研究者詳細 - 長谷　耕二

論文 - 長谷　耕二

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Protective effect of mesenchymal stromal cells in diabetic nephropathy: the In vitro and In vivo role of the M-Sec-tunneling nanotubes

Barutta F., Corbetta B., Bellini S., Gambino R., Bruno S., Kimura S., Hase K., Ohno H., Gruden G.

Clinical Science 138 （ 23 ） 1537 - 1559 2024年12月

研究論文（学術雑誌）, 査読有り

Mitochondrial dysfunction plays an important role in the development of podocyte injury in diabetic nephropathy (DN). Tunnelling nanotubes (TNTs) are long channels that connect cells and allow organelle exchange. Mesenchymal stromal cells (MSCs) can transfer mitochondria to other cells through the M-Sec-TNTs system. However, it remains unexplored whether MSCs can form heterotypic TNTs with podocytes, thereby enabling the replacement of diabetes-damaged mitochondria. In this study, we analysed TNT formation, mitochondrial transfer, and markers of cell injury in podocytes that were pre-exposed to diabetes-related insults and then co-cultured with diabetic or non-diabetic MSCs. Furthermore, to assess the in vivo relevance, we treated DN mice with exogenous MSCs, either expressing or lacking M-Sec, carrying fluorescent-tagged mitochondria. MSCs formed heterotypic TNTs with podocytes, allowing mitochondrial transfer, via a M-Sec-dependent mechanism. This ameliorated mitochondrial function, nephrin expression, and reduced apoptosis in recipient podocytes. However, MSCs isolated from diabetic mice failed to confer cytoprotection due to Miro-1 down-regulation. In experimental DN, treatment with exogenous MSCs significantly improved DN, but no benefit was observed in mice treated with MSCs lacking M-Sec. Mitochondrial transfer from exogenous MSCs to podocytes occurred in vivo in a M-Sec-dependent manner. These findings demonstrate that the M-Sec-TNT-mediated transfer of mitochondria from healthy MSCs to diabetes-injured podocytes can ameliorate podocyte damage. Moreover, M-Sec expression in exogenous MSCs is essential for providing renoprotection in vivo in experimental DN.

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Intestinal epithelium dysfunctions cause IgA deposition in the kidney glomeruli of intestine-specific Ap1m2-deficient mice

Kinashi Y., Tanaka K., Kimura S., Hirota M., Komiyama S., Shindo T., Hashiguchi A., Takahashi D., Shibata S., Karaki S.I., Ohno H., Hase K.

EBioMedicine 106 2024年08月

研究論文（学術雑誌）, 最終著者, 責任著者, 査読有り

Background: Intestinal epithelial cells (IECs) serve as robust barriers against potentially hostile luminal antigens and commensal microbiota. Epithelial barrier dysfunction enhances intestinal permeability, leading to leaky gut syndrome (LGS) associated with autoimmune and chronic inflammatory disorders. However, a causal relationship between LGS and systemic disorders remains unclear. Ap1m2 encodes clathrin adaptor protein complex 1 subunit mu 2, which facilitates polarized protein trafficking toward the basolateral membrane and contributes to the establishment of epithelial barrier functions. Methods: We generated IEC-specific Ap1m2-deficient (Ap1m2ΔIEC) mice with low intestinal barrier integrity as an LSG model and examined the systemic impact. Findings: Ap1m2ΔIEC mice spontaneously developed IgA nephropathy (IgAN)-like features characterized by the deposition of IgA–IgG immune complexes and complement factors in the kidney glomeruli. Ap1m2 deficiency markedly enhanced aberrantly glycosylated IgA in the serum owing to downregulation and mis-sorting of polymeric immunoglobulin receptors in IECs. Furthermore, Ap1m2 deficiency caused intestinal dysbiosis by attenuating IL-22-STAT3 signaling. Intestinal dysbiosis contributed to the pathogenesis of IgAN because antibiotic treatment reduced aberrantly glycosylated IgA production and renal IgA deposition in Ap1m2ΔIEC mice. Interpretation: IEC barrier dysfunction and subsequent dysbiosis by AP-1B deficiency provoke IgA deposition in the mouse kidney. Our findings provide experimental evidence of a pathological link between LGS and IgAN. Funding: AMED, AMED-CREST, JSPS Grants-in-Aid for Scientific Research, JST CREST, Fuji Foundation for Protein Research, and Keio University Program for the Advancement of Next Generation Research Projects.

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Sugar and arginine facilitate oral tolerance by ensuring the functionality of tolerogenic immune cell subsets in the intestine

Nagai M., Okawa T., Nakata K., Takahashi D., Miyajima R., Shiratori H., Yamanaka D., Nakamura A., Oyama C., Takahashi S.I., Toyama-Sorimachi N., Suzuki K., Ohashi W., Dohi T., Kawamura Y.I., Hase K.

Cell Reports 43 （ 7 ） 2024年07月

研究論文（学術雑誌）, 最終著者, 責任著者, 査読有り

Although oral tolerance is a critical system in regulating allergic disorders, the mechanisms by which dietary factors regulate the induction and maintenance of oral tolerance remain unclear. To address this, we explored the differentiation and function of various immune cells in the intestinal immune system under fasting and ad libitum-fed conditions before oral ovalbumin (OVA) administration. Fasting mitigated OVA-specific Treg expansion, which is essential for oral tolerance induction. This abnormality mainly resulted from functional defects in the CX3CR1+ cells responsible for the uptake of luminal OVA and reduction of tolerogenic CD103+ dendritic cells. Eventually, fasting impaired the preventive effect of oral OVA administration on asthma and allergic rhinitis development. Specific food ingredients, namely carbohydrates and arginine, were indispensable for oral tolerance induction by activating glycolysis and mTOR signaling. Overall, prior food intake and nutritional signals are critical for maintaining immune homeostasis by inducing tolerance to ingested food antigens.

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A purified diet affects intestinal epithelial proliferation and barrier functions through gut microbial alterations

Shiratori H., Hattori K.M., Nakata K., Okawa T., Komiyama S., Kinashi Y., Kabumoto Y., Kaneko Y., Nagai M., Shindo T., Moritoki N., Kawamura Y.I., Dohi T., Takahashi D., Kimura S., Hase K.

International Immunology 36 （ 5 ） 223 - 240 2024年05月

ISSN 09538178

The gut microbiota plays a crucial role in maintaining epithelial barrier function. Although multiple studies have demonstrated the significance of dietary factors on the gut microbiota and mucosal barrier function, the impact of a purified diet, which has long been used in various animal experiments, on intestinal homeostasis remains to be elucidated. Here, we compared the impact of two different types of diets, a crude diet and an AIN-93G-formula purified diet, on epithelial integrity and the gut microbiota. Purified diet-fed mice exhibited shorter villi and crypt lengths and slower epithelial turnover, particularly in the ileum. In addition, antimicrobial products, including REG3γ, were substantially decreased in purified diet-fed mice. Purified diet feeding also suppressed α1,2-fucosylation on the epithelial surface. Furthermore, the purified diet induced metabolic rewiring to fatty acid oxidation and ketogenesis. 16S ribosomal RNA gene sequencing of the ileal contents and mucus layer revealed distinct gut microbiota compositions between the purified and crude diet-fed mice. Purified diet feeding reduced the abundance of segmented filamentous bacteria (SFB), which potently upregulate REG3γand fucosyltransferase 2 (Fut2) by stimulating group 3 innate lymphoid cells (ILC3s) to produce IL-22. These observations illustrate that the intake of a crude diet secures epithelial barrier function by facilitating SFB colonization, whereas a purified diet insufficiently establishes the epithelial barrier, at least partly owing to the loss of SFB. Our data suggest that the influence of purified diets on the epithelial barrier integrity should be considered in experiments using purified diets.

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MZB1-mediated IgA secretion suppresses the development and progression of colorectal cancer triggered by gut inflammation

Tang Y., Feng X., Lu Q., Cui C., Yu M., Wen Z., Luan Y., Dong L., Hu Z., Zhang R., Lu C., Liu J., Shinkura R., Hase K., Wang J.Y.

Mucosal Immunology 17 （ 3 ） 450 - 460 2023年12月

研究論文（学術雑誌）, 査読有り, ISSN 19330219

Colorectal cancer (CRC) ranks among the top causes of mortality globally. Gut inflammation is one crucial risk factor that augments CRC development since patients suffering from inflammatory bowel disease have an increased incidence of CRC. The role of immunoglobulin (Ig)A in maintaining gut homeostasis and preventing inflammation has been well established. Our earlier work demonstrated that the marginal zone and B1 cell-specific protein (MZB1) promotes gut IgA secretion and its absence results in pronounced dextran sulfate sodium salt (DSS)-induced colitis. In the present study, we explored the role of MZB1 in CRC development using the azoxymethane (AOM)/DSS-induced CRC model. We observed an increase in both the number and size of the tumor nodules in Mzb1−/− mice compared with Mzb1+/+ mice. The increase in CRC development and progression in Mzb1−/− mice was associated with reduced intestinal IgA levels, altered gut flora, and more severe gut and systemic inflammation. Oral administration of the monoclonal IgA, W27, alleviated both the gut inflammation and AOM/DSS-induced CRC. Notably, cohousing Mzb1+/+ and Mzb1−/− mice from the 10th day after birth led to similar CRC development. Our findings underscore the pivotal role of MZB1-mediated IgA secretion in suppressing the onset and progression of CRC triggered by gut inflammation. Moreover, our study highlights the profound impact of microbiota composition, modulated by gut IgA levels, on gut inflammation. Nonetheless, establishing a direct correlation between the severity of colitis and subsequent CRC development and the presence or absence of a particular microbiota is challenging.

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α-Glucosidase inhibitors boost gut immunity by inducing IgA responses in Peyer’s patches

Hattori-Muroi K., Naganawa-Asaoka H., Kabumoto Y., Tsukamoto K., Fujisaki Y., Fujimura Y., Komiyama S., Kinashi Y., Kato M., Sato S., Takahashi D., Hase K.

Frontiers in Immunology 14 1277637 2023年11月

研究論文（学術雑誌）, 最終著者, 責任著者, 査読有り

Peyer’s patches (PPs) are specialized gut-associated lymphoid tissues that initiate follicular helper T (Tfh)-mediated immunoglobulin A (IgA) response to luminal antigens derived from commensal symbionts, pathobionts, and dietary sources. IgA-producing B cells migrate from PPs to the small intestinal lamina propria and secrete IgA across the epithelium, modulating the ecological balance of the commensal microbiota and neutralizing pathogenic microorganisms. α-glucosidase inhibitors (α-GIs) are antidiabetic drugs that inhibit carbohydrate digestion in the small intestinal epithelium, leading to alterations in the commensal microbiota composition and metabolic activity. The commensal microbiota and IgA responses exhibit bidirectional interactions that modulate intestinal homeostasis and immunity. However, the effect of α-GIs on the intestinal IgA response remains unclear. We investigated whether α-GIs affect IgA responses by administering voglibose and acarbose to mice via drinking water. We analyzed Tfh cells, germinal center (GC) B cells, and IgA-producing B cells in PPs by flow cytometry. We also assessed pathogen-specific IgA responses. We discovered that voglibose and acarbose induced Tfh cells, GCB cells, and IgA-producing B cells in the PPs of the proximal small intestine in mice. This effect was attributed to the modification of the microbiota rather than a shortage of monosaccharides. Furthermore, voglibose enhanced secretory IgA (S-IgA) production against attenuated Salmonella Typhimurium. Our findings reveal a novel mechanism by which α-GIs augment antigen-specific IgA responses by stimulating Tfh-GCB responses in PPs, and suggest a potential therapeutic application as an adjuvant for augmenting mucosal vaccines.

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Safe and efficient oral allergy immunotherapy using one-pot-prepared mannan-coated allergen nanoparticles

Li S., Toriumi H., Takahashi D., Kamasaki T., Fujioka Y., Nagatoishi S., Li J., Liu Y., Hosokawa T., Tsumoto K., Ohba Y., Katayama Y., Murakami D., Hase K., Mori T.

Biomaterials 303 122381 2023年10月

研究論文（学術雑誌）, 責任著者, 査読有り, ISSN 01429612

Allergen immunotherapy (AIT) is the only curative treatment for allergic diseases. However, AIT has many disadvantages related to efficiency, safety, long-term duration, and patient compliance. Dendritic cells (DCs) have an important role in antigen-specific tolerance induction; thus, DC-targeting strategies to treat allergies such as glutaraldehyde crosslinked antigen to mannoprotein (MAN) have been established. However, glutaraldehyde crosslinking may reduce the antigen presentation efficiency of DCs. To overcome this, we developed a MAN-coated ovalbumin (OVA) nanoparticle (MDO), which uses intermolecular disulfide bond to crosslink OVA and MAN. MDO effectively targeted DCs resulting in tolerogenic DCs, and promoted higher antigen presentation efficiency by DCs compared with OVA or glutaraldehyde crosslinked nanoparticles. In vitro and in vivo experiments showed that DCs exposed to MDO induced Treg cells. Moreover, MDO had low reactivity with anti-OVA antibodies and did not induce anaphylaxis in allergic mice, demonstrating its high safety profile. In a mouse model of allergic asthma, MDO had significant preventative and therapeutic effects when administered orally or subcutaneously. Therefore, MDO represents a promising new approach for the efficient and safe treatment of allergies.

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Gut microbiota-derived lipid metabolites facilitate regulatory T cell differentiation

Shiratori H., Oguchi H., Isobe Y., Han K.H., Sen A., Yakebe K., Takahashi D., Fukushima M., Arita M., Hase K.

Scientific Reports 13 （ 1 ） 8903 2023年06月

研究論文（学術雑誌）, 最終著者, 責任著者, 査読有り

Commensal bacteria-derived metabolites are critical in regulating the host immune system. Although the impact of gut microbiota-derived hydrophilic metabolites, such as short-chain fatty acids, on immune cell functions and development has been well documented, the immunomodulatory effects of gut microbiota-derived lipids are still of interest. Here, we report that lipid extracts from the feces of specific-pathogen-free (SPF), but not germ-free (GF), mice showed regulatory T (Treg)-cell-inducing activity. We conducted RP-HPLC-based fractionation and liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based lipidome profiling and identified two bioactive lipids, 9,10-dihydroxy-12Z-octadecenoic acid (9,10-DiHOME) and all-trans retinoic acid (atRA), with Treg-inducing activity in vitro. The luminal abundance of 9,10-DiHOME in the large intestine was significantly decreased by dextran sulfate sodium (DSS)-induced colitis, indicating that 9,10-DiHOME may be a potential biomarker of colitis. These observations implied that commensal bacteria-derived lipophilic metabolites might contribute to Treg development in the large intestine.

