{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:41:05Z","timestamp":1768506065313,"version":"3.49.0"},"posted":{"date-parts":[[2023,4,22]]},"group-title":"Molecular Biology","reference-count":39,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,4,22]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background and aims<\/jats:title>\n                  <jats:p>WNT2B is a canonical Wnt ligand previously thought to be fully redundant with other Wnts in the intestinal epithelium. However, humans with WNT2B deficiency have severe intestinal disease, highlighting a critical role for WNT2B. We sought to understand how WNT2B contributes to intestinal homeostasis.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    We investigated the intestinal health of\n                    <jats:italic>Wnt2b<\/jats:italic>\n                    knock out (KO) mice. We assessed the impact of inflammatory challenge to the small intestine, using anti-CD3\u03c7 antibody, and to the colon, using dextran sodium sulfate (DSS). In addition, we generated human intestinal organoids (HIOs) from WNT2B-deficient human iPSCs for transcriptional and histological analyses.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Mice with WNT2B deficiency had significantly decreased\n                    <jats:italic>Lgr5<\/jats:italic>\n                    expression in the small intestine and profoundly decreased expression in the colon, but normal baseline histology. The small intestinal response to anti-CD3\u03c7 antibody was similar in\n                    <jats:italic>Wnt2b<\/jats:italic>\n                    KO and wild type (WT) mice. In contrast, the colonic response to DSS in\n                    <jats:italic>Wnt2b<\/jats:italic>\n                    KO mice showed an accelerated rate of injury, featuring earlier immune cell infiltration and loss of differentiated epithelium compared to WT. WNT2B-deficient HIOs showed abnormal epithelial organization and an increased mesenchymal gene signature.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>WNT2B contributes to maintenance of the intestinal stem cell pool in mice and humans. WNT2B deficient mice, which do not have a developmental phenotype, show increased susceptibility to colonic injury but not small intestinal injury, potentially due to a higher reliance on WNT2B in the colon compared to the small intestine.<\/jats:p>\n                  <jats:p>WNT2B deficiency causes a developmental phenotype in human intestine with HIOs showing a decrease in their mesenchymal component and WNT2B-deficient patients showing epithelial disorganization.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Data Transparency Statement<\/jats:title>\n                  <jats:p>All RNA-Seq data will be available through online repository as indicated in Transcript profiling. Any other data will be made available upon request by emailing the study authors.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.04.21.537894","type":"posted-content","created":{"date-parts":[[2023,4,23]],"date-time":"2023-04-23T01:50:13Z","timestamp":1682214613000},"source":"Crossref","is-referenced-by-count":1,"title":["WNT2B Deficiency Causes Increased Susceptibility to Colitis in Mice and Impairs Intestinal Epithelial Development in Humans"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9298-6991","authenticated-orcid":false,"given":"Amy E.","family":"O\u2019Connell","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sathuwarman","family":"Raveenthiraraj","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Comfort","family":"Adegboye","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanshu","family":"Qi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Radhika S.","family":"Khetani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akaljot","family":"Singh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nambirajam","family":"Sundaram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chidera","family":"Emeonye","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jasmine","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey D.","family":"Goldsmith","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jay R.","family":"Thiagarajah","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Diana L.","family":"Carlone","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0627-9455","authenticated-orcid":false,"given":"Jerrold R.","family":"Turner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pankaj B.","family":"Agrawal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Helmrath","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David