex virus type-1 thymidine kinase (HSVtk) and ganciclovir (GCV) system. In this study, we extended this in vitro findings to in vivo. First, the laryngeal cancer cell line (HEp-2) and HeLa were treated with AAVtk/GCV, the number of surviving cells was reduced as the concentration of GCV increased. Furthermore, the 4 Gy irradiation enhanced the killing effects of AAVtk/GCV by four-fold on HeLa cells and 15-fold on HEp-2 cells. Following the in vitro experiments, we evaluated the transgene expression and the antitumor activity of the AA V vectors in combination with gamma-ray in nude mice inoculated with HEp-2 subcutaneously. The LacZ expression was observed in the xenografted tumors and significantly increased by gamma-ray. The AAVtk/GCV system suppressed the tumors growth, and gamma-ray augmented the antitumor activity by five-fold. These findings suggest that the combination of AAVtk/GCV system with radiotherapy is significantly effective in the treatment of cancers and may lead to reduction of the potential toxicity of both AAVtk/GCV and gamma-ray.

Recently, we reported that a recurrent translocation, t(1;3)(p36;p21) is closely associated with prior chemotherapy including alkylating agents, assessing eight patients with various hematologic malignancies (Genes, Chromosomes and Cancer 34:186192), 2002). Furthermore, we delineated the 1p36 breakpoint in two patients lying between RP11-BAC47P3 and RP5PAC963K15 at 1p36.3 with a small deletion near the breakpoint. In one of them, we also found deletion at 3p21.3 with cosNRL9 probe, which is included in a 370-kb lung cancer homologous deletion region. However, due to scantiness of the patient materials at that time, we could not determine the precise breakpoint at 1p36 or 3p21 in any of the patients. In this report, we identified the 1p36 and 3p21 breakpoints of an AML (M3) patient who is included in the previous patient series. The patient showed t(1;3)(p36;p21) together with t(15; 17) at the third relapse. With FISH using BAC/PAC probes, we determined the I p36 breakpoint within RP11-295B1 at 1p36.2 and the 3p21 breakpoint between RP11-3B7 and RP11-901L6 at 3p21.3. There was no deletion around the two breakpoints in this patient. To the best of our knowledge, this is the first report that has identified the precise breakpoint of t(1;3)(p36;p21) translocation. It is obvious that the 1p36.2 and 3p21.3 breakpoints of this patient are different from those of the previous patients, suggesting that the genes and the molecular event is different from those of the previous patients. The patients with t(1;3)(p36;p21) should be subclassified according to the precise breakpoints or the genes involved. (C) 2002 Wiley-Liss, Inc.

Recently, we reported that a recurrent translocation, t(1;3)(p36;p21) is closely associated with prior chemotherapy including alkylating agents, assessing eight patients with various hematologic malignancies (Genes, Chromosomes and Cancer 34:186192), 2002). Furthermore, we delineated the 1p36 breakpoint in two patients lying between RP11-BAC47P3 and RP5PAC963K15 at 1p36.3 with a small deletion near the breakpoint. In one of them, we also found deletion at 3p21.3 with cosNRL9 probe, which is included in a 370-kb lung cancer homologous deletion region. However, due to scantiness of the patient materials at that time, we could not determine the precise breakpoint at 1p36 or 3p21 in any of the patients. In this report, we identified the 1p36 and 3p21 breakpoints of an AML (M3) patient who is included in the previous patient series. The patient showed t(1;3)(p36;p21) together with t(15; 17) at the third relapse. With FISH using BAC/PAC probes, we determined the I p36 breakpoint within RP11-295B1 at 1p36.2 and the 3p21 breakpoint between RP11-3B7 and RP11-901L6 at 3p21.3. There was no deletion around the two breakpoints in this patient. To the best of our knowledge, this is the first report that has identified the precise breakpoint of t(1;3)(p36;p21) translocation. It is obvious that the 1p36.2 and 3p21.3 breakpoints of this patient are different from those of the previous patients, suggesting that the genes and the molecular event is different from those of the previous patients. The patients with t(1;3)(p36;p21) should be subclassified according to the precise breakpoints or the genes involved. (C) 2002 Wiley-Liss, Inc.

The usefulness of two-color flow cytometry with a CD19 gate (CD19-FCM) for detecting clonal B-cells in the bone marrow was evaluated. Since kappa and lambda ratios analyzed in the bone marrow from healthy adults were within the range of 0.5-3.0, outside of this range was regarded as the existence of clonal B-cells. Mixing experiments in vitro showed that 10% or more of clonal B-cells contaminating normal bone marrow cells could be detected using CD19-FCM. Ninety-nine bone marrow samples from 60 patients with B-cell lymphoma were simultaneously analyzed using CD19-FCM and histologic examination of both bone marrow aspiration and core needle biopsy samples. The agreement of results obtained by both examinations was 82% (81/99). Discrepant results were found in 18 samples of 10 patients. All except one sample were positive for CD19-FCM analysis but negative for histologic examination and the conversion of CD19-FCM-positivity to CD19-FCM-negativity was associated with chemotherapy. These results suggest that CD19-FCM is useful for the evaluation of bone marrow involvement of B-cell lymphoma.

The usefulness of two-color flow cytometry with a CD19 gate (CD19-FCM) for detecting clonal B-cells in the bone marrow was evaluated. Since kappa and lambda ratios analyzed in the bone marrow from healthy adults were within the range of 0.5-3.0, outside of this range was regarded as the existence of clonal B-cells. Mixing experiments in vitro showed that 10% or more of clonal B-cells contaminating normal bone marrow cells could be detected using CD19-FCM. Ninety-nine bone marrow samples from 60 patients with B-cell lymphoma were simultaneously analyzed using CD19-FCM and histologic examination of both bone marrow aspiration and core needle biopsy samples. The agreement of results obtained by both examinations was 82% (81/99). Discrepant results were found in 18 samples of 10 patients. All except one sample were positive for CD19-FCM analysis but negative for histologic examination and the conversion of CD19-FCM-positivity to CD19-FCM-negativity was associated with chemotherapy. These results suggest that CD19-FCM is useful for the evaluation of bone marrow involvement of B-cell lymphoma.

