基本情報

所属

• 保健科学研究院　保健科学部門　医用生体理工学分野

職名

• 助教

学位

• 博士（工学）（北海道大学）

科研費研究者番号

• 80344505

J-Global ID

• 200901060185645755

研究キーワード

• ATP   エネルギー代謝   ミトコンドリア   リン酸化   石灰化   DNA損傷   放射線   ポリリン酸   放射線治療   腫瘍   細胞生物学   

研究分野

• ライフサイエンス / 放射線科学
• ライフサイエンス / 医化学
• ライフサイエンス / 細胞生物学
• ライフサイエンス / 分子生物学

担当教育組織

•  学士課程　医学部（保健学科）
•  修士課程　保健科学院
•  博士課程　保健科学院

職歴

• 2007年 - 北海道大学医学部保健学科助手（放射線技術科学専攻） 助教
• 2003年 - 北海道大学医学部保健学科助手（放射線技術科学専攻） 助手
• 2003年 - Research Associate
• 2002年 - 北海道大学医療技術短期大学部助手（診療放射線技術学科） 助手
• 2002年 - Research Associate

学歴

•         - 2001年   北海道大学   工学研究科   分子化学専攻
•         - 2001年   北海道大学
•         - 1998年   北海道大学   工学研究科   分子化学専攻
•         - 1998年   北海道大学
•         - 1996年   室蘭工業大学   工学部   応用化学科
•         - 1996年   室蘭工業大学
•         - 1993年   北海道大学医療技術短期大学部   医療技術学部
•         - 1993年   北海道大学医療技術短期大学部

所属学協会

• 日本放射線影響学会   日本放射線技術学会   分子生物学会   

研究活動情報

論文

• The dynamic relationship between inorganic polyphosphate and adenosine triphosphate in human non‐small cell lung cancer H1299 cells

  Kaori Tsutsumi, Thititip Tippayamontri, Mari Hayashi, Nobuto Matsuda, Yusaku Goto

  FEBS Open Bio 2024年01月01日 [査読有り]
   

  Inorganic polyphosphate (polyP) plays a vital role in cellular energy metabolism and signaling, owing to its structure and high‐energy phosphate bonds. Intracellular ATP functions both as a cellular energy source and a key factor in cell death, and ATP dynamics in tumor cells are crucial for advancing cancer therapy. In this study, we explored the interplay between polyP and ATP in cellular energy metabolism. Treatment with polyP did not affect cell proliferation of human non‐small cell lung cancer H1299 and human glioblastoma T98G cell lines as compared to their respective control cells until 72 h post‐treatment. The mitochondrial membrane potential (MMP) in polyP‐treated cells was low, contrasting with the time‐dependent increase observed in control cells. While the ATP content increased over time in untreated and Na‐phosphate‐treated control cells, it remained unchanged in polyP‐treated cells. Furthermore, the addition of cyclosporine A, a mitochondrial permeability transition pore (mPTP) inhibitor, failed to restore ATP levels in polyP‐treated cells. We performed lactate assays and western blot analysis to evaluate the effect of polyP on glucose metabolism and found no significant differences in lactate secretion or glucose‐6‐phosphate dehydrogenase (G6PD) activity between polyP‐treated and control cells. Additional pyruvate restored MMP but had no effect on the cellular ATP content in polyP‐treated cells. Moreover, we observed no correlation between the Warburg effect and glucose metabolism during ATP depletion in polyP‐treated cells. Further investigation is warranted to explore the roles of polyP and ATP in cancer cell energy metabolism, which might offer potential avenues for therapeutic interventions.
• Evaluation of Image Classification for Quantifying Mitochondrial Morphology using Deep Learning.

  Kaori Tsutsumi, Keima Tokunaga, Shun Saito, Tatsuya Sasase, Hiroyuki Sugimori

  Endocrine, metabolic & immune disorders drug targets 2022年07月01日 [査読有り]
   

  BACKGROUND: Mitochondrial morphology reversibly changes between division and fusion. As these changes (mitochondrial dynamics) reflect the cellular condition, they are one of the simplest indicators of cell state and predictors of cell fate. However, it is currently difficult to classify them using a simple and objective method. OBJECTIVE: The present study aimed to evaluate mitochondrial morphology using Deep Learning (DL) technique. METHODS: Mitochondrial images stained by MitoTracker were acquired from HeLa and MC3T3-E1 cells using fluorescent microscopy and visually classified into four groups based on fission or fusion. The intra- and inter-rater reliabilities for visual classification were excellent [(ICC(1,3), 0.961 for rater 1; and 0.981 for rater 2) and ICC(1,3), respectively]. The images were divided into test and train images, and a 50-layer ResNet CNN architecture (ResNet-50) using MATLAB software was used to train the images. The datasets were trained five times based on five-fold cross-validation. RESULT: The mean of the overall accuracy for classifying mitochondrial morphology was 0.73±0.10 in HeLa. For the classification of mixed images containing two types of cell lines, the overall accuracy using mixed images of both cell lines for training was higher (0.74±0.01) than that using different cell lines for training. CONCLUSION: We developed a classifier to categorize mitochondrial morphology using DL.
• Feasibility of an Ultrasound-Based Method for Measuring Talar Displacement during the Anterior Drawer Stress Test Using a Telos Device: A Preliminary Study.

  Kaori Tsutsumi, Utayo Nakaya, Yuta Koshino, Mari Tateno, Kazuhisa Matsumoto, Mai Tanaka, Mika Yokoyama, Tatsunori Horie, Mina Samukawa, Tamotsu Kamishima, Harukazu Tohyama

  International journal of environmental research and public health 19 4 2022年02月18日 [査読有り]
   

  This study was conducted to measured talar displacement using ultrasound during an anterior drawer test (ADT) with a Telos device. Five adults (3 men and 2 women; 8 ankles; mean age: 23.2 y) with a history of ankle sprain and eight adults (5 men and 3 women; 16 ankles; mean age: 22.1 y) without a history of ankle sprain were recruited into a history of ankle sprain (HAS) and a control group, respectively. Talar displacement was observed in response to load forces applied by a Telos device during the ultrasound stress imaging test. The ultrasound probe was placed 5 mm inside from the center of the Achilles tendon on the posterior ankle along the direction of the major axis. The inter-rater reliability for the present method was classified as good and excellent (ICC(2,2) = 0.858 and 0.957 at 120 N and 150 N, respectively) in the control group and excellent (ICC(2,2) = 0.940 and 0.905 at 120 N and 150 N, respectively) in the HAS group, according to specific intraclass correlation coefficient values. We found that talar displacement during the ADT was lower in the HAS group than in the control group. Analysis of the receiver operating characteristic curve revealed that the quantitative ultrasound-based ADT using a Telos device was superior to the X-ray-based test in detecting reduced ankle joint mobility during the ADT (area under the curve of 0.905 and 0.726 at a force of 150 N using ultrasound-based and X-ray-based tests, respectively). Further investigation is needed; nevertheless, this preliminary study suggests that the ultrasound-based quantitative ADT using a Telos device might detect talar displacement more sensitively than the conventional stress X-ray.
• Contribution of Neuropilin-1 in Radiation-Survived Subclones of NSCLC Cell Line H1299

  Kaori Tsutsumi, Ayaka Chiba, Yuta Tadaki, Shima Minaki, Takahito Ooshima, Haruka Takahashi

  Current Issues in Molecular Biology 2021年09月 [査読有り]
  
• Chromosomal Location of Genes Differentially Expressed in Tumor Cells Surviving High-Dose X-ray Irradiation: A Preliminary Study on Radio-Fragile Sites

  Kaori Tsutsumi, Moe Masuda, Hiroyuki Date

  Current Issues in Molecular Biology 43 2 1133 - 1141 2021年09月 [査読有り]
   

  Altered gene expression is a common feature of tumor cells after irradiation. Our previous study showed that this phenomenon is not only an acute response to cytotoxic stress, instead, it was persistently detected in tumor cells that survived 10 Gy irradiation (IR cells). The current understanding is that DNA double-strand breaks (DSBs) are recognized by the phosphorylation of histone H2AX (H2AX) and triggers the ataxia-telangiectasia mutated (ATM) protein or the ATM- and Rad3-related (ATR) pathway, which activate or inactivate the DNA repair or apoptotic or senescence related molecules and causes the expression of genes in many instances. However, because changes in gene expression persist after passaging in IR cells, it may be due to the different pathways from these transient intracellular signaling pathways caused by DSBs. We performed microarray analysis of 30,000 genes in radiation-surviving cells (H1299-IR and MCF7-IR) and found an interesting relation between altered genes and their chromosomal loci. These loci formed a cluster on the chromosome, especially on 1q21 and 6p21-p22 in both irradiated cell lines. These chromosome sites might be regarded as “radio-fragile” sites.
• Association between Physical Activity Levels and Body Composition among Healthy Older Japanese Adults during a Snowy Winter: A Cross-Sectional Study.

