Abstract P456: The Association Between Clonal Hematopoiesis Of Indeterminate Potential And Inflammatory Biomarkers Among Chronic Kidney Disease Patients

Clonal hematopoiesis (CH) of indeterminate potential (CHIP), driven by somatic mutations in leukemia-associated genes, confers increased risk of hematologic malignancies, cardiovascular disease, and all-cause mortality. In blood of healthy individuals, small CH clones can expand over time to reach 2% variant allele frequency (VAF), the current threshold for CHIP. Nevertheless, reliable detection of low-VAF CHIP mutations is challenging, often relying on deep targeted sequencing. Here, we present UNISOM, a streamlined workflow for enhancing CHIP detection from whole-genome and whole-exome sequencing data that are underpowered, especially for low VAFs. UNISOM utilizes a meta-caller for variant detection, in couple with machine learning models which classify variants into CHIP, germline, and artifact. In whole-exome sequencing data, UNISOM recovered nearly 80% of the CHIP mutations identified via deep targeted sequencing in the same cohort. Applied to whole-genome sequencing data from Mayo Clinic Biobank, it recapitulated the patterns previously established in much larger cohorts, including the most frequently mutated CHIP genes and predominant mutation types and signatures, as well as strong associations of CHIP with age and smoking status. Notably, 30% of the identified CHIP mutations had < 5% VAFs, demonstrating its high sensitivity toward small mutant clones. This workflow is applicable to CHIP screening in population genomic studies. The UNISOM pipeline is freely available at https://github.com/shulanmayo/UNISOM and https://ngdc.cncb.ac.cn/biocode/tool/7816.

The Growth Kinetics of Clonal Hematopoiesis Inform Risk of Blood Cancer Progression

Clonal hematopoiesis of indeterminate potential (CHIP) is defined by the age-dependent accumulation of somatic leukemia-associated driver mutations in hematopoietic stem cells, in individuals with normal blood counts and with absence of an underlying myeloid neoplasm (MN).1, 2 CHIP is associated with an increased risk of developing MN and an increased all-cause mortality, largely due to cardiovascular disease.3 The presence of CHIP prior to receiving chemotherapy and radiation has been associated with therapy related MN (T-MN), such as myelodysplastic syndromes (MDS) and acute myeloid leukemia.4 Autologous stem cell transplantation (ASCT) is an effective treatment modality for managing higher-risk patients with non-Hodgkin's lymphoma (NHL) and multiple myeloma (MM). In a seminal NHL study, 30% of patients were found to have CHIP at the time of ASCT, with the presence of CHIP being associated with an increased rate of T-MN (10-year cumulative incidence of 14.1% vs 4.3%) and an inferior overall survival (10 year OS 30.4% vs 60.9%).5 In MM, targeted sequencing of 629 patients prior to ASCT detected CHIP in 21.6% of patients, with the presence of CHIP strongly associating with inferior OS (HR 1.34, p = .02) and an inferior progression-free survival (PFS, HR 1.45, p < .001). Interestingly, in this study, adverse CHIP-associations were apparently overcome by lenalidomide maintenance therapy.6 Unlike in NHL, CHIP in MM was not associated with T-MN; while lenalidomide maintenance therapy, independent of the presence or absence of CHIP, was associated with T-MN (p = .047) and second primary malignancies (SPM).6 We carried out this study to assess the prevalence and prognostic impact of CHIP in a relatively uniform cohort of MM patients at the time of ASCT, with all patients going on to receive lenalidomide maintenance therapy. Successive MM patients who consented to have their pre-ASCT bone marrow (BM) sample collected and who underwent first ASCT at Mayo Clinic, followed by lenalidomide maintenance therapy, were included in the study. The BM mononuclear cell DNA from pre-ASCT samples was extracted after excluding CD38/CD138+ (negative selection) plasma cells and then subjected to targeted NGS testing (42-myeloid related genes) by previously described methods.7 All patients were closely followed for the development of T-MN as defined by the 2016 WHO criteria, arterial and venous thromboembolism (VTE) and SPM.8, 9 Response to therapy was assessed using the international myeloma working group (IMWG) consensus criteria 2016.10 Statistical methods are highlighted in the supplemental