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Missense substitutions in the GAS1 protein present in holoprosencephaly patients reduce the affinity for its ligand, SHH

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Abstract

Holprosencephaly (HPE) is the most common disorder of the developing forebrain in humans, and is characterized by varying degrees of abnormal union of the cerebral hemispheres. These defects are typically co-associated with midline craniofacial anomalies. The combination of forebrain and craniofacial defects that comprise HPE can present along a broad and variable phenotypic spectrum. Both the SHH and NODAL signaling pathways play important roles in the pathogenesis of this disorder. Disruption of these pathways by chromosomal rearrangements, mutations in pathway-related genes and/or biochemical alterations are proposed to contribute to HPE in a large number of patients. Additional factors that are not yet fully delineated are also very likely to be involved in the pathogenesis and phenotypic heterogeneity of the disorder. Genetic loss of GAS1, a cell membrane receptor and positive regulator of SHH, has been demonstrated to contribute to the HPE phenotypic spectrum in animal models. We have evaluated the coding and flanking sequence of GAS1 in 394 patients who have clinical findings within the HPE phenotypic spectrum, and now report five novel missense sequence variants among five unrelated HPE probands. Finally, we tested the effect of these variants (as well as previously reported GAS1 variants) on the ability of GAS1 to bind to SHH. Here, we demonstrate that sequence variants in GAS1 can impair its physical interaction with SHH, suggesting a decrease in the SHH downstream signaling cascade as a pathogenic mechanism of disease.

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Acknowledgments

We thank the families who participated in these research studies, and the National Institute of Neurological Disorders and Stroke’s (NINDS) DNA Sequencing Facility for their technical support with DNA sequencing. This work was supported in part by the Division of Intramural Research (DIR) of the National Human Genome Research Institute (MM) and RO1 DK084963 (C-M F).

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Authors and Affiliations

  1. Medical Genetics Branch, National Human Genome Research Institute (NHGRI), National Institutes of Health, 35 Convent Drive, Building 35, Room 1B403, Bethesda, ML, 20892-3717, USA

    Daniel E. Pineda-Alvarez, Erich Roessler, Ping Hu, Kshitij Srivastava, Benjamin D. Solomon & Maximilian Muenke

  2. Department of Embryology, Carnegie Institution for Science, Baltimore, ML, 21218, USA

    C. Evan Siple & Chen-Ming Fan

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  1. Daniel E. Pineda-Alvarez
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  2. Erich Roessler
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  3. Ping Hu
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  4. Kshitij Srivastava
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  5. Benjamin D. Solomon
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  6. C. Evan Siple
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  7. Chen-Ming Fan
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  8. Maximilian Muenke
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Correspondence to Maximilian Muenke.

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D. E. Pineda-Alvarez and E. Roessler contributed equally to this research work.

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Pineda-Alvarez, D.E., Roessler, E., Hu, P. et al. Missense substitutions in the GAS1 protein present in holoprosencephaly patients reduce the affinity for its ligand, SHH. Hum Genet 131, 301–310 (2012). https://doi.org/10.1007/s00439-011-1078-6

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