ARTICLE A Multiplex Human Syndrome Implicates a Key Role for Intestinal Cell Kinase in Development of Central Nervous, Skeletal, and Endocrine Systems Piya Lahiry, 1,2 Jian Wang, 1 John F. Robinson, 1 Jacob P. Turowec, 2 David W. Litchfield, 2 Matthew B. Lanktree, 1,2 Gregory B. Gloor, 2 Erik G. Puffenberger, 3 Kevin A. Strauss, 3 Mildred B. Martens, 4 David A. Ramsay, 4 C. Anthony Rupar, 2,5,6 Victoria Siu, 2,5,6 and Robert A. Hegele 1,2, * Six infants in an Old Order Amish pedigree were observed to be affected with endocrine-cerebro-osteodysplasia (ECO). ECO is a previ- ously unidentified neonatal lethal recessive disorder with multiple anomalies involving the endocrine, cerebral, and skeletal systems. Autozygosity mapping and sequencing identified a previously unknown missense mutation, R272Q, in ICK, encoding intestinal cell kinase (ICK). Our results established that R272 is conserved across species and among ethnicities, and three-dimensional analysis of the protein structure suggests protein instability due to the R272Q mutation. We also demonstrate that the R272Q mutant fails to localize at the nucleus and has diminished kinase activity. These findings suggest that ICK plays a key role in the development of multiple organ systems. Introduction Protein kinases belong to one of the largest and most func- tionally diverse gene families in eukaryotes, constituting approximately 1.7% of all human genes. 1 By phosphory- lating substrates, kinases can direct activity, localization, and overall function of numerous target proteins. Kinases are particularly important for signal transduction and coor- dination of complex cellular functions, as seen with the mitogen-activated protein (MAP) kinases and cyclin-depen- dent kinases (CDKs), which play a central role in regulating mammalian cell proliferation and division. 2 Altered kinase activity or abnormal substrate phosphorylation has been implicated in monogenic diseases—‘‘kinasopathies’’— including endocrine disorders, 3 cancers, 4 immunodefi- ciencies, 5 and cardiovascular diseases. 6 Inherited skeletal dysplasias or osteochondrodysplasias are characterized by abnormal development, growth, and maintenance of the skeleton. 7 Manifestations of skeletal dysplasias range from clinically undetectable to severe deformities and lethality. 7 A group of lethal autosomal- recessive skeletal dysplasias are the short rib-polydactyly (SRP) syndromes, 8 which include SRP type II or Majewski Syndrome (MIM: 263520). 9 Herein we report a new syndrome, to our knowledge, comprising osteodysplasia, cerebral anomalies, and endo- crine gland hypoplasia. A pedigree from an Old Order Amish community was identified as having six affected infants with this previously unreported multisystem, neonatal lethal condition, designated here as the endocrine-cere- bro-osteodysplasia (ECO) syndrome. Because the Old Order Amish population reportedly have a high degree of consan- guinity, 10 this pedigree was ideal for autozygosity mapping of the putative molecular defect. 11,12 We delineate the clinical features of ECO and report the first mutation, R272Q (c.1305G/A), within ICK, encod- ing intestinal cell kinase (ICK). Biochemical and immuno- cytochemical studies indicate function and localization deficits, implicating ICK as a key player in development of the central nervous, skeletal, and endocrine systems. Material and Methods Patients and Biological Materials Two families from an Old Order Amish community were referred for genetic assessment and counseling, and all six affected infants were examined by one of the authors (V.S.). Photos, blood, and tissue samples were provided for research purposes, with ethics approval (from the Office of Research Ethics at the University of Western Ontario) and informed consent from participating parents. Peripheral blood and skin biopsy for DNA extraction was collected from five affected children, three parents, and two unaf- fected siblings and used for autozygosity mapping. The genealogy of the families affected with the disease was prepared through inter- views and local Amish community records. DNA from umbilical cord blood and buccal swabs was extracted from 257 ECO-unaf- fected individuals born into the Old Order Amish community for determining the mutant-allele frequency in the community. Autopsies were performed on three patients (IV-1, IV-2, and IV-8 from pedigree) after 33, 29, and 23 weeks of gestation, respectively. Histology Routine tissue samples including liver, kidney, and central nervous system (CNS) from each of the three autopsy patients were collected 1 Robarts Research Institute, London, Ontario N6A 5K8, Canada; 2 Department of Biochemistry, University of Western Ontario, London, Ontario N6A 5C1, Canada; 3 Clinic for Special Children, Strasburg, PA 17579, USA; 4 Department of Pathology, London Health Sciences Centre, London, Ontario N6A 5A5, Canada; 5 Medical Genetics Program, London Health Sciences Centre, London, Ontario N6C 2V5, Canada; 6 Children’s Health Research Institute, London, Ontario N6C 2V5, Canada *Correspondence: hegele@robarts.ca DOI 10.1016/j.ajhg.2008.12.017. ª2009 by The American Society of Human Genetics. All rights reserved. 134 The American Journal of Human Genetics 84, 134–147, February 13, 2009