7 results on '"Yui Hiraguri"'
Search Results
2. Zranb1-mutant mice display abnormal colonic mucus production and exacerbation of DSS-induced colitis
- Author
-
Akiko Tamura, Go Ito, Hiroki Matsuda, Yoichi Nibe-Shirakihara, Yuichi Hiraoka, Sayuki Kitagawa, Yui Hiraguri, Sayaka Nagata, Emi Aonuma, Kana Otsubo, Yasuhiro Nemoto, Takashi Nagaishi, Mamoru Watanabe, Ryuichi Okamoto, and Shigeru Oshima
- Subjects
Inflammation ,Deubiquitinating Enzymes ,Dextran Sulfate ,Mucins ,Biophysics ,Cell Biology ,Colitis ,Biochemistry ,Mice ,Mucus ,Serine ,Animals ,Cysteine ,RNA, Messenger ,Ubiquitin-Specific Proteases ,Intestinal Mucosa ,Molecular Biology - Abstract
Expression of mucin MUC2, a component of the colonic mucus layer, plays a crucial role in intestinal homeostasis. Here, we describe a new regulator of MUC2 expression, the deubiquitinase ZRANB1 (Trabid). A ZRANB1 mutation changing cysteine to serine in amino acid position 443, affects ubiquitination. To analyze ZRANB1 function in the intestine, we generated Zranb1 C443S mutant knock-in (Zranb1
- Published
- 2022
3. Collagen type I-mediated mechanotransduction controls epithelial cell fate conversion during intestinal inflammation
- Author
-
Sakurako Kobayashi, Nobuhiko Ogasawara, Satoshi Watanabe, Yosuke Yoneyama, Sakura Kirino, Yui Hiraguri, Masami Inoue, Sayaka Nagata, Yoshimi Okamoto-Uchida, Satoshi Kofuji, Hiromichi Shimizu, Go Ito, Tomohiro Mizutani, Shinichi Yamauchi, Yusuke Kinugasa, Yoshihito Kano, Yasuhiro Nemoto, Mamoru Watanabe, Kiichiro Tsuchiya, Hiroshi Nishina, Ryuichi Okamoto, and Shiro Yui
- Subjects
Immunology ,Immunology and Allergy - Abstract
Background The emerging concepts of fetal-like reprogramming following tissue injury have been well recognized as an important cue for resolving regenerative mechanisms of intestinal epithelium during inflammation. We previously revealed that the remodeling of mesenchyme with collagen fibril induces YAP/TAZ-dependent fate conversion of intestinal/colonic epithelial cells covering the wound bed towards fetal-like progenitors. To fully elucidate the mechanisms underlying the link between extracellular matrix (ECM) remodeling of mesenchyme and fetal-like reprogramming of epithelial cells, it is critical to understand how collagen type I influence the phenotype of epithelial cells. In this study, we utilize collagen sphere, which is the epithelial organoids cultured in purified collagen type I, to understand the mechanisms of the inflammatory associated reprogramming. Resolving the entire landscape of regulatory networks of the collagen sphere is useful to dissect the reprogrammed signature of the intestinal epithelium. Methods We performed microarray, RNA-seq, and ATAC-seq analyses of the murine collagen sphere in comparison with Matrigel organoid and fetal enterosphere (FEnS). We subsequently cultured human colon epithelium in collagen type I and performed RNA-seq analysis. The enriched genes were validated by gene expression comparison between published gene sets and immunofluorescence in pathological specimens of ulcerative colitis (UC). Results The murine collagen sphere was confirmed to have inflammatory and regenerative signatures from RNA-seq analysis. ATAC-seq analysis confirmed that the YAP/TAZ-TEAD axis plays a central role in the induction of the distinctive signature. Among them, TAZ has implied its relevant role in the process of reprogramming and the ATAC-based motif analysis demonstrated not only Tead proteins, but also Fra1 and Runx2, which are highly enriched in the collagen sphere. Additionally, the human collagen sphere also showed a highly significant enrichment of both inflammatory and fetal-like signatures. Immunofluorescence staining confirmed that the representative genes in the human collagen sphere were highly expressed in the inflammatory region of ulcerative colitis. Conclusions Collagen type I showed a significant influence in the acquisition of the reprogrammed inflammatory signature in both mice and humans. Dissection of the cell fate conversion and its mechanisms shown in this study can enhance our understanding of how the epithelial signature of inflammation is influenced by the ECM niche.
