{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T06:44:43Z","timestamp":1790750683169,"version":"4.1.0"},"reference-count":35,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,7,16]],"date-time":"2018-07-16T00:00:00Z","timestamp":1531699200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2018,7,16]],"date-time":"2018-07-16T00:00:00Z","timestamp":1531699200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The R-Spondin (R-Spo) family regulates WNT signaling and stimulates the proliferation and differentiation of intestinal stem cells (ISCs). R-Spo plays a critical role in maintaining intestinal homeostasis, but endogenous producers of R-Spo in the intestine remain to be investigated. We found that R-Spo3 was the major R-Spo family member produced in the intestine and it was predominantly produced by CD45<jats:sup>\u2212<\/jats:sup>CD90<jats:sup>+<\/jats:sup>CD31<jats:sup>+<\/jats:sup> lymphatic endothelial cells (LECs) in the lamina propria of the intestinal mucosa. Transcriptome analysis demonstrated that LECs highly expressed R-Spo receptor, <jats:italic>Lgr5<\/jats:italic>, suggesting an autocrine stimulatory loop in LECs. LECs were significantly reduced in number, and their R-Spo3 production was impaired in intestinal graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation. The impaired production of R-Spo3 in the intestine may be a novel mechanism of delayed tissue repair and defective mucosal defense in intestinal GVHD. We demonstrate a novel role of intestinal LECs in producing R-Spondin3 to maintain intestinal homeostasis.<\/jats:p>","DOI":"10.1038\/s41598-018-29100-7","type":"journal-article","created":{"date-parts":[[2018,7,10]],"date-time":"2018-07-10T11:41:53Z","timestamp":1531222913000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":44,"title":["Intestinal Lymphatic Endothelial Cells Produce R-Spondin3"],"prefix":"10.1038","volume":"8","author":[{"given":"Reiki","family":"Ogasawara","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9489-9704","authenticated-orcid":false,"given":"Daigo","family":"Hashimoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shunsuke","family":"Kimura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eiko","family":"Hayase","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahide","family":"Ara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuichiro","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyuki","family":"Ohigashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kosuke","family":"Yoshioka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takahiro","family":"Tateno","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emi","family":"Yokoyama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ko","family":"Ebata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takeshi","family":"Kondo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junichi","family":"Sugita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masahiro","family":"Onozawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toshihiko","family":"Iwanaga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0941-271X","authenticated-orcid":false,"given":"Takanori","family":"Teshima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,7,16]]},"reference":[{"key":"29100_CR1","doi-asserted-by":"publisher","first-page":"262","DOI":"10.1038\/nature07935","volume":"459","author":"T Sato","year":"2009","unstructured":"Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262\u2013265, https:\/\/doi.org\/10.1038\/nature07935 (2009).","journal-title":"Nature"},{"key":"29100_CR2","doi-asserted-by":"publisher","first-page":"1003","DOI":"10.1038\/nature06196","volume":"449","author":"N Barker","year":"2007","unstructured":"Barker, N. et al. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449, 1003\u20131007, https:\/\/doi.org\/10.1038\/nature06196 (2007).","journal-title":"Nature"},{"key":"29100_CR3","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1038\/nrm3721","volume":"15","author":"N Barker","year":"2014","unstructured":"Barker, N. Adult intestinal stem cells: critical drivers of epithelial homeostasis and regeneration. Nat Rev Mol Cell Biol 15, 19\u201333, https:\/\/doi.org\/10.1038\/nrm3721 (2014).","journal-title":"Nat Rev Mol Cell Biol"},{"key":"29100_CR4","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1084\/jem.20101559","volume":"208","author":"S Takashima","year":"2011","unstructured":"Takashima, S. et al. The Wnt agonist R-spondin1 regulates systemic