研究等業績 - 原著論文 - 脇　裕典

Robust and highly efficient hiPSC generation from patient non-mobilized peripheral blood-derived CD34+ cells using the auto-erasable Sendai virus vector.

Takashi Okumura, Yumi Horie, Chen-Yi Lai, Huan-Ting Lin, Hirofumi Shoda, Bunki Natsumoto, Keishi Fujio, Eri Kumaki, Tsubasa Okano, Shintaro Ono, Kay Tanita, Tomohiro Morio, Hirokazu Kanegane, Hisanori Hasegawa, Fumitaka Mizoguchi, Kimito Kawahata, Hitoshi Kohsaka, Hiroshi Moritake, Hiroyuki Nunoi, Hironori Waki, Shin-Ichi Tamaru, Takayoshi Sasako, Toshimasa Yamauchi, Takashi Kadowaki, Hiroyuki Tanaka, Sachiko Kitanaka, Ken Nishimura, Manami Ohtaka, Mahito Nakanishi, Makoto Otsu

Stem cell research & therapy   10 ( 1 ) 185 - 185   2019年06月  [査読有り]

研究論文（学術雑誌）  

BACKGROUND: Disease modeling with patient-derived induced pluripotent stem cells (iPSCs) is a powerful tool for elucidating the mechanisms underlying disease pathogenesis and developing safe and effective treatments. Patient peripheral blood (PB) cells are used for iPSC generation in many cases since they can be collected with minimum invasiveness. To derive iPSCs that lack immunoreceptor gene rearrangements, hematopoietic stem and progenitor cells (HSPCs) are often targeted as the reprogramming source. However, the current protocols generally require HSPC mobilization and/or ex vivo expansion owing to their sparsity at the steady state and low reprogramming efficiencies, making the overall procedure costly, laborious, and time-consuming. METHODS: We have established a highly efficient method for generating iPSCs from non-mobilized PB-derived CD34+ HSPCs. The source PB mononuclear cells were obtained from 1 healthy donor and 15 patients and were kept frozen until the scheduled iPSC generation. CD34+ HSPC enrichment was done using immunomagnetic beads, with no ex vivo expansion culture. To reprogram the CD34+-rich cells to pluripotency, the Sendai virus vector SeVdp-302L was used to transfer four transcription factors: KLF4, OCT4, SOX2, and c-MYC. In this iPSC generation series, the reprogramming efficiencies, success rates of iPSC line establishment, and progression time were recorded. After generating the iPSC frozen stocks, the cell recovery and their residual transgenes, karyotypes, T cell receptor gene rearrangement, pluripotency markers, and differentiation capability were examined. RESULTS: We succeeded in establishing 223 iPSC lines with high reprogramming efficiencies from 15 patients with 8 different disease types. Our method allowed the rapid appearance of primary colonies (~ 8 days), all of which were expandable under feeder-free conditions, enabling robust establishment steps with less workload. After thawing, the established iPSC lines were verified to be pluripotency marker-positive and of non-T cell origin. A majority of the iPSC lines were confirmed to be transgene-free, with normal karyotypes. Their trilineage differentiation capability was also verified in a defined in vitro assay. CONCLUSION: This robust and highly efficient method enables the rapid and cost-effective establishment of transgene-free iPSC lines from a small volume of PB, thus facilitating the biobanking of patient-derived iPSCs and their use for the modeling of various diseases.

DOI PubMed

The RNA Methyltransferase Complex of WTAP, METTL3, and METTL14 Regulates Mitotic Clonal Expansion in Adipogenesis

Masatoshi Kobayashi, Mitsuru Ohsugi, Takayoshi Sasako, Motoharu Awazawa, Toshihiro Umehara, Aya Iwane, Naoki Kobayashi, Yukiko Okazaki, Naoto Kubota, Ryo Suzuki, Hironori Waki, Keiko Horiuchi, Takao Hamakubo, Tatsuhiko Kodama, Seiichiro Aoe, Kazuyuki Tobe, Takashi Kadowaki, Kohjiro Ueki

Molecular and Cellular Biology ( American Society for Microbiology )  38 ( 16 ) e00116 - 18   2018年06月  [査読有り]