T.","family":"Breault","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023042608202874000_2023.04.21.537894v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.05.012"},{"key":"2023042608202874000_2023.04.21.537894v1.2","doi-asserted-by":"publisher","DOI":"10.1101\/gad.235473.113"},{"key":"2023042608202874000_2023.04.21.537894v1.3","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.stem.2020.01.008","article-title":"Distinct Mesenchymal Cell Populations Generate the Essential Intestinal BMP Signaling Gradient","volume":"26","year":"2020","journal-title":"Cell Stem Cell"},{"key":"2023042608202874000_2023.04.21.537894v1.4","doi-asserted-by":"crossref","unstructured":"Shoshkes-Carmel M , Wang YJ , Wangensteen KJ , et al. Subepithelial telocytes are an important source of Wnts that supports intestinal crypts. Nature 2018.","DOI":"10.1038\/s41586-018-0084-4"},{"key":"2023042608202874000_2023.04.21.537894v1.5","doi-asserted-by":"crossref","unstructured":"Br\u00fcgger MD , Valenta T , Fazilaty H , et al. Distinct populations of crypt-associated fibroblasts act as signaling hubs to control colon homeostasis. PLOS Biology 2020;18:e3001032.","DOI":"10.1371\/journal.pbio.3001032"},{"key":"2023042608202874000_2023.04.21.537894v1.6","doi-asserted-by":"crossref","unstructured":"McCarthy N , Tie G , Madha S , et al. Smooth muscle contributes to the development and function of a layered intestinal stem cell niche. Dev Cell 2023.","DOI":"10.2139\/ssrn.4149949"},{"key":"2023042608202874000_2023.04.21.537894v1.7","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1016\/j.stem.2023.03.004","article-title":"Graded BMP signaling within intestinal crypt architecture directs self-organization of the Wnt-secreting stem cell niche","volume":"30","year":"2023","journal-title":"Cell Stem Cell"},{"key":"2023042608202874000_2023.04.21.537894v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.gastro.2005.06.007"},{"key":"2023042608202874000_2023.04.21.537894v1.9","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2012.08.031"},{"key":"2023042608202874000_2023.04.21.537894v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2016.03.088"},{"key":"2023042608202874000_2023.04.21.537894v1.11","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.stemcr.2013.12.012","article-title":"Wnt secretion from epithelial cells and subepithelial myofibroblasts is not required in the mouse intestinal stem cell niche in vivo","volume":"2","year":"2014","journal-title":"Stem Cell Reports"},{"key":"2023042608202874000_2023.04.21.537894v1.12","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.ajhg.2018.05.007","article-title":"Neonatal-Onset Chronic Diarrhea Caused by Homozygous Nonsense WNT2B Mutations","volume":"103","year":"2018","journal-title":"Am J Hum Genet"},{"key":"2023042608202874000_2023.04.21.537894v1.13","doi-asserted-by":"crossref","first-page":"998","DOI":"10.1038\/s41431-021-00812-1","article-title":"Novel variants in the stem cell niche factor WNT2B define the disease phenotype as a congenital enteropathy with ocular dysgenesis","volume":"29","year":"2021","journal-title":"Eur J Hum Genet"},{"key":"2023042608202874000_2023.04.21.537894v1.14","doi-asserted-by":"publisher","DOI":"10.1002\/9780470942390.mo130179"},{"key":"2023042608202874000_2023.04.21.537894v1.15","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2011.07.050"},{"key":"2023042608202874000_2023.04.21.537894v1.16","doi-asserted-by":"publisher","DOI":"10.1038\/nature07935"},{"key":"2023042608202874000_2023.04.21.537894v1.17","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1007\/978-1-4939-7021-6_7","article-title":"Establishment of 3D Intestinal Organoid Cultures from Intestinal Stem Cells","volume":"1612","year":"2017","journal-title":"Methods Mol Biol"},{"key":"2023042608202874000_2023.04.21.537894v1.18","doi-asserted-by":"publisher","DOI":"10.1016\/j.stem.2017.05.020"},{"key":"2023042608202874000_2023.04.21.537894v1.19","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1007\/978-1-4939-6949-4_12","article-title":"Generation of Gastrointestinal Organoids from Human Pluripotent Stem Cells","volume":"1597","year":"2017","journal-title":"Methods Mol Biol"},{"key":"2023042608202874000_2023.04.21.537894v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/nature09691"},{"key":"2023042608202874000_2023.04.21.537894v1.21","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4197"},{"key":"2023042608202874000_2023.04.21.537894v1.22","doi-asserted-by":"crossref","unstructured":"Love MI , Huber W , Anders S . Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 2014;15:550.","DOI":"10.1186\/s13059-014-0550-8"},{"key":"2023042608202874000_2023.04.21.537894v1.23","doi-asserted-by":"publisher","DOI":"10.1089\/omi.2011.0118"},{"key":"2023042608202874000_2023.04.21.537894v1.24","doi-asserted-by":"crossref","unstructured":"Wu T , Hu E , Xu S , et al. clusterProfiler 4.0: A universal enrichment tool for interpreting omics data. Innovation (Camb) 2021;2:100141.","DOI":"10.1016\/j.xinn.2021.100141"},{"key":"2023042608202874000_2023.04.21.537894v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.10.118"},{"key":"2023042608202874000_2023.04.21.537894v1.26","doi-asserted-by":"crossref","unstructured":"Eriguchi Y , Nakamura K , Yokoi Y , et al. Essential role of IFN-gamma in T cell- associated intestinal inflammation. JCI Insight 2018;3.","DOI":"10.1172\/jci.insight.121886"},{"key":"2023042608202874000_2023.04.21.537894v1.27","doi-asserted-by":"crossref","unstructured":"In JG , Yin J , Atanga R , et al. Epithelial WNT2B and Desert Hedgehog Are Necessary for Human Colonoid Regeneration after Bacterial Cytotoxin Injury. iScience 2020;23:101618.","DOI":"10.1016\/j.isci.2020.101618"},{"key":"2023042608202874000_2023.04.21.537894v1.28","doi-asserted-by":"publisher","DOI":"10.1002\/eji.1830200308"},{"key":"2023042608202874000_2023.04.21.537894v1.29","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.stemcr.2017.11.015","article-title":"JAK\/STAT-1 Signaling Is Required for Reserve Intestinal Stem Cell Activation during Intestinal Regeneration Following Acute Inflammation","volume":"10","year":"2018","journal-title":"Stem Cell Reports"},{"key":"2023042608202874000_2023.04.21.537894v1.30","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1093\/ecco-jcc\/jjy035","article-title":"Development of Reliable, Valid and Responsive Scoring Systems for Endoscopy and Histology in Animal Models for Inflammatory Bowel Disease","volume":"12","year":"2018","journal-title":"Journal of Crohn\u2019s and Colitis"},{"key":"2023042608202874000_2023.04.21.537894v1.31","doi-asserted-by":"crossref","unstructured":"Castillo-Azofeifa D , Fazio EN , Nattiv R , et al. Atoh1+ secretory progenitors possess renewal capacity independent of Lgr5+ cells during colonic regeneration. The EMBO Journal 2019;38:e99984.","DOI":"10.15252\/embj.201899984"},{"key":"2023042608202874000_2023.04.21.537894v1.32","doi-asserted-by":"crossref","unstructured":"Hu L , Xu J , Wang X , et al. Bone Morphogenetic Protein 4 Alleviates DSS- Induced Ulcerative Colitis Through Activating Intestinal Stem Cell by Target ID3. Front Cell Dev Biol 2021;9:700864.","DOI":"10.3389\/fcell.2021.700864"},{"key":"2023042608202874000_2023.04.21.537894v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2012.01.062"},{"key":"2023042608202874000_2023.04.21.537894v1.34","first-page":"230","article-title":"WNT2b Activates Epithelial- mesenchymal Transition Through FZD4: Relevance in Penetrating Croh\u0144s Disease","volume":"14","year":"2019","journal-title":"Journal of Crohn\u2019s and Colitis"},{"key":"2023042608202874000_2023.04.21.537894v1.35","doi-asserted-by":"publisher","DOI":"10.1158\/1541-7786.MCR-10-0568"},{"key":"2023042608202874000_2023.04.21.537894v1.36","doi-asserted-by":"publisher","DOI":"10.15252\/embj.201591517"},{"key":"2023042608202874000_2023.04.21.537894v1.37","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.cell.2022.11.001","article-title":"Hallmarks of aging: An expanding universe","volume":"186","year":"2023","journal-title":"Cell"},{"key":"2023042608202874000_2023.04.21.537894v1.38","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1016\/j.stem.2022.06.012","article-title":"Inflammatory exposure drives long- lived impairment of hematopoietic stem cell self-renewal activity and accelerated aging","volume":"29","year":"2022","journal-title":"Cell Stem Cell"},{"key":"2023042608202874000_2023.04.21.537894v1.39","doi-asserted-by":"crossref","unstructured":"He D , Wu H , Xiang J , et al. Gut stem cell aging is driven by mTORC1 via a p38 MAPK-p53 pathway. Nature Communications 2020;11:37.","DOI":"10.1038\/s41467-019-13911-x"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.04.21.537894","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T04:34:56Z","timestamp":1768451696000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.04.21.537894"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,22]]},"references-count":39,"URL":"https:\/\/doi.org\/10.1101\/2023.04.21.537894","relation":{},"subject":[],"published":{"date-parts":[[2023,4,22]]},"subtype":"preprint"}}