material. Clonal hematopoiesis was detectable in 23 (23%) of 101 MM patients assessed in the study (Table S1, Figure S1). Clinical characteristics, MM risk stratification, median number of prior therapies, response to therapy, ASCT conditioning regimens, engraftment data, day +100 post ASCT outcomes and median duration of lenalidomide maintenance are outlined in Table 1. Except for a higher median age at MM diagnosis in MM patient with CHIP (p = .002), there were no other significant differences between the two groups (Figure S2). Ten (43.5%) patients in the MM CHIP group and 30 (38.0%) in the MM no CHIP group, received alkylatingagent-based induction therapy prior to ASCT (p = .66). Melphalan 200 mg/m2 conditioning was used in 87.1% of patients (95.7% in the CHIP vs 84.6% in the no CHIP group, p = .45), while the remainder received melphalan 140 mg/m2 conditioning. The median duration of lenalidomide maintenance therapy was 21 months (10–36); 16 months in MM patients with CHIP and 22 months in MM patients without CHIP (p = .76), with the median lenalidomide dose being 15 mg (range 10–15 mg; 10 mg in the CHIP group and 15 mg in the no CHIP group, p = .08). The most frequent CHIP mutations encountered included DNMT3A [52%; median variant allele frequency (VAF) 9.0%, range 2.0%–29.2%], TET2 (26%; median VAF 3.0%, range 2.0%–4.8%), followed by TP53, PPM1D and BRAF (10.0% each), respectively (Figures 1(A) and S1). Sixteen patients (69.6%) had one mutation, while seven (30.4%) had >1 mutation and two patients had four mutations each (Figure 1(B)). Eight (66.6%) of 12 patients with DNMT3A mutations had nonsynonymous missense mutations, while four had deletion variants: with no patient harboring the commonly mutated DNMT3A R882 hot spot. There were no statistically significant differences in CHIP mutation distribution, including TP53 and PPM1D mutations, between MM CHIP patients that received alkylating-agent based induction therapy prior to ASCT, vs MM CHIP patients that did not (Figure 1(C)). At last follow up, 70 (69.3%) relapses after ASCT and 41 (40.6%) deaths were documented. Twenty-nine patients (28.7%) were on salvage therapy whereas 13 (12.9%) were on lenalidomide maintenance and 14 (13.8%) were on observation alone, with no statistically significant differences between MM patients with CHIP vs MM without CHIP. Rates of VTE were similar between MM patients with CHIP (30%) and those without CHIP (24%), with similar rates of provoked thromboses (33% vs 29%, p = .4). Thromboses were diagnosed in typical locations in individuals with CHIP including seven lower extremity deep vein thromboses (DVT) and two pulmonary emboli (PE). In contrast, a variety of VTE locations were diagnosed in those without CHIP, including 10 lower extremity DVT, three PE, one DVT with PE, two upper extremity DVT and one portal vein thrombosis. There was a distinction in VTE timing with regards to lenalidomide use between MM patients with CHIP and those without CHIP. For MM patients with CHIP, 2/9 (22.2%) VTE occurred while on lenalidomide, 2/9 (22.2%) occurred prior to lenalidomide and 4/9 (44.4%) occurred at least 3 months after discontinuation of lenalidomide therapy. For MM patients without CHIP, majority (13/17, 76.5%) of VTE occurred while on lenalidomide, with a minority (11%) occurring either before or at least 3 months after discontinuation of lenalidomide. While lenalidomide is a known risk factor for thrombosis in MM, the fact that 44% of VTE in MM with CHIP occurred >3 months after discontinuing lenalidomide, suggests that CHIP might increase VTE risk in this setting (P = .04). Median OS from the time of diagnosis of the entire cohort was 124.6 months (95%CI 97.5-N/A months) with a corresponding 5 year OS of 82.0% (95% CI 74.8%–89.9%). There was no difference in median OS between MM with CHIP vs those without CHIP (100.2 months; 95%CI 76.2-NA months vs 135.6 months; 95%CI 106.3-N/A months, p = .27) (Figure 1(D)), including assessments with individual CHIP-mutations. The median EFS after ASCT was 36.4 months (95%CI 30.5–48.5 months), with there being no difference in median PFS between MM CHIP (36.4 months, 95%CI 24.1–58.5 months) patients vs MM patients without CHIP (36.4 months, 95%CI 29.9–52.4 months) (p = .34) (Figure 1(E)), including TP53 mutations (Figure S3). There were also no differences between the two groups with regards to non-relapse mortality (Figure S4), cumulative