ORIGINAL ARTICLE No evidence of HAND2 involvement in nonsyndromic cleft lip with or without cleft palate Marcella Martinelli & Ambra Girardi & Francesca Farinella & Francesco Carinci & Furio Pezzetti & Elisabetta Caramelli & Luca Scapoli Received: 29 January 2010 / Accepted: 24 February 2011 / Published online: 24 March 2011 # Springer-Verlag 2011 Abstract Craniofacial morphogenesis is determined by multistep processes involving signalling molecules and transcription factors, which are organised into highly coordinated pathways. Derailment from this intricate network can lead to congenital malformations. Cells migrate from neural crests to populate different structures, such as branchial arches, involved in embryonal orofacial development. The EDN1 pathway is involved in branchial arch development. Gene knockout and knockdown experi- ments on EDN1 or its downstream effector dHAND resulted in mice that were characterised by craniofacial defects and cleft palate. Our aim was to evaluate whether the transcription factor HAND2 could be implicated in non- syndromic cleft lip with or without cleft palate (CL/P) aetiology. A sample study composed of 39 multiplex Italian pedigrees was enrolled to test linkage between two micro- satellite flanking HAND2 locus and CL/P. No evidence of linkage between HAND2 and CL/P was obtained. Indeed, formal levels of exclusion were obtained with different inheritance models. Investigation results did not support a role of HAND2 in CL/P aetiology. Nevertheless a minor contribute of the gene in clefting could not be ruled out. Keywords Cleft lip with or without cleft palate . HAND2 . Linkage . Microsatellite Introduction A variety of specific transcription factors act synergistically with perfectly orchestrated timing to give birth to the craniofacial morphogenesis. It is, therefore, conceivable that a failure in this delicate web could determine developmental alterations, such as cleft lip with or without cleft palate (CL/P). Certain transcription factors that are possibly involved in CL/ P aetiology have been the subject of different investigations, considering the wide geographical distribution of this congenital malformation, with an average birth prevalence of 1/700. Association has been reported between IRF6 (interferon regulatory factor-6) and CL/P and confirmed by different authors in different populations worldwide [1–3]. TP63 gene, that encodes a member of the p53 family of transcription factors, has been investigated on the basis of its involvement in ankyloblepharon-ectodermal dysplasia- clefting (AEC), a rare syndrome which is characterised by congenital ectodermal dysplasia and cleft lip and/or cleft palate [4, 5]. Demonstration of the crucial role of both p63 and IRF6 in palate development has been recently provided in a murine model, showing cooperation between the two transcription factors [6]. Leoyklang [7] supported a role for this gene in non-syndromic CL/P, establishing three novel non-synonymous changes (S90L, R313G, and D564H) in addition to 21 variant sites. The MSX1 gene has been investigated because of its involvement in tooth agenesis with various combinations of cleft lip and cleft palate [8]. The role of this encoded protein, which functions as a transcriptional repressor during embryogenesis, was confirmed M. Martinelli (*) : A. Girardi : F. Pezzetti : E. Caramelli : L. Scapoli Department of Histology, Embryology and Applied Biology, Centre of Molecular Genetics, University of Bologna, Via Belmeloro 8, 40126 Bologna, Italy e-mail: marcella.martinelli@unibo.it F. Farinella : F. Carinci Department of D.M.C.C.C., Section of Maxillo-Facial and Plastic Surgery, University of Ferrara, Corso Giovecca 203, 44121 Ferrara, Italy Clin Oral Invest (2012) 16:619–623 DOI 10.1007/s00784-011-0539-6