研究論文（学術雑誌）  

<title>ABSTRACT</title>
Adipocyte differentiation is regulated by various mechanisms, of which mitotic clonal expansion (MCE) is a key step. Although this process is known to be regulated by cell cycle modulators, the precise mechanism remains unclear. <italic>N</italic>⁶-Methyladenosine (m⁶A) posttranscriptional RNA modification, whose methylation and demethylation are performed by respective enzyme molecules, has recently been suggested to be involved in the regulation of adipogenesis. Here, we show that an RNA <italic>N</italic>⁶-adenosine methyltransferase complex consisting of Wilms' tumor 1-associating protein (WTAP), methyltransferase like 3 (METTL3), and METTL14 positively controls adipogenesis by promoting cell cycle transition in MCE during adipogenesis. WTAP, coupled with METTL3 and METTL14, is increased and distributed in nucleus by the induction of adipogenesis dependently on RNA <italic>in vitro</italic>. Knockdown of each of these three proteins leads to cell cycle arrest and impaired adipogenesis associated with suppression of cyclin A2 upregulation during MCE, whose knockdown also impairs adipogenesis. Consistent with this, <italic>Wtap</italic> heterozygous knockout mice are protected from diet-induced obesity with smaller size and number of adipocytes, leading to improved insulin sensitivity. These data provide a mechanism for adipogenesis through the WTAP-METTL3-METTL14 complex and a potential strategy for treatment of obesity and associated disorders.

DOI PubMed

NFIA co-localizes with PPARγ and transcriptionally controls the brown fat gene program.

Yuta Hiraike, Hironori Waki, Jing Yu, Masahiro Nakamura, Kana Miyake, Gaku Nagano, Ryo Nakaki, Ken Suzuki, Hirofumi Kobayashi, Shogo Yamamoto, Wei Sun, Tomohisa Aoyama, Yusuke Hirota, Haruya Ohno, Kenji Oki, Masayasu Yoneda, Andrew P White, Yu-Hua Tseng, Aaron M Cypess, Therese J Larsen, Naja Z Jespersen, Camilla Scheele, Shuichi Tsutsumi, Hiroyuki Aburatani, Toshimasa Yamauchi, Takashi Kadowaki

Nature cell biology   19 ( 9 ) 1081 - 1092   2017年09月  [査読有り]

研究論文（学術雑誌）  

Brown fat dissipates energy as heat and protects against obesity. Here, we identified nuclear factor I-A (NFIA) as a transcriptional regulator of brown fat by a genome-wide open chromatin analysis of murine brown and white fat followed by motif analysis of brown-fat-specific open chromatin regions. NFIA and the master transcriptional regulator of adipogenesis, PPARγ, co-localize at the brown-fat-specific enhancers. Moreover, the binding of NFIA precedes and facilitates the binding of PPARγ, leading to increased chromatin accessibility and active transcription. Introduction of NFIA into myoblasts results in brown adipocyte differentiation. Conversely, the brown fat of NFIA-knockout mice displays impaired expression of the brown-fat-specific genes and reciprocal elevation of muscle genes. Finally, expression of NFIA and the brown-fat-specific genes is positively correlated in human brown fat. These results indicate that NFIA activates the cell-type-specific enhancers and facilitates the binding of PPARγ to control the brown fat gene program.

DOI PubMed

CDK5 Regulatory Subunit-Associated Protein 1-like 1 Negatively Regulates Adipocyte Differentiation through Activation of Wnt Signaling Pathway.

Kazumi Take, Hironori Waki, Wei Sun, Takahito Wada, Jing Yu, Masahiro Nakamura, Tomohisa Aoyama, Toshimasa Yamauchi, Takashi Kadowaki

Scientific reports   7 ( 1 ) 7326 - 7326   2017年08月  [査読有り]

研究論文（学術雑誌）  

CDK5 Regulatory Subunit-Associated Protein 1-like 1 (CDKAL1) was identified as a susceptibility gene for type 2 diabetes and body mass index in genome-wide association studies. Although it was reported that CDKAL1 is a methylthiotransferase essential for tRNALys(UUU) and faithful translation of proinsulin generated in pancreatic β cells, the role of CDKAL1 in adipocytes has not been understood well. In this study, we found that CDKAL1 is expressed in adipose tissue and its expression is increased during differentiation. Stable overexpression of CDKAL1, however, inhibited adipocyte differentiation of 3T3-L1 cells, whereas knockdown of CDKAL1 promoted differentiation. CDKAL1 increased protein levels of β-catenin and its active unphosphorylated form in the nucleus, thereby promoting Wnt target gene expression, suggesting that CDKAL1 activated the Wnt/β-catenin pathway-a well-characterized inhibitory regulator of adipocyte differentiation. Mutant experiments show that conserved cysteine residues of Fe-S clusters of CDKAL1 are essential for its anti-adipogenic action. Our results identify CDKAL1 as novel negative regulator of adipocyte differentiation and provide insights into the link between CDKAL1 and metabolic diseases such as type 2 diabetes and obesity.