  Tomoko Shimoda, Teppei Suzuki, Kaori Tsutsumi, Mina Samukawa, Sadako Yoshimura, Katsuhiko Ogasawara

  International journal of environmental research and public health 17 15 2020年07月23日 [査読有り]
   

  BACKGROUND: Despite a long average lifespan, increased life expectancy does not guarantee higher quality of life. METHODS: To contribute in understanding some determinants of healthy life expectancies in older Japanese individuals in a snowy winter region, we investigated the indicators of health. Local residents (n = 124) in the city of Iwamizawa volunteered for health examinations from January 2016 to March 2016. We recorded activity via daily steps for 2-week periods. In addition, we measured body composition, grip strength, and assessed nutritional status. RESULTS: Analysis of body composition and daily activity indicated that women who walked more than 4000 steps had lower fat mass and increased muscle mass. Men with >3.0 metabolic equivalents (METs) when walking had lower body fat. CONCLUSION: For healthy older Japanese individuals in this snowy winter region, walking >4000 steps daily for women and exercise of >3.0 METs for men may indicate health-promoting activities.
• Cellular calcification induced by inorganic polyphosphate involves ATP depletion and opening of the mitochondrial permeability transition pore (mPTP).

  Kaori Tsutsumi, Tatsuya Sasase

  FEBS open bio 9 9 1617 - 1622 2019年09月 [査読有り]
   

  Inorganic polyphosphate (polyP) is a linear polymer containing tens to hundreds of orthophosphate residues linked by high-energy phosphoanhydride bonds. PolyP promotes osteocalcification and bone mineralization in both mouse and human osteoblastic cells. In the present study, we examined the molecular mechanism by which polyP affects mitochondrial metabolism to promote cellular calcification in MC3T3-E1 osteoblastic cells. The cellular content of adenosine triphosphate (ATP) was diminished one day after polyP treatment, and this was accompanied by increased conversion to adenosine diphosphate. Furthermore, mitochondrial membrane potential was significantly decreased in polyP-treated cells. These results suggest that the depletion of intracellular ATP and the decrease in mitochondrial membrane potential induced by polyP treatment may be a trigger to promote cell calcification.
• A Peptide Derived from Phosphoinositide 3-kinase Inhibits Endocytosis and Influenza Virus Infection.

  Yoichiro Fujioka, Aya O Satoh, Kosui Horiuchi, Mari Fujioka, Kaori Tsutsumi, Junko Sasaki, Prabha Nepal, Sayaka Kashiwagi, Sarad Paudel, Shinya Nishide, Asuka Nanbo, Takehiko Sasaki, Yusuke Ohba

  Cell structure and function 44 1 61 - 74 2019年04月25日 [査読有り][通常論文]
   

  Endocytosis mediates the internalization and ingestion of a variety of endogenous or exogenous substances, including virus particles, under the control of intracellular signaling pathways. We have previously reported that the complex formed between the small GTPase Ras and phosphoinositide 3-kinase (PI3K) translocates from the plasma membrane to endosomes, signaling from which thereby regulates clathrin-independent endocytosis, endosome maturation, influenza virus internalization, and infection. However, the molecular mechanism by which the Ras-PI3K complex is recruited to endosomes remains unclear. Here, we have identified the amino acid sequence responsible for endosomal localization of the Ras-PI3K complex. PI3K lacking this sequence failed to translocate to endosomes, and expression of the peptide comprising this PI3K-derived sequence inhibited clathrin-independent endocytosis, influenza virus internalization, and infection. Moreover, treatment of cells with this peptide in an arginine-rich, cell-penetrating form successfully suppressed influenza virus infection in vitro and ex vivo, making this peptide a potential therapeutic agent against influenza virus infection.Key words: signal transduction, endocytosis, endosome, imaging, influenza virus.
• Forefoot and hindfoot kinematics in subjects with medial tibial stress syndrome during walking and running.

  Takumi Okunuki, Yuta Koshino, Masanori Yamanaka, Kaori Tsutsumi, Masato Igarashi, Mina Samukawa, Hiroshi Saitoh, Harukazu Tohyama

  Journal of orthopaedic research : official publication of the Orthopaedic Research Society 37 4 927 - 932 2019年04月 [査読有り][通常論文]
   

  Excessive foot pronation during static standing, walking and running has been reported as a contributing factor for the development of medial tibial stress syndrome (MTSS). The motion of foot pronation consists of hindfoot and forefoot motion. However, no previous studies have investigated forefoot and hindfoot kinematics during walking and running in subjects with MTSS. The current study sought to compare hindfoot and forefoot kinematics between subjects with and without MTSS while walking and running. Eleven subjects with MTSS and 11 healthy controls (each group containing 10 males and one female) participated in the current study. Segment angles of the hindfoot and forefoot during walking and running barefoot on a treadmill were recorded using three-dimensional kinematic analysis. An independent t-test was used to compare kinematic data between groups. Subjects with MTSS exhibited significantly greater hindfoot eversion and abduction (p < 0.05) during walking and running than subjects without MTSS, significantly greater forefoot eversion and abduction (p < 0.05) during walking, and significantly greater forefoot abduction during running (p < 0.05). Hindfoot and forefoot kinematics during walking and running were significantly different between subjects with and without MTSS. For prevention and rehabilitation of MTSS, it may be important to focus on not only hindfoot but also forefoot kinematics during both running and walking. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.
• Investigation of dose-rate effects and cell-cycle distribution under protracted exposure to ionizing radiation for various dose-rates.

  Yusuke Matsuya, Stephen J McMahon, Kaori Tsutsumi, Kohei Sasaki, Go Okuyama, Yuji Yoshii, Ryosuke Mori, Joma Oikawa, Kevin M Prise, Hiroyuki Date

  Scientific reports 8 1 8287 - 8287 2018年05月29日 [査読有り][通常論文]
   

  During exposure to ionizing radiation, sub-lethal damage repair (SLDR) competes with DNA damage induction in cultured cells. By virtue of SLDR, cell survival increases with decrease of dose-rate, so-called dose-rate effects (DREs). Here, we focused on a wide dose-rate range and investigated the change of cell-cycle distribution during X-ray protracted exposure and dose-response curves via hybrid analysis with a combination of in vitro experiments and mathematical modelling. In the course of flow-cytometric cell-cycle analysis and clonogenic assays, we found the following responses in CHO-K1 cells: (1) The fraction of cells in S phase gradually increases during 6 h exposure at 3.0 Gy/h, which leads to radio-resistance. (2) Slight cell accumulation in S and G2/M phases is observed after exposure at 6.0 Gy/h for more than 10 hours. This suggests that an increase of SLDR rate for cells in S phase during irradiation may be a reproducible factor to describe changes in the dose-response curve at dose-rates of 3.0 and 6.0 Gy/h. By re-evaluating cell survival for various dose-rates of 0.186-60.0 Gy/h considering experimental-based DNA content and SLDR, it is suggested that the change of S phase fraction during irradiation modulates the dose-response curve and is possibly responsible for some inverse DREs.
• Factors affecting bone mineral density among snowy region residents in Japan: Analysis using multiple linear regression and Bayesian network model

  Teppei Suzuki, Tomoko Shimoda, Noriko Takahashi, Kaori Tsutsumi, Mina Samukawa, Sadako Yoshimura, Katsuhiko Ogasawara

  Journal of Medical Internet Research 20 5 e10  2018年05月01日 [査読有り][通常論文]
   

  Background: As the onset of osteoporosis leads to reduced activities of daily living and may result in patients being bedridden, efforts to prevent decreased bone density are necessary. Various studies on the relationship between sex, age, nutrients, and exercise habits and bone mineral density have been conducted to date. However, for snowy region residents, the magnitude of influence of various factors affecting bone mineral density and the influence level have not been clarified. Objective: This study aimed to clarify the degree of influence and factors influencing bone mineral density based on survey results on health conditions and lifestyle habits in heavy snow areas. Methods: A total of 354 citizens who visited a drugstore in the target area were included in a study that included using the brief-type self-administered diet history questionnaire on lifestyle and exercise habits. Height, weight, body composition, and bone densitometer values were analyzed using multiple regression to calculate their association with bone mineral density. In addition, a Bayesian network model was used to determine the influence level of each factor as a conditional probability. Results: Multiple regression analysis revealed that age, sex, fracture, and calcium intake significantly influenced bone mineral density. In addition, the result of Bayesian network analysis suggested that age and sex affected bone mineral density, whereas nutrients and exercise habits might not have a direct impact. However, calcium intake and the T-score were significant factors affecting the presence or absence of fracture experiences, suggesting that adequate calcium intake is essential for preventing fractures. Conclusions: In the multiple regression analysis, age, sex, fracture, and calcium intake were selected as factors however, in the Bayesian analysis, only age and sex affected bone mineral density while nutrients did not. In addition, the fact that calcium intake and the T-score were shown to affect bone fracture history suggests that calcium intake is an important measure that can prevent bone fractures. Overall, these results suggest that measures such as ensuring a bone fracture-free environment and providing nutritional advice for calcium intake can be effective in preventing bone loss.
• Modeling cell survival and change in amount of DNA during protracted irradiation

