Deficiency of ARHGAP21 alters megakaryocytic cell lineage responses and enhances platelet hemostatic function

Author(s)

Bernusso, Vanessa Aline ; Vieira, Karla P. ; Duarte, Adriana S. S. ; Lescano, Caroline Honaiser ; Monica, Fabiola Zakia ; Vicente, Cristina Pontes ; De Paula, Erich Vinicius ; Olalla Saad, Sara Teresinha ; Lazarini, Mariana

• Document type: Journal article
• Source: BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH; v. 1868, n. 6, p. 13-pg., 2021-03-25.

Abstract

Background: Microtubules, actin and Rho GTPase proteins are essential players in the megakaryocyte biology for platelet formation and function. Objectives: To investigate the role of ARHGAP21, a RhoGAP protein, in megakaryocytic differentiation and platelet function. Methods: Cytoskeletal proteins were investigated in HEL cells silenced for ARHGAP21 and submitted to megakaryocyte differentiation. The role of Arhgap21 in platelet function was accessed using haploinsufficient (Arhgap21(+/-)) mice. Arhgap21(+/-) platelet aggregation and p-selectin exposure were evaluated in response to thrombin. Vessel occlusion time and thrombus formation were detected after injury of the carotid artery. Platelet morphology was accessed by electronic microscopy. Results: ARHGAP21 was upregulated during megakaryocytic differentiation of the cell line and primary mouse cells. In the HEL cell model, ARHGAP21 was detected in the cytoplasmic protrusions, colocalized and associated with alpha-tubulin and was mostly detected in the protein cell fraction containing the polymerized tubulin. Silencing of ARHGAP21 decreased the expression of Glu-tubulin, suggesting microtubule instability, and enhanced cell spreading. Platelets from Arhgap21(+/-) mice presented enhanced thrombin-induced aggregation and p-selectin exposure associated with increased size of alpha-granules. Arhgap21(+/-) mice also showed increased CDC42 and RHOA activities, shorter tail-bleeding and accelerated thrombus formation. Conclusions: Our results indicate that ARHGAP21 may be a critical protein in the regulation of platelet production and function through the control of cytoskeletal rearrangement. (AU)

• FAPESP's process: 17/19674-2 - Study of Rho family of GTPases in Myelodysplastic Syndromes and Acute Myeloid Leukemia
• Grantee: Mariana Lazarini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/13022-2 - Study of the role of ARHGAP21 protein in platelet formation and angiogenesis
• Grantee: Vanessa Aline Bernusso
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/21801-2 - Predictors of severity and new treatments for bone marrow neoplasias
• Grantee: Sara Teresinha Olalla Saad
• Support Opportunities: Research Projects - Thematic Grants