DOI PubMed

Echinomycin inhibits adipogenesis in 3T3-L1 cells in a HIF-independent manner.

Junna Yamaguchi, Tetsuhiro Tanaka, Hisako Saito, Seitaro Nomura, Hiroyuki Aburatani, Hironori Waki, Takashi Kadowaki, Masaomi Nangaku

Scientific reports   7 ( 1 ) 6516 - 6516   2017年07月  [査読有り]

研究論文（学術雑誌）  

Obesity is a risk factor for many diseases including diabetes, cancer, cardiovascular disease, and chronic kidney disease. Obesity is characterized by the expansion of white adipose tissue (WAT). Hypertrophy and hyperplasia of adipocytes cause tissue hypoxia followed by inflammation and fibrosis. Its trigger, preadipocyte differentiation into mature adipocytes, is finely regulated by transcription factors, signal molecules, and cofactors. We found that echinomycin, a potent HIF-1 inhibitor, completely inhibited adipogenesis in 3T3-L1 WAT preadipocytes by affecting the early phase of mitotic clonal expansion. The dose required to exert the effect was surprisingly low and the time was short. Interestingly, its inhibitory effect was independent of HIF-1 pathways. Time-course DNA microarray analysis of drug-treated and untreated preadipocytes extracted a major transcription factor, CCAAT/enhancer-protein β, as a key target of echinomycin. Echinomycin also inhibited adipogenesis and body weight gain in high fat diet mice. These findings highlight a novel role of echinomycin in suppressing adipocyte differentiation and offer a new therapeutic strategy against obesity and diabetes.

DOI PubMed

Previous dropout from diabetic care as a predictor of patients' willingness to use mobile applications for self-management: A cross-sectional study.

Satoko Yamaguchi, Kayo Waki, Nobuko Tomizawa, Hironori Waki, Yasuhito Nannya, Masaomi Nangaku, Takashi Kadowaki, Kazuhiko Ohe

Journal of diabetes investigation   8 ( 4 ) 542 - 549   2017年07月  [査読有り]

研究論文（学術雑誌）  

AIMS/INTRODUCTION: Preventing dropout is crucial in managing diabetes. Accordingly, we investigated whether patients who had dropped out of diabetic care are suitable candidates for the use of mobile technologies - such as smartphone applications - to support self-management (mHealth), which might help prevent dropout. MATERIALS AND METHODS: We carried out a cross-sectional study in Tokyo, Japan. Patients aged 20 years or older who were clinically diagnosed as diabetic and who regularly visited the outpatient unit at the University of Tokyo Hospital were recruited between August 2014 and March 2015. Data were collected through face-to-face structured interviews, physical measurements and medical records. Participants were asked whether they were willing to use mHealth after being shown DialBetics - an mHealth application for diabetics - as an example, and about their history of dropout and previous mHealth experience. Data were analyzed by multivariate logistic regression models. RESULTS: Of 307 patients with type 1 and type 2 diabetes, 34 (11.1%) had previously dropped out from diabetic care. Multivariate analysis identified previous mHealth experience as a negative predictor of dropout (odds ratio 0.211, P = 0.023). Of those 34 patients, 27 (79.4%) expressed willingness to use mHealth, a significantly higher percentage than for those who had never dropped out (51.5%, P = 0.002). After adjusting for confounders, history of dropout remained a strong predictor of willingness (odds ratio 3.870, P = 0.004). CONCLUSIONS: Patients who previously dropped out of diabetic care are suitable candidates for mHealth. Future studies must evaluate whether mHealth is effective for preventing repeated dropout and improving glycemic control among this population.

DOI PubMed

RNA-binding protein PSPC1 promotes the differentiation-dependent nuclear export of adipocyte RNAs.

