Go to the content. | Move to the navigation | Go to the site search | Go to the menu | Contacts | Accessibility

| Create Account

Vettore, Silvia (2008) Caratterizzazione Molecolare delle Macrotrombocitopenie Ereditarie a Trasmissione Autosomica Dominante. [Ph.D. thesis]

Full text disponibile come:

[img]
Preview
Documento PDF
675Kb

Abstract (english)

Patients with an autosomal dominant inheritance of macrothrombocytopenia and not fitting any other known classification such as Myosin Related Diseases (MRD) or Bernard-Soulier Syndrome (BSS), were in the past years assigned to the group of Mediterranean forms, that we also named as Chronic Isolated Macrothrombocytopenia (CHMT). We performed a systematic review of patients with Autosomal Dominant Macrothrombocytopenia in order to better characterize the disease and investigate the molecular defects responsible for the unclassified forms of macrothrombocytopenia. This work was carried out with the new methods (flow cytometry, genetic tests) and diagnostic tools (specific monoclonal antibodies - mAbs) now available also in our laboratory.
Here we described the results obtained from the investigations performed on 5 families (TP1, TP2, TP3, TP4, TP5).

Family TP1.
The propositus was a 49 years old man, whose thrombocytopenia was at the beginning misdiagnosed as ITP. Some year later, his daughter also was recognized as affected. May-Grunwald staining of peripheral blood and Ristocetin aggregation of both patients didn't showed any defect, and they were fist assigned to the CHMT group. When the new methods were implemented, the immunofluorescent staining with mAbs anti Non Muscular Myosin Heavy Chain (NMMCH-IIA) revealed in patients' neutrophil granulocytes an abnormal cytoplasmic distribution of the protein, which is a typical sign of MRD, one of the most common hereditary macrothrombocytopenia.
Genetic analysis revealed in these patients a heterozygous mutation R1162T in the exon 25 of the myh9 gene (coding for the NMMCH-IIA) never described before.
After these results, the diagnosis for TP1 family was changed in MRD.

Family TP2.
The proposita, her father and her sister showed lifelong story of macrothrombocytopenia without spontaneous bleeding. Only the proposita refers metrorrhagia without other hemorrhagic symptoms.
The immunostaining of blood smears with anti NMMCH-IIA mAbs showed normal distribution in the proposita, while in her sister, different experiments revealed an ambiguous distribution of the protein. Sequencing analysis of the myh9 exons seat of known mutation gave negative results.
So we performed on both patients flow cytometry tests in order to study platelets surface glycoproteins expression, and exclude BSS deriving from their anomalies, but these tests also didn't revealed any abnormality. Sequence analysis of GPIba chain of GPIb/V/IX complex (von Willebrand - vWF - Receptor), in which are located the most of the mutations responsible for BSS was normal.
TP2 at the moment remains assigned to the group of uncharacterized forms of CHMT, and the investigations will continue with the sequencing of the remaining exons of myh9 gene and the other chains of vWF receptor in order to definitively exclude MRD and BSS.

Family TP3.
The propositus was a 64 year-old man with no hemorrhagic history. His mother and his son was thrombocytopenic too. In the first test performed in the propositus, Ristocetin aggregation (RIPA) at 1,5 mg/ml was normal, as well as MRD-oriented tests. This lead us toward a provisional diagnosis of CHMT. Conversely, RIPA was reduced lowering the agonist at 1,2 mg/ml. Moreover, in propositus'affected son this test resulted completely impaired both at 1,5 and 1,2 mg/ml. This suggested for this family the diagnosis of BSS, which was confirmed by flow cytometry data on GPIba mAbs binding, reduced in the father and completely absent in the son.
Genetic analysis showed a four bases deletion (TGAG) in gpIba gene already described in a case of compound heterozygosis, bearing a putative truncated protein of 50 aminoacids. Our patient showing absence of GPIba was the first exemple described of homozygous state of this TGAG deletion. It is interesting to note that after genetical screening, also the unaffected mother of the homozygous subject (she and her husband are consanguineous of 3rd generation) carried the mutation at heterozygous state. This result underlines that the autosomal dominance of the trait shows an incomplete penetrance in one of the heterozygous and consanguineous carriers.

Family TP4 and TP5.
Following the previous scheme, we found that the affected members of this two unrelated families were actually heterozygous BSS patients. Sequencing analysis in this cases revealed a novel mutation, a heterozygous A>C transversion at nucleotide +169, resulting in an N41H substitution in the GPI? protein sequence. The amino acid substitution, named Padova variant of BSS, is located in the first leucine-rich repeat (LRR) of the protein. Replacement of the asparagine 41 with a histidine (N41H) drastically disturbs the conformational behavior of the first portion of the N-terminal region of GPIb?, which is directly involved in vWF binding. In fact, the mutant N41H lost two of the three stabilizing interactions during the molecular dynamics simulation.
The two unrelated families described here represent a form of heterozygous BSS with an autosomal dominant inheritance never described before.