graft-versus-host disease by protecting intestinal stem cells. The Journal of experimental medicine 208, 285\u2013294, https:\/\/doi.org\/10.1084\/jem.20101559 (2011).","journal-title":"The Journal of experimental medicine"},{"key":"29100_CR5","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1016\/j.immuni.2012.05.028","volume":"37","author":"AM Hanash","year":"2012","unstructured":"Hanash, A. M. et al. Interleukin-22 protects intestinal stem cells from immune-mediated tissue damage and regulates sensitivity to graft versus host disease. Immunity 37, 339\u2013350, https:\/\/doi.org\/10.1016\/j.immuni.2012.05.028 (2012).","journal-title":"Immunity"},{"key":"29100_CR6","doi-asserted-by":"publisher","first-page":"767","DOI":"10.1038\/nrm3470","volume":"13","author":"C Niehrs","year":"2012","unstructured":"Niehrs, C. The complex world of WNT receptor signalling. Nat Rev Mol Cell Biol 13, 767\u2013779, https:\/\/doi.org\/10.1038\/nrm3470 (2012).","journal-title":"Nat Rev Mol Cell Biol"},{"key":"29100_CR7","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2012-13-3-242","volume":"13","author":"WB de Lau","year":"2012","unstructured":"de Lau, W. B., Snel, B. & Clevers, H. C. The R-spondin protein family. Genome Biol 13, 242, https:\/\/doi.org\/10.1186\/gb-2012-13-3-242 (2012).","journal-title":"Genome Biol"},{"key":"29100_CR8","doi-asserted-by":"publisher","first-page":"1256","DOI":"10.1126\/science.1112521","volume":"309","author":"KA Kim","year":"2005","unstructured":"Kim, K. A. et al. Mitogenic influence of human R-spondin1 on the intestinal epithelium. Science 309, 1256\u20131259 (2005).","journal-title":"Science"},{"key":"29100_CR9","doi-asserted-by":"publisher","first-page":"3507","DOI":"10.1084\/jem.20170418","volume":"214","author":"E Hayase","year":"2017","unstructured":"Hayase, E. et al. R-Spondin1 expands Paneth cells and prevents dysbiosis induced by graft-versus-host disease. The Journal of experimental medicine 214, 3507\u20133518, https:\/\/doi.org\/10.1084\/jem.20170418 (2017).","journal-title":"The Journal of experimental medicine"},{"key":"29100_CR10","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1038\/nature16466","volume":"529","author":"EE Storm","year":"2016","unstructured":"Storm, E. E. et al. Targeting PTPRK-RSPO3 colon tumours promotes differentiation and loss of stem-cell function. Nature 529, 97\u2013100, https:\/\/doi.org\/10.1038\/nature16466 (2016).","journal-title":"Nature"},{"key":"29100_CR11","doi-asserted-by":"publisher","first-page":"310","DOI":"10.1007\/s11894-010-0135-y","volume":"12","author":"IV Pinchuk","year":"2010","unstructured":"Pinchuk, I. V., Mifflin, R. C., Saada, J. I. & Powell, D. W. Intestinal mesenchymal cells. Curr Gastroenterol Rep 12, 310\u2013318, https:\/\/doi.org\/10.1007\/s11894-010-0135-y (2010).","journal-title":"Curr Gastroenterol Rep"},{"key":"29100_CR12","doi-asserted-by":"publisher","first-page":"2982","DOI":"10.1016\/j.yexcr.2010.06.013","volume":"316","author":"G Jurisic","year":"2010","unstructured":"Jurisic, G., Iolyeva, M., Proulx, S. T., Halin, C. & Detmar, M. Thymus cell antigen 1 (Thy1, CD90) is expressed by lymphatic vessels and mediates cell adhesion to lymphatic endothelium. Exp Cell Res 316, 2982\u20132992, https:\/\/doi.org\/10.1016\/j.yexcr.2010.06.013 (2010).","journal-title":"Exp Cell Res"},{"key":"29100_CR13","doi-asserted-by":"publisher","first-page":"1819","DOI":"10.1002\/eji.1830181125","volume":"18","author":"MA Jutila","year":"1988","unstructured":"Jutila, M. A. et al. Ly-6C is a monocyte\/macrophage and endothelial cell differentiation antigen regulated by interferon-gamma. Eur J Immunol 18, 1819\u20131826, https:\/\/doi.org\/10.1002\/eji.1830181125 (1988).","journal-title":"Eur J Immunol"},{"key":"29100_CR14","doi-asserted-by":"publisher","first-page":"238","DOI":"10.1038\/nature22313","volume":"545","author":"KS Yan","year":"2017","unstructured":"Yan, K. S. et al. Non-equivalence of Wnt and R-spondin ligands during Lgr5(+) intestinal stem-cell self-renewal. Nature 545, 238\u2013242, https:\/\/doi.org\/10.1038\/nature22313 (2017).","journal-title":"Nature"},{"key":"29100_CR15","doi-asserted-by":"publisher","first-page":"2522","DOI":"10.15252\/embj.201490583","volume":"34","author":"ES Demitrack","year":"2015","unstructured":"Demitrack, E. S. et al. Notch signaling regulates gastric antral LGR5 stem cell function. EMBO J 34, 2522\u20132536, https:\/\/doi.org\/10.15252\/embj.201490583 (2015).","journal-title":"EMBO