Supplementary Material for: TGIF Mutations in Human Holoprosencephaly: Correlation between Genotype and Phenotype

Holoprosencephaly (HPE), which results from failed or incomplete midline forebrain division early in gestation, is the most common forebrain malformation. The etiology of HPE is complex and multifactorial. To date, at least 12 HPE-associated genes have been identified, including <i>TGIF</i> (transforming growth factor beta-induced factor), located on chromosome 18p11.3. <i>TGIF</i> encodes a transcriptional repressor of retinoid responses involved in TGF-β signaling regulation, including Nodal signaling. <i>TGIF</i> mutations are reported in approximately 1–2% of patients with non-syndromic, non-chromosomal HPE.We combined data from our comprehensive studies of HPE with a literature search for all individuals with HPE and evidence of mutations affecting <i>TGIF</i> in order to establish the genotypic and phenotypic range. We describe 2 groups of patients: 34 with intragenic mutations and 21 with deletions of <i>TGIF</i>. These individuals, which were ascertained from our research group, in collaboration with other centers, and through a literature search, include 38 probands and 17 mutation-positive relatives. The majority of intragenic mutations occur in the TGIF homeodomain. Patients with mutations affecting <i>TGIF</i>recapitulate the entire phenotypic spectrum observed in non-chromosomal, non-syndromic HPE. We identified a statistically significant difference between the 2 groups with respect to inheritance, as <i>TGIF</i> deletions were more likely to be de novo in comparison to <i>TGIF</i> mutations (χ<sup>2</sup><sub>(2)</sub> = 6.97, p<sub>permutated</sub> = 0.0356). In addition, patients with <i>TGIF</i> deletions were also found to more commonly present with manifestations beyond the craniofacial and neuroanatomical features associated with HPE (p = 0.0030). These findings highlight differences in patients with intragenic mutations versus deletions affecting <i>TGIF</i>, and draw attention to the homeodomain region, which appears to be particularly relevant to HPE. These results may be useful for genetic counseling of affected patients.