Familial thrombocytosis caused by the novel germ-line mutation p.Pro106Leu in the MPL gene
El-Harith Abdelghaffar El-Harith
Institute of Human Genetics
These authors contributed equally to the study.
Search for more papers by this authorCornelia Roesl
Institute of Human Genetics
These authors contributed equally to the study.
Search for more papers by this authorMatthias Ballmaier
Department of Paediatric Haematology and Oncology
Search for more papers by this authorManuela Germeshausen
Department of Paediatric Haematology and Oncology
Search for more papers by this authorNils Von Neuhoff
Department of Cell and Molecular Pathology, Medical University of Hannover, Hannover
Search for more papers by this authorChristian Becker
Cologne Centre for Genomics and Institute for Genetics
Search for more papers by this authorGudrun Nürnberg
Cologne Centre for Genomics and Institute for Genetics
Search for more papers by this authorPeter Nürnberg
Cologne Excellence Cluster on Cellular Stress Responses in Ageing-associated Diseases (CECAD), University of Cologne, Cologne, Germany
Search for more papers by this authorMirghani Ali Mohamed Ahmed
College of Medicine, University of Khartoum, Khartoum, Sudan
Search for more papers by this authorJeannette Hübener
Institute of Human Genetics
Present address: J. Hübener, Department of Medical Genetics, University Tuebingen, Germany.
Search for more papers by this authorEl-Harith Abdelghaffar El-Harith
Institute of Human Genetics
These authors contributed equally to the study.
Search for more papers by this authorCornelia Roesl
Institute of Human Genetics
These authors contributed equally to the study.
Search for more papers by this authorMatthias Ballmaier
Department of Paediatric Haematology and Oncology
Search for more papers by this authorManuela Germeshausen
Department of Paediatric Haematology and Oncology
Search for more papers by this authorNils Von Neuhoff
Department of Cell and Molecular Pathology, Medical University of Hannover, Hannover
Search for more papers by this authorChristian Becker
Cologne Centre for Genomics and Institute for Genetics
Search for more papers by this authorGudrun Nürnberg
Cologne Centre for Genomics and Institute for Genetics
Search for more papers by this authorPeter Nürnberg
Cologne Excellence Cluster on Cellular Stress Responses in Ageing-associated Diseases (CECAD), University of Cologne, Cologne, Germany
Search for more papers by this authorMirghani Ali Mohamed Ahmed
College of Medicine, University of Khartoum, Khartoum, Sudan
Search for more papers by this authorJeannette Hübener
Institute of Human Genetics
Present address: J. Hübener, Department of Medical Genetics, University Tuebingen, Germany.
Search for more papers by this authorSummary
Familial thrombosis (FT) has been described as a rare autosomal-dominant disorder, mostly caused by activating mutations of the thrombopoietin gene (THPO). Other cases of FT have been linked to one of two different germline mutations in the myeloproliferative leukaemia virus oncogene gene (MPL), which codes for the thrombopoietin receptor MPL. We studied an Arab family with two siblings with severe thrombocytosis by linkage analysis and obtained evidence for linkage to MPL. Sequencing revealed homozygosity for the novel MPL germline mutation p.Pro106Leu (c.317C > T) in the two siblings. Subsequently, homozygosity for p.Pro106Leu was identified in six further FT patients from three other Arab families. Of 18 heterozygous carriers, 14 had normal platelet counts, while four had mild thrombocytosis. Strong support for association of the novel MPL mutation p.Pro106Leu with development of familial thrombocytosis has been obtained. Overall, p.Pro106Leu was absent on 386 alleles of 193 healthy German controls and present on 14 of 426 alleles (3·3%) of 213 unrelated Arabs, which was statistically significantly different (P < 0·001, Fisher’s exact test). We assume that p.Pro106Leu is a frequent MPL mutation in the Arab population, leading to severe thrombocytosis in homozygotes and occasionally to mild thrombocytosis in heterozygotes. In the families described the mode of inheritance could be regarded as autosomal-recessive with possible mild heterozygote manifestation rather than autosomal-dominant with high penetrance as usually seen in FT.
Supporting Information
Figure S1. LOD score for the MPL region on chromosome 1. Coordinate 0 cM (centi Morgan) at the X-axis equals 52.1 cM at the deCODE genetic map (Kong et al, 2002). A LOD (logarithm of the odds) score of 2.18 was calculated for a candidate region of 27.4 cM (54.9 to 82.3 cM at the deCODE genetic map). MPL is located within the candidate region at 67.8 cM (approximately at coordinate 16.6 cM at the X-axis).
Figure S2. Surface expression of MPL on platelets. Platelets of a normal donor with MPL wild type sequence (ND), of patients VI3 and VI5 with homozygous p.Pro106Leu mutation and their heterozygously mutated parents (V8 and V9) were stained either with a monoclonal antibody (clone AHM21, upper row) or a polyclonal rabbit antiserum against the extracellular domain of human MPL. Grey curves: unspecific control, Black curves: specific antibody.
Table SI. Primer sequences, fragment lengths and annealing temperatures for microsatellite markers linked to the candidate genes THPO, MPL and JAK2. The forward primers (above) were 5′end labelled with fluorescent dye fluorescein.
Table SII. Primer sequences, fragment lengths and annealing temperatures for MPL PCR and sequencing.
Table SIII. Marker analysis at the MPL locus. ID-numbers of patients with FT are written in bold. MPL extends from 43,576 to 43,592 kB (NC_000001.9 NCBI Reference assembly). Markers D1S545 and D1S447 were informative. The affected family members VI:3 and VI:5 were concordant for both parental alleles.
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