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Molecular and Functional Characterization of a Cohort of Spanish Patients with Ataxia-Telangiectasia

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Abstract

Ataxia-telangiectasia is a multisystemic disease with severe neurological affectation, immunodeficiency and telangiectasia. The disorder is caused by alterations in the ATM gene, whose size and complexity make molecular diagnosis difficult. We designed a target-enrichment next-generation sequencing strategy to characterize 28 patients from several regions of Spain. This approach allowed us to identify gene variants affecting function in 54 out of the 56 alleles analyzed, although the two unresolved alleles belong to brothers. We found 28 ATM gene mutations, of which 10 have not been reported. A total of 171 gene variants not affecting function were also found, of which 22 are reported to predispose to disease. Interestingly, all Roma (Spanish Gypsies) patients are homozygous for the same mutation and share the H3 ATM haplotype, which is strong evidence of a founder effect in this population. In addition, we generated a panel of 27 primary T cell lines from A-T patients, which revealed significant expression of ATM in two patients and traces of the protein in nine more. None of them retained residual ATM activity, and almost all T cell lines show increased or intermediate radiosensitivity.

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Acknowledgments

We are indebted to Mr. Patxi Villén, President of the Spanish Association of Families with Ataxia-Telangiectasia (AEFAT) for his invaluable help in recruiting this cohort. We also thank all members of the association who eagerly contributed samples and the patient’s caring physicians for their support. We acknowledge the continuous and generous supply of rIL-2 provided by the National Institutes of Health AIDS Research and Reference Reagent Program (Rockville, MD). This work was funded by Grant 12UDG01-ATF from Sparks, The Children’s Medical Charity, London, UK, in coordination with Action for A-T and the Ataxia-Telangiectasia Society, UK. STR had a research contract funded by the Sparks grant.

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    Authors and Affiliations

    1. Institute of Biopathology and Regenerative Medicine, Center for Biomedical Research, University of Granada, Health Sciences Technology Park, Avda. del Conocimiento s/n, 18016, Armilla, Granada, Spain

      Diana Carranza, Ana Karina Vega, Sara Torres-Rusillo, Enrique Montero & Ignacio J. Molina

    2. Genomic Unit, Center Pfizer-University of Granada-Junta de Andalucia for Genomics and Oncological Research (GENYO), Health Sciences Technology Park, Granada, Spain

      Luis Javier Martinez

    3. Service of Immunology and Department of Cell Biology and Immunology, “Reina Sofia” University Hospital, University of Córdoba, Córdoba, Spain

      Manuel Santamaría

    4. Service of Pediatrics, “Virgen de las Nieves” University Hospital, University of Granada, Granada, Spain

      Juan Luis Santos

    5. Instituto de Investigación Biosanitaria ibs.GRANADA, Granada University Hospitals, University of Granada, Granada, Spain

      Ignacio J. Molina

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    1. Diana Carranza
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    Correspondence to Ignacio J. Molina.

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    Diana Carranza and Ana Karina Vega have equally contributed to this work.

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    Figure S1

    DNA sequencing chromatographs for patients carrying non-annotated sequence variants affecting ATM function (TIFF 904 kb)

    Figure S2

    Sequence alignment and phylogeny of p.(Leu1046Pro) (TIFF 2225 kb)

    Figure S3

    Sequence alignment and phylogeny of p.(Tyr2080Asp) (TIFF 2089 kb)

    Figure S4

    Sequence alignment and phylogeny of and p.(Phe2834Leu) (TIFF 1919 kb)

    Table SI

    Sequences of primers used for PCR amplification and confirmation of new gene variants by Sanger sequencing (DOCX 15 kb)

    Table SII

    Single nucleotide polymorphisms associated with disease identified in the cohort of A-T patients, with indication of references (DOCX 51 kb)

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    Carranza, D., Vega, A.K., Torres-Rusillo, S. et al. Molecular and Functional Characterization of a Cohort of Spanish Patients with Ataxia-Telangiectasia. Neuromol Med 19, 161–174 (2017). https://doi.org/10.1007/s12017-016-8440-8

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    • DOI: https://doi.org/10.1007/s12017-016-8440-8

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