{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:59:19Z","timestamp":1760237959179,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,7,23]],"date-time":"2022-07-23T00:00:00Z","timestamp":1658534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of Scientific Research","award":["RC 2005"],"award-info":[{"award-number":["RC 2005"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Children"],"abstract":"<jats:p>TITF1 (Thyroid Transcription Factor-1) is a homeodomain-containing transcription factor. Previous studies showed that Titf1 null mice are characterized by failure of tracheo-oesophageal separation and impaired lung morphogenesis resulting in Pulmonary Hypoplasia (PH). In this study, we aim to evaluate the role of TITF1 in the pathogenesis of congenital diaphragmatic hernia (CDH) in humans. We investigated TITF1 expression in human trachea and lungs and performed direct mutation analysis in a CDH population. We studied 13 human fetuses at 14 to 24 weeks of gestation. Five \u03bcm sections were fixed in paraformaldehyde and incubated with anti-TITF1 primary antibody. Positive staining was visualized by biotinylated secondary antibody. We also performed TITF1 screening on genomic DNA extracted from peripheral blood of 16 patients affected by CDH and different degrees of PH, searching for mutations, insertions, and\/or deletions, by sequencing the exonic regions of the gene. Histochemical studies showed positive brown staining of fetal follicular thyroid epithelium, normal fetal trachea, and normal fetal lung bronchial epithelium. Fetal esophageal wall was immunohistochemically negative. Molecular genetic analysis showed complete identity between the sequences obtained and the Wild Type (WT) form of the gene in all cases. No mutation, insertion and\/or deletion was detected. Although TITF1 is expressed in the human fetal lung and has been considered to have a role in the pathogenesis of PH in CDH, the results of our study do not support the hypothesis that TITF1 mutations play a key role in the etiopathogenesis of CDH.<\/jats:p>","DOI":"10.3390\/children9081108","type":"journal-article","created":{"date-parts":[[2022,7,24]],"date-time":"2022-07-24T22:49:02Z","timestamp":1658702942000},"page":"1108","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["TITF1 Screening in Human Congenital Diaphragmatic Hernia (CDH)"],"prefix":"10.3390","volume":"9","author":[{"given":"Maria Eugenia","family":"Gulino","sequence":"first","affiliation":[{"name":"UOC di Chirurgia Pediatrica, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7709-8937","authenticated-orcid":false,"given":"Giuseppe","family":"Martucciello","sequence":"additional","affiliation":[{"name":"Pediatric Surgery DiNOGMI, University of Genova, 16147 Genoa, Italy"},{"name":"Department of Paediatric Surgery, IRCCS Giannina Gaslini Children\u2019s Hospital, 16147 Genoa, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elio","family":"Biffali","sequence":"additional","affiliation":[{"name":"Stazione Zoologica \u201cA. Dohrn\u201d, 80121 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5012-5790","authenticated-orcid":false,"given":"Patrizia","family":"Morbini","sequence":"additional","affiliation":[{"name":"UOC Anatomia Patologica, Fondazione IRCCS Policlinico San Matteo, 27100 Pavia, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Roberta","family":"Patti","sequence":"additional","affiliation":[{"name":"UOC di Chirurgia Pediatrica, Ospedale Policlinico San Marco, 95121 Catania, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marco","family":"Borra","sequence":"additional","affiliation":[{"name":"Stazione Zoologica \u201cA. Dohrn\u201d, 80121 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Grazia","family":"Scuderi","sequence":"additional","affiliation":[{"name":"UOC di Chirurgia Pediatrica, Ospedale Policlinico San Marco, 95121 Catania, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1002\/tera.1420460605","article-title":"A population-based study of congenital diaphragmatic hernia","volume":"46","author":"Torfs","year":"1992","journal-title":"Teratology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"F137","DOI":"10.1136\/archdischild-2014-306174","article-title":"Epidemiology of congenital diaphragmatic hernia in Europe: A register-based study","volume":"100","author":"McGivern","year":"2015","journal-title":"Arch. Dis. Child.-Fetal Neonatal Ed."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1002\/bdra.20230","article-title":"Epidemiologic characteristics of congenital diaphragmatic hernia among 2.5 million California births, 1989\u20131997","volume":"76","author":"Yang","year":"2006","journal-title":"Birth Defects Res. Part A Clin. Mol. Teratol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1002\/uog.3863","article-title":"Prenatal detection and outcome of congenital diaphragmatic hernia: A French registry-based study","volume":"29","author":"Gallot","year":"2007","journal-title":"Ultrasound Obstet. Gynecol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e356","DOI":"10.1542\/peds.2004-2845","article-title":"Outcomes of congenital diaphragmatic hernia: A population-based study in Western Australia","volume":"116","author":"Colvin","year":"2005","journal-title":"Pediatrics"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1186\/s40748-017-0045-1","article-title":"Congenital Diaphragmatic hernia\u2014A review","volume":"3","author":"Chandrasekharan","year":"2017","journal-title":"Matern. Health Neonatol. Perinatol."