Platelet Activation and Reactivity in a Large Cohort of Patients with Gaucher Disease

Shoshana Revel-Vilk; Mira Naamad; Dafna Frydman; Michael R. Freund; Tama Dinur; Majdolen Istaiti; Michal Becker-Cohen; Roni Falk; Eti Broide; Alan D. Michelson; Andrew L. Frelinger III; Ari Zimran

doi:10.1055/a-1642-4206

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2022; 122(06): 951-960
DOI: 10.1055/a-1642-4206

Cellular Haemostasis and Platelets

Platelet Activation and Reactivity in a Large Cohort of Patients with Gaucher Disease

Shoshana Revel-Vilk

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

²Pediatric Hematology/Oncology Unit, Shaare Zedek Medical Center, Jerusalem, Israel

³Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel

,

Mira Naamad

⁴Flow Cytometry Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Dafna Frydman

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Michael R. Freund

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Tama Dinur

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Majdolen Istaiti

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Michal Becker-Cohen

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Roni Falk

⁴Flow Cytometry Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Eti Broide

⁴Flow Cytometry Unit, Shaare Zedek Medical Center, Jerusalem, Israel

,

Alan D. Michelson

⁵Center for Platelet Research Studies, Dana-Farber/Boston Children's Cancer and Blood Disorders Center, Harvard Medical School, Boston, United States

,

Andrew L. Frelinger III

⁵Center for Platelet Research Studies, Dana-Farber/Boston Children's Cancer and Blood Disorders Center, Harvard Medical School, Boston, United States

,

Ari Zimran

¹Gaucher Unit, Shaare Zedek Medical Center, Jerusalem, Israel

³Faculty of Medicine, Hebrew University of Jerusalem, Jerusalem, Israel

› Author Affiliations

Abstract

Objectives Patients with Gaucher disease (GD) are at increased risk of bleeding and have varying degrees of thrombocytopenia, making the analysis of platelet function difficult. This study aimed to provide a clinically relevant quantitative assessment of platelet function and determine its relationship with bleeding and GD-related data.

Methods Unstimulated and stimulated platelet function was measured by whole blood flow cytometry of platelet surface-activated αIIbβ3 integrin (detected with monoclonal antibody PAC1), P-selectin (CD62P), and lysosomal-associated membrane protein (LAMP3/CD63) in 149 GD patients.

Results GD patients had a higher level of unstimulated CD63 expression than healthy subjects, which was mildly correlated with glucosylsphingosine (lyso-Gb1) levels (r = 0.17, p-value = 0.042). Splenectomized GD patients had a higher level of unstimulated αIIbβ3 integrin and P-selectin expression. Reduced platelet reactivity (−2 standard deviation of reference range) was found in 79 (53%, 95% confidence interval [CI]: 44–61%) patients, of whom 10 (6.7%, 95% CI: 3.3–12%) had more severe platelet dysfunction. In a multivariate model, only lyso-Gb1 levels were associated with the more severe platelet dysfunction. Fifty-four (49%) of 128 adult patients who completed the bleeding tendency questionnaire reported positive bleeding history. In a multivariate logistic model, older age (odds ratio [OR]: 1.05, 95% CI: 1.01–1.1) and low P-selectin reactivity (OR: 2.03, 95% CI: 1.25–3.35) were associated with more than one bleeding manifestation.

Conclusion Flow cytometry enables the study of platelet function in thrombocytopenic GD patients. A platelet degranulation defect, but not αIIbβ3 integrin activation defect, is associated with clinical bleeding. In vivo increased CD63 expression may be related to GD-related inflammation.

Keywords

bleeding - flow cytometry - Gaucher disease - platelet activation - platelet function