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Dysfunction of Foxp3<sup>+</sup> Regulatory T Cells Induces Dysbiosis of Gut Microbiota via Aberrant Binding of Immunoglobulins to Microbes in the Intestinal Lumen

Koshida K., Ito M., Yakabe K., Takahashi Y., Tai Y., Akasako R., Kimizuka T., Takano S., Sakamoto N., Haniuda K., Ogawa S., Kimura S., Kim Y.G., Hase K., Harada Y.

International Journal of Molecular Sciences 24 （ 10 ） 2023年05月

ISSN 16616596

Foxp3+ regulatory T (Treg) cells prevent excessive immune responses against dietary antigens and commensal bacteria in the intestine. Moreover, Treg cells contribute to the establishment of a symbiotic relationship between the host and gut microbes, partly through immunoglobulin A. However, the mechanism by which Treg cell dysfunction disturbs the balanced intestinal microbiota remains unclear. In this study, we used Foxp3 conditional knockout mice to conditionally ablate the Foxp3 gene in adult mice and examine the relationship between Treg cells and intestinal bacterial communities. Deletion of Foxp3 reduced the relative abundance of Clostridia, suggesting that Treg cells have a role in maintaining Treg-inducing microbes. Additionally, the knockout increased the levels of fecal immunoglobulins and immunoglobulin-coated bacteria. This increase was due to immunoglobulin leakage into the gut lumen as a result of loss of mucosal integrity, which is dependent on the gut microbiota. Our findings suggest that Treg cell dysfunction leads to gut dysbiosis via aberrant antibody binding to the intestinal microbes.

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Dietary iodine attenuates allergic rhinitis by inducing ferroptosis in activated B cells

Nakamura Y., Fuse Y., Komiyama S., Nagatake T., Kunisawa J., Hase K.

Scientific Reports 13 （ 1 ） 5398 2023年04月

研究論文（学術雑誌）, 共著, 最終著者, 責任著者, 査読有り

Iodine-containing formulations have been widely used to treat iodine deficiency and as antiseptics. Lecithin-bound iodine (LBI) has been approved to treat allergic diseases in Japan; however, its underlying mechanism remains unknown. In this study, we show that LBI ameliorated disease symptoms in an ovalbumin (OVA)-induced allergic rhinitis mouse model. LBI suppressed OVA-specific IgE production by attenuating germinal center (GC) reaction in the draining lymph nodes. The antiallergic effect of LBI is most likely attributed to increased serum iodine levels but not thyroid hormone levels. In vitro treatment of activated B cells with potassium iodide induced ferroptosis by increasing intracellular reactive oxygen species (ROS) and ferrous iron in a concentration-dependent manner. Accordingly, LBI diets increased ROS levels in GC B cells of the draining lymph nodes. This study suggests that iodine directly promotes ferroptosis in activated B cells and attenuates GC reactions, leading to the alleviation of allergic symptoms.

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Protective effect of the tunneling nanotube-TNFAIP2/M-sec system on podocyte autophagy in diabetic nephropathy

Barutta F., Bellini S., Kimura S., Hase K., Corbetta B., Corbelli A., Fiordaliso F., Bruno S., Biancone L., Barreca A., Papotti M.G., Hirsh E., Martini M., Gambino R., Durazzo M., Ohno H., Gruden G.

Autophagy 19 （ 2 ） 505 - 524 2023年02月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 15548627

Podocyte injury leading to albuminuria is a characteristic feature of diabetic nephropathy (DN). Hyperglycemia and advanced glycation end products (AGEs) are major determinants of DN. However, the underlying mechanisms of podocyte injury remain poorly understood. The cytosolic protein TNFAIP2/M-Sec is required for tunneling nanotubes (TNTs) formation, which are membrane channels that transiently connect cells, allowing organelle transfer. Podocytes express TNFAIP2 and form TNTs, but the potential relevance of the TNFAIP2-TNT system in DN is unknown. We studied TNFAIP2 expression in both human and experimental DN and the renal effect of tnfaip2 deletion in streptozotocin-induced DN. Moreover, we explored the role of the TNFAIP2-TNT system in podocytes exposed to diabetes-related insults. TNFAIP2 was overexpressed by podocytes in both human and experimental DN and exposre of podocytes to high glucose and AGEs induced the TNFAIP2-TNT system. In diabetic mice, tnfaip2 deletion exacerbated albuminuria, renal function loss, podocyte injury, and mesangial expansion. Moreover, blockade of the autophagic flux due to lysosomal dysfunction was observed in diabetes-injured podocytes both in vitro and in vivo and exacerbated by tnfaip2 deletion. TNTs allowed autophagosome and lysosome exchange between podocytes, thereby ameliorating AGE-induced lysosomal dysfunction and apoptosis. This protective effect was abolished by tnfaip2 deletion, TNT inhibition, and donor cell lysosome damage. By contrast, Tnfaip2 overexpression enhanced TNT-mediated transfer and prevented AGE-induced autophagy and lysosome dysfunction and apoptosis. In conclusion, TNFAIP2 plays an important protective role in podocytes in the context of DN by allowing TNT-mediated autophagosome and lysosome exchange and may represent a novel druggable target. Abbreviations: AGEs: advanced glycation end products; AKT1: AKT serine/threonine kinase 1; AO: acridine orange; ALs: autolysosomes; APs: autophagosomes; BM: bone marrow; BSA: bovine serum albumin; CTSD: cathepsin D; DIC: differential interference contrast; DN: diabetic nephropathy; FSGS: focal segmental glomerulosclerosis; HG: high glucose; KO: knockout; LAMP1: lysosomal-associated membrane protein 1; LMP: lysosomal membrane permeabilization; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; PI3K: phosphoinositide 3-kinase; STZ: streptozotocin; TNF: tumor necrosis factor; TNFAIP2: tumor necrosis factor, alpha-induced protein 2; TNTs: tunneling nanotubes; WT: wild type.

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Medium-chain fatty acids suppress lipotoxicity-induced hepatic fibrosis via the immunomodulating receptor GPR84

Ohue-Kitano R., Nonaka H., Nishida A., Masujima Y., Takahashi D., Ikeda T., Uwamizu A., Tanaka M., Kohjima M., Igarashi M., Katoh H., Tanaka T., Inoue A., Suganami T., Hase K., Ogawa Y., Aoki J., Kimura I.

JCI Insight （JCI Insight） 8 （ 2 ） 2023年01月

Medium-chain triglycerides (MCTs), which consist of medium-chain fatty acids (MCFAs), are unique forms of dietary fat with various health benefits. G protein–coupled 84 (GPR84) acts as a receptor for MCFAs (especially C10:0 and C12:0); however, GPR84 is still considered an orphan receptor, and the nutritional signaling of endogenous and dietary MCFAs via GPR84 remains unclear. Here, we showed that endogenous MCFA-mediated GPR84 signaling protected hepatic functions from diet-induced lipotoxicity. Under high-fat diet (HFD) conditions, GPR84-deficient mice exhibited nonalcoholic steatohepatitis (NASH) and the progression of hepatic fibrosis but not steatosis. With markedly increased hepatic MCFA levels under HFD, GPR84 suppressed lipotoxicity-induced macrophage overactivation. Thus, GPR84 is an immunomodulating receptor that suppresses excessive dietary fat intake–induced toxicity by sensing increases in MCFAs. Additionally, administering MCTs, MCFAs (C10:0 or C12:0, but not C8:0), or GPR84 agonists effectively improved NASH in mouse models. Therefore, exogenous GPR84 stimulation is a potential strategy for treating NASH.

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Dietary-protein sources modulate host susceptibility to Clostridioides difficile infection through the gut microbiota

Yakabe K., Higashi S., Akiyama M., Mori H., Murakami T., Toyoda A., Sugiyama Y., Kishino S., Okano K., Hirayama A., Gotoh A., Li S., Mori T., Katayama T., Ogawa J., Fukuda S., Hase K., Kim Y.G.

Cell Reports （Cell Reports） 40 （ 11 ） 2022年09月

Clostridioides difficile causes nosocomial antibiotic-associated diarrhea on a global scale. Susceptibility to C. difficile infection (CDI) is influenced by the composition and metabolism of gut microbiota, which in turn are affected by diet. However, the mechanism underlying the interplay between diet and gut microbiota that modulates susceptibility to CDI remains unclear. Here, we show that a soy protein diet increases the mortality of antibiotic-treated, C. difficile-infected mice while also enhancing the intestinal levels of amino acids (aas) and relative abundance of Lactobacillus genus. Indeed, Ligilactobacillus murinus-mediated fermentation of soy protein results in the generation of aas, thereby promoting C. difficile growth, and the process involves the anchored cell wall proteinase PrtP. Thus, mutual interaction between dietary protein and the gut microbiota is a critical factor affecting host susceptibility to CDI, suggesting that dietary protein sources can be an important determinant in controlling the disease.

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A Novel HDAC1-Selective Inhibitor Attenuates Autoimmune Arthritis by Inhibiting Inflammatory Cytokine Production

Zhe W., Hoshina N., Itoh Y., Tojo T., Suzuki T., Hase K., Takahashi D.

Biological and Pharmaceutical Bulletin （Biological and Pharmaceutical Bulletin） 45 （ 9 ） 1364 - 1372 2022年09月

ISSN 09186158

Rheumatoid arthritis (RA) is systemic autoimmune arthritis that causes joint inflammation and destruction. Accumulating evidence has shown that inhibitors of class I histone deacetylases (HDACs) (i.e., HDAC1, 2, 3, and 8) are potential therapeutic candidates as targeted synthetic disease-modifying antirheumatic drugs (tsDMARDs). Nevertheless, the inhibition of class I HDACs has severe adverse effects because of their broad spectrum. We evaluated the therapeutic effect of a novel selective HDAC1 inhibitor TTA03-107 for collagen-induced arthritis (CIA) and collagen antibody-induced arthritis (CAIA) models in mice. We also examined the effect of TTA03-107 in bone marrow-derived macrophages (BMDMs) and T helper 17 (Th17) cells in vitro. Here, we delineate that TTA03-107 reduced the severity of autoimmune arthritis without obvious adverse effects in CIA and CAIA models. Moreover, TTA03-107 suppressed the production of inflammatory cytokines, such as interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-17A, in serum and joint tissue. In vitro treatment of BMDMs with TTA03-107 dampened the M1 differentiation and inflammatory cytokine production. TTA03-107 also suppressed the differentiation of Th17 cells. These results demonstrate that TTA03-107 can attenuate the development of arthritis in experimental RA models by inhibiting the differentiation and activation of macrophages and Th17 cells. Therefore, TTA03-107 is a potential tsDMARD candidate.

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<inf>D</inf>-Tryptophan suppresses enteric pathogen and pathobionts and prevents colitis by modulating microbial tryptophan metabolism

Seki N., Kimizuka T., Gondo M., Yamaguchi G., Sugiura Y., Akiyama M., Yakabe K., Uchiyama J., Higashi S., Haneda T., Suematsu M., Hase K., Kim Y.G.

iScience （iScience） 25 （ 8 ） 2022年08月

D-Amino acids (D-AAs) have various functions in mammals and microbes. D-AAs are produced by gut microbiota and can act as potent bactericidal molecules. Thus, D-AAs regulate the ecological niche of the intestine; however, the actual impacts of D-AAs in the gut remain unknown. In this study, we show that D-Tryptophan (D-Trp) inhibits the growth of enteric pathogen and colitogenic pathobionts. The growth of Citrobacter rodentium in vitro is strongly inhibited by D-Trp treatment. Moreover, D-Trp protects mice from lethal C. rodentium infection via reduction of the pathogen. Additionally, D-Trp prevents the development of experimental colitis by the depletion of specific microbes in the intestine. D-Trp increases the intracellular level of indole acrylic acid (IA), a key molecule that determines the susceptibility of enteric microbes to D-Trp. Treatment with IA improves the survival of mice infected with C. rodentium. Hence, D-Trp could act as a gut environmental modulator that regulates intestinal homeostasis.

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Cooperative action of gut-microbiota-accessible carbohydrates improves host metabolic function

Tomioka S., Seki N., Sugiura Y., Akiyama M., Uchiyama J., Yamaguchi G., Yakabe K., Ejima R., Hattori K., Kimizuka T., Fujimura Y., Sato H., Gondo M., Ozaki S., Honme Y., Suematsu M., Kimura I., Inohara N., Núñez G., Hase K., Kim Y.G.

Cell Reports （Cell Reports） 40 （ 3 ） 2022年07月

Microbiota-accessible carbohydrates (MACs) exert health-promoting effects, but how each MAC impacts gut microbiota and regulates host physiology remains unclear. Here, we show that L-arabinose and sucrose cooperatively act on gut microbiota and exert anti-obesogenic effects. Specifically, L-arabinose, a monosaccharide that is poorly absorbed in the gut and inhibits intestinal sucrase, suppresses diet-induced obesity in mice in the presence of sucrose. Additionally, the suppressive effect of L-arabinose on adiposity is abrogated in mice lacking the short-chain fatty acid (SCFA) receptors GPR43 and GPR41. Mechanistically, L-arabinose increases the relative abundance of acetate and propionate producers (e.g., Bacteroides), while sucrose enhances SCFA production. Furthermore, L-arabinose and sucrose activate the glycolytic and pentose phosphate pathways of Bacteroides, respectively, indicating that they synergistically promote acetate production through distinct pathways. These findings suggest that each MAC has a unique property and thus may serve as a precision gut-microbiota modulator to promote host homeostasis.

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Thymic epithelial cells co-opt lineage-defining transcription factors to eliminate autoreactive T cells

Michelson D.A., Hase K., Kaisho T., Benoist C., Mathis D.

Cell （Cell） 185 （ 14 ） 2542 - 2558.e18 2022年07月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 00928674

Medullary thymic epithelial cells (mTECs) ectopically express thousands of peripheral-tissue antigens (PTAs), which drive deletion or phenotypic diversion of self-reactive immature T cells during thymic differentiation. Failure of PTA expression causes multiorgan autoimmunity. By assaying chromatin accessibility in individual mTECs, we uncovered signatures of lineage-defining transcription factors (TFs) for skin, lung, liver, and intestinal cells—including Grhl, FoxA, FoxJ1, Hnf4, Sox8, and SpiB—in distinct mTEC subtypes. Transcriptomic and histologic analyses showed that these subtypes, which we collectively term mimetic cells, expressed PTAs in a biologically logical fashion, mirroring extra-thymic cell types while maintaining mTEC identity. Lineage-defining TFs bound to mimetic-cell open chromatin regions and were required for mimetic cell accumulation, whereas the tolerogenic factor Aire was partially and variably required. Expression of a model antigen in mimetic cells sufficed to induce cognate T cell tolerance. Thus, mTECs co-opt lineage-defining TFs to drive mimetic cell accumulation, PTA expression, and self-tolerance.