incidence of relapse after ASCT (Figure S5) and time to next treatment. Nineteen (18.8%) SPM were documented, 7 (30.4%) in MM CHIP group vs 12 (15.3%) in the MM no CHIP group (Figure 1(F), p = .13), with corresponding 5-year cumulative incidence rates of 22% and 13%, respectively. These SPM included five (4.9%) hematological malignancies (two in MM CHIP vs three in MM no CHIP), seven (6.9%) skin cancers (three in MM CHIP vs four in MM no CHIP) and seven visceral malignancies (two in MM CHIP vs five in MM no CHIP) (Table S2, Figure S6). The two MM CHIP patients who developed T-MN/MDS had TP53 and PPM1D mutations, respectively. In the MM no CHIP group, there was one patient with B-acute lymphoblastic leukemia and two patients with T-MDS with monosomal karyotypes. In summary, we define the CHIP landscape in MM patients at the time of ASCT, with mutations in epigenetic regulator genes being most common (57%), followed by tumor suppressor genes (17%). Unlike in NHL, presence of CHIP at time of ASCT in MM did not impact OS, PFS and incidence of T-MN; a finding potentially attributable to immunomodulatory properties of lenalidomide, or to the use of maintenance therapy in general.6, 11 While the presence of CHIP and lenalidomide therapy have individually been associated with increased risk of thromboses,1-3, 9 we did not see synergy in MM patients with CHIP, although the timing and patterns of thromboses suggest that CHIP might negatively influence thrombotic risk. While SPM have been well described with lenalidomide maintenance therapy,9 we did not see any differences in SPM and hematological malignancies between the two groups. The findings of this study independently confirm a prior observation on the potential ability of lenalidomide maintenance to mitigate the expected adverse effects of CHIP on OS and PFS in MM patients' post-ASCT6; an important consideration given that approximately 13 000 MM patients undergo ASCT in the US annually, with lenalidomide maintenance considered as standard of care.12, 13 Given the smaller sample size and the inherent flaws of a retrospective analysis, future clinical trials evaluating therapies in MM patients' post-ASCT should consider accounting for the presence of CHIP and its impact on outcomes. The authors would like to acknowledge the “Henry Predolin Leukemia Foundation”, Mayo Clinic, Rochester, MN, USA. Mrinal Patnaik has served on the advisory board of Kura Oncology. A Keith Stewart has served on the advisory board for Celgene. Rafael Fonseca has the following disclosures: Consulting: Amgen, BMS, Celgene, Takeda, Bayer, Janssen, Novartis, Pharmacyclics, Sanofi, Karyopharm, Merck, Juno, Kite, Aduro, OncoTracker, Oncopeptides, GSK, AbbVie. Scientific Advisory Board: Adaptive Biotechnologies, Caris Life Sciences and OncoTracker. Gene mutations annotated in the study have been provided in the supplementary material. Raw sequencing data can be made available on request. Appendix S1 Supporting Information. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Emerging evidence on the role of clonal hematopoiesis of indeterminate potential in chronic kidney disease

Clonal hematopoiesis of indeterminate potential (CHIP) and dementia disproportionately burden patients with chronic kidney disease (CKD). The association between CHIP and cognitive impairment in CKD patients is unknown. We conducted time-to-event analyses in up to 1452 older adults with CKD from the Chronic Renal Insufficiency Cohort who underwent CHIP gene sequencing. Cognition was assessed using four validated tests in up to 6 years mean follow-up time. Incident cognitive impairment was defined as a test score one standard deviation below the baseline mean. Compared to non-carriers, CHIP carriers were markedly less likely to experience impairment in attention (adjusted hazard ratio [HR] [95% confidence interval {CI}]=0.44 [0.26, 0.76], p=0.003) and executive function (adjusted HR [95% CI]=0.60 [0.37, 0.97], p=0.04). There were no significant associations between CHIP and impairment in global cognition or verbal memory. CHIP was associated with lower risks of impairment in attention and executive function among CKD patients. Our study is the first to examine the role of CHIP in cognitive decline in CKD. CHIP markedly decreased the risk of impairment in attention and executive function. CHIP was not associated with impairment in global cognition or verbal memory.

