Recent progress of mutation breeding in modern genome science era -Ion-beam mutagenesis-

Tomoko Abe 한국육종학회 2012 한국육종학회 심포지엄 Vol.2012 No.07

The world is facing a serious food and energy crisis. Plant mutation breeding has played an important role in overcoming this crisis and maintaining world stability. New techniques are required to achieve faster and more effective breeding. At RIKEN, we have developed a unique technology for mutation induction by using heavy-ion beams from particle accelerators at the RI Beam Factory. This development was achieved through an efficient synergistic link between agricultural science and accelerator physics. The use of ion beams for mutagenesis has a number of advantages: the approach has low exposure levels and high survival rates with high mutation rates, and it creates a wide variety of different mutations. Because heavy-ion beams provide a very high amount of energy, even a single ion is enough to significantly damage a gene – in fact, the beams have enough energy to break the double strand of the DNA. The technique is also very useful in producing mutants that lack just a single gene; multiple propagation technology can be used to convert these mutants into new cultivars. Examples of such breeds include ‘Safinia Rose’ (petunia), ‘Temari Bright Pink’ (vervena) and ‘Olivia Pure White’ (dianthus). The development period for producing new varieties is only 2 years. Over last decade, molecular biology has made great advancements through technological innovation. We use high-throughput DNA sequencing techniques such as next-generation sequence instruments and microarray technologies for analysis of gene mutations. Mutants have become more and more useful and important in modern genetic studies, enabling the discovery of genes that control important traits, and revealing the functions and mechanisms underlying their operations. The discovery of genes using mutants may lead to the emergence of a new field in biology, ‘Mutagenomics’.

중이온 빔 조사에 의한 담배의 돌연변이 유도와 내염성 식물의 선발

배창휴,Abe, Tomoko,민경수,김동철,정재성,이춘환,임용표,이효연 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

중이온 빔을 이용한 방사선 돌연변이의 기초자료를 얻고자 담배의 수분·수정 직후의 원배에 100 Gy의 중이온빔을 조사하여 M_1세대의 종자를 얻었다. NaCI 2.0과 2.5%를 포함된 MS배지에 M_1 세대의 종자를 파종하여 17개통의 NaCI 저항성 식물체를 선발하였다. 저항성 식물체는 BY-4품종에 ^14N빔을 조사한 처리구에서만 관찰되었다. M_1 세대의 저항성 식물체의 경우 영양생장기간 중에는 외형적 변이가 관찰되지 않았으나 생식생장기의 화기에 있어서는 다양한 변이가 관찰되었다. 특히 암술 수술의 길이의 변화, 꽃잎의 변화, 불완전 화분립, 줄기당 꽃의 착생수 감소 등의 변이가 발생하였다. M_1 세대에서 17개의 NaCI저항성 계통 중에서 8계통은 M_2 세대에도 저항성형질이 발현되었고, 또한 M_2 세대의 3계통은 Mannitol이 포함된 배지에서 저항성을 보여주었다. Tobacco proembryos were irradiated with 100 Gy of heavy-ion beams (^14N, ^20Ne : 135 Mev/u) after 24 to 96 hours of pollination as a mutagen and screened M_1 generation for morphological mutants and salt-tolerant plants. Morphological and physilolgical characteristics of the salt-tolerant plants derived from the irradiated proembryo are discussed in this report. Mutants irradiated porembryos with the beams after pollination produced various kinds of morphological variation. A total of 17 salt-tolerant plants were selected from tobacco cultivar (BY-4) by treatment with ^14N beam. Shapes of filament and pollen grain of most salt-tolerant plants were abnormal compared with non-irradiated wild type, and seeds weight and fertility obviously decreased. The germination rates of the several M_2 lines on the saline and the mannitol condition were higher than that of wild type.

Safety Assessment of Intravenous Administration of Trastuzumab in 100ml Saline for the Treatment of HER2-Positive Breast Cancer Patients

Abe, Hajime,Mori, Tsuyoshi,Kawai, Yuki,Tomida, Kaori,Yamazaki, Keiichi,Kubota, Yoshihiro,Umeda, Tomoko,Tani, Tohru Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.8

