Platelet transcriptome analysis in patients with germline RUNX1 mutations

• Published in: Journal of thrombosis and haemostasis Vol. 21; no. 5; pp. 1352 - 1365
• Main Authors: Palma-Barqueros, Verónica, Bastida, José María, López Andreo, María José, Zámora-Cánovas, Ana, Zaninetti, Carlo, Ruiz-Pividal, Juan Francisco, Bohdan, Natalia, Padilla, José, Teruel-Montoya, Raúl, Marín-Quilez, Ana, Revilla, Nuria, Sánchez-Fuentes, Ana, Rodriguez-Alen, Agustín, Benito, Rocío, Vicente, Vicente, Iturbe, Teodoro, Greinacher, Andreas, Lozano, María Luisa, Rivera, José
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.05.2023
• Subjects: Blood Platelet Disorders - complications; Core Binding Factor Alpha 2 Subunit - genetics; Core Binding Factor Alpha 2 Subunit - metabolism; Gene Expression Profiling; Germ Cells - metabolism; Germ-Line Mutation; high-throughput sequencing; Humans; inherited thrombocytopenias; leukemia; Leukemia, Myeloid, Acute - genetics; Mutation; myelodysplastic syndrome; Myelodysplastic Syndromes; platelet transcriptomics; RUNX1-familial platelet disorder; Thrombocytopenia - complications; Thrombocytopenia - genetics; leukemia; inherited thrombocytopenias; platelet transcriptomics; myelodysplastic syndrome; high-throughput sequencing; RUNX1-familial platelet disorder
• DOI: 10.1016/j.jtha.2023.01.023
• ISSN: 1538-7836

Germline mutations in RUNX1 can cause a familial platelet disorder that may lead to acute myeloid leukemia, an autosomal dominant disorder characterized by moderate thrombocytopenia, platelet dysfunction, and a high risk of developing acute myeloid leukemia or myelodysplastic syndrome. Discerning the pathogenicity of novel RUNX1 variants is critical for patient management. To extend the characterization of RUNX1 variants and evaluate their effects by transcriptome analysis. Three unrelated patients with long-standing thrombocytopenia carrying heterozygous RUNX1 variants were included: P1, who is a subject with recent development of myelodysplastic syndrome, with c.802 C>T[p.Gln268∗] de novo; P2 with c.586A>G[p.Thr196Ala], a variant that segregates with thrombocytopenia and myeloid neoplasia in the family; and P3 with c.476A>G[p.Asn159Ser], which did not segregate with thrombocytopenia or neoplasia. Baseline platelet evaluations were performed. Ultrapure platelets were prepared for platelet transcriptome analysis. In P1 and P2, but not in P3, transcriptome analysis confirmed aberrant expression of genes recognized as RUNX1 targets. Data allowed grouping patients by distinct gene expression profiles, which were partitioned with clinical parameters. Functional studies and platelet mRNA expression identified alterations in the actin cytoskeleton, downregulation of GFI1B, defective GPVI downstream signaling, and reduction of alpha granule proteins, such as thrombospondin-1, as features likely implicated in thrombocytopenia and platelet dysfunction. Platelet phenotype, familial segregation, and platelet transcriptomics support the pathogenicity of RUNX1 variants p.Gln268∗ and p.Thr196Ala, but not p.Asn159Ser. This study is an additional proof of concept that platelet RNA analysis could be a tool to help classify pathogenic RUNX1 variants and identify novel RUNX1 targets. •Germinal RUNX1 variants may cause hematological malignancies at a young age.•Discerning the pathogenicity of novel RUNX1 variants is critical for patient management.•Gene expression analysis demonstrates the deleterious effects of Gln268∗ and Thr196Ala variants.•Platelet transcriptomics is a useful tool for pathogenicity assessment of novel RUNX1 variants.