  Yusuke Matsuya, Kaori Tsutsumi, Kohei Sasaki, Yuji Yoshii, Takaaki Kimura, Hiroyuki Date

  JOURNAL OF RADIATION RESEARCH 58 3 302 - 312 2017年05月 [査読有り][通常論文]
   

  Hyper-radiosensitivity (HRS) is a well-known bioresponse under low-dose or low-dose-rate exposures. Although disorder of the DNA repair function, non-targeted effects and accumulation of cells in G(2) have been experimentally observed, the mechanism for inducing HRS by long-term irradiation is still unclear. On the basis of biological experiments and a theoretical study, we have shown that change in the amount of DNA associated with accumulation of cells in G(2) enhances radiosensitivity. To demonstrate continuous irradiation with 250 kVp X-rays, we adopted a fractionated regimen of 0.186 or 1.00 Gy per fraction at intervals of 1 h (i.e. 0.186 Gy/h, 1.00 Gy/h on average) to Chinese Hamster Ovary (CHO)-K1 cells. The change in the amount of DNA during irradiation was quantified by flow cytometric analysis with propidium iodide (PI). Concurrently, we attempted a theoretical evaluation of the DNA damage by using a microdosimetric-kinetic (MK) model that was modified to incorporate the change in the amount of DNA. Our experimental results showed that the fraction of the cells in G(2)/M phase increased by 6.7% with 0.186 Gy/h and by 22.1% with 1.00 Gy/h after the 12th irradiation. The MK model considering the change in amount of DNA during the irradiation exhibited a higher radiosensitivity at a high dose range, which could account for the experimental clonogenic survival. The theoretical results suggest that HRS in the high dose range is associated with an increase in the total amount of DNA during irradiation.
• Nutritional Status and Body Composition of Independently Living Older Adults in a Snowy Region of Japan.

  Shimoda T, Suzuki T, Takahashi N, Tsutsumi K, Samukawa M, Yoshimachi S, Goto T, Enomoto H, Kise N, Ogasawara K, Yoshimura S

  Gerontology & geriatric medicine 3 2333721417706854  2017年01月 [査読有り][通常論文]
   

  Lifestyle diseases, which are associated with nutrition, account for 30% of elderly requiring long-term care. To increase health expectancy among Japan's rapidly aging population, we investigated the nutritional status and body composition of elderly adults living in a region subject to heavy snowfall, to identify pertinent health indicators. The dietary habits of 288 local residents aged >= 50 years were analyzed using body composition and a brief-type self-administered diet history questionnaire. Body mass index of all residents was normal. Basal metabolic rate (BMR) and muscle mass were reduced in the older group. Dietary habits did not differ with age among men, but older women had significantly higher dietary intake. BMR and muscle mass declined with age, even when dietary intake was sustained. Despite sufficient dietary intake, independently living older adults demonstrate less efficient use of food with age. Interventions to reduce excessive sodium and protein intake are required.
• Inorganic polyphosphate enhances radio-sensitivity in a human non-small cell lung cancer cell line, H1299

  Kaori Tsutsumi, Yusuke Matsuya, Tomoki Sugahara, Manami Tamura, Satoshi Sawada, Sagiri Fukura, Hisashi Nakano, Hiroyuki Date

  Tumor Biology 39 6 1010428317705033  2017年01月01日 [査読有り][通常論文]
   

  Inorganic polyphosphate is a linear polymer containing tens to hundreds of orthophosphate residues linked by highenergy phosphoanhydride bonds. Polyphosphate has been recognized as a potent anti-metastasis reagent. However, the molecular mechanism underlying polyphosphate action on cancer cells is poorly understood. In this study, we investigated the involvement of polyphosphate in radio-sensitivity using a human non-small cell lung cancer cell line, H1299. We found that polyphosphate treatment decreases cellular adenosine triphosphate levels, suggesting a disruption of energy metabolism. We also found that the induction of DNA double-strand breaks was enhanced in polyphosphatetreated cells after X-ray irradiation and colony formation assay revealed that cell survival decreased compared with that of the control groups. These findings suggest that polyphosphate is a promising radio-sensitizer for cancer cells. Therefore, we hypothesized that polyphosphate treatment disrupts adenosine triphosphate-mediated energy transfer for cellular survival and DNA repair, thereby reducing the cellular capability to resist X-ray irradiation.
• Detection and measurement of rheumatoid bone and joint lesions of fingers by tomosynthesis: a phantom study for reconstruction filter setting optimization

  Yohei Ono, Tamotsu Kamishima, Nobutoshi Yasojima, Kenichi Tamura, Kaori Tsutsumi

  Radiological Physics and Technology 9 1 6 - 14 2016年01月01日 [査読有り][通常論文]
   

  Rheumatoid arthritis (RA) is a systemic disease that is caused by autoimmunity. RA causes synovial proliferation, which may result in bone erosion and joint space narrowing in the affected joint. Tomosynthesis is a promising modality which may detect early bone lesions such as small bone erosion and slight joint space narrowing. Nevertheless, so far, the optimal reconstruction filter for detection of early bone lesions of fingers on tomosynthesis has not yet been known. Our purpose in this study was to determine an optimal reconstruction filter setting by using a bone phantom. We obtained images of a cylindrical phantom with holes simulating bone erosions (diameters of 0.6, 0.8, 1.0, 1.2, and 1.4 mm) and joint spaces by aligning two phantoms (space widths from 0.5 to 5.0 mm with 0.5 mm intervals), examining six reconstruction filters by using tomosynthesis. We carried out an accuracy test of the bone erosion size and joint space width, done by one radiological technologist, and a test to assess the visibility of bone erosion, done by five radiological technologists. No statistically significant difference was observed in the measured bone erosion size and joint space width among all of the reconstruction filters. In the visibility assessment test, reconstruction filters of Thickness+− and Thickness−− were among the best statistically in all characteristics except the signal-to-noise ratio. The Thickness+− and Thickness−− reconstruction filter may be optimal for evaluation of RA bone lesions of small joints in tomosynthesis.
• Tomosynthesis can facilitate accurate measurement of joint space width under the condition of the oblique incidence of X-rays in patients with rheumatoid arthritis

  Yohei Ono, Rina Kashihara, Nobutoshi Yasojima, Hideki Kasahara, Yuka Shimizu, Kenichi Tamura, Kaori Tsutsumi, Kenneth Sutherland, Takao Koike, Tamotsu Kamishima

  BRITISH JOURNAL OF RADIOLOGY 89 1062 20150967  2016年 [査読有り][通常論文]
   

  Objective: Accurate evaluation of joint space width (JSW) is important in the assessment of rheumatoid arthritis (RA). In clinical radiography of bilateral hands, the oblique incidence of X-rays is unavoidable, which may cause perceptional or measurement error of JSW. The objective of this study was to examine whether tomosynthesis, a recently developed modality, can facilitate a more accurate evaluation of JSW than radiography under the condition of oblique incidence of X-rays. Methods: We investigated quantitative errors derived from the oblique incidence of X-rays by imaging phantoms simulating various finger joint spaces using radiographs and tomosynthesis images. We then compared the qualitative results of the modified total Sharp score of a total of 320 joints from 20 patients with RA between these modalities. Results: A quantitative error was prominent when the location of the phantom was shifted along the JSW direction. Modified total Sharp scores of tomosynthesis images were significantly higher than those of radiography, that is to say JSW was regarded as narrower in tomosynthesis than in radiography when finger joints were located where the oblique incidence of X-rays is expected in the JSW direction. Conclusion: Tomosynthesis can facilitate accurate evaluation of JSW in finger joints of patients with RA, even with oblique incidence of X-rays. Advances in knowledge: Accurate evaluation of JSW is necessary for the management of patients with RA. Through phantom and clinical studies, we demonstrate that tomosynthesis may achieve more accurate evaluation of JSW.
• Evaluation of the cell survival curve under radiation exposure based on the kinetics of lesions in relation to dose-delivery time