J"},{"key":"29100_CR16","doi-asserted-by":"publisher","first-page":"2471","DOI":"10.1038\/nprot.2013.153","volume":"8","author":"H Miyoshi","year":"2013","unstructured":"Miyoshi, H. & Stappenbeck, T. S. In vitro expansion and genetic modification of gastrointestinal stem cells in spheroid culture. Nat Protoc 8, 2471\u20132482, https:\/\/doi.org\/10.1038\/nprot.2013.153 (2013).","journal-title":"Nat Protoc"},{"key":"29100_CR17","doi-asserted-by":"publisher","first-page":"2206","DOI":"10.1242\/dev.104976","volume":"141","author":"Z Kabiri","year":"2014","unstructured":"Kabiri, Z. et al. Stroma provides an intestinal stem cell niche in the absence of epithelial Wnts. Development 141, 2206\u20132215, https:\/\/doi.org\/10.1242\/dev.104976 (2014).","journal-title":"Development"},{"key":"29100_CR18","doi-asserted-by":"publisher","first-page":"e84651","DOI":"10.1371\/journal.pone.0084651","volume":"9","author":"NY Lei","year":"2014","unstructured":"Lei, N. Y. et al. Intestinal subepithelial myofibroblasts support the growth of intestinal epithelial stem cells. PloS one 9, e84651, https:\/\/doi.org\/10.1371\/journal.pone.0084651 (2014).","journal-title":"PloS one"},{"key":"29100_CR19","doi-asserted-by":"publisher","first-page":"e26898","DOI":"10.1371\/journal.pone.0026898","volume":"6","author":"N Lahar","year":"2011","unstructured":"Lahar, N. et al. Intestinal subepithelial myofibroblasts support in vitro and in vivo growth of human small intestinal epithelium. PloS one 6, e26898, https:\/\/doi.org\/10.1371\/journal.pone.0026898 (2011).","journal-title":"PloS one"},{"key":"29100_CR20","doi-asserted-by":"publisher","first-page":"e0152859","DOI":"10.1371\/journal.pone.0152859","volume":"11","author":"E Kang","year":"2016","unstructured":"Kang, E., Yousefi, M. & Gruenheid, S. R-Spondins Are Expressed by the Intestinal Stroma and are Differentially Regulated during Citrobacter rodentium- and DSS-Induced Colitis in Mice. PloS one 11, e0152859, https:\/\/doi.org\/10.1371\/journal.pone.0152859 (2016).","journal-title":"PloS one"},{"key":"29100_CR21","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1038\/nature23642","volume":"548","author":"M Sigal","year":"2017","unstructured":"Sigal, M. et al. Stromal R-spondin orchestrates gastric epithelial stem cells and gland homeostasis. Nature 548, 451\u2013455, https:\/\/doi.org\/10.1038\/nature23642 (2017).","journal-title":"Nature"},{"key":"29100_CR22","doi-asserted-by":"publisher","first-page":"14348","DOI":"10.1073\/pnas.1309393110","volume":"110","author":"L Kannan","year":"2013","unstructured":"Kannan, L. et al. R-spondin3 prevents mesenteric ischemia\/reperfusion-induced tissue damage by tightening endothelium and preventing vascular leakage. Proceedings of the National Academy of Sciences of the United States of America 110, 14348\u201314353, https:\/\/doi.org\/10.1073\/pnas.1309393110 (2013).","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"29100_CR23","doi-asserted-by":"publisher","first-page":"1757","DOI":"10.1016\/j.celrep.2015.10.049","volume":"13","author":"AS Rocha","year":"2015","unstructured":"Rocha, A. S. et al. The Angiocrine Factor Rspondin3 Is a Key Determinant of Liver Zonation. Cell Rep 13, 1757\u20131764, https:\/\/doi.org\/10.1016\/j.celrep.2015.10.049 (2015).","journal-title":"Cell Rep"},{"key":"29100_CR24","doi-asserted-by":"publisher","first-page":"2320","DOI":"10.1016\/j.celrep.2017.02.036","volume":"18","author":"F Lacour","year":"2017","unstructured":"Lacour, F. et al. R-spondin1 Controls Muscle Cell Fusion through Dual Regulation of Antagonistic Wnt Signaling Pathways. Cell Rep 18, 2320\u20132330, https:\/\/doi.org\/10.1016\/j.celrep.2017.02.036 (2017).","journal-title":"Cell Rep"},{"key":"29100_CR25","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1016\/j.cell.2017.07.028","volume":"170","author":"JH Lee","year":"2017","unstructured":"Lee, J. H. et al. Anatomically and Functionally Distinct Lung Mesenchymal Populations Marked by Lgr5 and Lgr6. Cell 170, 1149\u20131163 e1112, https:\/\/doi.org\/10.1016\/j.cell.2017.07.028 (2017).","journal-title":"Cell"},{"key":"29100_CR26","doi-asserted-by":"publisher","first-page":"2151","DOI":"10.1182\/blood-2013-01-478941","volume":"122","author":"JY Jang","year":"2013","unstructured":"Jang, J. Y. et al. Conditional ablation of LYVE-1\u2009+\u2009cells unveils defensive roles of lymphatic vessels in intestine and lymph nodes. Blood 122, 2151\u20132161, https:\/\/doi.org\/10.1182\/blood-2013-01-478941 (2013).","journal-title":"Blood"},{"key":"29100_CR27","doi-asserted-by":"publisher","first-page":"3863","DOI":"10.1172\/JCI72189","volume":"124","author":"S D\u2019Alessio","year":"2014","unstructured":"D\u2019Alessio, S. et al. VEGF-C-dependent stimulation of lymphatic function ameliorates experimental inflammatory bowel disease. The Journal of clinical investigation 124, 3863\u20133878, https:\/\/doi.org\/10.1172\/JCI72189 (2014).","journal-title":"The Journal of clinical investigation"},{"key":"29100_CR28","doi-asserted-by":"publisher","first-page":"1865","DOI":"10.1182\/blood-2016-08-734210","volume":"129","author":"S Mertlitz","year":"2017","unstructured":"Mertlitz, S. et al. Lymphangiogenesis is a feature of acute GVHD, and VEGFR-3 inhibition protects against experimental GVHD. Blood 129, 1865\u20131875, https:\/\/doi.org\/10.1182\/blood-2016-08-734210 (2017).","journal-title":"Blood"},{"key":"29100_CR29","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1182\/blood-2011-12-401166","volume":"120","author":"Y Eriguchi","year":"2012","unstructured":"Eriguchi, Y. et al. Graft-versus-host disease disrupts intestinal microbial ecology by inhibiting Paneth cell production of alpha-defensins. Blood 120, 223\u2013231, https:\/\/doi.org\/10.1182\/blood-2011-12-401166 (2012).","journal-title":"Blood"},{"key":"29100_CR30","doi-asserted-by":"publisher","first-page":"e8014","DOI":"10.1371\/journal.pone.0008014","volume":"4","author":"P Bhanja","year":"2009","unstructured":"Bhanja, P. et al. Protective role of R-spondin1, an intestinal stem cell growth factor, against radiation-induced gastrointestinal syndrome in mice. PloS one 4, e8014, https:\/\/doi.org\/10.1371\/journal.pone.0008014 (2009).","journal-title":"PloS one"},{"key":"29100_CR31","doi-asserted-by":"publisher","first-page":"1331","DOI":"10.1053\/j.gastro.2007.02.001","volume":"132","author":"J Zhao","year":"2007","unstructured":"Zhao, J. et al. R-spondin1, a novel intestinotrophic mitogen, ameliorates experimental colitis in mice. Gastroenterology 132, 1331\u20131343, https:\/\/doi.org\/10.1053\/j.gastro.2007.02.001 (2007).","journal-title":"Gastroenterology"},{"key":"29100_CR32","doi-asserted-by":"publisher","first-page":"807","DOI":"10.1038\/bmt.2008.261","volume":"42","author":"B Sadeghi","year":"2008","unstructured":"Sadeghi, B. et al. GVHD after chemotherapy conditioning in allogeneic transplanted mice. Bone Marrow Transplant 42, 807\u2013818, https:\/\/doi.org\/10.1038\/bmt.2008.261 (2008).","journal-title":"Bone Marrow Transplant"},{"key":"29100_CR33","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1016\/j.immuni.2009.08.010","volume":"31","author":"M Bogunovic","year":"2009","unstructured":"Bogunovic, M. et al. Origin of the lamina propria dendritic cell network. Immunity 31, 513\u2013525, https:\/\/doi.org\/10.1016\/j.immuni.2009.08.010 (2009).","journal-title":"Immunity"},{"key":"29100_CR34","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1016\/j.cell.2014.04.050","volume":"158","author":"PA Muller","year":"2014","unstructured":"Muller, P. A. et al. Crosstalk between muscularis macrophages and enteric neurons regulates gastrointestinal motility. Cell 158, 300\u2013313, https:\/\/doi.org\/10.1016\/j.cell.2014.04.050 (2014).","journal-title":"Cell"},{"key":"29100_CR35","doi-asserted-by":"publisher","first-page":"200","DOI":"10.1111\/cas.13129","volume":"108","author":"M Ibata","year":"2017","unstructured":"Ibata, M. et al. Leukemogenic kinase FIP1L1-PDGFRA and a small ubiquitin-like modifier E3 ligase, PIAS1, form a positive cross-talk through their enzymatic activities. Cancer Sci 108, 200\u2013207, https:\/\/doi.org\/10.1111\/cas.13129 (2017).","journal-title":"Cancer Sci"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-29100-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-29100-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-29100-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T07:38:37Z","timestamp":1671608317000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-29100-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,7,16]]},"references-count":35,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["29100"],"URL":"https:\/\/doi.org\/10.1038\/s41598-018-29100-7","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,7,16]]},"assertion":[{"value":"8 January 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"3 July 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"16 July 2018","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"10719"}}