Jiexin Wang, Prashant Rajbhandari, Andrey Damianov, Areum Han, Tamer Sallam, Hironori Waki, Claudio J Villanueva, Stephen D Lee, Ronni Nielsen, Susanne Mandrup, Karen Reue, Stephen G Young, Julian Whitelegge, Enrique Saez, Douglas L Black, Peter Tontonoz

The Journal of clinical investigation   127 ( 3 ) 987 - 1004   2017年03月  [査読有り]

研究論文（学術雑誌）  

A highly orchestrated gene expression program establishes the properties that define mature adipocytes, but the contribution of posttranscriptional factors to the adipocyte phenotype is poorly understood. Here we have shown that the RNA-binding protein PSPC1, a component of the paraspeckle complex, promotes adipogenesis in vitro and is important for mature adipocyte function in vivo. Cross-linking and immunoprecipitation followed by RNA sequencing revealed that PSPC1 binds to intronic and 3'-untranslated regions of a number of adipocyte RNAs, including the RNA encoding the transcriptional regulator EBF1. Purification of the paraspeckle complex from adipocytes further showed that PSPC1 associates with the RNA export factor DDX3X in a differentiation-dependent manner. Remarkably, PSPC1 relocates from the nucleus to the cytoplasm during differentiation, coinciding with enhanced export of adipogenic RNAs. Mice lacking PSPC1 in fat displayed reduced lipid storage and adipose tissue mass and were resistant to diet-induced obesity and insulin resistance due to a compensatory increase in energy expenditure. These findings highlight a role for PSPC1-dependent RNA maturation in the posttranscriptional control of adipose development and function.

DOI PubMed

Willingness of patients with diabetes to use an ICT-based self-management tool: a cross-sectional study.

Tomomi Shibuta, Kayo Waki, Nobuko Tomizawa, Ayumi Igarashi, Noriko Yamamoto-Mitani, Satoko Yamaguchi, Hideo Fujita, Shigeko Kimura, Katsuhito Fujiu, Hironori Waki, Yoshihiko Izumida, Takayoshi Sasako, Masatoshi Kobayashi, Ryo Suzuki, Toshimasa Yamauchi, Takashi Kadowaki, Kazuhiko Ohe

BMJ open diabetes research & care   5 ( 1 ) e000322   2017年  [査読有り]

研究論文（学術雑誌）  

OBJECTIVES: To examine the prevalence of the willingness of patients with diabetes to use a self-management tool based on information and communication technology (ICT) such as personal computers, smartphones, and mobile phones; and to examine the patient characteristics associated with that willingness. RESEARCH DESIGN AND METHODS: We conducted a cross-sectional interview survey of 312 adults with diabetes at a university hospital in an urban area in Japan. Participants were classified into 2 groups: those who were willing to use an ICT-based self-management tool and those who were unwilling. Multiple logistic regression analysis was used to identify factors associated with the willingness, including clinical and social factors, current use of ICT, self-management practices, self-efficacy, and diabetes-related emotional distress. RESULTS: The mean age of the 312 participants was 66.3 years (SD=11.5) and 198 (63%) were male. Most of the participants (93%) had type 2 diabetes. Although only 51 (16%) currently used ICT-based self-management tools, a total of 157 (50%) expressed the willingness to use such a tool. Factors associated with the willingness included: not having nephropathy (OR=2.02, 95% CI 1.14 to 3.58); outpatient visits once a month or more (vs less than once a month, OR=2.13, 95% CI 1.13 to 3.99); current use of personal computers and/or smartphones (OR=4.91, 95% CI 2.69 to 8.98); and having greater diabetes-related emotional distress (OR=1.10, 95% CI 1.01 to 1.20). CONCLUSIONS: Approximately half of the patients showed interest in using an ICT-based self-management tool. Willing patients may expect ICT-based self-management tools to complement outpatient visits and to make self-management easier. Starting with patients who display the willingness factors might optimize programs based on such tools.

DOI PubMed

Small Molecule-Induced Complement Factor D (Adipsin) Promotes Lipid Accumulation and Adipocyte Differentiation.

No-Joon Song, Suji Kim, Byung-Hyun Jang, Seo-Hyuk Chang, Ui Jeong Yun, Ki-Moon Park, Hironori Waki, Dean Y Li, Peter Tontonoz, Kye Won Park

PloS one   11 ( 9 ) e0162228   2016年  [査読有り]

研究論文（学術雑誌）  

The epigenome and its role in diabetes.

Hironori Waki, Toshimasa Yamauchi, Takashi Kadowaki

Current diabetes reports   12 ( 6 ) 673 - 85   2012年12月  [査読有り]

研究論文（学術雑誌）  

Global mapping of cell type-specific open chromatin by FAIRE-seq reveals the regulatory role of the NFI family in adipocyte differentiation.

Hironori Waki, Masahiro Nakamura, Toshimasa Yamauchi, Ken-ichi Wakabayashi, Jing Yu, Lisa Hirose-Yotsuya, Kazumi Take, Wei Sun, Masato Iwabu, Miki Okada-Iwabu, Takanori Fujita, Tomohisa Aoyama, Shuichi Tsutsumi, Kohjiro Ueki, Tatsuhiko Kodama, Juro Sakai, Hiroyuki Aburatani, Takashi Kadowaki

PLoS genetics   7 ( 10 ) e1002311   2011年10月  [査読有り]

研究論文（学術雑誌）  

TLE3 is a dual-function transcriptional coregulator of adipogenesis.

