Multiple clinical forms of dehydrated hereditary stomatocytosis arise from mutations in PIEZO1.

Autosomal dominant dehydrated hereditary stomatocytosis (DHSt) usually presents as a compensated hemolytic anemia with macrocytosis and abnormally shaped red blood cells (RBCs). DHSt is part of a pleiotropic syndrome that may also exhibit pseudohyperkalemia and perinatal edema. We identified PIEZO1 as the disease gene for pleiotropic DHSt in a large kindred by exome sequencing analysis within the previously mapped 16q23-q24 interval. In 26 affected individuals among 7 multigenerational DHSt families with the pleiotropic syndrome, 11 heterozygous PIEZO1 missense mutations cosegregated with disease. PIEZO1 is expressed in the plasma membranes of RBCs and its messenger RNA, and protein levels increase during in vitro erythroid differentiation of CD34(+) cells. PIEZO1 is also expressed in liver and bone marrow during human and mouse development. We suggest for the first time a correlation between a PIEZO1 mutation and perinatal edema. DHSt patient red cells with the R2456H mutation exhibit increased ion-channel activity. Functional studies of PIEZO1 mutant R2488Q expressed in Xenopus oocytes demonstrated changes in ion-channel activity consistent with the altered cation content of DHSt patient red cells. Our findings provide direct evidence that R2456H and R2488Q mutations in PIEZO1 alter mechanosensitive channel regulation, leading to increased cation transport in erythroid cells.

[1]  A. Iolascon,et al.  K-CL co-transport plays an important role in normal and β thalassemic erythropoiesis , 2007, Haematologica.

[2]  Jana Marie Schwarz,et al.  MutationTaster evaluates disease-causing potential of sequence alterations , 2010, Nature Methods.

[3]  Tchernia,et al.  A genetic syndrome associating dehydrated hereditary stomatocytosis, pseudohyperkalaemia and perinatal oedema , 1998, British journal of haematology.

[4]  S. Brunak,et al.  Quantitative Phosphoproteomics Reveals Widespread Full Phosphorylation Site Occupancy During Mitosis , 2010, Science Signaling.

[5]  K. Kosaka,et al.  A novel membrane protein, encoded by the gene covering KIAA0233, is transcriptionally induced in senile plaque-associated astrocytes , 2006, Brain Research.

[6]  S. Perrotta,et al.  Familial Pseudohyperkalemia Maps to the Same Locus as Dehydrated Hereditary Stomatocytosis (Hereditary Xerocytosis) , 1999 .

[7]  M. Mann,et al.  Lysine Acetylation Targets Protein Complexes and Co-Regulates Major Cellular Functions , 2009, Science.

[8]  Manuela Schmidt,et al.  Inflammatory signals enhance piezo2-mediated mechanosensitive currents. , 2012, Cell reports.

[9]  J. Whaun,et al.  Congenital hemolytic anemia with high-sodium, low-potassium red cells. Studies of three generations of a family with a new variant. , 1969, The New England journal of medicine.

[10]  G. Stewart,et al.  FAMILIAL PSEUDOHYPERKALÆMIA A New Syndrome , 1979, The Lancet.

[11]  Helga Thorvaldsdóttir,et al.  Integrative Genomics Viewer , 2011, Nature Biotechnology.

[12]  S. Henikoff,et al.  Predicting the effects of amino acid substitutions on protein function. , 2006, Annual review of genomics and human genetics.

[13]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[14]  A. Iolascon,et al.  Missense mutations in the ABCB6 transporter cause dominant familialpseudohyperkalemia , 2013, American journal of hematology.

[15]  J. Delaunay.  The molecular basis of hereditary red cell membrane disorders. , 2007, Blood reviews.

[16]  Frederick Sachs,et al.  Gating the mechanical channel Piezo1 , 2012, Channels.

[17]  J. Ayoubi,et al.  Dehydrated hereditary stomatocytosis: a cause of prenatal ascites , 2001, Prenatal diagnosis.

[18]  A. Basu,et al.  Dehydrated hereditary stomatocytosis with transient perinatal ascites , 2003, Archives of disease in childhood. Fetal and neonatal edition.

[19]  S. Baldwin,et al.  Stomatin-deficient cryohydrocytosis results from mutations in SLC2A1: a novel form of GLUT1 deficiency syndrome. , 2011, Blood.

[20]  J. Adelman,et al.  cDNA Cloning and Functional Characterization of the Mouse Ca2+-gated K+ Channel, mIK1 , 1998, The Journal of Biological Chemistry.

[21]  C. Beaumont,et al.  Mild dehydrated hereditary stomatocytosis revealed by marked hepatosiderosis. , 2006, Clinical and laboratory haematology.

[22]  J. Delaunay.  The hereditary stomatocytoses: genetic disorders of the red cell membrane permeability to monovalent cations. , 2004, Seminars in hematology.