  Yusuke Matsuya, Kaori Tsutsumi, Kohei Sasaki, Hiroyuki Date

  JOURNAL OF RADIATION RESEARCH 56 1 90 - 99 2015年01月 [査読有り][通常論文]
   

  We have investigated the dose rate effects on cell damage caused by photon-beam irradiation. During a relatively long dose-delivery time with a low dose rate, lesions created in cells may undergo some reactions, such as DNA repair. In order to investigate these reactions quantitatively, we adopted the microdosimetric-kinetic (MK) model and deduced a cell surviving fraction (SF) formula for continuous irradiation. This model enabled us to estimate the SF from dose and dose rate. The parameters in the MK model were determined so as to generate the SF, and we attempted to evaluate the dose rate effects on the SF. To deduce the cell-specific parameters in the SF formula, including the dose rate, we performed a split-dose experiment and a single-dose experiment with a constant dose-delivery time (10 min) (to retain the condition for equivalent behavior of cell lesions) by means of a clonogenic assay. Then, using the MK model parameters, the SFs were reproduced for a variety of dose rates (1.0, 0.31, 0.18, 0.025 and 0.0031 Gy/min) and were compared with reported experimental data. The SF curves predicted by the MK model agreed well with the experimental data, suggesting that the dose rate effects appear in the kinetics of cell lesions during the dose-delivery time. From fitting the analysis of the model formula to the experimental data, it was shown that the MK model could illustrate the characteristics of log-SF in a rectilinear form at a high dose range with a relatively low dose rate.
• Quantitative estimation of DNA damage by photon irradiation based on the microdosimetric-kinetic model

  Yusuke Matsuya, Yosuke Ohtsubo, Kaori Tsutsumi, Kohei Sasaki, Rie Yamazaki, Hiroyuki Date

  JOURNAL OF RADIATION RESEARCH 55 3 484 - 493 2014年05月 [査読有り][通常論文]
   

  The microdosimetric-kinetic (MK) model is one of the models that can describe the fraction of cells surviving after exposure to ionizing radiation. In the MK model, there are specific parameters, k and y(D), where k is an inherent parameter to represent the number of potentially lethal lesions (PLLs) and y(D) indicates the dose-mean lineal energy in keV/mu m. Assuming the PLLs to be DNA double-strand breaks (DSBs), the rate equations are derived for evaluating the DSB number in the cell nucleus. In this study, we estimated the ratio of DSBs for two types of photon irradiation (6 MV and 200 kVp X-rays) in Chinese hamster ovary (CHO-K1) cells and human non-small cell lung cancer (H1299) cells by observing the surviving fraction. The estimated ratio was then compared with the ratio of gamma-H2AX foci using immunofluorescent staining. For making a comparison of the number of DSBs among a variety of radiation energy cases, we next utilized the survival data in the literature for both cells exposed to other photon types, such as Co-60 gamma-rays, Cs-137 gamma-rays and 100 kVp X-rays. The ratio of DSBs based on the MK model with conventional data was consistent with the ratio of gamma-H2AX foci numbers, confirming that the gamma-H2AX focus is indicative of DSBs. It was also shown that the larger y(D) is, the larger the DSB number is. These results suggest that k and y(D) represent the characteristics of the surviving fraction and the biological effects for photon irradiation.
• Morphogenetic Study on the Maturation of Osteoblastic Cell as Induced by Inorganic Polyphosphate

  Kaori Tsutsumi, Nagahito Saito, Yumi Kawazoe, Hong-Kean Ooi, Toshikazu Shiba

  PLOS ONE 9 2 e86834  2014年02月 [査読有り][通常論文]
   

  Since inorganic polyphosphates [poly(P)] have an activity to induce bone differenciation in vitro and in vivo, we examined an effect of poly(P) on organelle by light microscopy and electron microscopy in Murine MC3T3-E1 osteoblastic cells. The MC3T3-E1 cells were ultrastructurally observed to possess morphological characteristics of osteoblasts. Cells cultured with poly(P) were strongly stained with an anti-collagen type I antibody but not in those cultured without poly(P). Ultrastructural analysis of cells cultured with poly(P) revealed a well-developed Golgi apparatus, swollen and elongated rough endoplasmic reticulum, large mitochondria and many coated pits. Since MC3T3-E1 cells can be transformed from a resting phase to an active blastic cell phase after supplementation with poly(P), it implies that poly(P) can be an effective material for bone regeneration.
• A Simulation Study of the Radiation-Induced Bystander Effect: Modeling with Stochastically Defined Signal Reemission

  Kohei Sasaki, Kosuke Wakui, Kaori Tsutsumi, Akio Itoh, Hiroyuki Date

  COMPUTATIONAL AND MATHEMATICAL METHODS IN MEDICINE 2012 389095  2012年 [査読有り][通常論文]
   

  The radiation-induced bystander effect (RIBE) has been experimentally observed for different types of radiation, cell types, and cell culture conditions. However, the behavior of signal transmission between unirradiated and irradiated cells is not well known. In this study, we have developed a new model for RIBE based on the diffusion of soluble factors in cell cultures using a Monte Carlo technique. The model involves the signal emission probability from bystander cells following Poisson statistics. Simulations with this model show that the spatial configuration of the bystander cells agrees well with that of corresponding experiments, where the optimal emission probability is estimated through a large number of simulation runs. It was suggested that the most likely probability falls within 0.63-0.92 for mean number of the emission signals ranging from 1.0 to 2.5.
• Visualization of Ras-PI3K interaction in the endosome using BiFC

  Kaori Tsutsumi, Yoichiro Fujioka, Masumi Tsuda, Hideaki Kawaguchi, Yusuke Ohba

  CELLULAR SIGNALLING 21 11 1672 - 1679 2009年11月 [査読有り][通常論文]
   

  Recent studies indicate the importance of spatiotemporal regulation in the diversity and specificity of intracellular signaling. Here, we show that Ras-PI3K signaling plays an important role in the local regulation of phosphatidylinositol metabolism in the endosome through live-cell imaging by using a bimolecular fluorescence complementation technique, in which molecular interaction is indicated by fluorescence emission. Using several possible combinations of Ras and the Ras-binding domain, we identified an optimal set of probe molecules that yielded the most significant increase in fluorescence intensity between the active and inactive forms of Ras. This combination revealed that, among the Ras effectors tested, phosphatidy-linositol 3-kinase (PI3K) was specifically implicated in signaling in the endosome. We also found that full length PI3K was recruited to the endosome in EGF- and Ras-dependent manners, which appears to be essential for the activation of PI3K in this compartment. Taken together, these findings demonstrate that the spatiotemporal regulation of Ras-PI3K signaling may dictate the activation of PI3K and subsequent downstream signaling in the endosome. (C) 2009 Elsevier Inc. All rights reserved.
• Increased Motility and Invasiveness in Tumor Cells That Survive 10 Gy Irradiation

  Kaori Tsutsumi, Masumi Tsuda, Natsuka Yazawa, Hirotaka Nakamura, Seiichiro Ishihara, Hisashi Haga, Motoaki Yasuda, Rie Yamazaki, Hiroki Shirato, Hideaki Kawaguchi, Takeshi Nishioka, Yusuke Ohba

  CELL STRUCTURE AND FUNCTION 34 2 89 - 96 2009年 [査読有り][通常論文]
   

  Radiotherapy is an important noninvasive treatment for many types of cancer. However, it has been reported that the proliferative, invasive, and metastatic capacities of tumor cells can be increased in the repopulated tumors that survive radiotherapy. We have previously established a radiation-surviving cell model for the human non-small cell lung cancer cell line H1299 by harvesting relic cells 14 days after irradiation (IR cells). Here, we report that cell invasion, cell migration, and cell adhesion are enhanced in these surviving cancer cells. The mRNA expression levels of matrix metalloproteinases (MMPs), including mmp1, mmp2, and mmp9, were upregulated in IR cells compared with parental cells. A gelatin zymogram, wound healing assay, and invasion assay showed increased MMP activity, cell motility, and invasiveness in IR cells, respectively. Moreover, IR cells adhered more tightly to collagen-coated dishes than parental cells. Consistently, paxillin, phosphorylated FAK, integrin beta 1, and vinculin were strongly localized at focal adhesions in IR cells, as visualized by immunofluorescence. In this report, we identify molecules responsible for the malignant properties of tumor cells that survive irradiation. These molecules may be important therapeutic targets for the control of repopulated tumors after radiotherapy.
• Matrixmetalloproteinases: Up-regulated in subclones that survived 10-Gy irradiation