- Published
- 2022
4. Functional analysis of isoflavones using patient-derived human colonic organoids
- Author
-
Minami Hama, Sho Anzai, Junichi Takahashi, Hady Yuki Sugihara, Shigeru Oshima, Tomohiro Mizutani, M. Tsuchiya, Hiromichi Shimizu, Sayaka Nagata, Go Ito, Mamoru Watanabe, Yui Hiraguri, Kiichiro Tsuchiya, Sayaka Takeoka, Ryuichi Okamoto, Shiro Yui, Reiko Kuno, Ami Kawamoto, and Kohei Suzuki
- Subjects
0301 basic medicine ,Biophysics ,Genistein ,Biochemistry ,Inflammatory bowel disease ,Receptor tyrosine kinase ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Intestinal mucosa ,Medicine ,Progenitor cell ,Molecular Biology ,biology ,business.industry ,Daidzein ,Cell Biology ,Isoflavones ,medicine.disease ,030104 developmental biology ,chemistry ,Hepatocyte Growth Factor Receptor ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,business - Abstract
Inflammatory bowel disease (IBD) comprises two major subtypes, ulcerative colitis (UC) and Crohn's disease, which are multifactorial diseases that may develop due to genetic susceptibility, dysbiosis, or environmental factors. Environmental triggers of IBD include food-borne factors, and a previous nationwide survey in Japan identified pre-illness consumption of isoflavones as a risk factor for UC. However, the precise mechanisms involved in the detrimental effects of isoflavones on the intestinal mucosa remain unclear. The present study employed human colonic organoids (hCOs) to investigate the functional effect of two representative isoflavones, genistein and daidzein, on human colonic epithelial cells. The addition of genistein to organoid reformation assays significantly decreased the number and size of reformed hCOs compared with control and daidzein treatment, indicating an inhibitory effect of genistein on colonic cell/progenitor cell function. Evaluation of the phosphorylation status of 49 different receptor tyrosine kinases showed that genistein selectively inhibited phosphorylation of epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor (HGFR). We established a two-dimensional wound-repair model using hCOs and showed that genistein significantly delayed the overall wound-repair response. Our results collectively show that genistein may exert its detrimental effects on the intestinal mucosa via negative regulation of stem/progenitor cell function, possibly leading to sustained mucosal injury and the development of UC.
- Published
- 2021
5. Suspension culture in a rotating bioreactor for efficient generation of human intestinal organoids
- Author
-
Junichi Takahashi, Tomohiro Mizutani, Hady Yuki Sugihara, Sayaka Nagata, Shu Kato, Yui Hiraguri, Sayaka Takeoka, Mao Tsuchiya, Reiko Kuno, Sei Kakinuma, Mamoru Watanabe, and Ryuichi Okamoto
- Subjects
Genetics ,Radiology, Nuclear Medicine and imaging ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Biochemistry ,Computer Science Applications ,Biotechnology - Abstract
Human intestinal organoids (HIOs) derived from human pluripotent stem cells (hPSCs) hold great promise for translational medical applications. A common method to obtain HIOs has been to harvest floating hindgut spheroids arising from hPSCs. As this technique is elegant but burdensome due to the complex protocol and line-to-line variability, a more feasible method is desired. Here, we establish a robust differentiation method into suspension-cultured HIOs (s-HIOs) by seeding dissociated cells on a spheroid-forming plate. This protocol realizes the reliable generation of size-controllable spheroids. Under optimized conditions in a rotating bioreactor, the generated spheroids quickly grow and mature into large s-HIOs with supporting mesenchyme. Upon mesenteric transplantation, s-HIOs further mature and develop complex tissue architecture
- Published
- 2022
6. TGF-β promotes fetal gene expression and cell migration velocity in a wound repair model of untransformed intestinal epithelial cells
- Author
-
Shiro Yui, Hiromichi Shimizu, Ami Kawamoto, Satoshi Watanabe, Mamoru Watanabe, Sayaka Nagata, Hady Yuki Sugihara, Junichi Takahashi, Shigeru Oshima, Sakurako Kobayashi, Yui Hiraguri, Sayaka Takeoka, Sho Anzai, Ryuichi Okamoto, Reiko Kuno, Kohei Suzuki, and Mao Kawai
- Subjects
0301 basic medicine ,Biophysics ,Models, Biological ,digestive system ,Biochemistry ,03 medical and health sciences ,APOA4 ,Fetus ,0302 clinical medicine ,Cell Movement ,Transforming Growth Factor beta ,WNT4 ,Gene expression ,Animals ,Molecular Biology ,Gene ,Wound Healing ,Chemistry ,Gene Expression Regulation, Developmental ,Epithelial Cells ,Cell migration ,Cell Biology ,Intestinal epithelium ,Up-Regulation ,Cell biology ,Intestines ,Mice, Inbred C57BL ,Organoids ,030104 developmental biology ,030220 oncology & carcinogenesis ,Transforming growth factor - Abstract
The early-phase wound repair response of the intestinal epithelium is characterized by rapid and organized cell migration. This response is regulated by several humoral factors, including TGF-β. However, due to a lack of appropriate models, the precise response of untransformed intestinal epithelial cells (IECs) to those factors is unclear. In this study, we established an in vitro wound repair model of untransformed IECs, based on native type-I collagen. In our system, IECs formed a uniform monolayer in a two-chamber culture insert and displayed a stable wound repair response. Gene expression analysis revealed significant induction of Apoa1, Apoa4, and Wnt4 during the collagen-guided wound repair response. The wound repair response was enhanced significantly by the addition of TGF-β. Surprisingly, addition of TGF-β induced a set of genes, including Slc28a2, Tubb2a, and Cpe, that were expressed preferentially in fetal IECs. Moreover, TGF-β significantly increased the peak velocity of migrating IECs and, conversely, reduced the time required to reach the peak velocity, as confirmed by the motion vector prediction (MVP) method. Our current in vitro system could be employed to assess other humoral factors involved in IEC migration and could contribute to a deeper understanding of the wound repair potentials of untransformed IECs.