Background Patients with chronic kidney disease (CKD) exhibit an extremely high prevalence of coronary artery disease. Clonal hematopoiesis of indeterminate potential (CHIP) and CKD share pathological features such as aging, chronic inflammation, and accelerated atherosclerosis. Their coexistence can synergistically exacerbate vascular damage and increase coronary risk. However, the association between CHIP and specific coronary lesions in CKD populations has not been reported, and its relationship with cardiovascular events remains controversial. Methods A total of 151 patients with CKD who underwent coronary angiography were prospectively included. To evaluate the status of CHIP, we utilized high-depth targeted sequencing, and measured serum inflammatory factor levels. Furthermore, we systematically followed up these patients to document the occurrence of adverse clinical events. Results CHIP was identified in 65 (43.0%) CKD patients, with the carrier rate steadily rising with age. The CHIP subjects had higher rates of left circumflex (LCx) stenosis, three-vessel disease, and Gensini scores than non-CHIP patients (all P &amp;lt; 0.05). After adjusting for relevant clinical risk factors, the presence of CHIP continued to show an independent association with three-vessel disease (OR 2.26, 95% CI 1.07–4.75; P = 0.032). The survival analysis indicated that CHIP, along with non-DNMT3A mutations and a larger clone size (variant allele frequency ≥ 0.10), correlated with the primary composite endpoint (P &amp;lt; 0.01). Even after controlling for various clinical variables, the CHIP status still demonstrated an independent association with the primary composite endpoint (HR 2.02, 95% CI 1.11–3.67; P = 0.022). Conclusions CHIP was associated with the severity of coronary lesions and unfavorable clinical outcomes in patients with CKD.

