Disease stage-specific mutational landscape in essential thrombocythemia and prognostic correlates: A Mayo Clinic study of 500 NGS-informative cases Free

In our first exploration of mutations in essential thrombocythemia (ET; N=183), we described somatic variants other than JAK2/CALR/MPL in 53% (N=96) of patients with the most frequent being TET2 (16%), ASXL1 (11%), DNMT3A (6%), and SF3B1 (5%) [Blood Adv 2016;1:21]. In a subsequent report (N=270; BJH 2020;189:291), we highlighted the prognostic relevance of SF3B1, SRSF2, and EZH2 mutations for overall survival (OS), SRSF2, EZH2, and TP53 for leukemia-free survival (LFS), and SF3B1 and U2AF1 for myelofibrosis-free survival (MFFS). We have now expanded our NGS-informative patient cohort to 500 and examined the distribution and frequencies of specific mutations at different stages of the disease and corresponding prognostic correlates.

Molecularly-annotated ET patients diagnosed at the Mayo Clinic between January 1980 and May 2025 were considered. Diagnoses of chronic phase ET (ET-CP), post-ET MF, and post-ET acute myeloid leukemia (AML) were retrospectively fitted to be in accordance with the International Consensus Classification criteria (Blood 2022;140:1200). Mutation and cytogenetic analyses were performed according to institutional protocols for clinical use that included multi-gene next-generation sequencing (NGS); in a subset of patients, historically archived samples were accessed for analysis in a research setting. Patients were monitored until death or time of last follow-up, determined through electronic medical records or direct communication with patients or healthcare providers. Conventional statistical methods were utilized (JMP Pro 17.1.0, SAS Institute, Cary, NC, USA).

A total of 500 ET patients (median 57 years; Females 61%) were informative for NGS-derived mutations that were obtained in the chronic (N=401; 80.2%; group A), post-ET MF (N=88; 17.6%; group B), or post-ET AML (N=11; 2.2%; group C) phase of the disease; group C included 9 (1.8%) patients who transformed directly to AML without transitioning through the MF phase.

The most frequent mutations with at least 1% (N=5) mutational frequency were (approximately) TET2 (10%), ASXL1 (10%), DNMT3A (7%), SF3B1 (4%), TP53 (5%), SRSF2 (3%), EZH2 (2%), U2AF1 (2%), CBL (1%), IDH1 (1%), IDH2 (1%), RUNX1 (1%), ZRSR2 (1%), and PPM1D (1%). Comparison of distribution of mutations and their frequencies based on timing of NGS (AML phase vs. MF phase vs. ET-CP) revealed significant differences for TP53^MUT (72.7% vs. 11.4% vs. 1%; p<0.01), ASXL1^MUT (18.2% vs. 25% vs. 7.2%; p<0.01), and EZH2^MUT (18.2% vs. 6.8% vs. 0.3%; p<0.01), respectively; borderline significance was also apparent for RUNX1^MUT (9.1% vs. 0% vs. 1.3%; p=0.10) and ZRSR2^MUT (9.1% vs. 2.3% vs. 0.5%; p=0.08). Notably, among 9 patients in whom NGS was performed at the time of direct transition from ET to AML, without an intermediary MF phase, 8 (88.9%) displayed TP53^MUT and only one (11.1%) EZH2^MUT and none (0%) ASXL1^MUT.

At a median follow-up of 66 months, 93 (18.6%) deaths, 26 (5.2%) leukemic transformations (LT), and 108 (21.6%) MF transformations were reported. In group A patients, age-adjusted multivariable analysis (MVA) of mutations identified ASXL1^MUT (N=29; HR 3.8, 1.9-7.7; p<0.01) and SF3B1^MUT (N=12; HR 3.9, 1.4-11.0; p=0.01) to be associated with inferior OS, ASXL1^MUT (HR 4.9, 1.01-24.2; p=0.048) and SRSF2^MUT (N=11; HR 11.9, 1.3-104.8; p=0.03) with inferior LFS, and ASXL1^MUT (HR 9.0, 2.8-29.5; p<0.01) and SF3B1^MUT (HR 15.5, 3.1-77.5; p<0.01) with inferior MFFS, all calculated from time of NGS. A similar analysis in group B patients identified ASXL1^MUT (N=22; HR 3.7, 1.2-11.7; p=0.03) to be associated with inferior OS and TP53^MUT (N=10; HR 13.4, 1.7-108.6; p=0.02) with inferior LFS. Age-adjusted MVA combining groups A and B (N=489) confirmed the association between inferior OS and ASXL1^MUT (N=51; p<0.01) and SF3B1^MUT (N=18; p<0.01) while inferior LFS was associated with TP53^MUT (N=14; p<0.01) and ASXL1^MUT (p=0.03).

The majority (⁓73%) of patients with post-ET AML harbor TP53^MUT, at the time of LT. By contrast, TP53^MUT are infrequent in ET-CP (⁓1%) and did not appear to predict LT, as was the case with post-ET MF. ASXL1^MUT was shown to be an independent risk factor for OS, MFFS, and LFS in ET-CP and for OS in post-ET MF. SF3B1^MUT was associated with inferior OS and MFFS in ET-CP and SRSF2 with inferior LFS in ET-CP. Identifying ET patients with adverse mutations allows for closer surveillance and potentially more tailored therapeutic interventions.