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Identification of novel TUBB1 variants in patients with macrothrombocytopenia

BACKGROUND/AIM: Macrothrombocytopenia is an autosomal-dominant disorder characterized by increased platelet size and a decreased number of circulating platelets. The membrane skeleton and the link between actin filaments of the skeleton and microtubules, which consist of alpha and beta tubulin [incl...

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Autores principales: ÇALIŞKANER, Zihni Onur, WAHEED, Abdullah Abdul, TUZLAKOĞLU ÖZTÜRK, Merve, OYMAK, Yeşim, TAZEBAY, Uygar Halis, AKAR, Nejat, KANDİLCİ, Ayten, TORUN ÖZKAN, Didem
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Scientific and Technological Research Council of Turkey 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8203157/
https://www.ncbi.nlm.nih.gov/pubmed/32892537
http://dx.doi.org/10.3906/sag-2003-259
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author ÇALIŞKANER, Zihni Onur
WAHEED, Abdullah Abdul
TUZLAKOĞLU ÖZTÜRK, Merve
OYMAK, Yeşim
TAZEBAY, Uygar Halis
AKAR, Nejat
KANDİLCİ, Ayten
TORUN ÖZKAN, Didem
author_facet ÇALIŞKANER, Zihni Onur
WAHEED, Abdullah Abdul
TUZLAKOĞLU ÖZTÜRK, Merve
OYMAK, Yeşim
TAZEBAY, Uygar Halis
AKAR, Nejat
KANDİLCİ, Ayten
TORUN ÖZKAN, Didem
author_sort ÇALIŞKANER, Zihni Onur
collection PubMed
description BACKGROUND/AIM: Macrothrombocytopenia is an autosomal-dominant disorder characterized by increased platelet size and a decreased number of circulating platelets. The membrane skeleton and the link between actin filaments of the skeleton and microtubules, which consist of alpha and beta tubulin [including the tubulin beta-1 chain (TUBB1)] heterodimers, are important for normal platelet morphology, and defects in these systems are associated with macrothrombocytopenia. MATERIALS AND METHODS: In this study, we sequenced the exons of the TUBB1 gene using DNA isolated from the peripheral blood samples of healthy controls (n = 47) and patients with macrothrombocytopenia (n = 37) from Turkey. The TUBB1 expression levels in fractioned blood samples from patients and healthy controls were analyzed by RT-qPCR and Western blot. Microtubule organization of the platelets in the peripheral blood smears of patients, and in mutant TUBB1 -transfected HeLa cells, were analyzed by immunofluorescence staining. RESULTS: A new TUBB1 c.803G>T (p.T178T) variant was detected in all of the control and patient samples. Importantly, we found 3 new heterozygous TUBB1 variants predicting amino acid substitutions: G146R (in 1 patient), E123Q (in 1 patient), and T274M (in 4 patients); the latter variant was associated with milder thrombocytopenia in cancer patients treated with paclitaxel. Ectopic expression of TUBB1 T274M/R307H variant in HeLa cells resulted in irregular microtubule organization. CONCLUSION: Further clinical and functional studies of the newly identified TUBB1 variants may offer important insights into their pathogenicity in macrothrombocytopenia.
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spelling pubmed-82031572021-06-24 Identification of novel TUBB1 variants in patients with macrothrombocytopenia ÇALIŞKANER, Zihni Onur WAHEED, Abdullah Abdul TUZLAKOĞLU ÖZTÜRK, Merve OYMAK, Yeşim TAZEBAY, Uygar Halis AKAR, Nejat KANDİLCİ, Ayten TORUN ÖZKAN, Didem Turk J Med Sci Article BACKGROUND/AIM: Macrothrombocytopenia is an autosomal-dominant disorder characterized by increased platelet size and a decreased number of circulating platelets. The membrane skeleton and the link between actin filaments of the skeleton and microtubules, which consist of alpha and beta tubulin [including the tubulin beta-1 chain (TUBB1)] heterodimers, are important for normal platelet morphology, and defects in these systems are associated with macrothrombocytopenia. MATERIALS AND METHODS: In this study, we sequenced the exons of the TUBB1 gene using DNA isolated from the peripheral blood samples of healthy controls (n = 47) and patients with macrothrombocytopenia (n = 37) from Turkey. The TUBB1 expression levels in fractioned blood samples from patients and healthy controls were analyzed by RT-qPCR and Western blot. Microtubule organization of the platelets in the peripheral blood smears of patients, and in mutant TUBB1 -transfected HeLa cells, were analyzed by immunofluorescence staining. RESULTS: A new TUBB1 c.803G>T (p.T178T) variant was detected in all of the control and patient samples. Importantly, we found 3 new heterozygous TUBB1 variants predicting amino acid substitutions: G146R (in 1 patient), E123Q (in 1 patient), and T274M (in 4 patients); the latter variant was associated with milder thrombocytopenia in cancer patients treated with paclitaxel. Ectopic expression of TUBB1 T274M/R307H variant in HeLa cells resulted in irregular microtubule organization. CONCLUSION: Further clinical and functional studies of the newly identified TUBB1 variants may offer important insights into their pathogenicity in macrothrombocytopenia. The Scientific and Technological Research Council of Turkey 2021-04-30 /pmc/articles/PMC8203157/ /pubmed/32892537 http://dx.doi.org/10.3906/sag-2003-259 Text en Copyright © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Article
ÇALIŞKANER, Zihni Onur
WAHEED, Abdullah Abdul
TUZLAKOĞLU ÖZTÜRK, Merve
OYMAK, Yeşim
TAZEBAY, Uygar Halis
AKAR, Nejat
KANDİLCİ, Ayten
TORUN ÖZKAN, Didem
Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title_full Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title_fullStr Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title_full_unstemmed Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title_short Identification of novel TUBB1 variants in patients with macrothrombocytopenia
title_sort identification of novel tubb1 variants in patients with macrothrombocytopenia
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8203157/
https://www.ncbi.nlm.nih.gov/pubmed/32892537
http://dx.doi.org/10.3906/sag-2003-259
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