- Published
- 2020
7. Ubiquitin D is Upregulated by Synergy of Notch Signalling and TNF-α in the Inflamed Intestinal Epithelia of IBD Patients
- Author
-
Ryuichi Okamoto, Reiko Kuno, Kiichiro Tsuchiya, Daichi Nogawa, Kouhei Yamamoto, Sayaka Nagata, Kohei Suzuki, Minami Hama, Hiromichi Shimizu, Junichi Takahashi, Shigeru Oshima, Masanobu Kitagawa, Mao Kawai, Yui Hiraguri, Ami Kawamoto, Tetsuya Nakamura, Shiro Yui, Kazuo Ohtsuka, Mamoru Watanabe, and Sho Anzai
- Subjects
0301 basic medicine ,Transcription, Genetic ,Notch signaling pathway ,Gene Expression ,digestive system ,Cell Line ,03 medical and health sciences ,0302 clinical medicine ,Gastrointestinal Agents ,Ubiquitin ,Downregulation and upregulation ,Gene expression ,Humans ,Medicine ,Intestinal Mucosa ,Ubiquitins ,Ubiquitin D ,Regulation of gene expression ,Receptors, Notch ,biology ,Tumor Necrosis Factor-alpha ,business.industry ,NF-kappa B ,Gastroenterology ,Drug Synergism ,Epithelial Cells ,General Medicine ,Inflammatory Bowel Diseases ,Intestinal epithelium ,Infliximab ,Anti-Bacterial Agents ,Up-Regulation ,Organoids ,030104 developmental biology ,Gene Expression Regulation ,Doxycycline ,biology.protein ,Cancer research ,030211 gastroenterology & hepatology ,Signal transduction ,Transcriptome ,business ,Signal Transduction - Abstract
Background and aims The intestinal epithelium of inflammatory bowel disease [IBD] patients is exposed to various pro-inflammatory cytokines, most notably tumour necrosis factor alpha [TNF-α]. We have previously shown that the Notch signalling pathway is also upregulated in such an epithelium, contributing to intestinal epithelial cell [IEC] proliferation and regeneration. We aimed to reproduce such environment in vitro and explore the gene regulation involved. Methods Human IEC cell lines or patient-derived organoids were used to analyse Notch- and TNF-α-dependent gene expression. Immunohistochemistry was performed to analyse expression of ubiquitin D [UBD] in various patient-derived intestinal tissues. Results In human IEC cell lines, we found that Notch signalling and TNF-α-induced NFκB signalling are reciprocally regulated to promote expression of a specific gene subset. Global gene expression analysis identified UBD to be one of the most highly upregulated genes, due to synergy of Notch and TNF-α. The synergistic expression of UBD was regulated at the transcriptional level, whereas the UBD protein had an extremely short half-life due to post-translational, proteasomal degradation. In uninflamed intestinal tissues from IBD patients, UBD expression was limited to IECs residing at the crypt bottom. In contrast, UBD-expressing IECs were seen throughout the crypt in inflamed tissues, indicating substantial induction by the local inflammatory environment. Analysis using patient-derived organoids consistently confirmed conserved Notch- and TNF-α-dependent expression of UBD. Notably, post-infliximab [IFX] downregulation of UBD reflected favourable outcome in IBD patients. Conclusion We propose that UBD is a novel inflammatory-phase protein expressed in IECs, with a highly rapid responsiveness to anti-TNF-α treatment.
- Published
- 2018
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.