BackgroundAutoimmune thyroid disease (AITD) is the most common organ-specific autoimmune disease, often remaining asymptomatic until the thyroid is significantly affected. Clonal hematopoiesis of indeterminate potential (CHIP) has been reported to drive many inflammatory diseases and autoimmune diseases. The association between CHIP and AITD is scarcely reported. This study aims to investigate whether CHIP is associated with the risk of AITD.MethodsWe conducted a prospective community-based cohort study at the UK Biobank. CHIP, defined as the exposure, was identified using whole-exome sequencing (WES) data. AITD was sourced from the inpatient hospitalization register, the death register, and the primary healthcare register. Cox regression models were utilized to estimate the hazard ratio (HR) and 95% confidence interval (CI) for the association between CHIP and AITD. Next, we conducted a subgroup analysis to investigate the role of specific gene mutations (DNMT3A, TET2, ASXL1, PPM1D, SRSF2, and JAK2) in the investigated association. Finally, we assessed the association across small CHIP clones (variant allele frequency, VAF: 2–10%) and large CHIP clones (VAF ≥ 10%). All models were adjusted for sex, age, ethnicity, education, Townsend deprivation index, body mass index, smoking status, and drinking status.ResultsA total of 454,618 individuals were included in the final analysis. We identified 14,059 (3.1%) participants with CHIP. Compared with individuals without CHIP, those with CHIP were generally older and more likely to be smokers. Over a median follow-up of 12.7 years (interquartile range, IQR: 11.9–13.5), 21,708 cases with AITD were diagnosed. CHIP was associated with an increased risk of AITD (HR 1.11, 95% CI 1.03–1.19). Specifically, individuals with TET2-mutant CHIP (HR 1.23, 95% CI 1.07–1.41) had an elevated risk of AITD. A large CHIP clone (HR 1.17, 95% CI 1.08–1.27) was associated with an increased risk of AITD. Focusing on large CHIP clone, we also observed an association between TET2-mutant (HR 1.27, 95% CI 1.10–1.47) and ASXL1-mutant (HR 1.33, 95% CI 1.02–1.73) CHIP and risk of AITD.ConclusionsIndividuals with CHIP were associated with a modestly increased risk of AITD, especially TET2-mutant CHIP. Future studies are needed to verify current findings and elaborate potential mechanisms.

Clonal haematopoiesis of indeterminate potential (CHIP), defined as a clonal expansion of age-related recurrent somatic mutations, has recently emerged as a novel cardiovascular risk factor. However, the precise role of CHIP in the development of atherosclerotic cardiovascular disease (ASCVD) remains unclear. Among 4300 asymptomatic Korean participants aged 40-79 years, we investigated the risk of ASCVD by CHIP and the interplay between CHIP and conventional risk factors in ASCVD development. Additionally, we assessed changes in coronary arteries based on the presence of CHIP using coronary computed tomography angiography (CCTA). CHIP was present in 363 participants (8.4%), and its prevalence increased with age. Commonly mutated genes were DNMT3A, TET2, and ASXL1, in order. During the follow-up (median 4.7 years), 18 ASCVD cases (5.0%) were observed in CHIP carriers vs. 62 (1.6%) in non-carriers (P < 0.001), indicating an elevated risk of ASCVD associated with CHIP [adjusted hazard ratio (HR) 2.49; 95% confidence interval (CI) 1.45-4.29; P < 0.001]. Notably, with high levels of LDL cholesterol, CHIP enhanced the risk of ASCVD (adjusted HR 6.20; 95% CI 3.14-12.23; P < 0.001), demonstrating synergism between CHIP and LDL cholesterol levels (S-index 4.94; 95% CI 1.08-22.53; P = 0.039). Serial CCTAs confirmed that CHIP, in conjunction with high LDL cholesterol levels, had a significant early impact on coronary arteries, revealing new measurable coronary atherosclerosis, mainly with unstable plaque, in proximal lesions. The presence of CHIP was significantly associated with the risk of ASCVD, promoting the early stage of atherosclerosis through synergy with high LDL cholesterol in the general population.

