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A Multiplex Kindred with Hennekam Syndrome due to Homozygosity for a CCBE1 Mutation that does not Prevent Protein Expression

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Abstract

Collagen and calcium-binding EGF domain-containing protein 1 (CCBE1) bi-allelic mutations have been associated with syndromes of widespread congenital lymphatic dysplasia, including Hennekam Syndrome (HS). HS is characterized by lymphedema, lymphangiectasia, and intellectual disability. CCBE1 encodes a putative extracellular matrix protein but the HS-causing mutations have not been studied biochemically. We report two HS siblings, born to consanguineous parents of Turkish ancestry, whose clinical phenotype also includes protein losing enteropathy, painful relapsing chylous ascites, and hypogammaglobulinemia. We identified by whole exome and Sanger sequencing the homozygous CCBE1 C174Y mutation in both siblings. This mutation had been previously reported in another HS kindred from the Netherlands. In over-expression studies, we found increased intracellular expression of all forms (monomers, dimers, trimers) of the CCBE1 C174Y mutant protein, by Western blot, despite mutant mRNA levels similar to wild-type (WT). In addition, we detected increased secretion of the mutant CCBE1 protein by ELISA. We further found the mutant and WT proteins to be evenly distributed in the cytoplasm, by immunofluorescence and confocal microscopy. Finally, we found a strong decrease of lymphatic vessels, with a corresponding diminished expression of CCBE1, by immunohistochemistry of the patients’ intestinal biopsies. In contrast, mucosal blood vessels and muscularis mucosae showed normal CCBE1 staining. Our findings show that the mutant CCBE1 C174Y protein is not loss-of-function by loss-of-expression.

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Acknowledgments

We warmly thank the patients for their participation. This work was supported by the National Center for Research Resources and the National Center for Advancing Sciences (NCATS) of the National Institutes of Health (NIH) (grant 8UL1TR000043), the Rockefeller University, INSERM, Paris Descartes University, and the St. Giles Foundation.

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Authors and Affiliations

  1. St. Giles Laboratory of Human Genetics of Infectious Diseases, Rockefeller Branch, The Rockefeller University, New York, NY, USA

    Carolyn C. Jackson, Lucy Best & Jean-Laurent Casanova

  2. Department of Pediatrics, The Memorial Sloan Kettering Cancer Center, New York, NY, USA

    Carolyn C. Jackson

  3. Laboratory of Human Genetics of Infectious Diseases, Necker Branch, INSERM U1163, Necker Hospital for Sick Children, INSERM, Paris, France

    Lazaro Lorenzo & Jean-Laurent Casanova

  4. Howard Hughes Medical Institute, New York, NY, USA

    Jean-Laurent Casanova

  5. Paris Descartes University, Imagine Institute, Paris, France

    Jean-Laurent Casanova

  6. Pediatric Hematology-Immunology Unit, Necker Hospital for Sick Children, Paris, France

    Jean-Laurent Casanova

  7. Department of Pathology, Universitätsklinikum Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany

    Jochen Wacker, Simone Bertz & Abbas Agaimy

  8. Department for Internal Medicine III, Universitätsklinikum Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany

    Thomas Harrer

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  1. Carolyn C. Jackson
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Correspondence to Carolyn C. Jackson.

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The authors declare they have no competing financial interests. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.

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Jackson, C.C., Best, L., Lorenzo, L. et al. A Multiplex Kindred with Hennekam Syndrome due to Homozygosity for a CCBE1 Mutation that does not Prevent Protein Expression. J Clin Immunol 36, 19–27 (2016). https://doi.org/10.1007/s10875-015-0225-6

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  • DOI: https://doi.org/10.1007/s10875-015-0225-6

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