We can conclude that the new diagnostic tools allowed to better characterize our patients affected by Autosomal Dominant Macrothrombocytopenia.
Four Families out of five were assigned to the more characterized areas of MRD (1 Family) and BSS (3 Families), but the defects found responsible for the disease are new mutations (families TP1, TP4 and TP5) or are present in an homozygous condition (Family TP3) never described before.
One Family (TP2) remains to be characterized and will be matter of further investigations.
The study also allowed to identified conformational mAbs able to differentiate the newly identified BSS-Padova Variant, from other BSS forms.


Statistiche Download - Aggiungi a RefWorks
EPrint type:Ph.D. thesis
Tutor:Fabris, Fabrizio
Ph.D. course:Ciclo 20 > Scuole per il 20simo ciclo > SCIENZE MEDICHE, CLINICHE E SPERIMENTALI > METODOLOGIA CLINICA ED EMATOLOGIA
Data di deposito della tesi:January 2008
Anno di Pubblicazione:January 2008
Key Words:Macrotrombocitopenia ereditaria, Bernard-Soulier Syndrome, Myosin Related Diseases
Settori scientifico-disciplinari MIUR:Area 06 - Scienze mediche > MED/15 Malattie del sangue
Struttura di riferimento:Dipartimenti > pre 2012 - Dipartimento di Scienze Mediche e Chirurgiche
Codice ID:455
Depositato il:13 Nov 2008
Simple Metadata
Full Metadata
EndNote Format

Bibliografia

I riferimenti della bibliografia possono essere cercati con Cerca la citazione di AIRE, copiando il titolo dell'articolo (o del libro) e la rivista (se presente) nei campi appositi di "Cerca la Citazione di AIRE".
Le url contenute in alcuni riferimenti sono raggiungibili cliccando sul link alla fine della citazione (Vai!) e tramite Google (Ricerca con Google). Il risultato dipende dalla formattazione della citazione.

- Afshar-Khargan V and Lopez JA. Bernard-Soulier Syndrome caused by a dinucleotide deletion and reading frameshift in the region encoding the glycoprotein Iba transmembrane domain. Blood 1997;90:2634-43 Cerca con Google

- Afshar-Khargan V, Craig FE, Lopez JA. Bernard-Soulier syndrome in a patient doubly heterozygous for two frameshift mutations in the glycoprotein Iba gene. Br. J. Haemat. 2000,110:919-924. Cerca con Google

- Afshar-Kharghan V, Gineys G, Schade AJ, Sun L, Li CQ, McIntire LV, Dong JF and Lopez JA. Necessity of conserved Asparagine residues in the Leucine-Rich Repeats of platelet glycoprotein Iba for the proper conformation and function of the ligand-binding region. Biochemistry; 2000,39:3384-3391 Cerca con Google

- Balduini CL, Cattaneo M, Fabris F, Gresele P, Iolascon A, Pulcinelli FM, Savoia A; Italian Gruppo di Studio delle Piastrine. Inherited thrombocytopenias: a proposed diagnostic algorithm from the Italian Gruppo di Studio delle Piastrine. Haematologica. 2003;88:582-92 Cerca con Google

- Balduini CL, Iolascon A, Savoia A. Inherited thrombocitopenias: from genes to therapy. Haematologica 2002, 87: 860-80 Cerca con Google

- Ballmaier M., Germeshausen M, Schulze H, Cherkaoui K, Lang S, Gaudig A, Krukemeier S, Eilers M, Strauß G, and Welte K. c-mpl mutations are the cause of congenital amegakaryocytic thrombocytopenia. Blood, 2001;97:139-146 Cerca con Google

- Celikel R, McClintock R.A., Roberts J.R., Mendolicchio G.L., Ware J., Varughese K.I., Ruggeri Z.M. Modulation of alpha-thrombin function by distinct interactions with platelet glycoprotein Ibalpha Science 2003, 301:218-221 Cerca con Google

- Cornell W.D.C., Bayly C.I., Gould I.R., Merz K.M., Ferguson D.M., Spellmeyer D.C., Fox T., Caldwell J.W., Kollman P.A. A second generation force field for the simulation of proteins, nucleic acids and organic molecules. J. Am. Chem. Soc. 1995, 117:5179-5196 Cerca con Google