  Takeshi Nishioka, Motoaki Yasuda, Kaori Tsutsumi, Hisashi Haga, Hiroki Shirato

  Radiation Medicine - Medical Imaging and Radiation Oncology 25 8 430 - 431 2007年10月 [査読無し][通常論文]
  
• X-ray irradiation altered chemosensitivity of a p53-null non-small cell lung cancer cell line

  Kaori Tsutsumi, Motoaki Yasuda, Takeshi Nishioka

  Cell Structure and Function 31 2 47 - 52 2006年 [査読有り][通常論文]
   

  Radiotherapy is an effective approach to treating many types of cancer. Recent progress in radiotherapy technology, such as intensity-modulated radiation therapy (IMRT) and three-dimensional (3D) radiotherapy, allow precise energy transfer to the tumor, which has improved local control rates. However, the emergence of tolerant cells during or after radiotherapy remains problematic. In the present study, we first established a cell population from H1299, the p53-null non-small cell lung cancer cell line, by 10 Gy irradiation using 6 MV X-rays. The radio- and chemosensitivity of this cell population (referred to as H1299-IR) was determined using colony formation analyses and MTS assays. Compared with the parental cell line, the radiosensitivity of H1299-IR was apparently the same. H1299 and H1299-IR were both more radio tolerant than the A549 cell line. However, H1299-IR became significantly more sensitive to cisplatin, an antitumor agent. After exposure to 25 μg/ml cisplatin for 2 h, parental cells steadily grew during the MTS assay, whereas the sensitivity of H1299-IR cells doubled both at 24 and 48 h. Microarray analysis of over 30,000 H1299-IR genes (Agilent Technology) revealed that 12 and 15 genes were up- (> 2.0) and down- (< 2.0) regulated, respectively. Rad51d (homologous recombination repair protein) gene was down-regulated 2.8-fold, whereas matrix metalloproteinase 1 (collagenase-1) gene was up-regulated 4.4-fold. These results indicated that some p53-null non-small cell lung cancers could be successfully treated when X-ray radiotherapy was administered with subsequent or concurrent cisplatin chemotherapy. © 2006 by Japan Society for Cell Biology.
• Induction of calcification in MC3T3-E1 cells by inorganic polyphosphate

  Y Kawazoe, T Shiba, R Nakamura, A Mizuno, K Tsutsumi, T Uematsu, M Yamaoka, M Shindoh, T Kohgo

  JOURNAL OF DENTAL RESEARCH 83 8 613 - 618 2004年08月 [査読有り][通常論文]
   

  Relatively large amounts of inorganic polyphosphate [poly(P)] (400 muM) have been found in normal osteoblasts. The effect of poly( P) with an average chain length of 65 phosphate residues on cell calcification was therefore investigated with the use of MC3T3-E1 cells. Expression of both osteopontin and osteocalcin was induced by poly(P) (0.1 similar to 1 mM), and cells treated with poly(P) were strongly stained by alizarin red. In addition, the level of alkaline phosphatase activity induced in poly(P)-treated cells was two-fold higher than that in either orthophosphate-treated or control cells but not higher than that in cells treated with beta-glycerophosphate and ascorbic acid. In contrast, however, polyphosphatase activities were activated by poly(P) treatment to levels up to sixfold greater than that in controls. MC3T3-E1 cells may utilize poly(P) as a phosphate source for calcification rather than phosphate sources that are mainly produced by ALPase. Poly(P)-dependent induction of polyphosphatase activities may therefore promote calcification in MC3T3-E1 cells.
• Modulation of mitogenic activity of fibroblast growth factors by inorganic polyphosphate

  T Shiba, D Nishimura, Y Kawazoe, Y Onodera, K Tsutsumi, R Nakamura, M Ohshiro

  JOURNAL OF BIOLOGICAL CHEMISTRY 278 29 26788 - 26792 2003年07月 [査読有り][通常論文]
   

  The proliferation of normal human fibroblast cells was enhanced by the addition of inorganic polyphosphate ( poly( P)) into culture media. The mitogenic activities of acidic fibroblast growth factor (FGF-1) and basic fibroblast growth factor (FGF-2) were also enhanced by poly( P). A physical interaction between poly( P) and FGF-2 was observed, and FGF-2 was both physically and functionally stabilized by poly( P). Furthermore, poly( P) facilitated the FGF-2 binding to its cell surface receptors. Because poly( P) is widely distributed in mammalian tissues, it may be a spontaneous modulator of FGFs.
• Involvement of inorganic polyphosphate in expression of SOS genes

  K Tsutsumi, M Munekata, T Shiba

  BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION 1493 1-2 73 - 81 2000年09月 [査読有り][通常論文]
   

  Inorganic polyphosphate (poly(P)) is a linear polymer that has been found in every organism so far examined. To elucidate the functions of poly(P) in the regulation of gene expression, the level of cellular poly(P) in Escherichia coli was reduced to a barely detectable concentration by overproduction of exopolyphosphatase (exopoly(P)ase) with a plasmid encoding yeast exopoly(P)ase (Shiba et al., Proc. Natl. Acad. Sci. USA 94 (1997) 11210-11215). It was found that exopoly(P)ase-overproducing cells were more sensitive to UV or mitomycin C (MMC) than were control cells. Poly(P) accumulation was observed after treatment with MMC, whereas the poly(P) level was below the detectable level in cells that overproduced exopoly(P)ase. When exopoly(P)ase-overproducing cells were transformed again by a multiple copy number plasmid that carries the polyphosphate kinase gene (ppk), the cells accumulated a great amount of poly(P) and restored the UV and MMC sensitivities to the level of control cells. In exopoly(P)ase-overproducing cells, the expression of recA and umuDC were not induced by MMC. In addition, a strain containing multiple copies of ppk accumulated not only a large amount of poly(P) but also recA mRNA. Since recA expression was induced in a reed-deletion strain harboring a plasmid with the ppk gene, poly(P) could be necessary for regulating the expression of SOS genes without depending on the RecA-LexA regulatory network. (C) 2000 Elsevier Science B.V. All rights reserved.
• Inorganic polyphosphate and polyphosphate kinase: Their novel biological functions and applications

  T Shiba, K Tsutsumi, K Ishige, T Noguchi

  BIOCHEMISTRY-MOSCOW 65 3 315 - 323 2000年03月 [査読無し][通常論文]
   

  In this review, we discuss the following two subjects: 1) the physiological function of polyphosphate (poly(P)) as a regulatory factor for gene expression in Escherichia coli, and 2) novel functions of E. coli polyphosphate kinase (PPK) and their applications. With regard to the first subject, it has been shown that E. coli cells in which yeast exopolyphosphatase (poly(P)ase), PPX1, was overproduced reduced resistance to H2O2 and heat shock as did a mutant whose polyphosphate kinase gene is disrupted. Sensitivity to H2O2 and heat shock evinced by cells that overproduce PPX1 is attributed to depressed levels of rpoS expression. Since rpoS is a central element in a regulatory network that governs the expression of stationary-phase-induced genes, poly(P) affects the expression of many genes through controlling rpoS expression. Furthermore, poly(P) is also involved in expression of other stress-inducible genes that are not directly regulated by rpoS. The second subject includes the application of novel functions of PPK for nucleoside triphosphate (NTP) regeneration. Recently E. coli PPK has been found to catalyze the kination of not only ADP but also other nucleoside diphosphates using poly(P) as a phospho-donor, yielding NTPs. This nucleoside diphosphate kinase-like activity of PPK was confirmed to be available for NTP regeneration essential for enzymatic oligosaccharide synthesis using the sugar nucleotide cycling method. PPK has also been found to express a poly(P):AMP phosphotransferase activity by coupling with adenylate kinase (ADK) in E. coli. The ATP-regeneration system consisting of ADK, PPK, and poly(P) was shown to be promising for practical utilization of poly(P) as ATP substitute.
• An Analysis of DNA-based Computing Process

  Hirayama, T, Shiba, T, Yamamoto, M, Tsutsumi, K, Takiya, S, Munekata, M, Suzuki, K, Ohuchi, A

  The Fourth International Symposium on Artificial Life and Robotics 358 - 361 1999年 [査読無し]
  
• Experimental Results in DNA Computation: solution of an HPP with eight vertices

  Hirayama, T, Shiba, T, Yamamoto, M, Tsutsumi, K, Suzuki, K, Takiya, S, Munekata, M, Ohuchi, A

  Preliminary Proceedings of DNA Based Computers 5 249  1999年 [査読無し]
  
• Inorganic polyphosphate and the induction of rpoS expression

  T Shiba, K Tsutsumi, H Yano, Y Ihara, A Kameda, K Tanaka, H Takahashi, M Munekata, NN Rao, A Kornberg

  PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 94 21 11210 - 11215 1997年10月 [査読有り][通常論文]
   

  Inorganic polyphosphate [poly(P)] levels in Escherichia coli were reduced to barely detectable concentrations by expression of the plasmid-borne gene for a potent yeast exopolyphosphatase [poly(P)ase]. As a consequence, resistance to H2O2 was greatly diminished, particularly in katG (catalase HPI) mutants, implying a major role for the other catalase, the stationary-phase KatE (HPII), which is rpoS dependent. Resistance was restored to wild-type levels by complementation with plasmids expressing ppk, the gene for PPK [the polyphosphate kinase that generates poly(P)], Induction of expression of both katE and rpoS (the stationary-phase sigma factor) was prevented in cells in which the poly(P)ase was overproduced, Inasmuch as this inhibition by poly(P)ase did not affect the levels of the stringent-response guanosine nucleotides (pppGpp and ppGpp) and in view of the capacity of additional rpoS expression to suppress the poly(P)ase inhibition of katE expression, a role is proposed for poly(P) in inducing the expression of rpoS.