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Gut microbiota reinforce host antioxidant capacity via the generation of reactive sulfur species

Uchiyama J., Akiyama M., Hase K., Kumagai Y., Kim Y.G.

Cell Reports （Cell Reports） 38 （ 10 ） 2022年03月

Gut microbiota act beyond the gastrointestinal tract to regulate the physiology of the host. However, their contribution to the antioxidant capacity of the host remains largely understudied. In this study, we observe that gut bacteria increase the steady-state plasma levels of high-antioxidant molecules, reactive sulfur species (RSS), such as hydrogen sulfide and cysteine persulfide (CysSSH), in the host. Moreover, gut bacteria utilize cystine as a substrate to enzymatically produce CysSSH. Administration of cystine to mice increases their plasma levels of RSS and suppresses the concanavalin-A-induced oxidative stress and liver damage in a gut-microbiota-dependent manner. We find that gut bacteria belonging to the Lachnospiraceae and Ruminococcaceae families have a high capacity to produce RSS, requiring pyridoxal 5′-phosphate for their enzymatic reactions. Collectively, our data demonstrate that gut microbiota enhance the antioxidant capacity of the host through the generation of RSS.

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Skate-skin mucin, rich in sulfated sugars and threonine, promotes proliferation of Akkermansia muciniphila in feeding tests in rats and in vitro fermentation using human feces

Miyata T., Mizushima T., Miyamoto N., Yamada T., Hase K., Fukushima M., Nishimura N., Hino S., Morita T.

Bioscience, Biotechnology and Biochemistry （Bioscience, Biotechnology and Biochemistry） 86 （ 3 ） 397 - 406 2022年03月

ISSN 09168451

Dietary factors, affect Akkermansia muciniphila (AM) abundance in the colon, have attracted attention, driven by the inverse correlation between AM abundance and metabolic disorders. We prepared skate-skin mucin (SM), porcine stomach mucin (PM), and rat gastrointestinal mucin (RM). SM contained more sulfated sugars and threonine than PM or RM. Rats were fed a control diet or diets including SM, PM, or RM (15 g/kg), or SM (12 g/kg) from 5 different threonine contents for 14 d. Cecal total bacteria and AM were less and more numerous, respectively, in SM-fed rats than the others, but SM did not affect microbial species richness. Low-Threonine SM did not induce AM proliferation. The in vitro fermentation with human feces showed that the rate of AM increase was greater with SM than PM. Collectively, heavy SM sulfation facilitates a priority supply of SM-derived amino sugars and threonine that promotes AM proliferation in rats and human feces.

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Glia maturation factor-γis involved in S1P-induced marginal zone B-cell chemotaxis and optimal IgM production to type II T-independent antigen

Li Y., Tang Y., Liu J., Meng X., Wang Y., Min Q., Hong R., Tsubata T., Hase K., Wang J.Y.

International Immunology （International Immunology） 34 （ 1 ） 35 - 43 2022年01月

ISSN 09538178

Marginal zone B cells (MZBs) represent a unique B-cell sub-population that rapidly differentiate into IgM-secreting plasma cells in response to T-independent (T-I) antigen. Sphingosine 1-phosphate (S1P) promotes MZB localization to the marginal zone. However, intracellular molecules involved in MZB localization and migration remain largely unknown. Here, we show that MZBs lacking the glia maturation factor-γ(GMFG) are impaired in chemotaxis toward S1P under both in vitro and in vivo conditions, suggesting that GMFG is an effector downstream of S1P receptors. GMFG undergoes serine phosphorylation upon S1P stimulation and is required for S1P-induced desensitization of S1P receptor 1 (S1PR1). Compared with wild-type mice, Gmfg-/- mice produce elevated levels of 4-hydroxy-3-nitrophenyl-acetyl (NP)-specific IgM against a T-I type II antigen, NP-Ficoll, accompanied by dysregulated MZB localization. These results identify GMFG as a regulator of S1P-induced MZB chemotaxis and reveal a role for MZB localization in the marginal zone for optimal IgM production against a T-I antigen.

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The role of gut microbiota in intestinal immune tolerance

Shiratori H., Hase K.

Drug Delivery System （Drug Delivery System） 37 （ 2 ） 159 - 167 2022年

ISSN 09135006

The intestinal mucosa is constantly exposed to foreign substances and thus owns several lines of defense machinery, including epithelial barrier and secretory IgA to prevent the invasion of pathogens. Meanwhile, the intestinal immune system elicits immune tolerance to food antigens and commensal bacteria. Dysregulation of the intestinal immunity often causes severe inflammatory disorders such as inflammatory bowel disease, allergies, and autoimmune diseases. Regulatory T（Treg） cells, one of the major T cell subsets in the intestine, play a critical role in maintaining immune homeostasis. Compelling evidence has demonstrated that commensal bacteria in the intestine are essential for the expansion of Treg cells in the intestinal lamina propria. This effect is at least partly mediated by metabolites like butyrate produced by commensal bacteria. Bacterial metabolites are transported to the bloodstream to affect the immune response in local and systemic tissues, especially in the respiratory system. Thus, the regulation of immune response by gut microbiota and its pathological relevance has drawn much attention. Furthermore, the development of therapies targeting the gut microbiota by fecal transplantation and probiotic administration is underway. This article will review the latest findings on the immune regulation by commensal bacteria and discuss therapeutic intervention strategies targeting the intestinal microbiota.

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Intestinal immunity: To be, or not to be, induced? That is the question

Takahashi D., Kimura S., Hase K.

International Immunology （International Immunology） 33 （ 12 ） 755 - 759 2021年12月

ISSN 09538178

The intestinal immune system maintains intestinal homeostasis in collaboration with diverse immune cell subsets residing at the epithelial layer, lamina propria and gut-associated lymphoid tissue (GALT). Bacterial components and their metabolites are essential for the establishment of the gut immune system. In addition, nutritional signals contribute to maintaining the mucosal immune response. Specialized epithelial microfold (M) cells in GALT facilitate immune surveillance on the mucosal surface by actively taking up external antigens to transport them into the lymphoid follicles. Because hyperplasia of M cells causes an excessive immune response in GALT, there is a self-regulatory mechanism to control the development of M cells appropriately. In this review, we will discuss the molecular mechanisms of mucosal immune regulation and their biological importance.

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Profiling of tumour-associated microbiota in human hepatocellular carcinoma

Komiyama S., Yamada T., Takemura N., Kokudo N., Hase K., Kawamura Y.I.

Scientific Reports （Scientific Reports） 11 （ 1 ） 2021年12月

Liver cancer is the fourth leading cause of cancer-related death. Hepatocellular carcinoma (HCC) is a primary liver cancer that results from chronic hepatitis caused by multiple predisposing factors such as viral infection, alcohol consumption, and non-alcoholic fatty liver disease. Accumulating studies have indicated that dysfunction of the gut epithelial barrier and hepatic translocation of gut microbes may be implicated in the pathogenesis of HCC. However, the translocated bacteria in HCC patients remains unclear. Here, we characterised tumour-associated microbiota in patients with liver cancer and focused on HCC. We observed that the number of amplicon sequence variants in tumour-associated microbiota was significantly higher compared with that in non-tumour regions of the liver. The tumour-associated microbiota consisted of Bacteroidetes, Firmicutes, and Proteobacteria as the dominant phyla. We identified an unclassified genus that belonged to the Bacteroides, Romboutsia, uncultured bacterium of Lachnospiraceae as a signature taxon for primary liver cancer. Additionally, we identified Ruminococcus gnavus as a signature taxon for HCC patients infected with hepatitis B and/or hepatitis C viruses. This study suggests that tumour microbiota may contribute to the pathology of HCC.

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Safety and tolerability of medicinal parasite ova (Trichuris suis) in healthy Japanese volunteers: A randomized, double-blind, placebo-controlled trial

Hoshina T., Sakurai T., Ichimura H., Ishiwata K., En S., Yamada T., Kwangyole L., Shimizu A., Hase K., Kanuka H.

Parasitology International （Parasitology International） 85 2021年12月

ISSN 13835769

Background: Trichuris suis ova (TSO), with the potential to modulate the human immune system, have been tested for therapeutic application in autoimmune and allergic diseases such as inflammatory bowel disease (IBD). Previous clinical studies were limited to European and American participants, whereas Asian populations have not been well documented. In this study, a clinical trial was conducted to examine the safety and tolerability of TSO administration among a healthy Japanese population. Methods: The study was a randomized, double-blind, placebo-controlled trial held at Jikei University Hospital, Tokyo. Twelve volunteers were stratified into three groups receiving different doses of TSO (TSO 1000, 2500, and 7500) and another into the control group. These cases were limited to healthy Japanese men aged over 20 years old. Single doses of medicinal TSO or placebo were given to three participants of each group. All participants were followed up to 56 days after ingestion. During the follow-up period, clinical practitioners checked each participant at the clinic at 7, 14, 28, and 56 days post-ingestion (dpi). Clinical symptoms were evaluated using questionnaire-based self-reporting, which participants filled at every visit. Blood samples were drawn at 7, 14, 28, and 56 dpi. Fecal samples were collected at 28 and 56 dpi. Results: During the study period, twelve healthy Japanese male volunteers were enrolled. All participants completed the follow-up period. No severe adverse events were observed during the study period in all groups. Three participants in the TSO 1000, 2500, and 7500 groups had mild to moderate abdominal symptoms, diarrhea, bloating, and appetite loss during the observation period. One participant in the placebo group presented with mild diarrhea. Microscopic examination identified no parasite ova in any fecal samples. Blood sample examination indicated elevated eosinophil count in several cases, especially in the groups with the higher dose of TSO. No extra-abdominal symptoms were present in all cases. Conclusions: Healthy Japanese people tolerated all doses of TSO without any severe adverse events. On the other hand, mild to moderate abdominal symptoms were observed in several participants. This study suggested that the medicinal use of TSO in Japan is relatively safe, and close follow-up is recommended for sustainable usage.

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Characterization of M Cells in Tear Duct-Associated Lymphoid Tissue of Mice: A Potential Role in Immunosurveillance on the Ocular Surface

Oya Y., Kimura S., Nakamura Y., Ishihara N., Takano S., Morita R., Endo M., Hase K.

Frontiers in Immunology （Frontiers in Immunology） 12 779709 2021年11月

研究論文（学術雑誌）, 査読有り

The ocular mucosal tissues are exposed to potentially harmful foreign antigens in the air and tear fluid. The tear duct-associated lymphoid tissue (TALT) may contribute to immune surveillance in the eye region. Follicle-associated epithelium (FAE) of TALTs is classified as stratified squamous epithelium and consists of squamous epithelial cells arranged in layers on the basement membrane. In contrast, most mucosa-associated lymphoid tissue is covered by a monolayer of epithelium containing microfold (M) cells. Therefore, antigen uptake and the presence of M cells in TALT are not fully understood. The present study found that a small population of FAE cells in the TALT expressed intestinal M-cell markers, namely Sox8, Tnfaip2, GP2, and OPG. This cell population was identified as functional M cells because of their uptake capacity of luminal nanoparticles. In addition, RANKL, which is essential for M-cell differentiation, was expressed by stroma-like cells at the subepithelial region and its receptor RANK by the FAE in the TALT. The administration of RANKL markedly increased the number of Sox8+ M cells. In contrast, deficiency in OPG, an endogenous inhibitor of RANKL, increased the number of M cells in the TALT. These data demonstrate that the RANKL-RANK axis is essential for M-cell differentiation in the TALT. Furthermore, immunization via eye drops elicited the production of antigen-specific antibodies in tears, which was enhanced by RANKL administration. Thus, TALT M cells play an important role in the immunosurveillance of the eye region.

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Identification of Novel Histone Deacetylase 6-Selective Inhibitors Bearing 3,3,3-Trifluorolactic Amide (TFLAM) Motif as a Zinc Binding Group

Kurohara T., Tanaka K., Takahashi D., Ueda S., Yamashita Y., Takada Y., Takeshima H., Yu S., Itoh Y., Hase K., Suzuki T.

ChemBioChem （ChemBioChem） 22 （ 22 ） 3158 - 3163 2021年11月

ISSN 14394227

Pharmacological inhibition of histone deacetylase 6 (HDAC6) is an effective therapeutic strategy for cancer and immunological diseases. Most of the previously reported HDAC6 inhibitors have a hydroxamate group as a zinc binding group (ZBG), which coordinates to the catalytic zinc ion of HDAC6. The hydroxamate group is liable to metabolically generate mutagenetic hydroxylamine; therefore, non-hydroxamate HDAC6 inhibitors would be advantageous. In this study, to identify novel non-hydroxamate HDAC6-selective inhibitors, screening of a chemical library and the subsequent structural optimization were performed, which led to the identification of HDAC6-selective inhibitors with 3,3,3-trifluorolactic amide (TFLAM) as a novel ZBG. The identified inhibitor showed potent and selective HDAC6-inhibitory activity in cells and induced regulatory T (Treg) cell differentiation.

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Specific adsorption of a β-lactam antibiotic: In vivo by an anion-exchange resin for protection of the intestinal microbiota

Li S., Yakabe K., Zai K., Liu Y., Kishimura A., Hase K., Kim Y.G., Mori T., Katayama Y.

Biomaterials Science （Biomaterials Science） 9 （ 21 ） 7219 - 7227 2021年11月

ISSN 20474830

The fraction of antibiotics that are excreted from the intestine during administration leads to disruption of commensal bacteria as well as resulting in dysbiosis and various diseases. To protect the gut microbiota during treatment with antibiotics, use of activated carbon (AC) has recently been reported as a method to adsorb antibiotics. However, the antibiotic adsorption by AC is nonspecific and may also result in the adsorption of essential biological molecules. In this work, we reported that an anion exchange resin (AER) has better specificity than AC for adsorbing the β-lactam antibiotic cefoperazone (CEF). Because CEF has a negatively charged carboxylate group and a conjugated system, the AER was used to adsorb CEF through electrostatic and π-π interactions. The AER was specific for CEF over biological molecules such as bile acids and vitamins in the intestine. The AER protected Escherichia coli from CEF in vitro. Furthermore, oral administration of the AER reduced the fecal free CEF concentration, and protected the gut microbiota from CEF-induced dysbiosis. This journal is

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Commensal microbe-derived acetate suppresses NAFLD/NASH development via hepatic FFAR2 signalling in mice

Aoki R., Onuki M., Hattori K., Ito M., Yamada T., Kamikado K., Kim Y.G., Nakamoto N., Kimura I., Clarke J.M., Kanai T., Hase K.