- De Marco L, Mazzuccato M, Fabris F, De Roia D, Coser P, Girolami A, Vicente V, Ruggeri ZM. Variant Bernard - Soulier Sindrome Type Bolzano. A Congenital Bleeding Disorder Due to a Structural and Functional Abnormality of the Platelet Glycoprotein Ib-IX Complex. J Clin Invest 1990, 86:25-31 Cerca con Google

- Drachman J.G. Inherited Thrombocytopenia: when a low platelet count does not mean ITP. Blood 2004; 103:390-398 Cerca con Google

- Epstein CJ, Shaud MA, Piel CF, Goodman JR, Bernfield MR, Kushner JH, Ablin AR. Hereditary macrothrombocytopenia, nephritis and deafness. Am J Med. 1972;52:299-310 Cerca con Google

- Fabris F, Cordiano I, Salvan F, Ramon R, Valente M, Luzzatto G, Girolami A. Chronic isolated macrothrombocytopenia with autosomal dominant transmission: a morphological and qualitative platelet disorder. Eur J Haematol 1997,58:40-45 Cerca con Google

- Fabris F, Cordiano I, Steffan A, Ramon R, Scandellari R, Nichol JL, Girolami A. A direct study of thrombopoiesis (TPO, reticulated platelets, glycocalicin) in patients with hereditary macrothrombocytopenia. Eur J Haematol 2000,64:151-156 Cerca con Google

- Fabris F, Fagioli F, Basso G, Girolami A. Autosomal dominant macrothrombocytopenia with ineffective thrombopoiesis. Haematologica 2002, 87: ELT27 Cerca con Google

- Frederiksen H, Schmidt K. The incidence of the idiopatic thrombocytopenic purpura in adults increases with age. Blood 1999;94:909-913 Cerca con Google

- Freson K, Devriendt K, Matthijs G, Van Hoof A, De Vos R, Thys C, Minner K, Hoylaerts MF, Vermylen J, Van Geet C. Platelet characteristics in patients with X-linked macrothrombocytopenia because of a novel GATA1 mutation.Blood. 2001, 98:85-92 Cerca con Google

- Girolami A.; Randi ML; Casonato A; Pasini L; Boccato C; Fabris F. A study of platelet function and morphology in a new family with May Hegglin anomaly. Folia Haemat 1980;2:256-268 Cerca con Google

- Gohda F; Uchiumi H; Handa H; Matsushima T; Tsukamoto N; Morita K; Amagai H; Murakami M; Murakami H; Nojima Y; Karasawa M. Identification of inherited thrombocytopenia based on mean platelet volume among patients diagnosed with idiopathic thrombocytopenia. Thrombosis Research, 2007;119:741-746. Cerca con Google

- Gonzalez-Manchon C, Larrucea S, Pastor AL, Butta N, Arias-Salgado EG, Ayuso MS, Parrilla R. Compound Heterozygosity of the gpIba gene associated with Bernard-Soulier Syndrome. Thromb Haemost 2001;86:1385-91 Cerca con Google

- Greinacher A, Nieuwenhuis HK, White JG. Sebastian platelet syndrome: a new variant of hereditary macrothrombocytopenia with leukocyte inclusions. Blut. 1990, 61: 282-288 Cerca con Google

- Heath KE, Campos-Barros A, Toren A, Rozennfeld-Granot G, Carlsson LE, Savige J, Denison JC, Gregory MC, White JG, Baker DF, Greinacher A, Epstein CJ, Gluksman MJ and Martignetti J. Nonmuscle myosin heavy chain IIA mutations define a spectrum of autosomal dominant macrothrombocytopenias; May-Hegglin anomaly and Fetchner, Sebastian, Epstein, and Alport Syndromes. Am J Hum Genet.2001;69:1033-1045 Cerca con Google

- Hegglin R. Gleichzeitige konstitutionelle Veranderungen an Neurtophilen und Thrombocyten. Helv. Med. Acta 1945, 12: 439-440 Cerca con Google

- Huizinga EG; Tsuji S; Romijn RA; Shiphorst ME; de Groot PG; Sixma JJ and Gros P.Structures of Glycoprotein Iba and its complex with von Willebrand Factor A1 domain.Science 2002; 297, 1176-1179 Cerca con Google

- Kenny D., Jonsson OG, Morateck PA, Montgomery RR. Naturally occurring mutations in glycoprotein Ib alpha that result in defective ligand binding and synthesis of a truncated protein. Blood 1998,92:175-183. Cerca con Google