書籍

• 人体のメカニズムから学ぶ 放射線生物学

  松本 義久 
  メジカルビュー社 2017年02月 (ISBN: 4758317259) 315

  

講演・口頭発表等

• “Challenge for identification of key molecules that characterize the property of tumor cells that survive radiotherapy"  [通常講演]

  堤 香織

  Cycle de conférences JSPS Strasbourg et MUFJ 2010年03月

その他活動・業績

• 超音波装置を用いたストレス荷重時の足関節前方移動距離計測の試み

  中谷 詩世, 館野 真理, 堀江 達則, 松本 和久, 寒川 美奈, 遠山 晴一, 堤 香織 超音波検査技術 44 (2) 277 -277 2019年04月 [査読有り][通常論文]
  
• 二次元画像を用いた脊柱アライメント評価法の妥当性検証

  石田 優子, 寒川 美奈, 谷 祐児, 堤 香織, 大場 健裕, 鶴喰 涼, 三上 兼太朗, 山中 正紀, 遠山 晴一 日本臨床スポーツ医学会誌 24 (4) S312 -S312 2016年10月 [査読無し][通常論文]
  
• 放射線の長時間照射中における細胞周期と放射線感受性の解析

  松谷 悠佑, 吉井 勇治, 堤 香織, 伊達 広行, 森 諒輔, 佐々木 恒平, 及川 青亮 北海道放射線技術雑誌 (81) 127 -127 2016年10月 [査読無し][通常論文]
  
• Medial tibial stress syndrome症例におけるトレッドミル走行時の膝および股関節運動に対する前・後足部運動の比較

  奥貫 拓実, 越野 裕太, 遠山 晴一, 堤 香織, 五十嵐 將斗, 江沢 侑也, 寒川 美奈, 山中 正紀 日本臨床スポーツ医学会誌 24 (4) S315 -S315 2016年10月 [査読無し][通常論文]
  
• Medial tibial stress syndrome症例における走行時の後・前足部kinematicsおよび内側縦アーチの動態の検討

  奥貫 拓実, 越野 裕太, 遠山 晴一, 堤 香織, 生田 亮平, 佐橋 健人, 横山 美翔, 江沢 侑也, 寒川 美奈, 齊藤 展士, 山中 正紀 理学療法学 43 (Suppl.2) O -SP 2016年10月 [査読無し][通常論文]
   

  【はじめに，目的】Medial tibial stress syndrome（MTSS）は運動時に脛骨後内側縁に疼痛が発生する症状とされ，ランニング障害の13～17%を占める下肢オーバーユース障害である。MTSSの危険因子として，荷重動作時の内側縦アーチ低下や後足部外反角度増大が報告されている。荷重動作時の内側縦アーチ低下や後足部運動は，前足部運動と関連することが知られているが，MTSS症例において前足部や内側縦アーチを含めた走行時の足部kinematicsは明らかではない。本研究の目的は，multi-segment foot modelを用いて，MTSS症例におけるトレッドミル走行時の後足部，前足部および内側縦アーチの動態の特徴を検討することとした。【方法】対象は，MTSS症例群11名（男性10名，女性1名，年齢：20.5±1.5歳，BMI：22.0±2.0kg/m²）および健常群11名（男性10名，女性1名，年齢：20.5±1.4歳，BMI：19.7±1.3 kg/m²）とした。MTSSの定義はYatesら（2004）の基準に従い，1）脛骨後内側縁の圧痛がある，2）圧痛部位が5cm以上である，3）X線画像上で脛骨疲労骨折の所見がない，こととした。Leardiniら（2007）のmulti-segment foot modelに準じて下腿と足部に反射マーカーを16個貼付し，赤外線カメラ6台（Motion Analysis社製：200 Hz）を用いて，裸足によるトレッドミル走行（12km/h）の5ストライドを記録・解析した。解析にはVisual 3D（C-motion社製）を用いて，下腿に対する後足部内外反・内外転，後足部に対する前足部内外反・内外転・底背屈および内側縦アーチ角（第1中足骨頭，舟状骨，踵骨下部のマーカーから成る角）を算出した。立脚相を100%に正規化し，対応のないt-検定を用いて各kinematicsを群間で比較した。有意水準はp＜0.05とした。【結果】MTSS症例群は健常群に比し，立脚相3～14%の区間で有意に後足部外反が大きく，立脚相0～28%および55～100%の2つの区間で有意に後足部外転が大きかった（p＜0.05）。さらに，MTSS症例群は立脚相21～35%の区間において健常群に比し，有意に前足部外転が大きかった（p＜0.05）。一方で，前足部内外反・底背屈および内側縦アーチ角では群間で差は認められなかった。【結論】本研究結果は，走行立脚相においてMTSS症例は後足部外反・外転および前足部外転が健常者に比べ，有意に大きいことを示した。したがって，MTSS症例では，後足部外反および外転の増大により足部内返し筋がより伸長された状態で遠心性収縮を要求され，脛骨後内側縁に牽引ストレスが生じる可能性がある。加えて，前足部外転および後足部外転の増大は，下腿に対し足部をより外側に向けた状態であることを示唆し，これは走行時の荷重応答および踏み切りに伴う足部内転フリーモーメントの発生に寄与し，脛骨に回旋ストレスが加わる可能性がある。したがって，MTSS症例に対する理学療法実施の際には，後足部だけでなく前足部運動にも着目する必要があることを示唆した。
• Medial tibial stress syndrome症例におけるトレッドミル走行時の足部kinematicsの特徴(第1報)

  奥貫 拓実, 山中 正紀, 越野 裕太, 五十嵐 将斗, 江沢 侑也, 寒川 美奈, 堤 香織, 遠山 晴一 日本臨床スポーツ医学会誌 23 (4) S273 -S273 2015年10月 [査読無し][通常論文]
  
• Morphological Alteration of Murine MC3T3-E1 Osteoblastic cells Induced by Inorganic Polyphosphate

  Kaori Tsutsumi, Nagahito Saito, Yumi Kawazoe, Hong-Kean Ooi, Toshikazu Shiba FASEB JOURNAL 29 2015年04月 [査読有り][通常論文]
  
• テロスストレス装置を用いたX線足関節前方引き出しテストにおける性差の検討

  横山 美翔, 寒川 美奈, 堤 香織, 田中 茉衣, 遠山 晴一 理学療法学 42 (Suppl.2) P1 -B 2015年04月 [査読無し][通常論文]
   