Background: The infusion rate is considered to affect incidence and severity of infusion reactions (IRs) caused by protein formulations. Trastuzumab (TRS) is approved for 90-minute infusion as the initial dose followed by 30-minute infusion with 250 ml saline. In the study, we evaluated the safety of TRS intravenously administered over 30 minutes with 100 ml saline to reduce burden of patients, safety of infusion with 250 ml saline already being established. Materials and Methods: Women with HER2 positive breast cancer, ${\geq}18$ years and ${\geq}55%$ left ventricular ejection fraction (LVEF), were registered in the study. Patients received 8mg/kg of TRS 250 ml over 90 minutes followed by 6mg/kg of TRS 100ml over 30 minutes in a three-week cycle. Results: A total of 31 patients were recruited, 24 for adjuvant therapy and seven with metastases. The median age was 59 years (range 39 to 82). The total number of TRS doses ranged from 5 to 17 with the median of 15. Mild IR occurred in two patients at the first dose. However, no IR was observed after reducing to 100 ml saline. No decrease of LVEF, increase of serum brain natriuretic peptide or any other adverse events were reported. Conclusions: Intravenous infusion of TRS with 100 ml saline over 30 minutes in breast cancer patients can be considered safe based on results from the study. It can be given on an outpatient basis as with the currently recommended dilution in 250 ml saline.

The Kampo Medicine Goshajinkigan Prevents Neuropathy in Breast Cancer Patients Treated with Docetaxel

Abe, Hajime,Kawai, Yuki,Mori, Tsuyoshi,Tomida, Kaori,Kubota, Yoshihiro,Umeda, Tomoko,Tani, Tohru Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Background: Goshajinkigan (GJG) is used for the treatment of several neurological symptoms. We investigated the efficacy of GJG and mecobalamin (B12) against neurotoxicity associated with docetaxel (DOC) in breast cancer patients. Materials and Methods: Sixty breast cancer patients were treated with DOC. Thirty-three patients (GJG group) received oral administration of 7.5 g/day GJG and 27 patients (B12 group) received oral administration of 1500 ${\mu}g/day$ B12. Neuropathy was evaluated according to DEB-NTC (Neurotoxicity Criteria of Debiopharm), Common Terminology Criteria for Adverse Events (NCI-CTC) ver. 3.0, and a visual analogue scale (VAS). This study employed a randomized open design. Results: The incidence of neuropathy was 39.3% in the GJG group, and 88.9% in the B12 group (p<0.01). In the GJG group, grade 1 DEB-NTC was observed in 2 cases, grade 2 in 5 cases and grade 3 in 5 cases. Grade 1 NCI-CTC was observed in 7 cases, grade 2 in 6 cases, and VAS was $2.7{\pm}2.2$. In the B12 group, grades 1, 2 and 3 DEB-NTC were observed in one case, 12 cases and 12 cases, respectively; and grades 1, 2 and 3 NCI-CTC were observed in 11 cases, 12 cases and one case, and VAS was $4.9{\pm}2.4$. Conclusions: Concomitant administration of GJG is useful in preventing neuropathy in breast cancer patients treated with a DOC regimen.

Feasibility Study of Docetaxel and Cyclophosphamide Six- Cycle Therapy as Adjuvant Chemotherapy for Japanese Human Epidermal Growth Factor Receptor 2-Negative Breast Cancer Patients

Abe, Hajime,Mori, Tsuyoshi,Kawai, Yuki,Tomida, Kaori,Kubota, Yoshihiro,Umeda, Tomoko,Tani, Tohru Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.8

Background: We compared treatment completion rates and safety of docetaxel and cyclophosphamide sixcycle therapy (TC6) with docetaxel followed by 5FU, epirubicin and cyclophosphamide (T-FEC) therapy in Japanese patients with human epidermal growth factor receptor 2 (HER2)-negative breast cancer. Materials and Methods: We administered TC6 q3w or T-FEC q3w to HER2-negative breast cancer patients. The primary endpoint of this trial was toxicity. As second endpoints, the treatment completion rate and relative dose intensity were evaluated. Results: The TC6 and T-FEC group consisted of 22 and 21 patients, respectively. Concerning hematological toxicity, grade 3 or higher adverse reactions included neutropenia and febrile neutropenia. As non-hematological adverse events, exanthema and peripheral neuropathy were frequently reported in the TC6 group, whereas more patients of the T-FEC group reported nausea and vomiting. In TC6, the treatment completion rate was 86.4% and the relative dose intensity of docetaxel was 93.2%. In T-FEC, the values were 95.2% and 98.9%, respectively. Conclusions: These results suggest that TC6 is tolerable in Japanese, and that this regimen can also be performed in outpatient clinics. However, with the TC6 regimen, the compliance was slightly lower than with the T-FEC regimen, and supportive therapy needs to be managed appropriately.