INTRO: Clonal hematopoiesis (CH) of indeterminate potential (CHIP) is associated with poor outcomes in those with non-hematologic cancers. CH mutated macrophages and neutrophils have increased capacity to home to peripheral tissues, driving local inflammation; they are also implicated in immunotherapy related toxicity. The relationship between CH and the microbiome in those treated with immunotherapy is not well understood. RESULTS: 35 patients with stage IV melanoma treated with upfront immunotherapy; treatment and cohort characteristics are below (Table 1). We performed error corrected duplex sequencing on peripheral blood mononuclear cells using a panel previously validated for CH mutations. With a cutoff of 0.5% variant allele frequency (VAF), among 10 CHIP positive patients, we found mutations in TET2 (8/10), DNMT3A (7/10), PPM1D (2/10), and JAK2 (1/10). There was no difference in either survival or response based on CHIP status (Log-Rank Chi-Square p=0.44); however, CHIP positivity was associated with greater microbiome richness as measured by alpha diversity (p=0.0012 Shannon diversity) and distinct structural/compositional diversity vs CHIP negative patients as measured by beta diversity (p=0.032, R Squared= 0.045). We inferred KEGG pathway activity from whole genome sequencing and identified that CHIP positive patients were enriched for exopolysaccharide biosynthesis while CHIP negative patients had higher expression of nucleotide sugars biosynthesis and amino acid metabolism pathways. DISCUSSION: In patients treated with immunotherapy for melanoma, microbiome diversity signatures correlate with presence of CHIP. The analysis suggests that there are distinct taxonomic and functional features defining CHIP positivity. The effect of these cells in the tumor microenvironment and their role in immunotherapy response requires further exploration. Citation Format: Eleanor A. Fallon, Samuel Urrutia, Reed Ayabe, Manoj Chelvanambi, Ashish Damania, Sarah Johnson, Tomoyuki Tanaka, Zongrui Li, Yongwoo David Seo, Samuel Cass, Matthew C. Wong, Nadim Ajami, Jennifer Wargo, Koichi Takahashi. Identifying gut microbial determinants of clonal hematopoiesis of indeterminate potential (CHIP) in immunotherapy treated melanoma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2801.

Premature menopause is an independent risk factor for cardiovascular disease in women, but mechanisms underlying this association remain unclear. Clonal hematopoiesis of indeterminate potential (CHIP), the age-related expansion of hematopoietic cells with leukemogenic mutations without detectable malignancy, is associated with accelerated atherosclerosis. Whether premature menopause is associated with CHIP is unknown. We included postmenopausal women from the UK Biobank (n=11 495) aged 40 to 70 years with whole exome sequences and from the Women's Health Initiative (n=8111) aged 50 to 79 years with whole genome sequences. Premature menopause was defined as natural or surgical menopause occurring before age 40 years. Co-primary outcomes were the presence of any CHIP and CHIP with variant allele frequency >0.1. Logistic regression tested the association of premature menopause with CHIP, adjusted for age, race, the first 10 principal components of ancestry, smoking, diabetes, and hormone therapy use. Secondary analyses considered natural versus surgical premature menopause and gene-specific CHIP subtypes. Multivariable-adjusted Cox models tested the association between CHIP and incident coronary artery disease. The sample included 19 606 women, including 418 (2.1%) with natural premature menopause and 887 (4.5%) with surgical premature menopause. Across cohorts, CHIP prevalence in postmenopausal women with versus without a history of premature menopause was 8.8% versus 5.5% (P<0.001), respectively. After multivariable adjustment, premature menopause was independently associated with CHIP (all CHIP: odds ratio, 1.36 [95% 1.10-1.68]; P=0.004; CHIP with variant allele frequency >0.1: odds ratio, 1.40 [95% CI, 1.10-1.79]; P=0.007). Associations were larger for natural premature menopause (all CHIP: odds ratio, 1.73 [95% CI, 1.23-2.44]; P=0.001; CHIP with variant allele frequency >0.1: odds ratio, 1.91 [95% CI, 1.30-2.80]; P<0.001) but smaller and nonsignificant for surgical premature menopause. In gene-specific analyses, only DNMT3A CHIP was significantly associated with premature menopause. Among postmenopausal middle-aged women, CHIP was independently associated with incident coronary artery disease (hazard ratio associated with all CHIP: 1.36 [95% CI, 1.07-1.73]; P=0.012; hazard ratio associated with CHIP with variant allele frequency >0.1: 1.48 [95% CI, 1.13-1.94]; P=0.005). Premature menopause, especially natural premature menopause, is independently associated with CHIP among postmenopausal women. Natural premature menopause may serve as a risk signal for predilection to develop CHIP and CHIP-associated cardiovascular disease.