Microbiome （Microbiome） 9 （ 1 ） 2021年09月

研究論文（学術雑誌）, 共著

Background: Non-alcoholic liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome, and it can progress to non-alcoholic steatohepatitis (NASH). Alterations in the gut microbiome have been implicated in the development of NAFLD/NASH, although the underlying mechanisms remain unclear. Results: We found that the consumption of the prebiotic inulin markedly ameliorated the phenotype of NAFLD/NASH, including hepatic steatosis and fibrosis, in mice. Inulin consumption resulted in global changes in the gut microbiome, including concomitant enrichment of the genera Bacteroides and Blautia, and increased concentrations of short-chain fatty acids, particularly acetate, in the gut lumen and portal blood. The consumption of acetate-releasing resistant starch protected against NAFLD development. Colonisation by Bacteroides acidifaciens and Blautia producta in germ-free mice resulted in synergetic effects on acetate production from inulin. Furthermore, the absence of free fatty acid receptor 2 (FFAR2), an acetate receptor, abolished the protective effect of inulin, as indicated by the more severe liver hypertrophy, hypercholesterolaemia and inflammation. These effects can be attributed to an exacerbation of insulin resistance in the liver, but not in muscle or adipose tissue. Conclusion: These findings demonstrated that the commensal microbiome–acetate–FFAR2 molecular circuit improves insulin sensitivity in the liver and prevents the development of NAFLD/NASH. [MediaObject not available: see fulltext.]

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A Retinoid X Receptor Agonist Directed to the Large Intestine Ameliorates T-Cell-Mediated Colitis in Mice

Matsumoto R., Takahashi D., Watanabe M., Nakatani S., Takamura Y., Kurosaki Y., Kakuta H., Hase K.

Frontiers in Pharmacology （Frontiers in Pharmacology） 12 2021年08月

Retinoid X receptor (RXR) is a nuclear receptor that heterodimerizes with several nuclear receptors, integrating ligand-mediated signals across the heterodimers. Synthetic RXR agonists have been developed to cure certain inflammatory diseases, including inflammatory bowel diseases (IBDs). However, pre-existing RXR agonists, which are lipophilic and readily absorbed in the upper intestine, cause considerable adverse effects such as hepatomegaly, hyperlipidemia, and hypothyroidism. To minimize these adverse effects, we have developed an RXR agonist, NEt-3IB, which has lipophilic and thus poorly absorptive properties. In this study, we evaluated the effects of NEt-3IB in an experimental murine colitis model induced through the adoptive transfer of CD45RBhighCD4+ T cells. Pharmacokinetic studies demonstrated that the major portion of NEt-3IB was successfully delivered to the large intestine after oral administration. Notably, NEt-3IB treatment suppressed the development of T cell-mediated chronic colitis, as indicated by improvement of wasting symptoms, inflammatory infiltration, and mucosal hyperplasia. The protective effect of NEt-3IB was mediated by the suppression of IFN-γ-producing Th1 cell expansion in the colon. In conclusion, NEt-3IB, a large intestine-directed RXR agonist, is a promising drug candidate for IBDs.

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Seaweed dietary fiber sodium alginate suppresses the migration of colonic inflammatory monocytes and diet-induced metabolic syndrome via the gut microbiota

Ejima R., Akiyama M., Sato H., Tomioka S., Yakabe K., Kimizuka T., Seki N., Fujimura Y., Hirayama A., Fukuda S., Hase K., Kim Y.G.

Nutrients （Nutrients） 13 （ 8 ） 2021年08月

Metabolic syndrome (MetS) is a multifactorial chronic metabolic disorder that affects ap-proximately one billion people worldwide. Recent studies have evaluated whether targeting the gut microbiota can prevent MetS. This study aimed to assess the ability of dietary fiber to control MetS by modulating gut microbiota composition. Sodium alginate (SA) is a seaweed-derived dietary fiber that suppresses high-fat diet (HFD)-induced MetS via an effect on the gut microbiota. We observed that SA supplementation significantly decreased body weight gain, cholesterol levels, and fat weight, while improving glucose tolerance in HFD-fed mice. SA changed the gut microbiota composition and significantly increased the abundance of Bacteroides. Antibiotic treatment completely abolished the suppressive effects of SA on MetS. Mechanistically, SA decreased the number of co-lonic inflammatory monocytes, which promote MetS development, in a gut microbiota-dependent manner. The abundance of Bacteroides was negatively correlated with that of inflammatory mono-cytes and positively correlated with the levels of several gut metabolites. The present study revealed a novel food function of SA in preventing HFD-induced MetS through its action on gut microbiota.

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Gut microbiota prevents sugar alcohol-induced diarrhea

Hattori K., Akiyama M., Seki N., Yakabe K., Hase K., Kim Y.G.

Nutrients （Nutrients） 13 （ 6 ） 2021年06月

While poorly-absorbed sugar alcohols such as sorbitol are widely used as sweeteners, they may induce diarrhea in some individuals. However, the factors which determine an individual’s susceptibility to sugar alcohol-induced diarrhea remain unknown. Here, we show that specific gut bacteria are involved in the suppression of sorbitol-induced diarrhea. Based on 16S rDNA analysis, the abundance of Enterobacteriaceae bacteria increased in response to sorbitol consumption. We found that Escherichia coli of the family Enterobacteriaceae degraded sorbitol and suppressed sorbitol-induced diarrhea. Finally, we showed that the metabolism of sorbitol by the E. coli sugar phosphotransferase system helped suppress sorbitol-induced diarrhea. Therefore, gut microbiota prevented sugar alcohol-induced diarrhea by degrading sorbitol in the gut. The identification of the gut bacteria which respond to and degrade sugar alcohols in the intestine has implications for microbiome science, processed food science, and public health.

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Amino acid-based diet prevents lethal infectious diarrhea by maintaining body water balance in a murine citrobacter rodentium infection model

Kimizuka T., Seki N., Yamaguchi G., Akiyama M., Higashi S., Hase K., Kim Y.G.

Nutrients （Nutrients） 13 （ 6 ） 2021年06月

Infectious diarrhea is one of the most important health problems worldwide. Although nutritional status influences the clinical manifestation of various enteric pathogen infections, the effect of diet on enteric infectious diseases remains unclear. Using a fatal infectious diarrheal model, we found that an amino acid-based diet (AD) protected susceptible mice infected with the enteric pathogen Citrobacter rodentium. While the mice fed other diets, including a regular diet, were highly susceptible to C. rodentium infection, AD-fed mice had an increased survival rate. An AD did not suppress C. rodentium colonization or intestinal damage; instead, it prevented diarrhea-induced dehydration by increasing water intake. An AD altered the plasma and fecal amino acid levels and changed the gut microbiota composition. Treatment with glutamate, whose level was increased in the plasma and feces of AD-fed mice, promoted water intake and improved the survival of C. ro-dentium-infected mice. Thus, an AD changes the systemic amino acid balance and protects against lethal infectious diarrhea by maintaining total body water content.

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Protective role of the M-sec-tunneling nanotube system in podocytes

Barutta F., Kimura S., Hase K., Bellini S., Corbetta B., Corbelli A., Fiordaliso F., Barreca A., Papotti M.G., Ghiggeri G.M., Salvidio G., Roccatello D., Audrito V., Deaglio S., Gambino R., Bruno S., Camussi G., Martini M., Hirsch E., Durazzo M., Ohno H., Gruden G.

Journal of the American Society of Nephrology （Journal of the American Society of Nephrology） 32 （ 5 ） 1114 - 1130 2021年05月

ISSN 10466673

Background Podocyte dysfunction and loss are major determinants in the development of proteinuria. FSGS is one of the most common causes of proteinuria, but the mechanisms leading to podocyte injury or conferring protection against FSGS remain poorly understood. The cytosolic protein M-Sec has been involved in the formation of tunneling nanotubes (TNTs), membrane channels that transiently connect cells and allow intercellular organelle transfer. Whether podocytes express M-Sec is unknown and the potential relevance of the M-Sec-TNT system in FSGS has not been explored. Methods We studied the role of the M-Sec-TNT system in cultured podocytes exposed to Adriamycin and in BALB/c M-Sec knockout mice. We also assessed M-Sec expression in both kidney biopsies from patients with FSGS and in experimental FSGS (Adriamycin-induced nephropathy). Results Podocytes can form TNTs in a M-Sec-dependent manner. Consistent with the notion that the M-Sec-TNT system is cytoprotective, podocytes overexpressed M-Sec in both human and experimental FSGS. Moreover, M-Sec deletion resulted in podocyte injury, with mitochondrial abnormalities and development of progressive FSGS. In vitro, M-Sec deletion abolished TNT-mediated mitochondria transfer between podocytes and altered mitochondrial bioenergetics. Re-expression of M-Sec reestablishes TNT formation and mitochondria exchange, rescued mitochondrial function, and partially reverted podocyte injury. Conclusions These findings indicate that the M-Sec-TNT system plays an important protective role in the glomeruli by rescuing podocytes via mitochondrial horizontal transfer. M-Sec may represent a promising therapeutic target in FSGS, and evidence that podocytes can be rescued via TNT-mediated horizontal transfer may open new avenues of research.

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Partners in Leaky Gut Syndrome: Intestinal Dysbiosis and Autoimmunity

Kinashi Y., Hase K.

Frontiers in Immunology （Frontiers in Immunology） 12 2021年04月

The intestinal surface is constitutively exposed to diverse antigens, such as food antigens, food-borne pathogens, and commensal microbes. Intestinal epithelial cells have developed unique barrier functions that prevent the translocation of potentially hostile antigens into the body. Disruption of the epithelial barrier increases intestinal permeability, resulting in leaky gut syndrome (LGS). Clinical reports have suggested that LGS contributes to autoimmune diseases such as type 1 diabetes, multiple sclerosis, rheumatoid arthritis, and celiac disease. Furthermore, the gut commensal microbiota plays a critical role in regulating host immunity; abnormalities of the microbial community, known as dysbiosis, are observed in patients with autoimmune diseases. However, the pathological links among intestinal dysbiosis, LGS, and autoimmune diseases have not been fully elucidated. This review discusses the current understanding of how commensal microbiota contributes to the pathogenesis of autoimmune diseases by modifying the epithelial barrier.

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Symbiotic polyamine metabolism regulates epithelial proliferation and macrophage differentiation in the colon

Nakamura A., Kurihara S., Takahashi D., Ohashi W., Nakamura Y., Kimura S., Onuki M., Kume A., Sasazawa Y., Furusawa Y., Obata Y., Fukuda S., Saiki S., Matsumoto M., Hase K.

Nature Communications （Nature Communications） 12 （ 1 ） 2105 2021年04月

研究論文（学術雑誌）, 共著, 査読有り

Intestinal microbiota-derived metabolites have biological importance for the host. Polyamines, such as putrescine and spermidine, are produced by the intestinal microbiota and regulate multiple biological processes. Increased colonic luminal polyamines promote longevity in mice. However, no direct evidence has shown that microbial polyamines are incorporated into host cells to regulate cellular responses. Here, we show that microbial polyamines reinforce colonic epithelial proliferation and regulate macrophage differentiation. Colonisation by wild-type, but not polyamine biosynthesis-deficient, Escherichia coli in germ-free mice raises intracellular polyamine levels in colonocytes, accelerating epithelial renewal. Commensal bacterium-derived putrescine increases the abundance of anti-inflammatory macrophages in the colon. The bacterial polyamines ameliorate symptoms of dextran sulfate sodium-induced colitis in mice. These effects mainly result from enhanced hypusination of eukaryotic initiation translation factor. We conclude that bacterial putrescine functions as a substrate for symbiotic metabolism and is further absorbed and metabolised by the host, thus helping maintain mucosal homoeostasis in the intestine.

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Pancreatic glycoprotein 2 is a first line of defense for mucosal protection in intestinal inflammation

Kurashima Y., Kigoshi T., Murasaki S., Arai F., Shimada K., Seki N., Kim Y.G., Hase K., Ohno H., Kawano K., Ashida H., Suzuki T., Morimoto M., Saito Y., Sasou A., Goda Y., Yuki Y., Inagaki Y., Iijima H., Suda W., Hattori M., Kiyono H.

Nature Communications （Nature Communications） 12 （ 1 ） 2021年02月

研究論文（学術雑誌）, 査読有り

Increases in adhesive and invasive commensal bacteria, such as Escherichia coli, and subsequent disruption of the epithelial barrier is implicated in the pathogenesis of inflammatory bowel disease (IBD). However, the protective systems against such barrier disruption are not fully understood. Here, we show that secretion of luminal glycoprotein 2 (GP2) from pancreatic acinar cells is induced in a TNF–dependent manner in mice with chemically induced colitis. Fecal GP2 concentration is also increased in Crohn’s diease patients. Furthermore, pancreas-specific GP2-deficient colitis mice have more severe intestinal inflammation and a larger mucosal E. coli population than do intact mice, indicating that digestive-tract GP2 binds commensal E. coli, preventing epithelial attachment and penetration. Thus, the pancreas–intestinal barrier axis and pancreatic GP2 are important as a first line of defense against adhesive and invasive commensal bacteria during intestinal inflammation.

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Polyvinyl Butyrate Nanoparticles as Butyrate Donors for Colitis Treatment

Mu Y., Kinashi Y., Li J., Yoshikawa T., Kishimura A., Tanaka M., Matsui T., Mori T., Hase K., Katayama Y.

ACS Applied Bio Materials （ACS Applied Bio Materials） 4 （ 3 ） 2335 - 2341 2021年02月

研究論文（学術雑誌）

Butyrate has been attracting attention for the suppression of inflammatory bowel disease (IBD). However, clinical trials of butyrate for IBD treatment have resulted in controversial outcomes, likely owing to the adverse effect of butyrate on the intestinal epithelium that was observed at high butyrate concentrations. Herein, we propose polyvinyl butyrate (PVBu) nanoparticles (NPs) as butyrate donors for delivery to the lower part of the intestine for the treatment of colitis. The PVBu NPs suppressed the inflammatory activation of macrophages in vitro, although sodium butyrate inversely further activated macrophages. Oral administration of NPs did not change the luminal concentration of free butyrate; however, NPs showed a therapeutic effect on a colitis mouse model. In addition, incorporation of vitamin D3 into the NPs enhanced the therapeutic effect on colitis. Hence, PVBu NPs are a promising therapeutic for IBD treatment, not only as a butyrate donor but also as a carrier for hydrophobic drugs like vitamin D3.

