CRISPR-Mediated Gene Editing to Assess the Roles of Tet2 and Dnmt3a in Clonal Hematopoiesis and Cardiovascular Disease
Open accessAbstract
RATIONALE: Clonal hematopoiesis has been associated with increased mortality and cardiovascular disease. This condition can arise from somatic mutations in preleukemic driver genes within hematopoietic stem/progenitor cells. Approximately 40 candidate driver genes have been identified, but mutations in only 1 of these genes, TET2 (ten-eleven translocation-2), has been shown to casually contribute to cardiovascular disease in murine models. OBJECTIVE: To develop a facile system to evaluate the disease characteristics of different clonal hematopoiesis driver genes using lentivirus vector and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/clustered regularly interspaced short palindromic repeat-associated 9) methodology. Using this methodology, evaluate whether Dnmt3a (DNA [cytosine-5]-methyltransferase 3a)-a commonly occurring clonal hematopoiesis driver gene-causally contributes to cardiovascular disease. METHODS AND RESULTS: Lentivirus vectors were used to deliver Cas9 and guide RNA to introduce inactivating mutations in Tet2 and Dnmt3a in lineage-negative bone marrow cells. After implantation into lethally irradiated mice, these cells were engrafted and gave rise to labeled blood cell progeny. When challenged with an infusion of Ang II (angiotensin II), mice with inactivating mutations in Tet2 or Dnmt3a displayed greater cardiac hypertrophy, diminished cardiac function, and greater cardiac and renal fibrosis. In comparison with Tet2, inactivation of Dnmt3a did not lead to detectable expansion of the mutant hematopoietic cells during the time course of these experiments. Tet2 inactivation promoted the expression of IL (interleukin) 1β, IL-6, and Ccl5, whereas Dnmt3a inactivation promoted the expression of Cxcl1 (CXC chemokine ligand), Cxcl2, IL-6, and Ccl5 in a lipopolysaccharide-stimulated macrophage cell line. CONCLUSIONS: Experiments using lentivirus vector/CRISPR methodology provided evidence suggesting that inactivating DNMT3A mutations in hematopoietic cells contributes to cardiovascular disease. Comparative analyses showed that inactivation of Tet2 and Dnmt3 was similar in their ability to promote Ang II-induced cardiac dysfunction and renal fibrosis in mice. However, gene-specific actions were indicated by differences in kinetics of hematopoietic stem/progenitor cell expansion and different patterns of inflammatory gene expression.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Loss of Tet2 or Dnmt3a led to worsening heart function in a mouse model of congestive heart failure, corroborating the human genetic association.
“Subsequent studies showed that loss of Tet2 or Dnmt3a led to worsening heart function in a mouse model of congestive heart failure, corroborating the human genetic association 87,88.”
From Clonal hematopoiesis in human aging and disease (2019) · cites this paper as [87]
CRISPR mutation of Dnmt3a in mouse RAW 264.7 macrophages led to increased expression of Cxcl1, Cxcl2, and Il6 in response to endotoxin.
“Another study used CRISPR to mutate Dnmt3a in mouse RAW 264.7 macrophages, and observed increased expression of Cxcl1, Cxcl2, and Il6 in response to endotoxin 87.”
From Clonal hematopoiesis in human aging and disease (2019) · cites this paper as [87]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Age-related clonal hematopoiesis associated with adverse outcomes
Jaiswal, S., Fontanillas, P., Flannick, J., Manning, A., Grauman, P. V., Mar, B. G., Lindsley, R. C., Mermel, C. H., Burtt, N., Chavez, A., et al. (2014) · New England Journal of Medicine · doi:10.1056/nejmoa1408617 · Cited by 4,841 · see citing works
Clonal hematopoiesis and blood-cancer risk inferred from blood DNA sequence
Genovese, G., Kähler, A. K., Handsaker, R. E., Lindberg, J., Rose, S. A., Bakhoum, S. F., Chambert, K., Mick, E., Neale, B. M., Fromer, M., et al. (2014) · New England Journal of Medicine · doi:10.1056/nejmoa1409405 · Cited by 3,531 · see citing works
Clonal hematopoiesis and risk of atherosclerotic cardiovascular disease
Jaiswal, S., Natarajan, P., Silver, A. J., Gibson, C. J., Bick, A. G., Shvartz, E., McConkey, M., Gupta, N., Gabriel, S., Ardissino, D., et al. (2017) · New England Journal of Medicine · doi:10.1056/nejmoa1701719 · Cited by 2,683 · see citing works
Clonal hematopoiesis of indeterminate potential and its distinction from myelodysplastic syndromes