Full Text

References

References
1 Revel-Vilk S, Szer J, Zimran A. Gaucher disease and related lysosomal storage diseases. In: Kaushansky K, Lichtman M, Prchal J. et al., eds. Williams Hematology. 10th ed.. New York, NY: McGraw-Hill; 2021: 1189-1202
2 Mitrovic M, Antic D, Elezovic I. et al. Haemostatic abnormalities in treatment-naïve patients with Type 1 Gaucher's disease. Platelets 2012; 23 (02) 143-149
3 Rosenbaum H. Hemorrhagic aspects of Gaucher disease. Rambam Maimonides Med J 2014; 5 (04) e0039
4 Simchen MJ, Oz R, Shenkman B, Zimran A, Elstein D, Kenet G. Impaired platelet function and peripartum bleeding in women with Gaucher disease. Thromb Haemost 2011; 105 (03) 509-514
5 Revel-Vilk S, Szer J, Zimran A. Hematological manifestations and complications of Gaucher disease. Expert Rev Hematol 2021; 14 (04) 347-354
6 Gillis S, Hyam E, Abrahamov A, Elstein D, Zimran A. Platelet function abnormalities in Gaucher disease patients. Am J Hematol 1999; 61 (02) 103-106
7 Giona F, Palumbo G, Amendola A, Santoro C, Mazzuconi MG. Platelet function and coagulation abnormalities in type 1 Gaucher disease patients: effects of enzyme replacement therapy (ERT). J Thromb Haemost 2006; 4 (08) 1831-1833
8 Spectre G, Roth B, Ronen G. et al. Platelet adhesion defect in type I Gaucher disease is associated with a risk of mucosal bleeding. Br J Haematol 2011; 153 (03) 372-378
9 Ramström S, Södergren AL, Tynngård N, Lindahl TL. Platelet function determined by flow cytometry: new perspectives?. Semin Thromb Hemost 2016; 42 (03) 268-281
10 Michelson AD. Flow cytometry: a clinical test of platelet function. Blood 1996; 87 (12) 4925-4936
11 He R, Chen D. Confirmatory testing for diagnosis of platelet disorders. In: Shaz BH, Hillyer CD, Reyes Gil M. eds. Transfusion Medicine and Hemostasis. 3rd ed.. Amsterdam: Elsevier; 2019: 841-848
12 Michelson AD, Barnard MR, Krueger LA, Frelinger III AL, Furman MI. Evaluation of platelet function by flow cytometry. Methods 2000; 21 (03) 259-270
13 Frelinger III AL, Grace RF, Gerrits AJ. et al. Platelet function tests, independent of platelet count, are associated with bleeding severity in ITP. Blood 2015; 126 (07) 873-879
14 Frelinger III AL, Grace RF, Gerrits AJ, Carmichael SL, Forde EE, Michelson AD. Platelet function in ITP, independent of platelet count, is consistent over time and is associated with both current and subsequent bleeding severity. Thromb Haemost 2018; 118 (01) 143-151
15 Boknäs N, Ramström S, Faxälv L, Lindahl TL. Flow cytometry-based platelet function testing is predictive of symptom burden in a cohort of bleeders. Platelets 2018; 29 (05) 512-519
16 Rodeghiero F, Tosetto A, Abshire T. et al; ISTH/SSC joint VWF and Perinatal/Pediatric Hemostasis Subcommittees Working Group. ISTH/SSC bleeding assessment tool: a standardized questionnaire and a proposal for a new bleeding score for inherited bleeding disorders. J Thromb Haemost 2010; 8 (09) 2063-2065
17 Zimran A, Sorge J, Gross E, Kubitz M, West C, Beutler E. Prediction of severity of Gaucher's disease by identification of mutations at DNA level. Lancet 1989; 2 (8659): 349-352
18 Revel-Vilk S, Fuller M, Zimran A. Value of glucosylsphingosine (Lyso-Gb1) as a biomarker in Gaucher disease: a systematic literature review. Int J Mol Sci 2020; 21 (19) E7159
19 Rolfs A, Giese AK, Grittner U. et al. Glucosylsphingosine is a highly sensitive and specific biomarker for primary diagnostic and follow-up monitoring in Gaucher disease in a non-Jewish, Caucasian cohort of Gaucher disease patients. PLoS One 2013; 8 (11) e79732
20 Huskens D, Sang Y, Konings J. et al. Standardization and reference ranges for whole blood platelet function measurements using a flow cytometric platelet activation test. PLoS One 2018; 13 (02) e0192079
