Supplementary Material for: Molecular and Gene Network Analysis of Thyroid Transcription Factor 1 <b><i>(TTF1)</i></b> and Enhanced at Puberty <b><i>(EAP1)</i></b> Genes in Patients with GnRH-Dependent Pubertal Disorders

<b><i>Background/Aim:</i></b> TTF1 and EAP1 are transcription factors that modulate gonadotropin-releasing hormone expression. We investigated the contribution of <i>TTF1</i> and <i>EAP1</i> genes to central pubertal disorders. <b><i>Patients and Methods:</i></b> 133 patients with central pubertal disorders were studied: 86 with central precocious puberty and 47 with normosmic isolated hypogonadotropic hypogonadism. The coding region of <i>TTF1</i> and <i>EAP1</i> were sequenced. Variations of polyglutamine and polyalanine repeats in EAP1 were analyzed by GeneScan software. Association of <i>TTF1</i> and <i>EAP1</i> to genes implicated in timing of puberty was investigated by meta-network framework GeneMANIA and Cytoscape software. <b><i>Results:</i></b> Direct sequencing of the <i>TTF1</i> did not reveal any mutation or polymorphisms. Four EAP1 synonymous variants were identified with similar frequencies among groups. The most common EAP1 5′-distal polyalanine genotype was the homozygous 12/12, but the genotype 12/9 was identified in 2 central precocious puberty sisters without functional alteration in EAP1 transcriptional activity. TTF1 and EAP1 were connected, via genetic networks, to genes implicated in the control of menarche. <b><i>Conclusion:</i></b> No <i>TTF1 </i>or <i>EAP1 </i>germline mutations were associated with central pubertal disorders. <i>TTF1 </i>and <i>EAP1 </i>may affect puberty by changing expression in response to other members of puberty-associated gene networks, or by differentially affecting the expression of gene components of these networks.