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Dietary Intervention Impacts Immune Cell Functions and Dynamics by Inducing Metabolic Rewiring

Okawa T., Nagai M., Hase K.

Frontiers in Immunology （Frontiers in Immunology） 11 2021年02月

Accumulating evidence has shown that nutrient metabolism is closely associated with the differentiation and functions of various immune cells. Cellular metabolism, including aerobic glycolysis, fatty acid oxidation, and oxidative phosphorylation, plays a key role in germinal center (GC) reaction, B-cell trafficking, and T-cell-fate decision. Furthermore, a quiescent metabolic status consolidates T-cell-dependent immunological memory. Therefore, dietary interventions such as calorie restriction, time-restricted feeding, and fasting potentially manipulate immune cell functions. For instance, intermittent fasting prevents the development of experimental autoimmune encephalomyelitis. Meanwhile, the fasting response diminishes the lymphocyte pool in gut-associated lymphoid tissue to minimize energy expenditure, leading to the attenuation of Immunoglobulin A (IgA) response. The nutritional status also influences the dynamics of several immune cell subsets. Here, we describe the current understanding of the significance of immunometabolism in the differentiation and functionality of lymphocytes and macrophages. The underlying molecular mechanisms also are discussed. These experimental observations could offer new therapeutic strategies for immunological disorders like autoimmunity.

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Polyamines polarized Th2/Th9 cell-fate decision by regulating GATA3 expression

Nakamura A., Takahashi D., Nakamura Y., Yamada T., Matsumoto M., Hase K.

Archives of Biochemistry and Biophysics （Archives of Biochemistry and Biophysics） 693 2020年10月

ISSN 00039861

© 2020 Elsevier Inc. Polyamines produced by both prokaryotes and eukaryotes are bioactive substances with pleiotropic effects. Accumulating evidence has demonstrated that polyamines contribute to anti-inflammatory responses by suppressing the expression of proinflammatory cytokines in mononuclear cells and macrophages. However, the effects of polyamines on CD4+ T cell responses remain to be elucidated. Here, we investigated the effect of polyamines on cell fate decisions of naïve CD4+ T cells in vitro. We found that endogenously generated polyamines are essential for the development of T helper 2 (Th2) cells. Treatment with DL-2-difluoromethylornithine (DFMO), an inhibitor of polyamine biosynthesis, diminished GATA3 expression in CD4+ T cells under Th2-skewed conditions. Supplementation of exogenous polyamines rescued GATA3 downregulation caused by DFMO treatment in CD4+ T cells. Transcriptome analysis revealed that deprivation of endogenous polyamines resulted in upregulated Th9-related genes, such as Il9, Irf4, and Batf3, even under the Th2-skewing conditions. Depletion of intracellular polyamines reduced GATA3 expression but increased IL-9-producing CD4+ T cells under both Th2 and Th9-skewing conditions. Furthermore, oral administration of DFMO increased IL-9-producing CD4+ T cells in small intestine in mice. Thus, our data indicate that polyamines play a critical role in the regulation of the Th2/Th9 balance.

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Mucin-Derived O-Glycans act as endogenous fiber and sustain mucosal immune homeostasis via short-Chain fatty acid production in rat cecum

Hino S., Mizushima T., Kaneko K., Kawai E., Kondo T., Genda T., Yamada T., Hase K., Nishimura N., Morita T.

Journal of Nutrition （Journal of Nutrition） 150 （ 10 ） 2656 - 2665 2020年10月

ISSN 00223166

Copyright © The Author(s) 2020. Background: Intestinal mucins escape digestion and enter the large bowel where they are degraded by the microbiota. To what extent and how mucins impact large-bowel physiology remain unclear. Objective: This study examined the large-bowel fermentation characteristics of mucins and mucin-derived O-glycan sugars and whether they affect gut immunity. Methods: Mucin secretion from the terminal ileum was determined from feces of ileorectostomized male Wistar rats (age 6 wk) fed an AIN76-based control diet (CD) for 15 d (experiment 1). Normal male Wistar rats (age 6 wk; 4 wk for experiment 4) were fed CD ± porcine stomach mucin (PM) at 6 or 12 g/kg diet, equivalent to 1.5 and 3 times the daily mucin secretion, for 14 d (experiment 2); CD ± N-acetylglucosamine (GlcNAc), fucose, or N-acetylneuraminic acid at 10 g/kg diet for 14 d (experiment 3); or CD ± PM (15 g/kg diet) or GlcNAc (10 g/kg diet) for 29 d (experiment 4). SCFAs, microbial composition, and cecal O-glycan content were assessed. IgA+ plasma cells and regulatory T cells and inflammatory cytokine expression in the cecum were evaluated (experiment 4). Results: Daily mucin secretion corresponded to 43.2 μmol of O-glycans. Cecal O-glycan contents were comparable between CD- and PM-fed rats. PM-fed rats harbored more mucin-degrading bacteria. Cecal concentrations of acetate (+37%) and n-butyrate (+73%) were higher in 12-g/kg PM diet–fed rats versus CD (P < 0.05). Among O-glycan sugars, only GlcNAc produced higher n-butyrate concentrations (+68%) versus CD (P < 0.05), with increased numbers of butyrate-producing bacteria. GlcNAc increased the abundance of IgA+ plasma cells (+29%) and regulatory T cells (+33%) versus CD, whereas PM increased IgA+ plasma cells (+25%) (all P < 0.05). GlcNAc and PM decreased expression of Tnfa (−30%, −40%) and Ifng (−30%, −70%) versus CD (all P < 0.05). Conclusions: Mucin-derived O-glycans act as endogenous fiber and maintain mucosal immune homeostasis via large-bowel SCFA production in rats. J Nutr 2020;150:2656–2665.

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Effect of polyvinyl butyrate nanoparticles incorporated with immune suppressing vitamins on alteration of population of intestinal immune cells

Mu Y., Kinashi Y., Kishimura A., Mori T., Hase K., Katayama Y.

Progress in Natural Science: Materials International （Progress in Natural Science: Materials International） 30 （ 5 ） 707 - 710 2020年10月

ISSN 10020071

© 2020 Chinese Materials Research Society The nanoparticles (NPs) from polyvinyl butyrate (PVBu) which can work as an orally applicable donor of butyrate for intestine were prepared. Immunotolerance inducing molecules such as vitamin D3 and all-trans retinoic acid (ATRA) were incorporated into PVBu NPs. The alteration in populations and numbers of DC103+ dendritic cells (DC) and regulatory T (Treg) cells in intestinal immune tissues were examined after oral administration of NPs. It was found that NPs reduced the population of CD103+ DC and Treg cells in Peyer's patched in lower part of the intestine and inversely increased the population of CD103+ DC in mesenteric lymph node (MLN), while the population of Treg cells in MLN was unchanged. These observations indicate that NPs may enhance the immunotolerance at MLN and lamina propria toward luminal antigens, indicating the promising features of PVBu NPs as therapeutics of allergy and autoimmune diseases.

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Microbiota-derived butyrate limits the autoimmune response by promoting the differentiation of follicular regulatory T cells

Takahashi D., Hoshina N., Kabumoto Y., Maeda Y., Suzuki A., Tanabe H., Isobe J., Yamada T., Muroi K., Yanagisawa Y., Nakamura A., Fujimura Y., Saeki A., Ueda M., Matsumoto R., Asaoka H., Clarke J.M., Harada Y., Umemoto E., Komatsu N., Okada T., Takayanagi H., Takeda K., Tomura M., Hase K.

EBioMedicine （EBioMedicine） 58 102913 2020年08月

研究論文（学術雑誌）, 共著, 査読有り

© 2020 The Author(s) Background: Rheumatoid arthritis (RA) is a chronic debilitating autoimmune disorder with a high prevalence, especially in industrialized countries. Dysbiosis of the intestinal microbiota has been observed in RA patients. For instance, new-onset untreated RA (NORA) is associated with the underrepresentation of the Clostridium cluster XIVa, including Lachnospiraceae, which are major butyrate producers, although the pathological relevance has remained obscure. Follicular regulatory T (TFR) cells play critical regulatory roles in the pathogenesis of autoimmune diseases, including RA. Reduced number of circulating TFR cells has been associated with the elevation of autoantibodies and disease severity in RA. However, the contribution of commensal microbe-derived butyrate in controlling TFR cell differentiation remains unknown. Methods: We examined the contribution of microbe-derived butyrate in controlling autoimmune arthritis using collagen-induced arthritis (CIA) and SKG arthritis models. We phenotyped autoimmune responses in the gut-associated lymphoid tissues (GALT) in the colon and joint-draining lymph nodes in the CIA model. We developed an in vitro CXCR5+Bcl-6+Foxp3+ TFR (iTFR) cell culture system and examined whether butyrate promotes the differentiation of iTFR cells. Findings: Microbe-derived butyrate suppressed the development of autoimmune arthritis. The immunization of type II collagen (CII) caused hypertrophy of the GALT in the colon by amplifying the GC reaction prior to the onset of the CIA. Butyrate mitigated these pathological events by promoting TFR cell differentiation. Butyrate directly induced the differentiation of functional TFR cells in vitro by enhancing histone acetylation in TFR cell marker genes. This effect was attributed to histone deacetylase (HDAC) inhibition by butyrate, leading to histone hyperacetylation in the promoter region of the TFR-cell marker genes. The adoptive transfer of the butyrate-treated iTFR cells reduced CII-specific autoantibody production and thus ameliorated the symptoms of arthritis. Interpretation: Accordingly, microbiota-derived butyrate serves as an environmental cue to enhance TFR cells, which suppress autoantibody production in the systemic lymphoid tissue, eventually ameliorating RA. Our findings provide mechanistic insights into the link between the gut environment and RA risk. Funding: This work was supported by AMED-Crest (16gm1010004h0101, 17gm1010004h0102, 18gm1010004h0103, and 19gm1010004s0104 to KH), the Japan Society for the Promotion of Science (JP17KT0055, JP16H01369, and JP18H04680 to KH; JP17K15734 to DT), Keio University Special Grant-in-Aid for Innovative Collaborative Research Projects (KH), Keio Gijuku Fukuzawa Memorial Fund for the Advancement of Education and Research (DT), the SECOM Science and Technology Foundation (KH), the Cell Science Research Foundation (KH), the Mochida Memorial Foundation for Medical and Pharmaceutical Research (DT), the Suzuken Memorial Foundation (KH and DT), the Takeda Science Foundation (KH and DT), The Science Research Promotion Fund, and The Promotion and Mutual Aid Corporation for Private Schools of Japan (KH).

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Protection of gut microbiome from antibiotics: Development of a vancomycin-specific adsorbent with high adsorption capacity

Yuzuriha K., Yakabe K., Nagai H., Li S., Zendo T., Zai K., Kishimura A., Hase K., Kim Y.G., Mori T., Katayama Y.

Bioscience of Microbiota, Food and Health （Bioscience of Microbiota, Food and Health） 39 （ 3 ） 128 - 136 2020年02月

研究論文（学術雑誌）, 共著, ISSN 21866953

The fraction of administered antibiotics that reach the cecum and colon causes dysbiosis of the gut microbiome, resulting in various diseases. Protection of the gut microbiome from antibiotics using antibiotic adsorbents in the cecum and colon is a promising method to overcome this issue. Previously, activated charcoal (AC) has been reported to protect the gut microbiome of host animals. AC is an adsorbent that is widely used to capture toxic compounds and overdosed drugs in the gastrointestinal tract. The specificity of adsorbents for antibiotics is critical to avoid the risk of unexpected side effects caused by nonspecific adsorption of biological compounds in the intestinal fluid, such as bile acids and essential micronutrients. Here, we have developed specific adsorbents for vancomycin (VCM), which is known to cause gut dysbiosis. The adsorbents were composed of polyethyleneglycolbased microparticles (MPs) in which a specific ligand for VCM, D-Ala-D-Ala-OH, was attached via dendrons of D-lysine to raise the content of the ligand in the MPs. The MPs successfully protected Staphylococcus lentus from VCM in vitro because of the adsorption of VCM in the culture media. Pre-administration of MPs to mice reduced the amount of free VCM in the feces to an undetectable level. This treatment minimized the effect of VCM on gut microbiota and provided protection against Clostridioides difficile infection after oral challenge with spores.

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Maternal gut microbiota in pregnancy influences offspring metabolic phenotype in mice

Kimura I., Miyamoto J., Ohue-Kitano R., Watanabe K., Yamada T., Onuki M., Aoki R., Isobe Y., Kashihara D., Inoue D., Inaba A., Takamura Y., Taira S., Kumaki S., Watanabe M., Ito M., Nakagawa F., Irie J., Kakuta H., Shinohara M., Iwatsuki K., Tsujimoto G., Ohno H., Arita M., Itoh H., Hase K.

Science （Science） 367 （ 6481 ） eaaw8429 2020年02月

研究論文（学術雑誌）, 査読有り, ISSN 00368075

Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works Antibiotics and dietary habits can affect the gut microbial community, thus influencing disease susceptibility. Although the effect of microbiota on the postnatal environment has been well documented, much less is known regarding the impact of gut microbiota at the embryonic stage. Here we show that maternal microbiota shapes the metabolic system of offspring in mice. During pregnancy, short-chain fatty acids produced by the maternal microbiota dictate the differentiation of neural, intestinal, and pancreatic cells through embryonic GPR41 and GPR43. This developmental process helps maintain postnatal energy homeostasis, as evidenced by the fact that offspring from germ-free mothers are highly susceptible to metabolic syndrome, even when reared under conventional conditions. Thus, our findings elaborate on a link between the maternal gut environment and the developmental origin of metabolic syndrome.

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Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity

Kimura S., Nakamura Y., Kobayashi N., Shiroguchi K., Kawakami E., Mutoh M., Takahashi-Iwanaga H., Yamada T., Hisamoto M., Nakamura M., Udagawa N., Sato S., Kaisho T., Iwanaga T., Hase K.

Nature Communications （Nature Communications） in press （ 1 ） 2020年01月

研究論文（学術雑誌）, 共著, 査読有り

© 2020, The Author(s). Microfold cells (M cells) are responsible for antigen uptake to initiate immune responses in the gut-associated lymphoid tissue (GALT). Receptor activator of nuclear factor-κB ligand (RANKL) is essential for M cell differentiation. Follicle-associated epithelium (FAE) covers the GALT and is continuously exposed to RANKL from stromal cells underneath the FAE, yet only a subset of FAE cells undergoes differentiation into M cells. Here, we show that M cells express osteoprotegerin (OPG), a soluble inhibitor of RANKL, which suppresses the differentiation of adjacent FAE cells into M cells. Notably, OPG deficiency increases M cell number in the GALT and enhances commensal bacterium-specific immunoglobulin production, resulting in the amelioration of disease symptoms in mice with experimental colitis. By contrast, OPG-deficient mice are highly susceptible to Salmonella infection. Thus, OPG-dependent self-regulation of M cell differentiation is essential for the balance between the infectious risk and the ability to perform immunosurveillance at the mucosal surface.