Introduction: Patients with chronic kidney disease (CKD) are at substantially increased risk of dementia. Clonal hematopoiesis of indeterminate potential (CHIP), caused by clonal expansion of blood cells with somatic mutations in certain driver genes, is an established risk factor for cardiovascular diseases, including cerebral vascular disease. In contrast, CHIP was recently shown to be associated with a decreased risk of Alzheimer’s disease. Given emerging evidence of a key role of vascular compared to neurodegenerative mechanisms in CKD-related dementia, the relation of CHIP with cognitive decline in CKD patients needs clarity. We investigated the prospective associations of CHIP with cognitive impairment in a cohort of patients with CKD. Methods: Our study included 1989 participants aged 65 years or older from the Chronic Renal Insufficiency Cohort. Participants underwent high-depth targeted gene sequencing to ascertain CHIP status. Cognitive function was assessed by four cognitive tests (see Table ). For each test, incident cognitive impairment was defined as a test score one standard deviation worse than the baseline group mean. Sequential Cox proportional hazards models tested associations between CHIP and cognitive impairment after adjusting for demographics (model 1), clinical measures (model 2), and APOE genotype in a subset with available data (model 3). Results: CHIP carriers were significantly less likely to experience impairment in attention (Trails A) and executive function (Trails B) in our fully adjusted models ( Table ). There were no significant associations between CHIP and impairment in global cognition (3MS) or verbal memory (Buschke). There were no differences in the associations of CHIP with cognitive impairment across sex or race groups (data not shown). Conclusions: In patients with CKD, we observed markedly reduced risks of impairment in cognitive domains of attention and executive function. Further studies are needed to confirm our findings and evaluate mechanisms of these relations.

Introduction: Clonal hematopoiesis of indeterminate potential (CHIP) occurs secondary to acquired mutations in bone marrow progenitor cells. CHIP confers a twofold risk of atherosclerotic cardiovascular disease (ASCVD). The two most common CHIP mutations are ten eleven translocase 2 (TET2) and DNA methyltransferase 3A (DNMT3A). However, there is limited data on specific cardiovascular phenotypes in this population. Methods: We recruited patients from the Vanderbilt University Medical Center cardiac catheterization laboratory and performed next generation sequencing to assign CHIP status. Multivariable logistic regression models and proportional odds models were used to assess the association between CHIP status and coronary angiography. Results: We sequenced 1,469 patients, and 1,149 had complete angiographic data. Of these, 214 (18.6%) had CHIP mutations with a variant allele frequency of &gt;2%. CHIP patients were significantly older (68 vs. 59 years, p = 7.7 x 10-16). CHIP carrier status was significantly correlated with any stenosis and degree of stenosis in the left main (LM) coronary artery (OR 1.87 [1.28-2.74]; P = 0.0013 and OR 2.12 [1.42-3.15]; P = 2.2 x 10-4). This association was stronger among patients with TET2 CHIP, both for any LM stenosis (OR 2.98 [1.48 - 5.96]; P = 0.0021) and the degree of LM stenosis (OR 3.52 [1.78 - 6.95]; P = 2.9 x 10-4) (Fig. 1). TET2 CHIP was also correlated with obstructive (&gt;70%) stenosis in any vessel (OR 2.28 [1.15 - 4.52]; P = 0.019), and the number of obstructive stenoses across coronary vessels (OR 1.83 [1.04 - 3.23]; P = 0.037). Conclusions: This is the first description of a specific atherosclerotic phenotype in CHIP and serves as a basis for understanding enhanced morbidity and mortality in CHIP. Our findings reveal a population enriched for CHIP carriers in the catheterization laboratory, an important observation for future studies focused on secondary prevention in CHIP-related ASCVD.