  【はじめに，目的】足関節前方引き出しテストは，足関節不安定性評価によく用いられている方法である。しかしながら徒手による前方引き出しテストに関しては信頼性が低い（Lee，2014）という報告もみられることから，臨床ではテロスストレス装置を用いたX線前方引き出しテストも診断に行われている。前方引き出しストレステストは，ストレスを加えながら距骨前方移動量を定量的に計測する検査である。身体構造における性差は，筋横断面積（加賀谷，1998），筋厚（Kubo, 2003），関節弛緩性（Shultz, 2009）などに関して報告されており，性差が存在することが示唆されている。これまで，ストレス負荷時の距骨傾斜角をX線で計測した研究では，健常男女間で結果に差がみられている（Wilkerson, 2000）が，距骨前方引き出しテストにおける性差に関しては報告がみられていない。これまでの身体構造における性差に関する報告から，前方引き出しテストにおいても性差が生じる可能性がある。したがって本研究の目的は，テロスストレス装置を用いたX線足関節前方引き出しテストにおける距骨前方移動量の性差を検討することとした。【方法】対象は，足部に整形外科的及び神経学的既往歴のない男性10名（年齢21.9±0.7歳），女性8名（年齢21.3±0.7歳）の両足部とした。足関節不安定性に関しては，「足関節機能的不安定性の主観的評価法（，1991）」日本語版を用いて聴取した。Karlssonの先行研究に基づき，合計が81ポイント以上の場合，足関節機能的不安定性なしと判断した。X線ストレステスト時の測定位肢位は，検側を下にした側臥位で，股関節及び膝関節軽度屈曲位，足関節中間位とした。テロスストレス装置（Telos Stress Device, Griesheim, Germany）を足部に当て，内果と外果を結ぶ線は検査台に対して垂直とした。テロスストレス装置加圧点は脛骨内果から5cm近位とし，加圧なし（0N）時と最大加圧（120N）時のX線写真を撮影した。得られたX線画像より距骨の前方移動量を，脛骨後縁から距骨の最も近い関節面までを測定した。各画像は3回測定し，その平均値を用いて，最大加圧時と加圧なし時の差を算出した。算出した値を対応のないt検定により解析した。有意水準は5%とした。【結果】足関節不安定性に関する質問紙では，男性（右足99.5点，左足99.5点），女性（右足99.6点，左足99.6点）であり，足関節機能的不安定症の被験者は含まれていないことが示された。X線距骨前方引き出しテスト時の両側における距骨前方移動量は，男性2.5±1.2 cm，女性2.6±1.3 cmであり，差はみられなかった。また，一側下肢毎の検討では，右足で男性2.5±1.2 cm，女性2.9±1.4 cm，左足で男性2.5±1.3cm，女性2.2±1.1cmと，有意差は検出されなかった。【考察】本研究では，テロスストレス装置を用いたX線足関節前方引き出しテストにおいて，性差は検出されなかった。足関節構造の性差については，距骨の傾斜角が男性と比べて女性で大きい（Wilkerson, 2000）こと，及び足関節上方及び内側のスペースが女性より男性で広い（Murphy, 2014）こと等解剖学的性差の存在は示唆されていたが，本研究ではこれらと異なる結果を示した。足関節に限らず靭帯は，男性と比べて女性でより伸張性がみられる（Shultz, 2009）という報告もある。今回，足関節前方引き出し試験において性差はみられなかったため，臨床で本テスト実施時には性差の影響は小さいことが示唆される結果となった。一方，徒手で行われる足関節前方引き出しテストでは，足関節前距腓靭帯損傷の検査として足関節を内反も加えて行われることがある。しかしながら，テロスストレス装置による足関節前方引き出しテストでは，距骨は構造上前方に牽引されるのみである。したがって，ストレス負荷のかけ方によって靭帯の伸張性に性差が生じる可能性もある。また，足関節不安定性評価については被験者の主観でのみ判断されていることや，質問紙の結果から足関節機能的不安定症の被験者は含まれていなかったが，整形外科医による診断は実施していなかったことも，本研究の限界として挙げられる。【理学療法学研究としての意義】身体構造の性差については数多くの研究がされているが，靭帯伸張性に統一した見解は得られていない。本研究により，臨床で行われているテロスストレス装置を用いたX線足関節前方引き出しテストにおいては性差がみられなかったため，本試験においては性差による影響が小さい可能性が示唆された。
• 線量率の違いを考慮した細胞生存率モデルの有用性

  松谷悠佑, 佐々木恒平, 堤香織, 伊達広行 北海道放射線技術雑誌 75 (75) 135 -135 2013年10月25日 [査読無し][通常論文]
  
• 放射線によるDNA損傷の定量的モデル解析と生物学的効果比

  松谷悠佑, 大坪洋輔, 堤香織, 山崎理衣, 佐々木恒平, 伊達広行 日本放射線影響学会大会講演要旨集 56th 97 -97 2013年10月01日 [査読無し][通常論文]
  
• コンフルエント単層細胞環境における放射線誘発バイスタンダー効果のシミュレーション

  佐々木恒平, 松谷悠佑, 堤香織, 伊藤秋男, 伊達広行 日本放射線影響学会大会講演要旨集 56th 111 -111 2013年10月01日 [査読無し][通常論文]
  
• ストレス負荷超音波撮影法による足関節不安定症の定量的評価の試み

  堤 香織, 寒川 美奈, 遠山 晴一 日本臨床スポーツ医学会誌 21 (4) S178 -S178 2013年10月 [査読有り][通常論文]
  
• 超音波装置を用いた足関節に対する前方引き出し負荷と移動距離計測の試み

  中谷 詩世, 福良 沙霧, 寒川 美奈, 神島 保, 遠山 晴一, 堤 香織 北海道放射線技術雑誌 (75) 116 -116 2013年10月 [査読無し][通常論文]
  
• 修復効果を含む新しい細胞生存率モデルの検討

  泉井宏介, 佐々木恒平, 堤香織, 伊達広行 北海道放射線技術雑誌 73 (73) 127 -127 2012年11月05日 [査読無し][通常論文]
  
• 光子線照射がもたらす細胞核内損傷に関する解析

  大坪洋輔, 泉井宏介, 佐々木恒平, 堤香織, 伊達広行 北海道放射線技術雑誌 71 (71) 126 -126 2011年10月25日 [査読無し][通常論文]
  
• Matrix Metalloproteinase's Are Up-regulated in Sub-clones That Survived 10Gy Irradiation: A Possible Role of Hypoxic-inducible Factor-alpha (HIF1a)

  T. Nishioka, K. Tsutsumi, T. Takeshima, H. Shirato, M. Yasuda INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS 78 (3) S631 -S631 2010年 [査読無し][通常論文]
  
• Cell Motility and Invasion of Surviving Tumor Cells after 10 Gy Irradiation

  K. Tsutsumi, M. Tsuda, N. Yazawa, H. Nakamura, M. Yasuda, R. Yamazaki, H. Shirato, H. Kawaguchi, Y. Ohba, T. Nishioka INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS 75 (3) S538 -S539 2009年 [査読無し][通常論文]
  
• Visualization of an interaction between Ras and Ras-binding domain in living cells by bimolecular fluorescence complementation

  Kaori Tsutsumi, Masumi Tsuda, Hideaki Kawaguchi, Yusuke Ohba FASEB JOURNAL 22 2008年04月 [査読有り][通常論文]
  
• Novel function of transcription factor ATF5: Blockade of p53-dependent apoptosis induced by irradiation

  T. Nishioka, M. Yasuda, H. Haga, R. Yamazaki, K. Tsutsumi, H. Shirato INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS 72 (1) S43 -S43 2008年 [査読無し][通常論文]
  
• Three-dimensional tumor cell movement in collagen gel: mesenchymal-to-amoeboid transition in a sub-clone that survived 10 Gy irradiation

  T. Nishioka, H. Haga, Y. Miyai, M. Yasuda, K. Tsutsumi, R. Yamazaki, H. Shirato, K. Kawabata INTERNATIONAL JOURNAL OF RADIATION ONCOLOGY BIOLOGY PHYSICS 69 (3) S148 -S148 2007年 [査読無し][通常論文]
  

共同研究・競争的資金等の研究課題

• 悪性神経膠芽腫の放射線抵抗性におけるニューロピリン1の役割と機能解明

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2023年04月 -2026年03月 

  代表者 : 堤 香織
• 高水素濃度下でＸ線被ばくが及ぼす生物学的影響の検証

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2021年04月 -2023年03月 

  代表者 : 伊達 広行, 堤 香織

   

  培養細胞液の高濃度水素条件を実現する上で必要な、バブリング方式の水素ガス発生装置と水素濃度測定器を購入した。純水を用いて高濃度水素水を生成し、水素濃度がどの程度になるかを測定した。この測定において、発生装置の初期状態や反復作業に依存して水素濃度の検出結果に比較的大きなバラツキがあることが判明したため、種々の条件にて繰り返し水素水生成試験を行った。水素水の抽出直後濃度とその後６時間程度までの水素濃度変化を調べ、最大１ppm程度の水素濃度が得られること、また時間とともに指数関数的に減少することを確認した。この水素濃度の経時変化を基に、細胞へＸ線を照射するタイミングと、免疫蛍光染色法による細胞核内ＤＮＡ二本鎖切断（ＤＳＢ）数の観測やコロニーアッセイによる細胞生存率測定などの手順について、対照実験のための幾つかの有効と考えられるプランを考案した。 一方、高濃度水素環境下での細胞の放射線によるダメージについて、高濃度水素条件下と対極にある高濃度酸素下での解析を踏襲したアプローチとして、照射面積による効果を観測する実験系や、損傷低減を組み入れた細胞生存率のモデル化の可能性について検討した。特に放射線の非標的効果について、高水素濃度下では低酸素環境下における状態と類似した結果をもたらす可能性があり、その検証に向けた正確な水素濃度環境における照射実験を模擬する微視的線量動力学（ＭＫ）モデルの改良を視野に入れて解析を進めている。
• ポリリン酸がミトコンドリア活性を調節して放射線感受性を制御するメカニズムの解明