},{"key":"ref_7","first-page":"372","article-title":"Pulmonary acinar development in diaphragmatic hernia","volume":"115","author":"Nakamura","year":"1991","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"195","DOI":"10.3389\/fped.2017.00195","article-title":"Treatment Strategies for Congenital Diaphragmatic Hernia: Change Sometimes Comes Bearing Gifts","volume":"5","author":"Morini","year":"2017","journal-title":"Front. Pediatr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/j.siny.2014.09.006","article-title":"Prenatal management of the fetus with isolated congenital diaphragmatic hernia in the era of the TOTAL trial","volume":"19","author":"Deprest","year":"2014","journal-title":"Semin. Fetal Neonatal Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.1016\/j.jpedsurg.2015.10.082","article-title":"Prenatally diagnosed severe CDH: Mortality and morbidity remain high","volume":"51","author":"Coughlin","year":"2016","journal-title":"J. Pediatr. Surg."},{"key":"ref_11","first-page":"428","article-title":"A study of closure of the pleuropericardial and pleuroperitoneal canals in the human embryo","volume":"97","author":"Wells","year":"1947","journal-title":"Anat. Rec."},{"key":"ref_12","first-page":"174","article-title":"Correction of congenital diaphragmatic hernia in utero. I. The model: Intrathoracic balloon produces fatal pulmonary hypoplasia","volume":"88","author":"Harrison","year":"1980","journal-title":"Surgery"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1007\/BF00303142","article-title":"Experimental study on embryogenesis of congenital diaphragmatic hernia","volume":"169","author":"Iritani","year":"1984","journal-title":"Anat. Embryol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"L1159","DOI":"10.1152\/ajplung.2000.279.6.L1159","article-title":"Lung hypoplasia in the nitrofen model of congenital diaphragmatic hernia occurs early in development","volume":"279","author":"Guilbert","year":"2000","journal-title":"Am. J. Physiol. Cell. Mol. Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/S0022-3468(00)80028-7","article-title":"Early lung malformations in congenital diaphragmatic hernia","volume":"35","author":"Jesudason","year":"2000","journal-title":"J. Pediatr. Surg."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.1016\/S0002-9440(10)65000-6","article-title":"Dual-hit hypotesis explains pulmonary hypoplasia in the nitrofen model of congenital diapragmatic hernia","volume":"156","author":"Keijzer","year":"2000","journal-title":"Am. J. Pathol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1002\/sctm.20-0199","article-title":"Human induced pluripotent stem cell-derived lung organoids in an ex\u2009vivo model of the congenital diaphragmatic hernia fetal lung","volume":"10","author":"Kunisaki","year":"2021","journal-title":"Stem Cells Transl. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"955","DOI":"10.1242\/dmm.028365","article-title":"Congenital diaphragmatic hernias: From genes to mechanisms to therapies","volume":"10","author":"Kardon","year":"2017","journal-title":"Dis. Model. Mech."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4764","DOI":"10.1093\/hmg\/ddv196","article-title":"Increased burden of de novo predicted deleterious variants in complex congenital diaphragmatic hernia","volume":"24","author":"Yu","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"712","DOI":"10.1542\/peds.2010-0521","article-title":"Retinol status of newborn infants is associated with congenital diaphragmatic hernia","volume":"126","author":"Beurskens","year":"2010","journal-title":"Pediatrics"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"719","DOI":"10.1111\/j.1753-4887.2009.00247.x","article-title":"Role of nutrition, lifestyle factors, and genes in the pathogenesis of congenital diaphragmatic hernia: Human and animal studies","volume":"67","author":"Beurskens","year":"2009","journal-title":"Nutr. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1242\/dev.113.4.1093","article-title":"The transcription factor TTF-1 is expressed at the onset of thyroid and lung morphogenesis and in restricted regions of the foetal brain","volume":"113","author":"Lazzaro","year":"1991","journal-title":"Development"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1101\/gad.10.1.60","article-title":"The T\/ebp null mouse: Thyroid-specific enhancer-binding protein is essential for the organogenesis of the thyroid, lung, ventral forebrain, and pituitary","volume":"10","author":"Kimura","year":"1996","journal-title":"Genes Dev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/S0300-9084(99)80077-7","article-title":"Thyroid-specific enhancer-binding protein\/thyroid transcription factor 1 is not required for the initial specification of the thyroid and lung primordia","volume":"81","author":"Kimura","year":"1999","journal-title":"Biochimie"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1006\/dbio.1995.8080","article-title":"TTF-1 regulates lung epithelial morphogenesis","volume":"172","author":"Minoo","year":"1995","journal-title":"Dev. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"135S","DOI":"10.1378\/chest.111.6_Supplement.135S","article-title":"TTF-1 is an epithelial morphoregulatory transcriptional factor","volume":"111","author":"Minoo","year":"1997","journal-title":"Chest"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1006\/dbio.1999.9234","article-title":"Defects in tracheoesophageal and lung morphogenesis in Nkx2.1(-\/-) mouse embryos","volume":"209","author":"Minoo","year":"1999","journal-title":"Dev. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.molcel.2013.02.018","article-title":"Nkx2-1 represses a latent gastric differentiation program in lung adenocarcinoma","volume":"50","author":"Snyder","year":"2013","journal-title":"Mol. Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"20545","DOI":"10.1073\/pnas.1906663116","article-title":"Transcriptional control of lung alveolar type 1 cell development and maintenance by NK homeobox 2-1","volume":"116","author":"Little","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"673","DOI":"10.1177\/44.7.8675988","article-title":"Expression of thyroid transcription factor-1(TTF-1) in fetal and neonatal human lung","volume":"44","author":"Stahlman","year":"1996","journal-title":"J. Histochem. Cytochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.ydbio.2015.08.009","article-title":"Wt1 and \u03b2-catenin cooperatively regulate diaphragm development in the mouse","volume":"407","author":"Paris","year":"2015","journal-title":"Dev. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.amjsurg.2021.07.016","article-title":"Exome sequencing of fetuses with congenital diaphragmatic hernia supports a causal role for NR2F2, PTPN11, and WT1 variants","volume":"223","author":"Schwab","year":"2022","journal-title":"Am. J. Surg."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1136\/jmg.2011.089680","article-title":"Genomic alterations that contribute to the development of isolated and non-isolated congenital diaphragmatic hernia","volume":"48","author":"Wat","year":"2011","journal-title":"J. Med. Genet."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1002\/aja.1000920202","article-title":"An analysis of the syndrome of malformations induced by maternal vitamin A deficiency. Effects of restoration of vitamin A at various times during gestation","volume":"92","author":"Wilson","year":"1953","journal-title":"Am. J. Anat."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2749","DOI":"10.1242\/dev.120.10.2749","article-title":"Function of the retinoic acid receptors (RARs) during development (II). Multiple abnormalities at various stages of organogenesis in RAR double mutants","volume":"120","author":"Mendelsohn","year":"1994","journal-title":"Development"},{"key":"ref_36","first-page":"229","article-title":"STRA6: Role in cellular retinol uptake and efflux","volume":"4","author":"Kelly","year":"2015","journal-title":"Hepatobiliary Surg. Nutr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"550","DOI":"10.1086\/512203","article-title":"Mutations in STRA6 cause a broad spectrum of malformations including anophthalmia, congenital heart defects, diaphragmatic hernia, alveolar capillary dysplasia, lung hypoplasia, and mental retardation","volume":"80","author":"Pasutto","year":"2007","journal-title":"Am. J. Hum. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1016\/j.ejmg.2009.02.003","article-title":"Congenital diaphragmatic hernia is part of the new 15q24 microdeletion syndrome","volume":"52","author":"Backx","year":"2009","journal-title":"Eur. J. Med. Genet."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1186\/1752-1947-7-287","article-title":"Duplication of the ALDH1A2 gene in association with pentalogy of Cantrell: A case report","volume":"7","author":"Steiner","year":"2013","journal-title":"J. Med. Case Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e29","DOI":"10.1016\/j.jpedsurg.2005.06.032","article-title":"Congenital diaphragmatic hernia in a patient with tetrasomy 9p","volume":"40","author":"Tibboel","year":"2005","journal-title":"Pediatr. Surg."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"877","DOI":"10.1086\/429842","article-title":"Congenital diaphragmatic hernia and chromosome 15q26: Determination of a candidate region by use of fluorescent in situ hybridization and array-based comparative genomic hybridization","volume":"76","author":"Klaassens","year":"2005","journal-title":"Am. J. Hum. Genet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"16351","DOI":"10.1073\/pnas.0507832102","article-title":"Mouse lacking COUP-TFII as an animal model of Bochdalek-type congenital diaphragmatic hernia","volume":"102","author":"You","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","first-page":"4153","article-title":"Chicken ovalbumin upstream promoter transcription factor (COUP-TF) dimers bind to different GGTCA response elements, allowing COUP-TF to repress hormonal induction of the vitamin D3, thyroid hormone, and retinoic acid receptors","volume":"12","author":"Cooney","year":"1992","journal-title":"Mol. Cell. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1002\/pd.4244","article-title":"Identification of dosage-sensitive genes in fetuses referred with severe isolated congenital diaphragmatic hernia","volume":"33","author":"Brady","year":"2013","journal-title":"Prenat. Diagn."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1729","DOI":"10.1038\/s41431-021-00972-0","article-title":"Genetics of diaphragmatic hernia","volume":"29","author":"Schreiner","year":"2021","journal-title":"Eur. J. Hum. Genet."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"L44","DOI":"10.1152\/ajplung.00342.2004","article-title":"Congenital diaphragmatic hernia, tracheal occlusion, thyroid transcription factor-1, and fetal pulmonary epithelial maturation","volume":"289","author":"Chapin","year":"2005","journal-title":"Am. J. Physiol. Cell. Mol. Physiol."}],"container-title":["Children"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2227-9067\/9\/8\/1108\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:55:35Z","timestamp":1760140535000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2227-9067\/9\/8\/1108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,23]]},"references-count":46,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["children9081108"],"URL":"https:\/\/doi.org\/10.3390\/children9081108","relation":{},"ISSN":["2227-9067"],"issn-type":[{"type":"electronic","value":"2227-9067"}],"subject":[],"published":{"date-parts":[[2022,7,23]]}}}