- Kunishima S, Imai T, Hamaguchi M, Saito H. Novel heterozygous missense mutation in the second leucine rich repeat of GPIba affects GPIb/IX/V expression and results in macrothrombocytopenia in a patient initially misdiagnosed with idiopathic thrombocytopenic purpura. Eur J Haematol 2006,74:348-355 Cerca con Google

- Kunishima S, Miura H, Fukutani H, Yoshida H, Osumi K, Kobayashi S, Ohno R, Naoe T. Bernard-Soulier syndrome Kagoshima: Ser 444>Stop mutation of GP Iba resulting in circulating truncated GPIba and surface expression of GP Ibb and GPIX. Blood 1994;84:3356-62 Cerca con Google

- Li C, Pasquale DN, Roth GJ. Bernard-Soulier syndrome with severe bleeding: absent platelet glycoprotein Ib alpha due to homozygous one-base deletion. Thromb Haemost 1996;76:670-4 Cerca con Google

- Lopez JA, Andrews RK, Afshar-Kharghan V, Berndt MC. Bernard-Soulier syndrome. Blood. 1998, 91:4397-418 Cerca con Google

- Lopez JA. The platelet glycoprotein Ib-IX complex. Blood coagul and Fibrinol 1994;5:97-119 Cerca con Google

- Lova P, Campus F, Mancini F, Balduini C, Torti M. Evidence for thrombin-induced platelet activation independent of PAR1 PAR4 and GPIb/V/IX. J Thromb Haemost 2007;5 Supplement 2:P-W-251 Cerca con Google

- Margaglione M, D'Andrea G, Grandone E, Brancaccio V, Amoriello A, Di Minno G. Compound heterozygosity (554-589 del, C515-T transition) in the platelet glycoprotein Ib alpha gene in a patient with a severe bleeding tendency. Thromb Haemost. 1999, 81:486-92 Cerca con Google

- Margaglione M, D'Andrea G, Grandone E, Brancaccio V, Amoriello A, Di Minno G. Compound heterozygosity (554-589 del, C515-T transition) in the platelet Glycoprotein Iba gene in a patient with a severe bleeding tendency. Thromb Haemost 1999;81:486-92. Cerca con Google

- Marigo V.; Nigro A.; Pecci A.; Montanaro D.; Di Stazio M.; Balduini C. & Savoia A. Correlation Between the clinical phenotype of MYH9-related disease and tissue distribution of class II non muscular myosine heavy chains Genomics 2004, 83:1125-1133 Cerca con Google

- Miller JL, Cunningham D, Lyle V., Finch CN. Mutation in the gene encoding the a chain of platelet glycoprotein Ib in platelet-type von Willebrand disease. PNAS,1991 88:4761-4765. Cerca con Google

- Miller JL, Lyle VA, Cunningam D. Mutation of leucine-57 to phenylalanine in a platelet glycoprotein Iba leucine tandem repeat occurring in patients with an autosomal dominant variant of Bernard-Soulier disease. Blood 1992;79:439-446 Cerca con Google

- Mitsui T, Yokoyama S, Yazaki N, HayashiT, Suzuki K, Shimizu Y, Kawakami T, Kanazawa C, Katsuura M, Ikegami T, Hayasaka K. Severe bleeding tendency in a patient with Bernard-Soulier syndrome associated with a homozygous single base pair deletion in the gene coding for the human platelet glycoprotein Ibalpha gene. J Pediatr Hematol Oncol. 1998;20:246-51 Cerca con Google

- Molecular Operating Environment, version 2006.08; Chemical Computing Group: Montreal, Canada, 2006 Cerca con Google

- Najean Y, Lecompte T. Genetic thrombocytopenia with autosomal dominant transmission: a review of 54 cases. Br J Haematol. 1990, 74:203-8 Cerca con Google

- Nichols KE, Crispino JD, Poncz M, White JG, Orkin SH, Maris JM, Weiss MJ. Familial dyserythropoietic anaemia and thrombocytopenia due to an inherited mutation in GATA 1. Nature Genetics 2000, 24:266-270 Cerca con Google

- Noda M, Fujimura K, Takafuta T, Shimomura T, Fujimoto T, Yamamoto N, Tanoue K, Arai M, Suehiro A, Kakishita E et al. Heterogeneous expression of glycoprotein Ib, IX and V in platelets from two patients with Bernard-Soulier syndrome caused by different genetic abnormalities. Thromb Haemost 1995;74:1411-15. Cerca con Google

- Noris P, Guidetti GF, Conti V, Ceresa IF, Di Pumpo M, Pecci A, Torti M, Savoia A, Balduini CL. Autosomal dominant thrombocytopenias with reduced expression of glycoprotein Ia. Thromb Haemost 2006; 95:483-9 Cerca con Google

