{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,1]],"date-time":"2026-02-01T00:41:58Z","timestamp":1769906518241,"version":"3.49.0"},"reference-count":45,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,15]],"date-time":"2022-12-15T00:00:00Z","timestamp":1671062400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81571396"],"award-info":[{"award-number":["81571396"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81771528"],"award-info":[{"award-number":["81771528"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["ZNJC202013"],"award-info":[{"award-number":["ZNJC202013"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["JCZN2022009"],"award-info":[{"award-number":["JCZN2022009"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Translational Medicine and Interdisciplinary joint funds of Wuhan University Zhongnan Hospital","award":["81571396"],"award-info":[{"award-number":["81571396"]}]},{"name":"Translational Medicine and Interdisciplinary joint funds of Wuhan University Zhongnan Hospital","award":["81771528"],"award-info":[{"award-number":["81771528"]}]},{"name":"Translational Medicine and Interdisciplinary joint funds of Wuhan University Zhongnan Hospital","award":["ZNJC202013"],"award-info":[{"award-number":["ZNJC202013"]}]},{"name":"Translational Medicine and Interdisciplinary joint funds of Wuhan University Zhongnan Hospital","award":["JCZN2022009"],"award-info":[{"award-number":["JCZN2022009"]}]},{"name":"Basic and clinical Medicine incubation funds of Wuhan University","award":["81571396"],"award-info":[{"award-number":["81571396"]}]},{"name":"Basic and clinical Medicine incubation funds of Wuhan University","award":["81771528"],"award-info":[{"award-number":["81771528"]}]},{"name":"Basic and clinical Medicine incubation funds of Wuhan University","award":["ZNJC202013"],"award-info":[{"award-number":["ZNJC202013"]}]},{"name":"Basic and clinical Medicine incubation funds of Wuhan University","award":["JCZN2022009"],"award-info":[{"award-number":["JCZN2022009"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Intrauterine adhesion (IUA) causes menstrual disturbance and infertility. There is no effective treatment available for moderate to severe IUA cases. Stem cell-based therapy has been investigated for treating IUA but is limited in clinical applications due to issues including the precise induction of differentiation, tumorigenesis, and unclear molecular mechanisms. In our recent study, we isolated and expanded the long-term cultures of conditional reprogrammed (CR) mouse endometrial epithelial cells. Treating IUA mice with these CR cells (CRCs) restored the morphology and structure of the endometrium and significantly improved the pregnancy rate. In this study, our data with high-throughput sequencing, CRISPR knockout Ihh\u2212\/\u2212CRCs, and transplantation identified for the first time that the Indian hedgehog (Ihh) gene plays a critical role in the regulation of endometrial epithelial cell proliferation. We also found that aberrant activated Ihh-kr\u00fcppel-like factor 9 (KLF9) signaling contributes to the inhibition of normal progesterone receptor (PR) function in IUA mice. Thus, we hypothesized that inhibition of the Ihh-KLF9 pathway may be a novel strategy to treat IUA. Our data demonstrated that treatment with the hedgehog signaling inhibitor Vismodegib restored the morphology, structure, and microenvironment of the endometrium, and greatly improved the pregnancy rate in IUA mice. This study suggests a promising application of hedgehog inhibitors as a targeted drug in the IUA clinic.<\/jats:p>","DOI":"10.3390\/cells11244053","type":"journal-article","created":{"date-parts":[[2022,12,15]],"date-time":"2022-12-15T02:20:57Z","timestamp":1671070857000},"page":"4053","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["CRC Therapy Identifies Indian Hedgehog Signaling in Mouse Endometrial Epithelial Cells and Inhibition of Ihh-KLF9 as a Novel Strategy for Treating IUA"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2003-5967","authenticated-orcid":false,"given":"Xinhao","family":"Zhou","sequence":"first","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"},{"name":"Clinical Laboratory, Wuhan Fourth Hospital, Wuhan 430030, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yiyi","family":"Kang","sequence":"additional","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuntzu","family":"Chang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siyu","family":"Xia","sequence":"additional","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Wu","sequence":"additional","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun","family":"Liu","sequence":"additional","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dirong","family":"Dong","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Zhongnan Hospital of Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3867-7685","authenticated-orcid":false,"given":"Hui","family":"Li","sequence":"additional","affiliation":[{"name":"Taikang Medical School (School of Basic Medical Sciences), Wuhan University, Wuhan 430071, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"568","DOI":"10.1038\/ncb3516","article-title":"Long-term, hormone-responsive organoid cultures of human endometrium in a chemically defined medium","volume":"19","author":"Turco","year":"2017","journal-title":"Nat. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1149","DOI":"10.1152\/physrev.00031.2019","article-title":"Physiology of the Endometrium and Regulation of Menstruation","volume":"100","author":"Critchley","year":"2020","journal-title":"Physiol. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"543","DOI":"10.2174\/0929867003374958","article-title":"Steroids and the endometrium","volume":"7","author":"Oehler","year":"2000","journal-title":"Curr. Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"T1","DOI":"10.1530\/JME-19-0212","article-title":"90 YEARS OF PROGESTERONE: New insights into progesterone receptor signaling in the endometrium required for embryo implantation","volume":"65","author":"DeMayo","year":"2020","journal-title":"J. Mol. Endocrinol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"191","DOI":"10.2147\/IJWH.S165474","article-title":"Asherman\u2019s syndrome: Current perspectives on diagnosis and management","volume":"11","author":"Dreisler","year":"2019","journal-title":"Int. J. Womens Health"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Alawadhi, F., Du, H., Cakmak, H., and Taylor, H.S. (2014). Bone Marrow-Derived Stem Cell (BMDSC) transplantation improves fertility in a murine model of Asherman\u2019s syndrome. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0096662"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1016\/j.fertnstert.2008.02.096","article-title":"Asherman syndrome--one century later","volume":"89","author":"Yu","year":"2008","journal-title":"Fertil. Steril."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.ajog.2016.05.001","article-title":"Intrauterine adhesion prevention after hysteroscopy: A systematic review and meta-analysis","volume":"215","author":"Healy","year":"2016","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_9","first-page":"Cd011110","article-title":"Anti-adhesion therapy following operative hysteroscopy for treatment of female subfertility","volume":"11","author":"Bosteels","year":"2017","journal-title":"Cochrane Database Syst. Rev."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.rbmo.2010.11.018","article-title":"Management of Asherman\u2019s syndrome","volume":"23","author":"March","year":"2011","journal-title":"Reprod. Biomed. Online"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3386","DOI":"10.1210\/en.2009-1334","article-title":"Reepithelialization of the uterine surface arises from endometrial glands: Evidence from a functional mouse model of breakdown and repair","volume":"151","author":"Ye","year":"2010","journal-title":"Endocrinology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1089\/ten.tea.2014.0052","article-title":"Regeneration of uterine horns in rats using collagen scaffolds loaded with human embryonic stem cell-derived endometrium-like cells","volume":"21","author":"Song","year":"2015","journal-title":"Tissue Eng. Part A"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"808","DOI":"10.1002\/stem.1337","article-title":"Estrogen and progesterone together expand murine endometrial epithelial progenitor cells","volume":"31","author":"Janzen","year":"2013","journal-title":"Stem Cells"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1186\/s13287-022-02860-w","article-title":"Treating intrauterine adhesion using conditionally reprogrammed physiological endometrial epithelial cells","volume":"13","author":"Xia","year":"2022","journal-title":"Stem Cell Res. Ther."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e13262","DOI":"10.1111\/aji.13262","article-title":"Transforming growth factor-beta1 in intrauterine adhesion","volume":"84","author":"Abudukeyoumu","year":"2020","journal-title":"Am. J. Reprod. Immunol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1038\/nprot.2016.174","article-title":"Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens","volume":"12","author":"Liu","year":"2017","journal-title":"Nat. Protoc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1007\/s00335-015-9583-x","article-title":"Creating reference gene annotation for the mouse C57BL6\/J. genome assembly","volume":"26","author":"Mudge","year":"2015","journal-title":"Mamm. Genome"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"D766","DOI":"10.1093\/nar\/gky955","article-title":"GENCODE reference annotation for the human and mouse genomes","volume":"47","author":"Frankish","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1038\/nprot.2016.095","article-title":"Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown","volume":"11","author":"Pertea","year":"2016","journal-title":"Nat. Protoc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: An R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"Omics"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"15267","DOI":"10.18632\/oncotarget.14840","article-title":"Ex vivo 2D and 3D HSV-2 infection model using human normal vaginal epithelial cells","volume":"8","author":"Zhu","year":"2017","journal-title":"Oncotarget"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4682","DOI":"10.1158\/1078-0432.CCR-11-0975","article-title":"Pharmacokinetic-pharmacodynamic analysis of vismodegib in preclinical models of mutational and ligand-dependent Hedgehog pathway activation","volume":"17","author":"Wong","year":"2011","journal-title":"Clin. Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1818","DOI":"10.1016\/j.ymthe.2020.05.015","article-title":"Perivascular stem cell-derived cyclophilin A improves uterine environment with Asherman\u2019s Syndrome via HIF1a-dependent angiogenesis","volume":"28","author":"Park","year":"2020","journal-title":"Mol. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/nrg2484","article-title":"RNA-Seq: A revolutionary tool for transcriptomics","volume":"10","author":"Wang","year":"2009","journal-title":"Nat. Rev. Genet."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1101\/gr.079558.108","article-title":"RNA-seq: An assessment of technical reproducibility and comparison with gene expression arrays","volume":"18","author":"Marioni","year":"2008","journal-title":"Genome Res."