Toyo-oka, T., Kawada, T., Xi, H., Nakazawa, M., Masui, F., Hemmi, C., Nakata, J., Tezuka, A., Iwasawa, K., Urabe, M., Monahan, J., and Ozawa, K.: Gene therapy prevents disruption of dystrophin-related proteins in a model of hereditary dilated cardiomyopathy in hamsters.

Ageyama, N., Hanazono, Y., Shibata, H., Ohto, K., Ono, F., Nagashima, T., Ueda, Y., Donahue, R.E., Hasegawa, M., Ozawa, K., Yoshikawa, Y., and Terao, K.: Safe and efficient methods of autologous hematopoietic stem cell transplantation for biomedical research in cynomolgus monkeys.

Tanaka, M., Borgeld, H. J., Zhang, J., Muramatsu, S., Gong, J. S., Yoneda, M., Maruyama, W., Naoi, M., Ibi, T., Sahashi, K., Shamoto, M., Fuku, N., Kurata, M., Yamada, Y., Nishizawa, K., Akao, Y., Ohishi, N., Miyabayashyi, S., Umemoto, H., Muramatus, T., Furukawa, K., Kikuchi, A., Nakano, I., Ozawa, K., and Yagi, K.: Gene therapy for mitochondrial disease by delivering restriction endonuclease SmaI into mitochondria<br /> <br />

Xin, K. Q., Ooki, T., Mizukami, H., Hamajima, K., Okudela, K., Hashimoto, K., Kojima, Y., Jounai, N., Kumamoto, Y., Sasaki, S., Klinman, D., Ozawa, K., and Okuda, K.: Oral administration of recombinant adeno-associated virus elicits human immunodeficiency virus-specific immune responses

Hanazono, Y. Terao, K., Shibata, H., Nagashima, T., Ageyama, N., Asano, T., Ueda, Y., Katao, I., Kume, A., Hasegawa, M., and Ozawa, K.: Introduction of the green fluorescent protein gene into hematopoietic stem cells results in prolonged discrepancy of in vivo transduction levels between bone marrow progenitors and peripheral blood cells in nonhuman primates

Wang, L.-J., Lu, Y.-Y., Muramatsu, S., Ikeguchi, K., Fujimoto, K., Okada, T., Mizukami, H., Matsushita, T., Hanazono, Y., Kume, A., Nagatsu, Tl, Ozawa, K., and Nakano, I.: Neuroprotective effects of glial cell line-derived neurotrophic factor meddiated by an adeno-associated virus vector in a transgenic animal model of amyotrophic lateral sclerosis

Mishima, Y., Terui, Y., Mishima, Y., Katsuyama, M., Mori, M., Tomizuka, H., Takizawa, T., Miyazato, A., Ueda, M., Yamada, M., Hayasawa, H., Mizunuma, N., Ishizaka, Y., Ikeda, K., Kato, T., Ozawa, K., and Hatake, K.: New human myelodysplastic cell line, TER-3: G-CSF specific downregulation of Ca&lt;sup&gt;2+&lt;/sup&gt;/calmodulin-dependent protein kinase Ⅳ

Wang, L., Muramatsu, S., Lu, Y., Ikeguchi, K., Fujimoto, K., Okada, T., Mizukami, H., Hanazono, Y., Kume, A., Urano, F., Ichinose, H., Nagatsu, T., Nakano, I., and Ozawa, K.: Delayed delivery of AAV-GDNF prevents nigral neurodegeneration and promotes functional recovery in a rat model of Parkinson&#039;s disease

Shimpo, M., Ikeda, U., Maeda, Y., Takahashi, M., Miyashita, H., Mizukami, H., Urabe, M., Kume, A., Takizawa, T., Shibuya, M., Ozawa, K., and Shimada, K.: AAV-mediated VEGF gene transfer into skeletal muscle stimulates angiogenesis and improves blood flow in a rat hindlimb ischemia model

Muramatsu, S., Fujimoto, K., Ikeguchi, K., Shizuma, N., Kawasaki, K., Ono, F., Shen, Y., Wang, L., Mizukami, H., Kume, K., Matsumura, M., Nagatsu, N., Urano, F., Ichinose, H., Nagatsu, T., Terao, T., Nakano, I., and Ozawa, K.: Behavioral recovery in a primate model of Parkinson&#039;s disease by triple transduction of striatal cells with adeno-associated viral vectors expressing dopamine-synthesizing enzymes

Kawada, T., Nakazawa, M., Nakauchi, S., Yamazaki, K., Shimamoto, R., Urabe, M., Nakata, J., Hemmi, C., Masui, F., Nakajima, T., Suzuki, J. I., Monahan, J., Sato, H., Masaki, T., Ozawa, K., and Toyo-Oka, T.; Rescue ofhereditary from of dilated cardiomyopathy by rAAV-mediated somatic gene therapy: Amelioration of morphological findings, carcolemmal permeability, cardiac performances, and the prognosis of TO-2 hamster

Toyo-oka, T., Kawada, T., Xi, H., Nakazawa, M., Masui, F., Hemmi, C., Nakata, J., Tezuka, A., Iwasawa, K., Urabe, M., Monahan, J., and Ozawa, K.: Gene therapy prevents disruption of dystrophin-related proteins in a model of hereditary dilated cardiomyopathy in hamsters.<br />