RNA-Seq Analysis of Clonal Hematopoiesis (CHIP) Blood Leukocytes Shows Dysregulation of Neutrophil / Innate Immunity-Related Genes

Clonal hematopoiesis of indeterminate potential (CHIP) is a common inflammatory condition of aging that causes myriad end-organ damage. We have recently shown associations for CHIP with acute kidney injury and with kidney function decline in the general population, with stronger associations for CHIP driven by mutations in genes other than DNMT3A (non- DNMT3A CHIP). Longitudinal kidney function endpoints in individuals with pre-existing chronic kidney disease (CKD) and CHIP have been examined in two previous studies, which reported conflicting findings and were limited by small sample sizes. In this study, we examined the prospective associations between CHIP and CKD progression events in four cohorts of CKD patients (total N = 5,772). The primary outcome was a composite of 50% kidney function decline or kidney failure. The slope of eGFR decline was examined as a secondary outcome. Mendelian randomization techniques were then used to investigate potential causal effects of CHIP on eGFR decline. Finally, kidney function was assessed in adenine-fed CKD model mice having received a bone marrow transplant recapitulating Tet2 -CHIP compared to controls transplanted wild-type bone marrow. Across all cohorts, the average age was 66.4 years, the average baseline eGFR was 42.6 ml/min/1.73m 2 , and 24% had CHIP. Upon meta-analysis, non- DNMT3A CHIP was associated with a 59% higher relative risk of incident CKD progression (HR 1.59, 95% CI: 1.02-2.47). This association was more pronounced among individuals with diabetes (HR 1.29, 95% CI: 1.03-1.62) and with baseline eGFR ≥ 30 ml/min/1.73m (HR 1.80, 95% CI: 1.11-2.90). Additionally, the annualized slope of eGFR decline was steeper among non- DNMT3A CHIP carriers, relative to non-carriers (β -0.61 ± 0.31 ml/min/1.73m 2 , p = 0.04). Mendelian randomization analyses suggested a causal role for CHIP in eGFR decline among individuals with diabetes. In a dietary adenine mouse model of CKD, Tet2 -CHIP was associated with lower GFR as well as greater kidney inflammation, tubular injury, and tubulointerstitial fibrosis. Non- DNMT3A CHIP is a potentially targetable novel risk factor for CKD progression.

Accumulation of somatic hematopoietic stem cell mutations with aging has been revealed by the recent genome-wide analysis. Clonal expansion, known as clonal hematopoiesis of indeterminate potential (CHIP), is a premalignant condition of hematological cancers. It is defined as the absence of definitive morphological evidence of a hematological neoplasm and occurrence of ≥2% of mutant allele fraction in the peripheral blood. In CHIP, the most frequently mutated genes are epigenetic regulators such as DNMT3A, TET2, and ASXL1. CHIP induces inflammation. CHIP is shown to be associated with not only hematological malignancy but also non-malignant disorders such as atherosclerosis, cardiovascular diseases and chronic liver disease. In addition, recent several large clinical trials have shown that CHIP is also the risk factor for developing chronic kidney disease (CKD). In this review article, we proposed novel findings about CHIP and CHIP related kidney disease based on the recent basic and clinical research. The possible mechanism of the kidney injury in CHIP is supposed to be due to the clonal expansion in both myeloid and lymphoid cell lines. In myeloid cell lines, the mutated macrophages increase the inflammatory cytokine level and induce chronic inflammation. It leads to epigenetic downregulation of kidney and macrophage klotho level. In lymphoid cell lines, CHIP might be related to monoclonal gammopathy of renal significance (MGRS). It describes any B cell or plasma cell clonal disorder that does not fulfill the criteria for cancer yet produces a nephrotoxic monoclonal immunoglobulin that leads to kidney injury or disease. MGRS causes M-protein related nephropathy frequently observed among aged CKD patients. It is important to consider the CHIP-related complications such as hematological malignancy, cardiovascular diseases and metabolic disorders in managing the elderly CKD patients. There are no established therapies for CHIP and CHIP-related CKD yet. However, recent studies have supported the development of effective CHIP therapies, such as blocking the expansion of aberrant HSCs and inhibiting chronic inflammation. In addition, drugs targeting the epigenetic regulation of Klotho in the kidney and macrophages might be therapeutic targets of CHIP in the kidney.