Claudio J Villanueva, Hironori Waki, Cristina Godio, Ronni Nielsen, Wen-Ling Chou, Leo Vargas, Kevin Wroblewski, Christian Schmedt, Lily C Chao, Rima Boyadjian, Susanne Mandrup, Andrea Hevener, Enrique Saez, Peter Tontonoz

Cell metabolism   13 ( 4 ) 413 - 427   2011年04月  [査読有り]

研究論文（学術雑誌）  

Adiponectin, adiponectin receptors, and epigenetic regulation of adipogenesis.

Takashi Kadowaki, Toshimasa Yamauchi, Hironori Waki, Masato Iwabu, Miki Okada-Iwabu, Masahiro Nakamura

Cold Spring Harbor symposia on quantitative biology   76   257 - 65   2011年  [査読有り]

研究論文（学術雑誌）  

The small molecule phenamil is a modulator of adipocyte differentiation and PPARgamma expression.

Kye Won Park, Hironori Waki, Sung-Pil Choi, Ki-Moon Park, Peter Tontonoz

Journal of lipid research   51 ( 9 ) 2775 - 84   2010年09月  [査読有り]

研究論文（学術雑誌）  

Adiponectin and AdipoR1 regulate PGC-1alpha and mitochondria by Ca(2+) and AMPK/SIRT1.

Masato Iwabu, Toshimasa Yamauchi, Miki Okada-Iwabu, Koji Sato, Tatsuro Nakagawa, Masaaki Funata, Mamiko Yamaguchi, Shigeyuki Namiki, Ryo Nakayama, Mitsuhisa Tabata, Hitomi Ogata, Naoto Kubota, Iseki Takamoto, Yukiko K Hayashi, Naoko Yamauchi, Hironori Waki, Masashi Fukayama, Ichizo Nishino, Kumpei Tokuyama, Kohjiro Ueki, Yuichi Oike, Satoshi Ishii, Kenzo Hirose, Takao Shimizu, Kazushige Touhara, Takashi Kadowaki

Nature   464 ( 7293 ) 1313 - 9   2010年04月  [査読有り]

研究論文（学術雑誌）  

The small molecule phenamil induces osteoblast differentiation and mineralization.

Kye Won Park, Hironori Waki, Woo-Kyun Kim, Brandon S J Davies, Stephen G Young, Farhad Parhami, Peter Tontonoz

Molecular and cellular biology   29 ( 14 ) 3905 - 14   2009年07月  [査読有り]

研究論文（学術雑誌）  

The expression of GPIHBP1, an endothelial cell binding site for lipoprotein lipase and chylomicrons, is induced by peroxisome proliferator-activated receptor-gamma.

Brandon S J Davies, Hironori Waki, Anne P Beigneux, Emily Farber, Michael M Weinstein, Damien C Wilpitz, Li-Jung Tai, Ronald M Evans, Loren G Fong, Peter Tontonoz, Stephen G Young

Molecular endocrinology   22 ( 11 ) 2496 - 504   2008年11月  [査読有り]

研究論文（学術雑誌）  

Inhibitor of DNA binding 2 is a small molecule-inducible modulator of peroxisome proliferator-activated receptor-gamma expression and adipocyte differentiation.

Kye Won Park, Hironori Waki, Claudio J Villanueva, Laurel A Monticelli, Cynthia Hong, Sona Kang, Ormond A MacDougald, Ananda W Goldrath, Peter Tontonoz

Molecular endocrinology   22 ( 9 ) 2038 - 48   2008年09月  [査読有り]

研究論文（学術雑誌）  

Selective purification and characterization of adiponectin multimer species from human plasma.

Yusuke Hada, Toshimasa Yamauchi, Hironori Waki, Atsushi Tsuchida, Kazuo Hara, Hirokazu Yago, Osamu Miyazaki, Hiroyuki Ebinuma, Takashi Kadowaki

Biochemical and biophysical research communications   356 ( 2 ) 487 - 93   2007年05月  [査読有り]

研究論文（学術雑誌）  

STAMPing out Inflammation.

Hironori Waki, Peter Tontonoz

Cell   129 ( 3 ) 451 - 2   2007年05月

研究論文（学術雑誌）