S-RNase Genotypes of Wild Apples

Junko Morita,Tomoko Koketsu,Kazuyuki Abe,Hideo Bessho,Shogo Matsumoto 한국원예학회 2008 한국원예학회 기타간행물 Vol.- No.-

Identification of Specific DNA Bands in Variegated Orchid Rhizomes Induced by Ion Beam Irradiation

Jae-il Lyu,Tomoko Abe,Hyo-Shim Han,Jae-sung Jung,Hyo-Yeon Lee,Dong-Young Shin,Chang-Hyu Bae 한국원예학회 2006 한국원예학회 기타간행물 Vol.- No.-

중이온 빔조사 담배(Nicotiana plumbaginifolia) 식물체의 생장과 DNA 변이

배창휴,양덕춘,이효연,류재일,김민수,Abe Tomoko 한국자원식물학회 2005 한국자원식물학회지 Vol.18 No.1

Effects of heavy-ion beam(20Ne) irradiation on growth and DNA alteration of tobaccoplants were investigated. Seed germination and plant height were decresed as the ion-beamintensity was increased. However, the bolting and flowering were promoted by the lowintensities of 5 Gy to 10 Gy treatment. Out of the 100 primers screened, 59 primersgenerated 336 DNA fragments by RAPD analysis, and one specific DNA fragment thatamplified in control but not in the ion-beam irradiated plants was observed. By AFLPanalysis, DNA fragment difference related to the ion-beam treatment was not detected butobserved among the plant bodys. . 중이온빔(20Ne)을 담배 식물체에 조사(irradiation) 하여 식물체의 생장과 DNA의 변이에 미치는 영향을 검토하였다. 발아율과 초장은 조사선량이 증가함에 따라 감소하였다. 반면 5Gy~10Gy의 저선량에서 추대와 개화가 촉진되어 발육은 오히려 촉진되었다. 총 100개의 primer를 이용하여 RAPD 분석한 결과, 59개의 primer에서 총 336개의 DNA 단편이 증폭되었고, 1개의 primer에서 중이온빔 조사처리구에서만 출현하는 DNA 단편이 나타났다. AFLP분석 결과, 개체 특이적인 DNA 단편은 나타난 반면 중이온빔 처리구 특이적인 DNA 단편은 관찰되지 않았다.

Floral mutant induction of asparagus by irradiation of Heavy-ion beam

So Young Kim,Kee Hwa Bae,Tomoko Abe,Chang Hyu Bae,Hyo Yeon Lee,Yong Pyo Lim,Toshiaki Kameya,Pil Yong Yun 한국육종학회 2009 한국육종학회 학술발표회 발표요지 Vol.41 No.1

Clinical significance of systematic retroperitoneal lymphadenectomy during interval debulking surgery in advanced ovarian cancer patients

Haruko Iwase,Toshio Takada,Chiaki Iitsuka,Hidetaka Nomura,Akiko Abe,Tomoko Taniguchi,Ken Takizawa 대한부인종양학회 2015 Journal of Gynecologic Oncology Vol.26 No.4

Objective: To investigate the clinical significance of systematic retroperitoneal lymphadenectomy during interval debulking surgery (IDS) in advanced epithelial ovarian cancer (EOC) patients. Methods: We retrospectively reviewed the medical records of 124 advanced EOC patients and analyzed the details of neoadjuvant chemotherapy (NACT), IDS, postoperative treatment, and prognoses. Results: Following IDS, 98 patients had no gross residual disease (NGRD), 15 had residual disease sized <1 cm (optimal), and 11 had residual disease sized ≥1 cm (suboptimal). Two-year overall survival (OS) and progression-free survival (PFS) rates were 88.8% and 39.8% in the NGRD group, 40.0% and 13.3% in the optimal group (p<0.001 vs. NGRD for both), and 36.3% and 0% in the suboptimal group, respectively. Five-year OS and 2-year PFS rates were 62% and 56.1% in the lymph node-negative (LN–) group and 26.2% and 24.5% in the lymph node-positive (LN+) group (p=0.0033 and p=0.0024 vs. LN–, respectively). Furthermore, survival in the LN+ group, despite surgical removal of positive nodes, was the same as that in the unknown LN status group, in which lymphadenectomy was not performed (p=0.616 and p=0.895, respectively). Multivariate analysis identified gross residual tumor during IDS (hazard ratio, 3.68; 95% confidence interval, 1.31 to 10.33 vs. NGRD) as the only independent predictor of poor OS. Conclusion: NGRD after IDS improved prognosis in advanced EOC patients treated with NACT-IDS. However, while systematic retroperitoneal lymphadenectomy during IDS may predict outcome, it does not confer therapeutic benefits.