Steensma, D. P., Bejar, R., Jaiswal, S., Lindsley, R. C., Sekeres, M. A., Hasserjian, R. P., & Ebert, B. L. (2015) · Blood · doi:10.1182/blood-2015-03-631747 · Cited by 1,971 · see citing works
Age-related mutations associated with clonal hematopoietic expansion and malignancies
Xie, M., Lu, C., Wang, J., McLellan, M. D., Johnson, K. J., Wendl, M. C., McMichael, J. F., Schmidt, H. K., Yellapantula, V., Miller, C. A., et al. (2014) · Nature Medicine · doi:10.1038/nm.3733 · Cited by 1,874 · see citing works
Clonal hematopoiesis associated with TET2 deficiency accelerates atherosclerosis development in mice
Fuster, J. J., MacLauchlan, S., Zuriaga, M. A., Polackal, M. N., Ostriker, A. C., Chakraborty, R., Wu, C.-L., Sano, S., Muralidharan, S., Rius, C., et al. (2017) · Science · doi:10.1126/science.aag1381 · Cited by 1,536 · see citing works
Self-citationEpelman, S., Lavine, K. J., Beaudin, A. E., Sojka, D. K., Carrero, J. A., Calderon, B., Brija, T., Gautier, E. L., Ivanov, S., Satpathy, A. T., et al. (2014) · Immunity · doi:10.1016/j.immuni.2013.11.019 · Cited by 1,494 · see citing works
Dnmt3a is essential for hematopoietic stem cell differentiation
Challen, G. A., Sun, D., Jeong, M., Luo, M., Jelinek, J., Berg, J. S., Bock, C., Vasanthakumar, A., Gu, H., Xi, Y., et al. (2012) · Nature Genetics · doi:10.1038/ng.1009 · Cited by 1,122 · see citing works
Clonal hematopoiesis, with and without candidate driver mutations, is common in the elderly
Zink, F., Stacey, S. N., Norddahl, G. L., Frigge, M. L., Magnusson, O. T., Jonsdottir, I., Thorgeirsson, T. E., Sigurdsson, A., Gudjonsson, S. A., Gudmundsson, J., et al. (2017) · Blood · doi:10.1182/blood-2017-02-769869 · Cited by 777 · see citing works
Sano, S., Oshima, K., Wang, Y., MacLauchlan, S., Katanasaka, Y., Sano, M., Zuriaga, M. A., Yoshiyama, M., Goukassian, D., Cooper, M. A., et al. (2018) · Journal of the American College of Cardiology · doi:10.1016/j.jacc.2017.12.037 · Cited by 699 · see citing works
Self-citationHeckl, D., Kowalczyk, M. S., Yudovich, D., Belizaire, R., Puram, R. V., McConkey, M. E., Thielke, A., Aster, J. C., Regev, A., & Ebert, B. L. (2014) · Nature Biotechnology · doi:10.1038/nbt.2951 · Cited by 555 · see citing works
DNMT3A in haematological malignancies
Yang, L., Rau, R. E., & Goodell, M. A. (2015) · Nature reviews. Cancer · doi:10.1038/nrc3895 · Cited by 509 · see citing works
Monocyte and macrophage contributions to cardiac remodeling
Hulsmans, M., Sam, F., & Nahrendorf, M. (2015) · Journal of Molecular and Cellular Cardiology · doi:10.1016/j.yjmcc.2015.11.015 · Cited by 279 · see citing works
Angiotensin-Converting Enzyme II in the Heart and the Kidney
Danilczyk, U. G., & Penninger, J. M. (2006) · Circulation Research · doi:10.1161/01.res.0000205761.22353.5f · Cited by 271 · see citing works
Zhang, X., Su, J., Jeong, M., Ko, M., Huang, Y., Park, H. J., Guzman, A., Lei, Y., Huang, Y.-H., Rao, A., et al. (2016) · Nature Genetics · doi:10.1038/ng.3610 · Cited by 258 · see citing works
Loss of Dnmt3a Immortalizes Hematopoietic Stem Cells In Vivo
Jeong, M., Park, H. J., Celik, H., Ostrander, E. L., Reyes, J. M., Guzman, A., Rodriguez, B., Lei, Y., Lee, Y., Ding, L., et al. (2018) · Cell Reports · doi:10.1016/j.celrep.2018.03.025 · Cited by 243 · see citing works
Li, X., Zhang, Q., Ding, Y., Liu, Y., Zhao, D., Zhao, K., Shen, Q., Liu, X., Zhu, X., Li, N., et al. (2016) · Nature Immunology · doi:10.1038/ni.3464 · Cited by 202 · see citing works
Somatic Mutations and Clonal Hematopoiesis
Fuster, J. J., & Walsh, K. (2018) · Circulation Research · doi:10.1161/circresaha.117.312115 · Cited by 163 · see citing works
Self-citation<i>Dnmt3a</i> restrains mast cell inflammatory responses
Leoni, C., Montagner, S., Rinaldi, A., Bertoni, F., Polletti, S., Balestrieri, C., & Monticelli, S. (2017) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1616420114 · Cited by 138 · see citing works
Somatic mutations, genome mosaicism, cancer and aging
Vijg, J. (2014) · Current Opinion in Genetics & Development · doi:10.1016/j.gde.2014.04.002 · Cited by 134 · see citing works
Cole, C. B., Russler-Germain, D. A., Ketkar, S., Verdoni, A. M., Smith, A. M., Bangert, C. V., Helton, N. M., Guo, M., Klco, J. M., O’Laughlin, S., et al. (2017) · Journal of Clinical Investigation · doi:10.1172/jci93041 · Cited by 84 · see citing works
Epigenetic modifications of interleukin-6 in synovial fibroblasts from osteoarthritis patients
Yang, F., Zhou, S., Wang, C., et al. (2017) · Scientific Reports · doi:10.1038/srep43592 · Cited by 69 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Why does COVID-19 disproportionately affect older people?