21 Achison M, Joel C, Hargreaves PG, Sage SO, Barnes MJ, Farndale RW. Signals elicited from human platelets by synthetic, triple helical, collagen-like peptides. Blood Coagul Fibrinolysis 1996; 7 (02) 149-152
22 Doyle EL, Ridger V, Ferraro F, Turmaine M, Saftig P, Cutler DF. CD63 is an essential cofactor to leukocyte recruitment by endothelial P-selectin. Blood 2011; 118 (15) 4265-4273
23 Kraft S, Jouvin MH, Kulkarni N. et al. The tetraspanin CD63 is required for efficient IgE-mediated mast cell degranulation and anaphylaxis. J Immunol 2013; 191 (06) 2871-2878
24 Pandey MK, Burrow TA, Rani R. et al. Complement drives glucosylceramide accumulation and tissue inflammation in Gaucher disease. Nature 2017; 543 (7643): 108-112
25 Rigante D, Cipolla C, Basile U, Gulli F, Savastano MC. Overview of immune abnormalities in lysosomal storage disorders. Immunol Lett 2017; 188: 79-85
26 Marquardt L, Ruf A, Mansmann U. et al. Course of platelet activation markers after ischemic stroke. Stroke 2002; 33 (11) 2570-2574
27 Soma P, Swanepoel AC, du Plooy JN, Mqoco T, Pretorius E. Flow cytometric analysis of platelets type 2 diabetes mellitus reveals ‘angry’ platelets. Cardiovasc Diabetol 2016; 15: 52
28 Arbesu I, Bucsaiova M, Fischer MB, Mannhalter C. Platelet-borne complement proteins and their role in platelet-bacteria interactions. J Thromb Haemost 2016; 14 (11) 2241-2252
29 Bongiovanni D, Klug M, Lazareva O. et al. SARS-CoV-2 infection is associated with a pro-thrombotic platelet phenotype. Cell Death Dis 2021; 12 (01) 50
30 Israels SJ, McMillan-Ward EM. CD63 modulates spreading and tyrosine phosphorylation of platelets on immobilized fibrinogen. Thromb Haemost 2005; 93 (02) 311-318
31 Metzelaar MJ, Wijngaard PL, Peters PJ, Sixma JJ, Nieuwenhuis HK, Clevers HC. CD63 antigen. A novel lysosomal membrane glycoprotein, cloned by a screening procedure for intracellular antigens in eukaryotic cells. J Biol Chem 1991; 266 (05) 3239-3245
32 Zimmer KP, le Coutre P, Aerts HM. et al. Intracellular transport of acid beta-glucosidase and lysosome-associated membrane proteins is affected in Gaucher's disease (G202R mutation). J Pathol 1999; 188 (04) 407-414
33 Chanpeng P, Svasti S, Paiboonsukwong K, Smith DR, Leecharoenkiat K. Platelet proteome reveals specific proteins associated with platelet activation and the hypercoagulable state in β-thalassmia/HbE patients. Sci Rep 2019; 9 (01) 6059
34 Pommerening MJ, Rahbar E, Minei K. et al. Splenectomy is associated with hypercoagulable thrombelastography values and increased risk of thromboembolism. Surgery 2015; 158 (03) 618-626
35 Zhang H, Zhang S, Zhang J. et al. Improvement of human platelet aggregation post-splenectomy with paraesophagogastric devascularization in chronic hepatitis B patients with cirrhotic hypersplenism. Platelets 2020; 31 (08) 1019-1027
36 Shitrit D, Rudensky B, Zimran A, Elstein D. D-dimer assay in Gaucher disease: correlation with severity of bone and lung involvement. Am J Hematol 2003; 73 (04) 236-239
37 Komninaka V, Repa K, Marinakis T. et al. Platelet function defects in patients with Gaucher disease on long term ERT- implications for evaluation at bleeding challenges. Blood Cells Mol Dis 2020; 80: 102371
38 Lukas J, Cozma C, Yang F. et al. Glucosylsphingosine causes hematological and visceral changes in mice-evidence for a pathophysiological role in Gaucher disease. Int J Mol Sci 2017; 18 (10) E2192
39 Yatziv S, White M, Eldor A. Lysosomal enzyme activities in platelets of normal individuals and of patients with Gaucher's disease. Thromb Diath Haemorrh 1974; 32 (2–3): 665-669
40 Beutler E, Kuhl W, Matsumoto F, Pangalis G. Acid hydrolases in leukocytes and platelets of normal subjects and in patients with Gaucher's and Fabry's disease. J Exp Med 1976; 143 (04) 975-980
41 Merten M, Thiagarajan P. P-selectin expression on platelets determines size and stability of platelet aggregates. Circulation 2000; 102 (16) 1931-1936