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Microfold cell-dependent antigen transport alleviates infectious colitis by inducing antigen-specific cellular immunity

Nakamura Y., Mimuro H., Kunisawa J., Furusawa Y., Takahashi D., Fujimura Y., Kaisho T., Kiyono H., Hase K.

Mucosal Immunology （Mucosal Immunology） 13 （ 4 ） 679 - 690 2020年01月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 19330219

© 2020, Society for Mucosal Immunology. Infectious colitis is one of the most common health issues worldwide. Microfold (M) cells actively transport luminal antigens to gut-associated lymphoid tissue to induce IgA responses; however, it remains unknown whether M cells contribute to the induction of cellular immune responses. Here we report that M cell-dependent antigen transport plays a critical role in the induction of Th1, Th17, and Th22 responses against gut commensals in the steady state. The establishment of commensal-specific cellular immunity was a prerequisite for preventing bacterial dissemination during enteropathogenic Citrobacter rodentium infection. Therefore, M cell-null mice developed severe colitis with increased bacterial dissemination. This abnormality was associated with mucosal barrier dysfunction. These observations suggest that antigen transport by M cells may help maintain gut immune homeostasis by eliciting antigen-specific cellular immune responses.

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Commensal-bacteria-derived butyrate promotes the t-cell-independent iga response in the colon

Isobe J., Maeda S., Obata Y., Iizuka K., Nakamura Y., Fujimura Y., Kimizuka T., Hattori K., Kim Y.G., Morita T., Kimura I., Offermanns S., Adachi T., Nakao A., Kiyono H., Takahashi D., Hase K.

International Immunology （International Immunology） 32 （ 4 ） 243 - 258 2019年12月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 09538178

© 2020 Oxford University Press. All rights reserved. Secretory immunoglobulin A (SIgA), the most abundant antibody isotype in the body, maintains a mutual relationship with commensal bacteria and acts as a primary barrier at the mucosal surface. Colonization by commensal bacteria induces an IgA response, at least partly through a T-cell-independent process. However, the mechanism underlying the commensal-bacteria-induced T-cell-independent IgA response has yet to be fully clarified. Here, we show that commensalbacteria-derived butyrate promotes T-cell-independent IgA class switching recombination (CSR) in the mouse colon. Notably, the butyrate concentration in human stools correlated positively with the amount of IgA. Butyrate up-regulated the production of transforming growth factor β1 and all-trans retinoic acid by CD103+CD11b+ dendritic cells, both of which are critical for T-cell-independent IgA CSR. This effect was mediated by G-protein-coupled receptor 41 (GPR41/FFA3) and GPR109a/HCA2, and the inhibition of histone deacetylase. The butyrate-induced IgA response reinforced the colonic barrier function, preventing systemic bacterial dissemination under inflammatory conditions. These observations demonstrate that commensal-bacteria-derived butyrate contributes to the maintenance of the gut immune homeostasis by facilitating the T-cell-independent IgA response in the colon.

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Immunological Consequences of Antigen Sampling at Mucosal Surfaces

Mabbott NA and Hase K

Front. Immuol. 10 2773 2019年11月

研究論文（学術雑誌）, 共著

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The Roles of Peyer's Patches and Microfold Cells in the Gut Immune System: Relevance to Autoimmune Diseases

Kobayashi N., Takahashi D., Takano S., Kimura S., Hase K.

Frontiers in Immunology （Frontiers in Immunology） 10 2345 2019年10月

研究論文（学術雑誌）, 共著, 査読有り

Microfold (M) cells are located in the epithelium covering mucosa-associated lymphoid tissues, such as the Peyer's patches (PPs) of the small intestine. M cells actively transport luminal antigens to the underlying lymphoid follicles to initiate an immune response. The molecular machinery of M-cell differentiation and function has been vigorously investigated over the last decade. Studies have shed light on the role of M cells in the mucosal immune system and have revealed that antigen uptake by M cells contributes to not only mucosal but also systemic immune responses. However, M-cell studies usually focus on infectious diseases; the contribution of M cells to autoimmune diseases has remained largely unexplored. Accumulating evidence suggests that dysbiosis of the intestinal microbiota is implicated in multiple systemic diseases, including autoimmune diseases. This implies that the uptake of microorganisms by M cells in PPs may play a role in the pathogenesis of autoimmune diseases. We provide an outline of the current understanding of M-cell biology and subsequently discuss the potential contribution of M cells and PPs to the induction of systemic autoimmunity, beyond the mucosal immune response.

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Fasting-Refeeding Impacts Immune Cell Dynamics and Mucosal Immune Responses

Nagai M., Noguchi R., Takahashi D., Morikawa T., Koshida K., Komiyama S., Ishihara N., Yamada T., Kawamura Y., Muroi K., Hattori K., Kobayashi N., Fujimura Y., Hirota M., Matsumoto R., Aoki R., Tamura-Nakano M., Sugiyama M., Katakai T., Sato S., Takubo K., Dohi T., Hase K.

Cell （Cell） 178 （ 5 ） 1072 - 1087.e14 2019年08月

ISSN 00928674

© 2019 Elsevier Inc. Nutritional status potentially influences immune responses; however, how nutritional signals regulate cellular dynamics and functionality remains obscure. Herein, we report that temporary fasting drastically reduces the number of lymphocytes by ∼50% in Peyer's patches (PPs), the inductive site of the gut immune response. Subsequent refeeding seemingly restored the number of lymphocytes, but whose cellular composition was conspicuously altered. A large portion of germinal center and IgA+ B cells were lost via apoptosis during fasting. Meanwhile, naive B cells migrated from PPs to the bone marrow during fasting and then back to PPs during refeeding when stromal cells sensed nutritional signals and upregulated CXCL13 expression to recruit naive B cells. Furthermore, temporal fasting before oral immunization with ovalbumin abolished the induction of antigen-specific IgA, failed to induce oral tolerance, and eventually exacerbated food antigen-induced diarrhea. Thus, nutritional signals are critical in maintaining gut immune homeostasis. Temporary fasting drastically reduces the levels of B cells in Peyer's patches, with germinal center B cells undergoing apoptosis and naive cells migrating to the bone marrow and only egressing upon refeeding.

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Mast cells play role in wound healing through the ZnT2/GPR39/IL-6 axis

Nishida K., Hasegawa A., Yamasaki S., Uchida R., Ohashi W., Kurashima Y., Kunisawa J., Kimura S., Iwanaga T., Watarai H., Hase K., Ogura H., Nakayama M., Kashiwakura J.i., Okayama Y., Kubo M., Ohara O., Kiyono H., Koseki H., Murakami M., Hirano T.

Scientific Reports （Scientific Reports） 9 （ 1 ） 10842 2019年07月

研究論文（学術雑誌）, 共著, 査読有り

© 2019, The Author(s). Zinc (Zn) is an essential nutrient and its deficiency causes immunodeficiency and skin disorders. Various cells including mast cells release Zn-containing granules when activated; however, the biological role of the released Zn is currently unclear. Here we report our findings that Zn transporter ZnT2 is required for the release of Zn from mast cells. In addition, we found that Zn and mast cells induce IL-6 production from inflammatory cells such as skin fibroblasts and promote wound healing, a process that involves inflammation. Zn induces the production of a variety of pro-inflammatory cytokines including IL-6 through signaling pathways mediated by the Zn receptor GPR39. Consistent with these findings, wound healing was impaired in mice lacking IL-6 or GPR39. Thus, our results show that Zn and mast cells play a critical role in wound healing through activation of the GPR39/IL-6 signaling axis.

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Sox8 is essential for M-cell maturation to accelerate IgA response at the early stage after weaning in mice

Kimura S, Kobayashi N, Nakamura Y, Kanaya T, Takahashi D, Fujiki R, Mutoh M, Obata Y, Iwanaga T, Nakagawa T, Kato N, Sato S, Kaisho T, Ohno H, and *HASE Koji

J. Exp. Med. 216 （ 4 ） 831 - 846 2019年04月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 00221007

Microfold (M) cells residing in the follicle-associated epithelium (FAE) of the gut-associated lymphoid tissue are specialized for antigen uptake to initiate mucosal immune responses. The molecular machinery and biological significance of M cell differentiation, however, remain to be fully elucidated. Here, we demonstrate that Sox8, a member of the SRY-related HMG box transcription factor family, is specifically expressed by M cells in the intestinal epithelium. The expression of Sox8 requires activation of RANKL-RelB signaling. Chromatin immunoprecipitation and luciferase assays revealed that Sox8 directly binds the promoter region of Gp2 to increase Gp2 expression, which is the hallmark of functionally mature M cells.Furthermore, genetic deletion of Sox8 causes a marked decrease in the number of mature M cells, resulting in reduced antigen uptake in Peyer's patches. Consequently, juvenile Sox8-deficient mice showed attenuated germinal center reactions and antigen-specific IgA responses. These findings indicate that Sox8 plays an essential role in the development of M cells to establish mucosal immune responses.

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MZB1 promotes the secretion of J-chain–containing dimeric IgA and is critical for the suppression of gut inflammation

Xiong E., Li Y., Min Q., Cui C., Liu J., Hong R., Lai N., Wang Y., Sun J., Matsumoto R., Takahashi D., Hase K., Shinkura R., Tsubata T., Wang J.

Proceedings of the National Academy of Sciences of the United States of America （Proceedings of the National Academy of Sciences of the United States of America） 116 （ 27 ） 13480 - 13489 2019年

ISSN 00278424

© 2019 National Academy of Sciences. All rights reserved. IgA is the most abundantly produced antibody in the body and plays a crucial role in gut homeostasis and mucosal immunity. IgA forms a dimer that covalently associates with the joining (J) chain, which is essential for IgA transport into the mucosa. Here, we demonstrate that the marginal zone B and B-1 cell-specific protein (MZB1) interacts with IgA through the α-heavy-chain tailpiece dependent on the penultimate cysteine residue and prevents the intracellular degradation of α-light-chain complexes. Moreover, MZB1 promotes J-chain binding to IgA and the secretion of dimeric IgA. MZB1-deficient mice are impaired in secreting large amounts of IgA into the gut in response to acute inflammation and develop severe colitis. Oral administration of a monoclonal IgA significantly ameliorated the colitis, accompanied by normalization of the gut microbiota composition. The present study identifies a molecular chaperone that promotes J-chain binding to IgA and reveals an important mechanism that controls the quantity, quality, and function of IgA.

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Mucin O-glycans facilitate symbiosynthesis to maintain gut immune homeostasis

Yamada T., Hino S., Iijima H., Genda T., Aoki R., Nagata R., Han K., Hirota M., Kinashi Y., Oguchi H., Suda W., Furusawa Y., Fujimura Y., Kunisawa J., Hattori M., Fukushima M., Morita T., Hase K.

EBioMedicine （EBioMedicine） 48 513 - 525 2019年

© 2019 Background: The dysbiosis of gut microbiota has been implicated in the pathogenesis of inflammatory bowel diseases; however, the underlying mechanisms have not yet been elucidated. Heavily glycosylated mucin establishes a first-line barrier against pathogens and serves as a niche for microbial growth. Methods: To elucidate relationships among dysbiosis, abnormal mucin utilisation, and microbial metabolic dysfunction, we analysed short-chain fatty acids (SCFAs) and mucin components in stool samples of 40 healthy subjects, 49 ulcerative colitis (UC) patients, and 44 Crohn's disease (CD) patients from Japan. Findings: Levels of n-butyrate were significantly lower in stools of both CD and UC patients than in stools of healthy subjects. Correlation analysis identified seven bacterial species positively correlated with n-butyrate levels; the major n-butyrate producer, Faecalibacterium prausnitzii, was particularly underrepresented in CD patients, but not in UC patients. In UC patients, there were inverse correlations between mucin O-glycan levels and the production of SCFAs, such as n-butyrate, suggesting that mucin O-glycans serve as an endogenous fermentation substrate for n-butyrate production. Indeed, mucin-fed rodents exhibited enhanced n-butyrate production, leading to the expansion of RORgt+Treg cells and IgA-producing cells in colonic lamina propria. Microbial utilisation of mucin-associated O-glycans was significantly reduced in n-butyrate-deficient UC patients. Interpretation: Mucin O-glycans facilitate symbiosynthesis of n-butyrate by gut microbiota. Abnormal mucin utilisation may lead to reduced n-butyrate production in UC patients. Fund: Japan Society for the Promotion of Science, Health Labour Sciences Research Grant, AMED-Crest, AMED, Yakult Foundation, Keio Gijuku Academic Development Funds, The Aashi Grass Foundation, and The Canon Foundation.

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Maintenance of Intestinal Epithelial Homeostasis by Zinc Transporters

Ohashi W., Hara T., Takagishi T., Hase K., Fukada T.

Digestive Diseases and Sciences （Digestive Diseases and Sciences） 64 （ 9 ） 2404 - 2415 2019年

ISSN 01632116

© 2019, Springer Science+Business Media, LLC, part of Springer Nature. Zinc is an essential micronutrient for normal organ function, and dysregulation of zinc metabolism has been implicated in a wide range of diseases. Emerging evidence has revealed that zinc transporters play diverse roles in cellular homeostasis and function by regulating zinc trafficking via organelles or the plasma membrane. In the gastrointestinal tract, zinc deficiency leads to diarrhea and dysfunction of intestinal epithelial cells. Studies also showed that zinc transporters are very important in intestinal epithelial homeostasis. In this review, we describe the physiological roles of zinc transporters in intestinal epithelial functions and relevance of zinc transporters in gastrointestinal diseases.

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Airway M Cells arise in the lower airway due to RANKL signaling and reside in the bronchiolar epithelium associated with iBALT in murine models of respiratory disease

Kimura S., Mutoh M., Hisamoto M., Saito H., Takahashi S., Asakura T., Ishii M., Nakamura Y., Iida J., Hase K., Iwanaga T.

