ASXL1-Mutant Clonal Hematopoiesis is Associated with Calcific Aortic Valve Disease and Promotes Valvular Calcification In Vitro
- 1Massachusetts General Hospital
Abstract
Background and AimsCalcific aortic valve disease (CAVD) is a common valvular heart condition and has no effective medical therapy. Clonal hematopoiesis of indeterminate potential (CHIP), characterized by the acquisition of somatic mutations in hematopoietic stem cells which provides a selection advantage, has been associated with incident aortic stenosis (AS), particularly when driven by TET2 mutations.
MethodsWe evaluated the association of CHIP and gene-specific CHIP subtypes with AS in the UK Biobank (UKB) and with aortic valve hemodynamics in the Atherosclerosis Risk in Communities (ARIC) study to prioritize gene driver mutations for mechanistic study. We then generated human ASXL1-mutant THP-1 derived macrophage like cells and evaluated the ability of THP-1 conditioned media to promote a calcific response in human valvular interstitial cells (VICs). We additionally performed mass spectrometry proteomics to characterize the ASXL1-mutant THP-1 macrophage secretome. Therapeutically relevant inflammatory pathways were inhibited with anakinra (interleukin [IL]-1) or tocilizumab (IL-6).
ResultsAmong 449,109 individuals in the UKB with whole exome sequencing, large CHIP clones and non-DNMT3A CHIP subtypes (e.g., TET2, ASXL1, and JAK2 CHIP) were independently associated with risk of incident AS. Among 1,963 individuals in ARIC with CHIP sequencing and echocardiographic data, only ASXL1 CHIP was associated with worse aortic valve hemodynamics. In vitro, ASXL1-mutant human THP-1-derived macrophage-like cells had increased AIM2 inflammasome activation. THP-1 media promoted VIC calcification, which was accelerated in ASXL1 mutant macrophages. Proteomic profiling identified inflammatory proteins linked to valvular calcification. Accelerated VIC calcification was reduced by IL-1 inhibition by anakinra or IL-6 inhibition by tocilizumab.
ConclusionsIntegrating human cohort data with experimental evidence using ASXL1-mutant human macrophage-like cells and human valve cells, these findings identify ASXL1-mutant CHIP as a distinct inflammatory subtype associated with valvular calcification and support IL-1 and IL-6 signaling as candidate therapeutic pathways for future investigation in CAVD.
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BibTeX
@misc{small2026asxl1mutant,
title = {{ASXL1-Mutant Clonal Hematopoiesis is Associated with Calcific Aortic Valve Disease and Promotes Valvular Calcification In Vitro}},
author = {A. Small and L. Xue and S. Ito and C. De Jeronimo Diaz and Y. Nakamura and N. Mohammadnia and H. Liao and Y. Maekawa and L. Li and S. A. Singh and T. Kasai and L. Weiss and M. M. Uddin and A. Schuermans and S. Flynn and E. Mackenzie and J. C. Engert and G. Thanassoulis and P. Libby and A. Shah and C. M. Ballantyne and P. Natarajan and T. Fidler and E. Aikawa and M. C. Honigberg},
howpublished = {medRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.23.26363798},
url = {https://www.medrxiv.org/content/10.64898/2026.09.23.26363798v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - ASXL1-Mutant Clonal Hematopoiesis is Associated with Calcific Aortic Valve Disease and Promotes Valvular Calcification In Vitro
AU - A. Small
AU - L. Xue
AU - S. Ito
AU - C. De Jeronimo Diaz
AU - Y. Nakamura
AU - N. Mohammadnia
AU - H. Liao
AU - Y. Maekawa
AU - L. Li
AU - S. A. Singh
AU - T. Kasai
AU - L. Weiss
AU - M. M. Uddin
AU - A. Schuermans
AU - S. Flynn
AU - E. Mackenzie
AU - J. C. Engert
AU - G. Thanassoulis
AU - P. Libby
AU - A. Shah
AU - C. M. Ballantyne
AU - P. Natarajan
AU - T. Fidler
AU - E. Aikawa
AU - M. C. Honigberg
PY - 2026
DA - 2026/09/23
PB - medRxiv
DO - 10.64898/2026.09.23.26363798
UR - https://www.medrxiv.org/content/10.64898/2026.09.23.26363798v1
AB - Background and AimsCalcific aortic valve disease (CAVD) is a common valvular heart condition and has no effective medical therapy. Clonal hematopoiesis of indeterminate potential (CHIP), characterized by the acquisition of somatic mutations in hematopoietic stem cells which provides a selection advantage, has been associated with incident aortic stenosis (AS), particularly when driven by TET2 mutations. MethodsWe evaluated the association of CHIP and gene-specific CHIP subtypes with AS in the UK Biobank (UKB) and with aortic valve hemodynamics in the Atherosclerosis Risk in Communities (ARIC) study to prioritize gene driver mutations for mechanistic study. We then generated human ASXL1-mutant THP-1 derived macrophage like cells and evaluated the ability of THP-1 conditioned media to promote a calcific response in human valvular interstitial cells (VICs). We additionally performed mass spectrometry proteomics to characterize the ASXL1-mutant THP-1 macrophage secretome. Therapeutically relevant inflammatory pathways were inhibited with anakinra (interleukin [IL]-1) or tocilizumab (IL-6). ResultsAmong 449,109 individuals in the UKB with whole exome sequencing, large CHIP clones and non-DNMT3A CHIP subtypes (e.g., TET2, ASXL1, and JAK2 CHIP) were independently associated with risk of incident AS. Among 1,963 individuals in ARIC with CHIP sequencing and echocardiographic data, only ASXL1 CHIP was associated with worse aortic valve hemodynamics. In vitro, ASXL1-mutant human THP-1-derived macrophage-like cells had increased AIM2 inflammasome activation. THP-1 media promoted VIC calcification, which was accelerated in ASXL1 mutant macrophages. Proteomic profiling identified inflammatory proteins linked to valvular calcification. Accelerated VIC calcification was reduced by IL-1 inhibition by anakinra or IL-6 inhibition by tocilizumab. ConclusionsIntegrating human cohort data with experimental evidence using ASXL1-mutant human macrophage-like cells and human valve cells, these findings identify ASXL1-mutant CHIP as a distinct inflammatory subtype associated with valvular calcification and support IL-1 and IL-6 signaling as candidate therapeutic pathways for future investigation in CAVD.
ER -