},{"key":"ref_26","first-page":"3331","article-title":"Hedgehog signaling regulates epithelial-mesenchymal transition during biliary fibrosis in rodents and humans","volume":"118","author":"Omenetti","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1152\/physrev.00058.2009","article-title":"Mammalian Kr\u00fcppel-like factors in health and diseases","volume":"90","author":"McConnell","year":"2010","journal-title":"Physiol. Rev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1038\/gt.2015.53","article-title":"CRISPR-Cas9-mediated gene knockout in primary human airway epithelial cells reveals a proinflammatory role for MUC18","volume":"22","author":"Chu","year":"2015","journal-title":"Gene Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1095\/biolreprod.109.080259","article-title":"Ablation of Indian hedgehog in the murine uterus results in decreased cell cycle progression, aberrant epidermal growth factor signaling, and increased estrogen signaling","volume":"82","author":"Franco","year":"2010","journal-title":"Biol. Reprod."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1204","DOI":"10.1038\/ng1874","article-title":"Indian hedgehog is a major mediator of progesterone signaling in the mouse uterus","volume":"38","author":"Lee","year":"2006","journal-title":"Nat. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Sigafoos, A.N., Paradise, B.D., and Fernandez-Zapico, M.E. (2021). Hedgehog\/GLI Signaling Pathway: Transduction, Regulation, and Implications for Disease. Cancers, 13.","DOI":"10.3390\/cancers13143410"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1210\/me.2007-0242","article-title":"Kruppel-like factor 9 is a negative regulator of ligand-dependent estrogen receptor alpha signaling in Ishikawa endometrial adenocarcinoma cells","volume":"21","author":"Velarde","year":"2007","journal-title":"Mol. Endocrinol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"5503","DOI":"10.1210\/en.2006-0040","article-title":"Overlapping and distinct expression of progesterone receptors A and B in mouse uterus and mammary gland during the estrous cycle","volume":"147","author":"Mote","year":"2006","journal-title":"Endocrinology"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/0303-7207(95)03674-V","article-title":"Differential gene regulation by estrogen and progesterone in the primate endometrium","volume":"115","author":"Ace","year":"1995","journal-title":"Mol. Cell Endocrinol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1055\/s-0034-1376358","article-title":"Endometrial receptivity and intrauterine adhesive disease","volume":"32","author":"Young","year":"2014","journal-title":"Semin. Reprod. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1002\/hep.26394","article-title":"Hedgehog signaling pathway regulates autophagy in human hepatocellular carcinoma cells","volume":"58","author":"Wang","year":"2013","journal-title":"Hepatology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1530\/REP-20-0172","article-title":"Decreased Indian hedgehog signaling activates autophagy in endometriosis and adenomyosis","volume":"161","author":"Zhou","year":"2021","journal-title":"Reproduction"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2338","DOI":"10.1210\/me.2001-0154","article-title":"Identification of Indian hedgehog as a progesterone-responsive gene in the murine uterus","volume":"16","author":"Takamoto","year":"2002","journal-title":"Mol. Endocrinol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1006\/dbio.2002.0645","article-title":"Indian hedgehog as a progesterone-responsive factor mediating epithelial-mesenchymal interactions in the mouse uterus","volume":"245","author":"Matsumoto","year":"2002","journal-title":"Dev. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"E427","DOI":"10.1210\/jc.2013-3717","article-title":"Suppression of COUP-TFII by proinflammatory cytokines contributes to the pathogenesis of endometriosis","volume":"99","author":"Lin","year":"2014","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"R89","DOI":"10.1530\/JME-14-0310","article-title":"The Kruppel-like factors in female reproductive system pathologies","volume":"54","author":"Simmen","year":"2015","journal-title":"J. Mol. Endocrinol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"29286","DOI":"10.1074\/jbc.M403139200","article-title":"Subfertility, uterine hypoplasia, and partial progesterone resistance in mice lacking the Kruppel-like factor 9\/basic transcription element-binding protein-1 (Bteb1) gene","volume":"279","author":"Simmen","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1210\/endo.143.1.8590","article-title":"Direct interaction of the Kruppel-like family (KLF) member, BTEB1, and PR mediates progesterone-responsive gene expression in endometrial epithelial cells","volume":"143","author":"Zhang","year":"2002","journal-title":"Endocrinology"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.jaad.2013.09.012","article-title":"Expanded access study of patients with advanced basal cell carcinoma treated with the Hedgehog pathway inhibitor, vismodegib","volume":"70","author":"Chang","year":"2014","journal-title":"J. Am.Acad. Dermatol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2502","DOI":"10.1158\/1078-0432.CCR-10-2745","article-title":"Phase I trial of hedgehog pathway inhibitor vismodegib (GDC-0449) in patients with refractory, locally advanced or metastatic solid tumors","volume":"17","author":"LoRusso","year":"2011","journal-title":"Clin. Cancer Res."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/24\/4053\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:41:34Z","timestamp":1760146894000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/24\/4053"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,15]]},"references-count":45,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["cells11244053"],"URL":"https:\/\/doi.org\/10.3390\/cells11244053","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,15]]}}}