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2020年04月 -2023年03月 

  代表者 : 堤 香織

   

  初年度の研究成果により、ヒト悪性神経膠芽腫由来細胞株T98Gではポリリン酸によるミトコンドリア膜電位の低下に大きな違いがみられなかったことから、R3年度はヒト肺非小細胞癌由来細胞株H1299にターゲットを絞って研究を進めた。また培養培地の組成の違いによる細胞代謝への影響を確かめるために、培地中のピルビン酸の有無によるポリリン酸の影響の違いにも焦点を当てた。以下のR3年度の研究遂行状況とその結果を記す。 培地中のピルビン酸の有無による（１）ミトコンドリア膜電位の変化、（２）細胞内ATP量の変化、（３）グルコース-6-リン酸脱水素酵素（G6PD）活性の変化をポリリン酸の有無によって観察した。（１）ミトコンドリア膜電位の変化では、Cell Meter™ JC-10 Mitochondrion Membrane Potential Assay Kit （AAT Bioquest）により、ポリリン酸処理の有無によるミトコンドリアの膜電位を比較した。その結果、ポリリン酸による細胞処理によって処理後3日後のミトコンドリア膜電位の低下がみられたが、培地中にピルビン酸を添加することによりポリリン酸処理細胞とコントロール細胞との間にミトコンドリア膜電位の差がみられなくなった。（２）細胞内ATP量の変化では、Cell Titer Gro（Promega）を用いて細胞内のATP量を測定した。ミトコンドリア膜電位において、ピルビン酸の添加によって膜電位の回復がみられた一方で、ATP量についてはポリリン酸処理による細胞内ATP量の低下は培地中のピルビン酸の有無に関わらず低下を維持した。（３）G6PD活性の変化では、ポリリン酸がペントースリン酸回路の促進する可能性を考えて、G6PD活性をwestern blottingによるリン酸化によりコントロールとの比較検討を行ったが、ピルビン酸の有無、ポリリン酸処理の有無に関わらず活性に差は認められなかた。
• 病院と連携する在宅遠隔相談看護師のアセスメント能力向上プログラムの開発と検証

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2015年04月 -2018年03月 

  代表者 : 良村 貞子, 下田 智子, 小笠原 克彦, 吉田 祐子, 鈴木 哲平, 堤 香織, 高橋 紀子, 寒川 美奈, 田村 菜穂美, 佐藤 ひとみ, 船木 典子, 新岡 郁子

   

  看護師のアセスメント能力の向上には、健康相談希望者に関する情報収集能力と的確な分析力の向上が必要である。本研究では、遠隔健康相談を希望する地域住民のＢＭＩ、食事の摂取量と内容、自己採血データ、筋肉量等の体組成、日常生活習慣などのデータを、相談担当の在宅看護師と共有できる、個人情報を保護したデータ管理システムを開発した。在宅看護師による各データの分析のシミュレーション研修は、健康相談者とデータを共有しながらの、在宅看護師による問診および視診において、効率的で効果的であった。
• ポリリン酸によって骨芽細胞の石灰化が促進する情報伝達分子メカニズムの解明

  文部科学省：科学研究費補助金基盤研究(C)

  研究期間 : 2015年04月 -2018年03月 

  代表者 : 堤 香織
• ＭＲＩ複素誘電率画像は細胞内水分子のネットワーク構造を反映するか？

  日本学術振興会：科学研究費助成事業

  研究期間 : 2012年04月 -2015年03月 

  代表者 : 山本 徹, 堤 香織, タ キンキン

   

  病気によるタンパク濃度変化のMRIによる非侵襲的検出の可能性を検証するため、MRIで求まる縦緩和率、横緩和時間に加え誘電率のタンパク濃度依存性を調べ、横緩和率が最もタンパク濃度変化検出精度が高いことを明らかにした。特に、脳脊髄液の流れの影響を受けない信号抽出を工夫することで、細菌性髄膜炎などによる脳脊髄液中のタンパク濃度変化をT2値測定により判別できる可能性があることを示した。
• MRI 複素電気伝導度画像は細胞内分子のネットワーク構造を反映するか？

  文部科学省：科学研究費補助金萌芽研究

  研究期間 : 2012年04月 -2014年03月 

  代表者 : 山本 徹
• 放射線を生残した腫瘍細胞におけるニューロピリンの機能解析

  文部科学省：科学研究費補助金若手研究(B)

  研究期間 : 2010年04月 -2012年03月 

  代表者 : 堤 香織
• 放射線被曝影響解析に向けた線量付与の統計的モデルの構築

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  研究期間 : 2010年 -2012年 

  代表者 : 伊達 広行, 堤 香織

   

  生体細胞に照射した放射線の影響を、モンテカルロ法に基づく電子線や生理活性物質の挙動に関するシミュレーションと、放射線照射後の細胞核損傷観測の両者によって解析した。その結果、放射線照射により生体内に発生する電子線のエネルギー付与過程が細胞核内のDNA損傷に強く影響すること、また細胞生存率が損傷修復を考慮した統計分布と致死確率を考慮したモデルにより説明できること等を明らかにした。
• 放射線治療後にみられる再増殖腫瘍細胞の運動特性の解析

  文部科学省：科学研究費補助金若手研究(B)

  研究期間 : 2008年04月 -2010年03月 

  代表者 : 堤 香織
• 放射線照射を生き延びた癌細胞に秘められた謎:癌根絶への多角的アプローチ

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2008年 -2010年 

  代表者 : 西岡 健, 安田 元昭, 芳賀 永, 堤香 織, 酒井 正春, 本間 明宏

   

  放射線後生き残った細胞(以下IRとをす)の遺伝子発現をcDNAアレイにより調べた。マウス線維肉腫細胞のセルラインQRsPを使用し10Gyの照射を施行した。コロニーアッセイとともに6つのクローンの樹立が行われた。照射を生き延びた細胞では、サイクリン依存性キナーゼ阻害剤(CKI)、p16 p57 の著明な発現低下;14.8および12.0の低下、また耐性クローン(IR)をマウスに移植したところ移植比100%がえられた。この細胞株にCD34がふくまれていることをかんがみると、がん幹細胞が放射線耐性に含まれていた可能性がある。
• 放射線治療中に生ずる高度増殖浸潤能クローンの封じ込め

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2006年 -2007年 

  代表者 : 西岡 健, 安田 元昭, 芳賀 永, 本間 明宏, 堤 香織

   

  マウス繊維芽肉腫細胞株QR s P細胞に10Gyの放射線照射を施行し、生存したコロニーから24株をクローニングした。これらに対しさらに10Gy照射したところコロニーの出現のしかたはまちまちであり、母集団の中には放射線高感受性のものもあれば抵抗性のものもあることが判明した。このうち最も抵抗性の株(10-1IRと命名)と最も高感受性の株10-5IRについて様々な角度から相違点を見つけ出した。まず親株との間でそれぞれc DNAアレイを行った。またIR群それぞれをC57BL/6マウスに細胞移植を行ったところ造腫瘍能には変わりがないことが判明した。この腫瘍から細胞株を再び作成し親株との間で2回目のc DNAアレイを施行した。この計4回のアレイデータから、それぞれ親株と似ているもの、異なるものを抽出し、さらにそれらの遺伝子群について10-1IRと10-5IRの間で異なる遺伝子を70個検出した。このなかには転写因子であるATF5が存在した。放射線照射をmimicするためp53を強制導入して親株と10-5IRの細胞死分画をフローサイトメーターみたところ、親株で7%、10-5IRで25%と明らかに10-5IR細胞においてp53によるアポトーシスが多く見られた。次にATF5を10-5IRに強制導入しコロニーアッセイによる10Gy照射後の生存コロニー数、フローサイトメーターによるアポトーシスの文画を調べたところいずれも、放射線に対する耐性を獲得した。次にATF5とp53を共導入しp53のレポーターアッセイをおこなったところp53の転写活性が有意に低下していることが観察された。以上の結果からATF5はなんらかの機構を介してp53に対し抑制的に働く遺伝子であることが判明した。
• Radiobiology

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