Ann Lab Med.  2023 Mar;43(2):145-152. 10.3343/alm.2023.43.2.145.

Clinical, Mutational, and Transcriptomic Characteristics in Elderly Korean Individuals With Clonal Hematopoiesis Driver Mutations

• Moon I
• Kong MG
• Ji YS
• Kim SH
• Park SK
• Suh J
• Jang MA

• KMID: 2551691
• DOI: http://doi.org/10.3343/alm.2023.43.2.145

Abstract

Background
Clonal hematopoiesis of indeterminate potential (CHIP), which is defined as the presence of blood cells originating from somatically mutated hematopoietic stem cells, is common among the elderly and is associated with an increased risk of hematologic malignancies. We investigated the clinical, mutational, and transcriptomic characteristics in elderly Korean individuals with CHIP mutations.
Methods
We investigated CHIP in 90 elderly individuals aged ≥60 years with normal complete blood counts at a tertiary-care hospital in Korea between June 2021 and February 2022. Clinical and laboratory data were prospectively obtained. Targeted next-generation sequencing of 49 myeloid malignancy driver genes and massively parallel RNA sequencing were performed to explore the molecular spectrum and transcriptomic characteristics of CHIP mutations.
Results
We detected 51 mutations in 10 genes in 37 (41%) of the study individuals. CHIP prevalence increased with age. CHIP mutations were observed with high prevalence in DNMT3A (26 individuals) and TET2 (eight individuals) and were also found in various other genes, including KDM6A, SMC3, TP53, BRAF, PPM1D, SRSF2, STAG1, and ZRSR2. Baseline characteristics, including age, confounding diseases, and blood cell parameters, showed no significant differences. Using mRNA sequencing, we characterized the altered gene expression profile, implicating neutrophil degranulation and innate immune system dysregulation.
Conclusions
Somatic CHIP driver mutations are common among the elderly in Korea and are detected in various genes, including DNMT3A and TET2. Our study highlights that chronic dysregulation of innate immune signaling is associated with the pathogenesis of various diseases, including hematologic malignancies.

Keyword

Clonal hematopoiesis; High-throughput nucleotide sequencing; Transcriptome; Prognosis; Korea

Figure

• 

  Fig. 1 Number and distribution of CHIP mutations in the study cohort. (A) Number of CHIP mutations identified using a targeted NGS panel of 49 myeloid malignancy driver genes. (B) The frequency of CHIP mutation increases with age. (C) Forest plot showing the odds ratio with 95% CI for the presence of CHIP mutation according to a 1-year increase in age. (D) Numbers of CHIP mutations per individual. (E) VAFs of CHIP mutations detected in the study cohort. Mutations in DNMT3A, TET2, and other genes are indicated by specific symbols. (F) Distribution of the VAF of CHIP mutations per gene. Dotted lines indicate median VAFs of 2.0%, 2.5%, and 2.0% mutations in DNMT3A, TET2, and other genes, respectively. Abbreviations: CI, confidence interval; CHIP, clonal hematopoiesis of indeterminate potential; NGS, next-generation sequencing; VAF, variant allele frequency.

• 

  Fig. 2 Heatmap and hierarchical clustering of the 31 most significant DEGs by CHIP status. CHIP-positive samples (purple; N=6) are segregated from CHIP-negative samples (light purple; N=6), except for an outlier. All genes listed had a log2 fold change >1 or <–1 and adjusted P<0.05 in DESeq2 (v3.5e) analysis. The heatmap color range is from positive (red) to negative (blue) Z-score values. Abbreviations: DEG, differentially expressed gene; CHIP, clonal hematopoiesis of indeterminate potential.

• 

  Fig. 3 Enrichment of the neutrophil degranulation pathway in CHIP-positive samples. (A) Differential expression of neutrophil/innate immunity-related genes, compared between individuals with CHIP mutations and those without. Data are presented as the median and interquartile range. (B) Upper panel: enrichment plot of the neutrophil degranulation pathway shown as a snapshot of the GSEA. Lower panel: heatmap showing the expression of genes belonging to the set with the annotation of genes ordered. *P<0.05, **P<0.01. Abbreviations: CHIP, clonal hematopoiesis of indeterminate potential; GSEA, gene set enrichment analysis.

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