[23]  C. Haslett,et al.  Integrin activation by Fam38A uses a novel mechanism of R-Ras targeting to the endoplasmic reticulum , 2010, Journal of Cell Science.

[24]  R. Frydman,et al.  First‐trimester nuchal abnormalities secondary to dehydrated hereditary stomatocytosis , 2009, Prenatal diagnosis.

[25]  Frederick Sachs,et al.  Piezo1: properties of a cation selective mechanical channel. , 2012, Channels.

[26]  F. Sachs,et al.  The mechanosensitive ion channel Piezo1 is inhibited by the peptide GsMTx4. , 2011, Biochemistry.

[27]  M. DePristo,et al.  The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. , 2010, Genome research.

[28]  M. Humbert,et al.  An Extreme Consequence of Splenectomy in Dehydrated Hereditary Stomatocytosis: Gradual Thrombo‐embolic Pulmonary Hypertension and Lung–Heart Transplantation , 2003, Hemoglobin.

[29]  P. Bork,et al.  A method and server for predicting damaging missense mutations , 2010, Nature Methods.

[30]  G. Stewart,et al.  THROMBO‐EMBOLIC DISEASE AFTER SPLENECTOMY FOR HEREDITARY STOMATOCYTOSIS , 1996, British journal of haematology.

[31]  M. Babron,et al.  Pleiotropic syndrome of dehydrated hereditary stomatocytosis, pseudohyperkalemia, and perinatal edema maps to 16q23-q24. , 2000, Blood.

[32]  S. Alper,et al.  Mouse K-Cl cotransporter KCC1: cloning, mapping, pathological expression, and functional regulation. , 1999, The American journal of physiology.

[33]  N. Nomura,et al.  Prediction of the coding sequences of unidentified human genes. VI. The coding sequences of 80 new genes (KIAA0201-KIAA0280) deduced by analysis of cDNA clones from cell line KG-1 and brain. , 1996, DNA research : an international journal for rapid publication of reports on genes and genomes.

[34]  Brian E. Smith,et al.  Mutations in the mechanotransduction protein PIEZO1 are associated with hereditary xerocytosis. , 2012, Blood.

[35]  Manuela Schmidt,et al.  Piezo1 and Piezo2 Are Essential Components of Distinct Mechanically Activated Cation Channels , 2010, Science.

[36]  X. Jeunemaître,et al.  Dehydrated hereditary stomatocytosis mimicking familial hyperkalaemic hypertension: clinical and genetic investigation , 2007, European journal of haematology.

[37]  K Kozlowski,et al.  A new syndrome? Unusual facies, hooked clavicles, 13 pairs of ribs, widened metaphyses, square shaped vertebral bodies and communicating hydrocephalus. , 1992, Pediatric radiology.

[38]  S. Perrotta,et al.  Genomewide search for dehydrated hereditary stomatocytosis (hereditary xerocytosis): mapping of locus to chromosome 16 (16q23-qter). , 1998, American journal of human genetics.

[39]  M. Mann,et al.  In-depth analysis of the membrane and cytosolic proteome of red blood cells. , 2006, Blood.

[40]  F. Oski,et al.  Congenital hemolytic anemia with high sodium, low potassium red cells. I. Studies of membrane permeability. , 1968, The New England journal of medicine.

[41]  Ruedi Aebersold,et al.  Mass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins , 2009, Nature Biotechnology.

[42]  A. Patapoutian,et al.  The role of Drosophila Piezo in mechanical nociception , 2011, Nature.

[43]  I. Laurendeau,et al.  Sub‐lethal hydrops as a manifestation of dehydrated hereditary stomatocytosis in two consecutive pregnancies , 2003, Prenatal diagnosis.

[44]  G. Stewart.  Hemolytic disease due to membrane ion channel disorders , 2004, Current opinion in hematology.

[45]  R. Becker,et al.  Xerocytosis with concomitant intrauterine ascites: first description and therapeutic approach. , 1996, Blood.

[46]  J. Arnaud,et al.  The over-expression of TRPC6 channels in HEK-293 cells favours the intracellular accumulation of zinc. , 2011, Biochimica et biophysica acta.

[47]  S. Agarwal,et al.  Letter: Alpha-fetoprotein in Indian childhood cirrhosis. , 1974, Lancet.

[48]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[49]  S. Israels,et al.  Refinement of the hereditary xerocytosis locus on chromosome 16q in a large Canadian kindred. , 2011, Blood cells, molecules & diseases.

[50]  J. Bates,et al.  A new variant of hereditary hemolytic anemia with stomatocytosis and erythrocyte cation abnormality. , 1971, Blood.

[51]  J. Vives-Corrons,et al.  Nonimmune Hydrops Fetalis due to Congenital Xerocytosis , 2005, Journal of Perinatology.