Clonal hematopoiesis as determined by the HUMARA assay is a marker for acquired mutations in epigenetic regulators in older women

Julia Erin Wiedmeier, Catherine Kato, Zhenzhen Zhang, Hyunjung Lee, Jennifer Dunlap, Eric Nutt, Rogan Rattray, Sarah McKay, Christopher Eide, Richard Press, Motomi (Tomi) Mori, Brian Druker, Kim-Hien Dao

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Abstract

Recent large cohort studies revealed that healthy older individuals harbor somatic mutations that increase their risk for hematologic malignancy and all-cause cardiovascular deaths. The majority of these mutations are in chromatin and epigenetic regulatory genes (CERGs). CERGs play a key role in regulation of DNA methylation (DNMT3A and TET2) and histone function (ASXL1) and in clonal proliferation of hematopoietic stem cells. We hypothesize that older women manifesting clonal hematopoiesis, defined here as a functional phenomenon in which a hematopoietic stem cell has acquired a survival and proliferative advantage, harbor a higher frequency of somatic mutations in CERGs. The human androgen receptor gene (HUMARA) assay was used in our study to detect the presence of nonrandom X inactivation in women, a marker for clonal hematopoiesis. In our pilot study, we tested 127 blood samples from women ≥65 years old without a history of invasive cancer or hematologic malignancies. Applying stringent qualitative criteria, we found that 26% displayed clonal hematopoiesis; 52.8% displayed polyclonal hematopoiesis; and 21.3% had indeterminate patterns (too close to call by qualitative assessment). Using Illumina MiSeq next-generation sequencing, we identified somatic mutations in CERGs in 15.2% of subjects displaying clonal hematopoiesis (three ASXL1 and two DNMT3A mutations with an average variant allele frequency of 15.7%, range: 6.3%–23.3%). In a more limited sequencing analysis, we evaluated the frequency of ASXL1 mutations by Sanger sequencing and found mutations in 9.7% of the clonal samples and 0% of the polyclonal samples. By comparing several recent studies (with some caveats as described), we determined the fold enrichment of detecting CERG mutations by using the HUMARA assay as a functional screen for clonal hematopoiesis. We conclude that a functional assay of clonal hematopoiesis is enriching for older women with somatic mutations in CERGs, particularly for ASXL1 and TET2 mutations and less so for DNMT3A mutations.

Original languageEnglish (US)
Pages (from-to)857-865.e5
JournalExperimental Hematology
Volume44
Issue number9
DOIs
StatePublished - Sep 1 2016

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Hematopoiesis
Epigenomics
Regulator Genes
Mutation
Chromatin
Hematologic Neoplasms
Mutation Rate
Hematopoietic Stem Cells
X Chromosome Inactivation
DNA Methylation
Gene Frequency
Histones
Cause of Death
Cohort Studies
Survival

ASJC Scopus subject areas

  • Hematology
  • Molecular Biology
  • Genetics
  • Cell Biology
  • Cancer Research

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Clonal hematopoiesis as determined by the HUMARA assay is a marker for acquired mutations in epigenetic regulators in older women. / Wiedmeier, Julia Erin; Kato, Catherine; Zhang, Zhenzhen; Lee, Hyunjung; Dunlap, Jennifer; Nutt, Eric; Rattray, Rogan; McKay, Sarah; Eide, Christopher; Press, Richard; Mori, Motomi (Tomi); Druker, Brian; Dao, Kim-Hien.

In: Experimental Hematology, Vol. 44, No. 9, 01.09.2016, p. 857-865.e5.

Research output: Contribution to journalArticle

Wiedmeier, Julia Erin ; Kato, Catherine ; Zhang, Zhenzhen ; Lee, Hyunjung ; Dunlap, Jennifer ; Nutt, Eric ; Rattray, Rogan ; McKay, Sarah ; Eide, Christopher ; Press, Richard ; Mori, Motomi (Tomi) ; Druker, Brian ; Dao, Kim-Hien. / Clonal hematopoiesis as determined by the HUMARA assay is a marker for acquired mutations in epigenetic regulators in older women. In: Experimental Hematology. 2016 ; Vol. 44, No. 9. pp. 857-865.e5.
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