- Noris P., Pecci A., Di Bari F., Di Stazio MT., Di Pumpo M., Ceresa IF., Arezzi N., Ambaglio C., Savoia A., Balduini CL. Application of a diagnostic algorithm for inherited thrombocytopenias to 46 consecutive patients. Haematologica 2004;89:1219-1225 Cerca con Google

- Peng Y, Shrimptom CN, Dong FJ and Lopez JA. Gain of von Willebrand factor binding function by mutagenesis of a species-conserved residue within the leucine-rich repeat region of platelet glycoprotein Iba. Blood. 2005;106:1982-1987 Cerca con Google

- Peterson LC, Rao KV, Crosson JT & White JG. Fechtner syndrome: a variant of Alport's syndrome with leukocyte inclusions and macrothrombocytopenia. Blood 1985, 65: 397-406 Cerca con Google

- Ryckaert J.P., Ciccotti G., Berendsen H.J.C. Numerical integration of the Cartesian equations of motion of a system with constraints: molecular dynamics of n-alkanes. J. Comput. Phys. 1977, 23:327-341 Cerca con Google

- Savoia A, Balduini CL, Savino M, Noris P, Del Vecchio M, Perrotta S, Belletti S, Poggi V, Iolascon A. Autosomal dominant macrothrombocytopenia in Italy is most frequently a type of heterozygous Bernard-Soulier sindrome. Blood 2001,97:1330-1335 Cerca con Google

- Seri M, Pecci A, Di Bari F, Cusano R, Savino M, Panza E, Nigro A, Noris P, Gangarossa S, Rocca B, Gresele P, Bizzaro N, Malatesta P, Koivisto PA, Longo I, Musso R, Pecoraro C, Iolascon A, Magrini U, Rodriguez Soriano J, Renieri A, Ghiggeri GM, Ravazzolo R, Balduini CL, Savoia. MYH9-related disease: May-Hegglin anomaly, Sebastian syndrome, Fechtner syndrome, and Epstein syndrome are not distinct entities but represent a variable expression of a single illness. Medicine 2003;82:203-15 Cerca con Google

- Shen Y, Romo GM, Domg J, Schade A, McIntire LV, Kenny D, Whisstock, Berndt MC, Lopez JA and Andrews RK. Requirements of leucine-rich repeats of glycoprotein GPIba for shear-dependent and static binding of von Willebrand factor to the platelet membrane GP Ib-IX-V complex. Blood. 2000;95:903-910 Cerca con Google

- Shvdasani RA. Molecular and transcriptional regulation of megacaryocyte differentiation. Stem Cells 2001; 19:397-407 Cerca con Google

- Tsui V., Case D.A. Theory and applications of the generalized Born solvation model in macromolecular simulations. Biopolymers 2001, 56:275-291 Cerca con Google

- Unkelbach K, Kalb R, Santoso S, Kroll H, Mueller-Eckhardt and V. Kiefel. Genomic RFLP typing of human platelet alloantigens Zw(PlA), Ko, Bak and Br (HPA-1,2,3,5). Br J of Haemat,1995;89:169-176 Cerca con Google

- Von Behrens WE Mediterranean macrotrombocytopenia. Blood 1975; 46:199-208 Cerca con Google

- Ward CM, Andrews RK, Smith AI and Berndt MC. Mocharagin, a novel cobra venom metalloproteinase, cleaves the platelet von Willebrand Factor Receptor Glycoprotein Iba. Identification of the sulfated Tyrosine/anionic sequence Tyr276-Glu282 of glycoprotein Iba as a binding site for von Willebrand Factor and a-thrombin. Biochemistry, 1996,35:4929-4938 Cerca con Google

- Ware J, Russell SR, Marchese P, Murata M, Mazzucato M, De Marco L, Ruggeri ZM. Point mutation in a leucine-rich repeat of platelet glycoprotein Ib alpha resulting in the Bernard-Soulier syndrome. J Clin Invest. 1993, 92:1213-20 Cerca con Google

- Ware J., Russel S.R., Vicente V., Scharf R., Tomer A., McMilland R., Ruggeri Z. Nonsense mutation in the glycoprotein Iba coding sequence associated with Bernard-Soulier syndrome. PNAS 1990; 87:2026-2030 Cerca con Google

- Whisstock JC; Shen Y; Lopez JA; Andrews RK; Berndt MC. Molecular modeling of the seven tandem Leucine-Rich Repeats within the ligand-binding region of platelet glycoprotein Iba. Thromb Haemost 2002; 87:329-33 Cerca con Google

Download statistics

Solo per lo Staff dell Archivio: Modifica questo record