Mueller, A. L., McNamara, M. S., & Sinclair, D. A. (2020) · Aging · doi:10.18632/aging.103344 · Cited by 1,128 · see citing works
Clonal hematopoiesis in human aging and disease
Jaiswal, S., & Ebert, B. L. (2019) · Science · doi:10.1126/science.aan4673 · Cited by 1,156 · see citing works
“7 macrophages and observed increased expression of Cxcl1, Cxcl2, and Il6 in response to endotoxin (88).”
BackgroundDorsheimer, L., Assmus, B., Rasper, T., Ortmann, C. A., Ecke, A., Abou-El-Ardat, K., Schmid, T., Brüne, B., Wagner, S., Serve, H., et al. (2019) · JAMA Cardiology · doi:10.1001/jamacardio.2018.3965 · Cited by 470 · see citing works
Increased stem cell proliferation in atherosclerosis accelerates clonal hematopoiesis
Heyde, A., Rohde, D., McAlpine, C. S., Zhang, S., Hoyer, F. F., Gerold, J. M., Cheek, D., Iwamoto, Y., Schloss, M. J., Vandoorne, K., et al. (2021) · Cell · doi:10.1016/j.cell.2021.01.049 · Cited by 267 · see citing works
Clonal hematopoiesis is associated with protection from Alzheimer’s disease
Bouzid, H., Belk, J. A., Jan, M., Qi, Y., Sarnowski, C., Wirth, S., Ma, L., Chrostek, M. R., Ahmad, H., Nachun, D., et al. (2023) · Nature Medicine · doi:10.1038/s41591-023-02397-2 · Cited by 169 · see citing works
Aging drives <i>Tet2</i> <i>+/−</i> clonal hematopoiesis via IL-1 signaling
Caiado, F., Kovtonyuk, L. V., Gonullu, N. G., Fullin, J., Boettcher, S., & Manz, M. G. (2023) · Blood · doi:10.1182/blood.2022016835 · Cited by 161 · see citing works
Clonal hematopoiesis: Mutation-specific adaptation to environmental change
Florez, M. A., Tran, B. T., Wathan, T. K., DeGregori, J., Pietras, E. M., & King, K. Y. (2022) · Cell Stem Cell · doi:10.1016/j.stem.2022.05.006 · Cited by 129 · see citing works
Kim, J.-S., Trzebanski, S., Shin, S.-H., Schori, L., Frumer Friedman, G. R., Ilani, N. C., Kadam, A., Vicario, R., Aust, O., Bugaeva, P., et al. (2025) · Cell Reports · doi:10.1016/j.celrep.2025.115609 · Cited by 33 · see citing works
- Authors
- Sano, S., Oshima, K., Wang, Y., Katanasaka, Y., Sano, M., & Walsh, K.
- Affiliations
- University of Virginia Medical Center (US)
- Boston University (US)
- Shizuoka University (JP)
- University of Virginia (US)
- Venue
- Circulation Research
- Published
- 2018-07-20
- Volume
- 123
- Issue
- 3
- Pages
- 335-341
- ISSN
- 0009-7330 (ISSN-L)
- 1524-4571
- PMC
- PMC6054544
- OpenAlex
- W2800927741
- Cited by
- 417
- References
- 22
- Citations by year
- 2026: 30 · 2025: 56 · 2024: 61 · 2023: 65 · 2022: 73 · 2021: 60 · 2020: 42 · 2019: 23 · 2018: 6 · 2012: 1
- FWCI
- 23.49
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Circulation Research (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Japan Society for the Promotion of Science (16K18876)
- National Heart, Lung, and Blood Institute (R01HL131006, R01HL138014, R01HL141256, R01HL132564)
- Topics
- Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Keywords
- clonal hematopoiesis, TET2, DNMT3A, cardiovascular disease, CRISPR/Cas9 gene inactivation, lentiviral vectors, hematopoietic stem and progenitor cells, angiotensin II, cardiac hypertrophy, myocardial fibrosis, renal fibrosis, chemokine expression, interleukin-6, RANTES
- MeSH terms
- DNA Methyltransferase 3A, Angiotensin II, Animals, Cardiovascular Diseases, Cells, Cultured, DNA (Cytosine-5-)-Methyltransferases, DNA-Binding Proteins, Hematopoiesis, Hematopoietic Stem Cells, Humans, Macrophages, Male, Mice, Inbred C57BL, Proto-Oncogene Proteins, Cytokines, 3T3 Cells, Gene Silencing, Cell Proliferation, Dioxygenases, Mice, HEK293 Cells, CRISPR-Cas Systems
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