Frontiers in Immunology （Frontiers in Immunology） 10 （ JUN ） 2019年

© 2019 Kimura, Mutoh, Hisamoto, Saito, Takahashi, Asakura, Ishii, Nakamura, Iida, Hase and Iwanaga. Microfold (M) cells residing in the follicle-associated epithelium of mucosa-associated lymphoid tissues are specialized for sampling luminal antigens to initiate mucosal immune responses. In the past decade, glycoprotein 2 (GP2) and Tnfaip2 were identified as reliable markers for M cells in the Peyer's patches of the intestine. Furthermore, RANKL-RANK signaling, as well as the canonical and non-canonical NFκB pathways downstream, is essential for M-cell differentiation from the intestinal stem cells. However, the molecular characterization and differentiation mechanisms of M cells in the lower respiratory tract, where organized lymphoid tissues exist rarely, remain to be fully elucidated. Therefore, this study aimed to explore M cells in the lower respiratory tract in terms of their specific molecular markers, differentiation mechanism, and functions. Immunofluorescence analysis revealed a small number of M cells expressing GP2, Tnfaip2, and RANK is present in the lower respiratory tract of healthy mice. The intraperitoneal administration of RANKL in mice effectively induced M cells, which have a high capacity to take up luminal substrates, in the lower respiratory epithelium. The airway M cells associated with lymphoid follicles were frequently detected in the pathologically induced bronchus-associated lymphoid tissue (iBALT) in the murine models of autoimmune disease as well as pulmonary emphysema. These findings demonstrate that RANKL is a common inducer of M cells in the airway and digestive tracts and that M cells are associated with the respiratory disease. We also established a two-dimensional culture method for airway M cells from the tracheal epithelium in the presence of RANKL successfully. This model may be useful for functional studies of M cells in the sampling of antigens at airway mucosal surfaces.

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A partial agonist for retinoid X receptor mitigates experimental colitis

Masayoshi O., Masaki W., Narumi I., Koichiro S., Kei T., Masato H., Takahiro Y., Aiko E., Osamu S., Midori N., Michiko F., Makoto M., Daisuke T., Yukihiro F., Hiroki K., Koji H.

International Immunology （International Immunology） [Epub ahead of print] （ 4 ） 251 - 262 2018年12月

共著, 査読有り, ISSN 09538178

Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease, is an intractable disease of the gastrointestinal tract. Multiple environmental factors, including food ingredients, have been implicated in the development of these diseases. For example, animal fat-rich diets are predisposing factors for ulcerative colitis, whereas n-3 unsaturated fatty acids such as docosahexaenoic acid (DHA) show protective effects in experimental colitis and are negatively correlated with the incidence of ulcerative colitis and Crohn's disease. Given that DHA exhibits agonistic activity on retinoid X receptor (RXR), activation of RXR could be a therapeutic strategy for IBD. However, conventional full RXR agonists are known to show considerable adverse effects. We therefore took advantage of a partial RXR agonist, CBt-PMN, to minimize the adverse effects, and evaluated its efficacy in dextran sodium sulfate-induced colitis. Administration of CBt-PMN efficiently ameliorated the symptoms of colitis. This effect was attributed to the down-regulation of pro-inflammatory cytokines such as TNF and Il6 in colon-infiltrating monocytes. Down-regulation of pro-inflammatory cytokines by CBt-PMN was also evident in lipopolysaccharide-stimulated bone marrow-derived macrophages (BMDMs). Among many RXR-associated nuclear receptors, activation of peroxisome proliferator-activated receptor δ(PPARd) and nuclear hormone receptor 77 (Nur77) suppressed cytokine production by BMDMs. These observations suggest that the activation of PPARδ/RXR and Nur77/RXR heterodimers by CBt-PMN through the permissive mechanism is responsible for diminishing the monocyte-mediated inflammatory response in the gut. Our data highlight the importance of RXR activation in the regulation of colitis.

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Oral administration of Porphyromonas gingivalis alters the gut microbiome and serum metabolome

Kato T., Yamazaki K., Nakajima M., Date Y., Kikuchi J., Hase K., Ohno H., Yamazaki K.

mSphere （mSphere） 3 （ 5 ） e00460-18 2018年10月

研究論文（学術雑誌）, 共著, 査読有り

© 2018 Kato et al. Periodontal disease induced by periodontopathic bacteria like Porphyromonas gingivalis is demonstrated to increase the risk of metabolic, inflammatory, and autoimmune disorders. Although precise mechanisms for this connection have not been elucidated, we have proposed mechanisms by which orally administered periodontopathic bacteria might induce changes in gut microbiota composition, barrier function, and immune system, resulting in an increased risk of diseases characterized by low-grade systemic inflammation. Accumulating evidence suggests a profound effect of altered gut metabolite profiles on overall host health. Therefore, it is possible that P. gingivalis can affect these metabolites. To test this, C57BL/6 mice were administered with P. gingivalis W83 orally twice a week for 5 weeks and compared with sham-inoculated mice. The gut microbial communities were analyzed by pyrosequencing the 16S rRNA genes. Inferred metagenomic analysis was used to determine the relative abundance of KEGG pathways encoded in the gut microbiota. Serum metabolites were analyzed using nuclear magnetic resonance (NMR)-based metabolomics coupled with multivariate statistical analyses. Oral administration of P. gingivalis induced a change in gut microbiota composition. The distributions of metabolic pathways differed between the two groups, including those related to amino acid metabolism and, in particular, the genes for phenylalanine, tyrosine, and tryptophan biosynthesis. Also, alanine, glutamine, histidine, tyrosine, and phenylalanine were significantly increased in the serum of P. gingivalis-administered mice. In addition to altering immune modulation and gut barrier function, oral administration of P. gingivalis affects the host's metabolic profile. This supports our hypothesis regarding a gut-mediated systemic pathology resulting from periodontal disease.

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Therapeutic effect of vitamin D(3)-containing nanostructured lipid carriers on inflammatory bowel disease.

Zai K, Hirota M, Yamada T, Ishihara N, Mori T, Kishimura A, Suzuki K, *HASE K, *Katayama Y

J Control Release. 286 94 - 102 2018年09月

研究論文（学術雑誌）, 共著, 査読有り

Therapeutic effect of vitamin D<inf>3</inf>-containing nanostructured lipid carriers on inflammatory bowel disease

Zai K., Hirota M., Yamada T., Ishihara N., Mori T., Kishimura A., Suzuki K., Hase K., Katayama Y.

Journal of Controlled Release （Journal of Controlled Release） 286 94 - 102 2018年09月

ISSN 01683659

© 2018 Elsevier B.V. The active form of vitamin D3, 1,25(OH)2D3 has been found to exert multiple effects on the suppression of progression of inflammatory bowel disease (IBD). Vitamin D3 has been gathering attention as a therapy for IBD. However, the clinical trials conducted to date revealed that a relatively high dosage of vitamin D3 was required to see a significant therapeutic effect. Thus, effective formulation and delivery of vitamin D3 to colonic inflammatory lesions will be required. Herein we describe the preparation of a nanostructured lipid carrier (NLC) for the encapsulation of 1,25(OH)2D3 for colonic delivery via oral administration. The optimized fabrication procedure enabled the incorporation of 1,25(OH)2D3 in the NLC by minimizing the destruction of chemically unstable 1,25(OH)2D3. The obtained NLCs orally delivered 1,25(OH)2D3 to the colon in mice and maintained a high concentration of 1,25(OH)2D3 in the colonic tissue for at least 12 h. The NLC showed multiple effects on the suppression of symptoms of colitis induced by dextran sodium sulfate, namely maintaining crypt structure, reducing the tissue concentration of inflammatory cytokines, suppressing the infiltration of polymorphonuclear leukocytes, and augmenting anti-inflammatory CX3CR1high macrophages. Our NLCs containing 1,25(OH)2D3 may be an alternative treatment for IBD therapy.

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Macrophages switch their phenotype by regulating maf expression during different phases of inflammation

Kikuchi K., Iida M., Ikeda N., Moriyama S., Hamada M., Takahashi S., Kitamura H., Watanabe T., Hasegawa Y., Hase K., Fukuhara T., Sato H., Kobayashi E., Suzuki T., Yamamoto M., Tanaka M., Asano K.

Journal of Immunology （Journal of Immunology） 201 （ 2 ） 635 - 651 2018年07月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 00221767

Copyright © 2018 by The American Association of Immunologists, Inc Macrophages manifest distinct phenotype according to the organs in which they reside. In addition, they flexibly switch their character in adaptation to the changing environment. However, the molecular basis that explains the conversion of the macrophage phenotype has so far been unexplored. We find that CD169+ macrophages change their phenotype by regulating the level of a transcription factor Maf both in vitro and in vivo in C57BL/6J mice. When CD169+ macrophages were exposed to bacterial components, they expressed an array of acute inflammatory response genes in Maf-dependent manner and simultaneously start to downregulate Maf. This Maf suppression is dependent on accelerated degradation through proteasome pathway and microRNA-mediated silencing. The downregulation of Maf unlocks the NF-E2–related factor 2–dominant, cytoprotective/ antioxidative program in the same macrophages. The present study provides new insights into the previously unanswered question of how macrophages initiate proinflammatory responses while retaining their capacity to repair injured tissues during inflammation. The Journal of Immunology, 2018, 201: 635–651.

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Regulation of inflammatory response of macrophages and induction of regulatory T cells by using retinoic acid-loaded nanostructured lipid carrier

Zai K., Ishihara N., Oguchi H., Hirota M., Kishimura A., Mori T., Hase K., Katayama Y.

Journal of Biomaterials Science, Polymer Edition （Journal of Biomaterials Science, Polymer Edition） in press （ 1 ） 1 - 11 2018年06月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 09205063

© 2019, © 2019 Informa UK Limited, trading as Taylor & Francis Group. Immunomodulatory function of all-trans retinoic acid (ATRA) has been gathering much attention for the therapy of autoimmune diseases. ATRA is a chemically unstable molecule which requires proper formulation for targeted delivery. Here we examined nanostructured lipid carrier (NLC) for the formulation of ATRA. NLC is a representative nanoparticle formulation especially suited for oral delivery. We established the preparation procedures of ATRA-containing NLC (NLC-RA) which minimizes the degradation of ATRA during the preparation process. NLC-RA thus obtained was taken up by macrophages and induced anti-inflammatory response via suppressing NF-κB signaling as well as via enhancing the production of anti-inflammatory cytokines. Moreover, NLC-RA enhanced differentiation of naïve T cells to regulatory T cells in the co-culture system with dendritic cells. These results suggest that NLC-RA is a promising alternative therapy for the autoimmune diseases especially intestinal bowel disease. (Figure presented.).

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Intestinal epithelial cell-specific deletion of α-mannosidase II ameliorates experimental colitis

Suzuki K., Yamada T., Yamazaki K., Hirota M., Ishihara N., Sakamoto M., Takahashi D., Iijima H., Hase K.

Cell Structure and Function （Cell Structure and Function） 43 （ 1 ） 25 - 39 2018年03月

研究論文（学術雑誌）, 共著, 査読有り, ISSN 03867196

© 2018 The Author(s). Inflammatory bowel disease (IBD) is a refractory disease of the gastrointestinal tract that is believed to develop in genetically susceptible individuals. Glycosylation, a type of post-translational modification, is involved in the development of a wide range of diseases, including IBD, by modulating the function of various glycoproteins. To identify novel genes contributing to the development of IBD, we analyzed single nucleotide polymorphisms (SNPs) of glycosylation-related genes in IBD patients and identified MAN2A1, encoding alphamannosidase II (α-MII), as a candidate gene. α-MII plays a crucial, but not exclusive, role in the maturation of Nglycans. We also observed that intestinal epithelial cells (IECs), which establish the first-line barrier and regulate gut immunity, selectively expressed α-MII with minimal expression of its isozyme, alpha-mannosidase IIx (α-MIIx). This led us to hypothesize that IEC-intrinsic α-MII is implicated in the pathogenesis of IBD. To test this hypothesis, we generated IEC-specific α-MII-deficient (α-MIIΔIEC) mice. Although α-MII deficiency has been shown to have a minimal effect on N-glycan maturation in most cell types due to the compensation by α-MIIx, ablation of α-MII impaired the maturation of N-glycans in IECs. α-MIIΔIEC mice were less susceptible to dextran sulfate sodium-induced colitis compared with control littermates. In accordance with this, neutrophil infiltration in the colonic mucosa was attenuated in α-MIIΔIEC mice. Furthermore, gene expression levels of neutrophilattracting chemokines were downregulated in the colonic tissue. These results suggest that IEC-intrinsic α-MII promotes intestinal inflammation by facilitating chemokine expression. We propose SNPs in MAN2A1 as a novel genetic factor for IBD.

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Macrophage extracellular trap formation promoted by platelet activation is a key mediator of rhabdomyolysis-induced acute kidney injury

Okubo K., Kurosawa M., Kamiya M., Urano Y., Suzuki A., Yamamoto K., Hase K., Homma K., Sasaki J., Miyauchi H., Hoshino T., Hayashi M., Mayadas T., Hirahashi J.

Nature Medicine （Nature Medicine） 24 （ 2 ） 232 - 238 2018年02月

ISSN 10788956

Rhabdomyolysis is a serious syndrome caused by skeletal muscle injury and the subsequent release of breakdown products from damaged muscle cells into systemic circulation. The muscle damage most often results from strenuous exercise, muscle hypoxia, medications, or drug abuse and can lead to life-threatening complications, such as acute kidney injury (AKI). Rhabdomyolysis and the AKI complication can also occur during crush syndrome, an emergency condition that commonly occurs in victims of natural disasters, such as earthquakes, and man-made disasters, such as wars and terrorism. Myoglobin released from damaged muscle is believed to trigger renal dysfunction in this form of AKI. Recently, macrophages were implicated in the disease pathogenesis of rhabdomyolysis-induced AKI, but the precise molecular mechanism remains unclear. In the present study, we show that macrophages released extracellular traps (ETs) comprising DNA fibers and granule proteins in a mouse model of rhabdomyolysis. Heme-activated platelets released from necrotic muscle cells during rhabdomyolysis enhanced the production of macrophage extracellular traps (METs) through increasing intracellular reactive oxygen species generation and histone citrullination. Here we report, for the first time to our knowledge, this unanticipated role for METs and platelets as a sensor of myoglobin-derived heme in rhabdomyolysis-induced AKI. This previously unknown mechanism might be targeted for treatment of the disease. Finally, we found a new therapeutic tool for prevention of AKI after rhabdomyolysis, which might rescue some sufferers of this pathology.

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Macrophage extracellular chromatin release induced by Mac-1 mediated platelet-macrophage interactions triggers acute kidney injury in rhabdomyolysis.

Okubo K, Kurosawa M, Kamiya M, Urano Y, Suzuki A, Yamamoto K, HASE K, Homma K, Sasaki J, Miyauchi H, Hoshino T, Hayashi M, Mayadas TN, Hirahashi J.

Nat. Med. 24 232 - 238 2018年01月

研究論文（学術雑誌）, 共著, 査読有り

Intestinal M cells contribute to the maintenance of mucosal barrier functions and immune homeostasis

Ishihara N., Kobayashi N., Hase K.

Japanese Journal of Allergology （Japanese Journal of Allergology） 67 （ 3 ） 171 - 178 2018年

ISSN 00214884

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NALT M cells are important for immune induction for the common mucosal immune system.

Date Y, Ebisawa M, Fukuda S, Shima H, Obata Y, Takahashi D, Kato T, Hanazato M, Nakato G, Williams IR, HASE K, Ohno H.

Int Immunol. 29 471 - 478 2017年12月

研究論文（学術雑誌）, 共著, 査読有り

Attenuation of CD4+CD25+regulatory T cells in the tumor microenvironment by metformin, a type 2 diabetes drug.

長谷耕二

EBioMedicine, 25:154-164, 2017. 25 154 - 164 2017年11月

研究論文（学術雑誌）, 共著, 査読有り

IL-22BP dictates characteristics of Peyer's patch follicle-associated epithelium for antigen uptake.

Jinnohara T, Kanaya T, HASE K, Sakakibara S, Kato T, Tachibana N, Sasaki T, Hashimoto Y, Sato T, Watarai H, Kunisawa J, Shibata N, Williams I, Kiyono H, and Ohno H.

J. Exp. Med. 214 1607 - 1618 2017年06月

研究論文（学術雑誌）, 共著, 査読有り

Zinc Transporter SLC39A7/ZIP7 Promotes Intestinal Epithelial Self-Renewal by Resolving ER Stress.

1 Ohashi W, Kimura S, Iwanaga T, Furusawa Y, Irié T, Izumi H, Watanabe T, Hijikata A, Hara T, Ohara O, Koseki H, Sato T, Robine S, Mori H, Hattori Y, Watarai H, Mishima K, Ohno H, Hase K, Fukada T.

PLoS Genet. 12 e1006349 2016年10月

研究論文（学術雑誌）, 共著, 査読有り

Distinct Roles for the N- and C-terminal Regions of M-Sec in Plasma Membrane Deformation during Tunneling Nanotube Formation.

2 Kimura S, Yamashita M, Yamakami-Kimura M, Sato Y, Yamagata A, Kobashigawa Y, Inagaki F, Amada T, Hase K, Iwanaga T, Ohno H, Fukai S.

Sci. Rep. 6 33548 2016年09月

研究論文（学術雑誌）, 共著, 査読有り

Gut commensal Bacteroides acidifaciens prevents obesity and improves insulin sensitivity in mice.

Yang JY, Lee YS, Kim Y, Lee SH, Ryu S, Fukuda S, #HHase K<U/>, Yang CS, Lim HS, Kim MS, Kim HM, Ahn SH, Kwon BE, Ko HJ, Kweon MN

Mucosal Immunol. 10 104 - 116 2016年04月

研究論文（学術雑誌）, 共著, 査読有り

EAF2 mediates germinal centre B-cell apoptosis to suppress excessive immune responses and prevent autoimmunity.

Li Y, Takahashi Y, Fujii S, Zhou Y, Hong R, Suzuki A, Tsubata T, HASE Koji, Wang JY.

Nature Commun. 7 10836 2016年03月

研究論文（学術雑誌）, 共著, 査読有り

Epithelium-Intrinsic MicroRNAs Contribute to Mucosal Immune Homeostasis by Promoting M-Cell Maturation.

Nakato G, HASE Koji, Sato T, Kimura S, Sakakibara S, Sugiyama M, Obata Y,Hanazato M, Iwanaga T, Ohno H

PLoS One 11 （ 3 ） e0150379 2016年03月

研究論文（学術雑誌）, 共著, 査読有り

Potential Role of the Formation of Tunneling Nanotubes in HIV-1 Spread in Macrophages

<5 Hashimoto M, Bhuyan F, Hiyoshi M, Noyori O, Nasser H, Miyazaki M, Saito T,Kondoh Y, Osada H, Kimura S, HASE Koji, Ohno H, Suzu S.

J. Immunol. 196 18321 - 1841 2016年02月

研究論文（学術雑誌）, 共著, 査読有り

Distinct microRNA expression profiles in follicle-associated epithelium and villous epithelium.

Genom Data 5 388 - 390 2015年07月

研究論文（学術雑誌）, 共著, 査読有り

The microbiota regulates type 2 immunity through RORγt+ T cells.

Ohnmacht C, Park JH, Cording S, Wing JB, Atarashi K, Obata Y, Gaboriau-Routhiau V, Marques R, Dulauroy S, Fedoseeva M, Busslinger M, Cerf-Bensussan N, Boneca IG, Voehringer D, Hase K, Honda K, Sakaguchi S, Eberl G.

Science 349 989 - 993 2015年07月

研究論文（学術雑誌）, 査読有り

Botulinum toxin A complex exploits intestinal M cells to enter the host and exert neurotoxicity.

Matsumura T, Sugawara Y, Yutani M, Amatsu S, Yagita H，Kohda T, Fukuoka S-I, Nakamura Y, Hase K, Ohno H and Fujinaga Y.

Nat. Commun. 6 6255 2015年02月

研究論文（学術雑誌）, 共著

Epithelial-stromal interaction via Notch signaling is essential for the full maturation of gut-associated lymphoid tissues.

10 Obata Y, Kimura S, Nakato G, Iizuka K, Miyagawa Y, Nakamura Y, Furusawa Y, Sugiyama M, Suzuki K, Ebisawa M, Fujimura Y, Yoshida H, Iwanaga T, Hase K, Ohno H

EMBO Rep. 15 1297 - 1304 2014年11月

研究論文（学術雑誌）, 共著, 査読有り

The Role of the Clathrin Adaptor AP-1: Polarized Sorting and Beyond.

Nakatsu F, Hase K and Ohno H

Membranes 4 747 - 763 2014年11月

研究論文（学術雑誌）, 共著, 査読有り

Visualization of the entire differentiation process of murine M cells: suppression of their maturation in cecal patches.

Kimura S, Yamakami-Kimura M, Obata Y, Hase K, Kitamura H, Ohno H, Iwanaga T.

Mucosal Immunol 8 650 - 660 2014年10月

研究論文（学術雑誌）, 共著, 査読有り

Pitfalls in global normalization of ChIP-seq data in CD4(+) T cells treated with butyrate: A possible solution strategy.

Furusawa Y, Endo TA, Obata Y, Ohara O, Ohno H and Hase K.

Genome Data 2 176 - 180 2014年07月

研究論文（学術雑誌）, 共著, 査読有り

Epigenetic regulator Uhrf1 is critical for functional expansion of colonic regulatory T cells.

#Obata Y, #Furusawa Y, Endo TA, Sharif J, Takahashi D, Atarashi K, Onawa S, Fujimura Y, Takahashi M, Ikawa T, Otsubo T, Kawamura YI, Dohi T, Tajima S, Ohara O, Honda K, Hori S, Ohno H, Koseki H and *Hase K.

Nat Immunol 15 571-579 2014年05月

研究論文（学術雑誌）, 共著, 査読有り

Gut microbiota-generated metabolites in animal health and disease.

*Lee WJ and *Hase K.

Nat. Chem. Biol. 10 416-424 2014年05月

研究論文（学術雑誌）, 共著, 査読有り

Treg induction by a rationally selected Clostridia cocktail from the human microbiota.

Atarashi K, Tanoue T, Oshima K, Suda W, Nagano Y, Nishikawa H, Fukuda S, Saito T, Narushima S, Hase K, Kim S, Fritz JV, Wilmes P, Ueha S, Matsushima K, Ohno H, Olle B, Sakaguchi S, Taniguchi T, Morita H, *Hattori M, *Honda K.

Nature 500 232-236 2014年03月

研究論文（学術雑誌）, 共著, 査読有り

Selection of an aptamer against mouse GP2 by SELEX.

Hanazato M, Nakato G, Nishikawa F, Hase K, Nishikawa S and Ohno H.

Cell Struct. Funct. 39 23 - 29 2014年01月

研究論文（学術雑誌）, 共著, 査読有り

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Ubiquitin-specific protease 2-69 in macrophages potentially modulates metainflammation.

Kitamura H, Kimura S, Shimamoto Y, Okabe J, Ito M, Miyamoto T, Naoe Y,Kikuguchi C, Meek B, Toda C, Okamoto S, Kanehira K, Hase K, Watarai H, IshizukaM, El-Osta A, Ohara O, Miyoshi I.

FASEB J. 27 4940 - 4953 2013年12月

研究論文（学術雑誌）, 共著, 査読有り

AP-1B-mediated protein sorting regulates polarity and proliferation of intestinal epithelial cells in mice.

Hase K, Nakatsu F, Ohmae M, Sugihara K, Shioda N, Takahashi D, Obata Y, Furusawa Y, Fujimura Y, Yamashita T, Fukuda S, Okamoto H, Asano M, Yonemura S and *Ohno H.

Gastroenterology 145 625-635 2013年08月

研究論文（学術雑誌）, 共著, 査読有り

Distinct Roles for CXCR6+ and CXCR6- CD4+ T Cells in the Pathogenesis of Chronic Colitis.

18 Mandai Y, Takahashi D, *Hase K, Obata Y, Furusawa Y, Ebisawa M, Nakagawa T, Sato T, Katsuno T, Saito Y, Shimaoka T, Yokosuka O, Yokote K, Ohno H.

PLoS One 19 e65488 2013年06月

研究論文（学術雑誌）, 共著, 査読有り

Commensal microbe-derived butyrate induces colonic regulatory T cells.

Furusawa Y, Obata Y, *Fukuda S, Endo TA, Nakato G, Takahashi D, Nakanishi Y, Uetake C, Kato K, Kato T, Takahashi M, Fukuda NR, Murakami M, Miyauchi E, Hino S, Atarashi K, Onawa S, Fujimura Y, Lockett T, Clarke JM, Topping DL, Tomita M, Hori S, Ohara O, Morita T, Koseki H, Kikuchi J, Honda K, *Hase K and *Ohno H.

Nature 500 232-236 2013年06月

研究論文（学術雑誌）, 共著, 査読有り

Microbiota-derived lactate accelerates colon epithelial cell turnover in starvation-refed mice.

Okada T, Fukuda S, Hase K, Nishiumi S, Izumi Y, Yoshida M, Hagiwara T, Kawashima R, Yamazaki M, Oshio T, Otsubo T, Inagaki-Ohara K, Kakimoto K, Higuchi K, Kawamura Y,I,, Ohno H and *Dohi H.

Nat Commun 4 1654 2013年03月

研究論文（学術雑誌）, 共著, 査読有り

The Ets Transcription factor Spi-B is essential for the differentiation of intestinal microfold (M) Cells.

Kanaya T, Hase K, Takahashi D, Fukuda S, Hoshino K, Sasaki I, Hemmi H, Knoop KA, Kumar N, Sato M, Katsuno T, Yokosuka O, Toyooka K, Nakai K, Sakamoto A, Kitahara Y, Jinnohara T, McSorley SJ, *Kaisho T, *Williams IR and *Ohno H.

Nat Immunol 13 729-736 2012年12月

研究論文（学術雑誌）, 共著, 査読有り

Critical role of the IgM Fc receptor in IgM homeostasis, B-cell survival, and humoral immune responses.

Ouchida R, Mori H, Hase K, Takatsu H, Kurosaki T, Tokuhisa T, *Ohno H and *Wang J-Y.

Proc Natl Acad Sci USA 109 E2699-E2706 2012年11月

研究論文（学術雑誌）, 共著, 査読有り

Cutting Edge: Brucella abortus exploits a cellular prion protein on intestinal M cells as an invasive receptor.

20 Nakato G, Hase K, Suzuki M, Kimura M, Ato M, Hanazato M, Tobiume M, Horiuchi M, Atarashi R, Nishida N, Watarai M, Imaoka K and Ohno H.

J. Immunol. 189 1540 - 1544 2012年08月

研究論文（学術雑誌）, 査読有り

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AP1B plays an important role in intestinal tumorigenesis with the truncating mutation of an APC gene.

25 Mimura M, Masuda A, Nishiumi S, Kawakami K, Fujishima Y, Yoshie T, Mizuno S, Miki I, Ohno H, Hase K, Minamoto T, Azuma T and Yoshida M.

Int. J. Cancer 130 1011 - 1020 2012年04月

研究論文（学術雑誌）, 共著

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Epithelial cell-intrinsic Notch signaling plays an essential role in the maintenance of gut immune homeostasis.

Obata Y, Takahashi D, Ebisawa M, Kakiguchi M, Yonemura S, Jinnohara T, Kanaya T, Fujimura Y, Ohmae M, Hase K and Ohno H

J. Immunol. 188 2427 - 2436 2012年01月

共著, 査読有り

The Epithelia-specific membrane trafficking factor AP-1B controls gut immune homeostasis in mice.

Takahashi D, *Hase K , Kimura S, Nakatsu F, Ohmae M, Mandai Y, Sato T, Date Y, Ebisawa M, Kato T, Obata Y, Fukuda S, Kawamura Y, I, Dohi T, Katsuno T, Yokosuka O, Waguri S and Ohno H.

Gastroenterology 141 621-632 2011年12月

研究論文（学術雑誌）, 共著, 査読有り

Bifidobacteria protect host from enteropathogenic infection through production of acetate.

Fukuda S, Toh H, Hase K, Oshima K, Nakanishi Y, Yoshimura K, Tobe T, Clarke JM, Topping DM, Suzuki T, Taylor TD, Itoh K, Kikuchi J, Morita H, *Hattori M, *Ohno H.

Nature 469 543-547 2011年12月

研究論文（学術雑誌）, 共著, 査読有り

Uptake through glycoprotein 2 of FimH(+) bacteria by M cells initiates mucosal immune response.

Hase K, Kawano K, Nochi T, Pontes GS, Fukuda S, Ebisawa M, Kadokura K, Tobe T, Fujimura Y, Kawano S, Yabashi A, Waguri S, Nakato G, Kimura S, Murakami T, Iimura M, Hamura K, Fukuoka S, Lowe AW, Itoh K, Kiyono H, *Ohno H.

Nature 462 226-230 2009年12月

研究論文（学術雑誌）, 共著, 査読有り

M-Sec promotes membrane nanotube formation by interacting with Ral and the exocyst complex.

Hase K, Kimura S, Takatsu H, Ohmae M, Kawano S, Kitamura H, Ito M, Watarai H, Hazelett CC, Yeaman C and *Ohno H.

Nat Cell Biol 11 1427-1432 2009年12月

研究論文（学術雑誌）, 共著, 査読有り

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