In Vitro Erythropoiesis from Bone Marrow-derived Progenitors Provides a Physiological Assay for Toxic and Mutagenic Compounds

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2007 May 16

PMID 17502613

Citations 31

Authors

J Shuga

J Zhang

L D Samson

H F Lodish

L G Griffith

Affiliations

Soon will be listed here.

Abstract

The goal of this study was to create an in vitro cell culture system that captures essential features of the in vivo erythroid micronucleus (MN) genotoxicity assay, thus enabling increased throughput and controlled studies of the hematopoietic DNA damage response. We show that adult bone marrow (BM) cultures respond to erythropoietin, the principal hormone that stimulates erythropoiesis, with physiological erythropoietic proliferation, differentiation, and enucleation. We then show that this in vitro erythropoietic system clearly signals exposure to genotoxicants through erythroid MN formation. Furthermore, we determined that DNA repair-deficient (MGMT(-/-)) BM displayed sensitivity to genotoxic exposure in vivo compared with WT BM and that this phenotypic response was reflected in erythropoietic cultures. These findings suggest that this in vitro erythroid MN assay is capable of screening for genotoxicity on BM in a physiologically reflective manner. Finally, responses to genotoxicants during erythroid differentiation varied with exposure time, demonstrating that this system can be used to study the effect of DNA damage at specific developmental stages.

Citing Articles

DDX41 haploinsufficiency causes inefficient hematopoiesis under stress and cooperates with p53 mutations to cause hematologic malignancy.

Stepanchick E, Wilson A, Sulentic A, Choi K, Hueneman K, Starczynowski D Leukemia. 2024; 38(8):1787-1798.

PMID: 38937548 PMC: 11286521. DOI: 10.1038/s41375-024-02304-9.

IL-18 Binding Protein-Producing Cells Attenuate Anemia in Murine Macrophage Activation Syndrome.

Harel M, Fauteux-Daniel S, Rodriguez E, Palmer G, Gabay C J Immunol. 2023; 210(11):1790-1803.

PMID: 37074208 PMC: 7614543. DOI: 10.4049/jimmunol.2300065.

Arsenic impairs the lineage commitment of hematopoietic progenitor cells through the attenuation of GATA-2 DNA binding activity.

Medina S, Zhang H, Santos-Medina L, Wan G, Bolt A, Zhou X Toxicol Appl Pharmacol. 2022; 452:116193.

PMID: 35961411 PMC: 9583771. DOI: 10.1016/j.taap.2022.116193.

eIF6 rebinding dynamically couples ribosome maturation and translation.

Jaako P, Faille A, Tan S, Wong C, Escudero-Urquijo N, Castro-Hartmann P Nat Commun. 2022; 13(1):1562.

PMID: 35322020 PMC: 8943182. DOI: 10.1038/s41467-022-29214-7.

Arsenite and monomethylarsonous acid disrupt erythropoiesis through combined effects on differentiation and survival pathways in early erythroid progenitors.

Medina S, Bolt A, Zhou X, Wan G, Xu H, Lauer F Toxicol Lett. 2021; 350:111-120.

PMID: 34274428 PMC: 8487637. DOI: 10.1016/j.toxlet.2021.07.008.

References

Zeiger E . Carcinogenicity of mutagens: predictive capability of the Salmonella mutagenesis assay for rodent carcinogenicity. Cancer Res. 1987; 47(5):1287-96. View

Cornetta K, Croop J, Dropcho E, Abonour R, Kieran M, Kreissman S . A pilot study of dose-intensified procarbazine, CCNU, vincristine for poor prognosis brain tumors utilizing fibronectin-assisted, retroviral-mediated modification of CD34+ peripheral blood cells with O6-methylguanine DNA methyltransferase. Cancer Gene Ther. 2006; 13(9):886-95. DOI: 10.1038/sj.cgt.7700963. View

ROMAGNA F, Staniforth C . The automated bone marrow micronucleus test. Mutat Res. 1989; 213(1):91-104. DOI: 10.1016/0027-5107(89)90035-3. View

LUDLUM D . DNA alkylation by the haloethylnitrosoureas: nature of modifications produced and their enzymatic repair or removal. Mutat Res. 1990; 233(1-2):117-26. DOI: 10.1016/0027-5107(90)90156-x. View

Hirai O, Miyamae Y, Fujino Y, Izumi H, Miyamoto A, Noguchi H . Prior bleeding enhances the sensitivity of the in vivo micronucleus test. Mutat Res. 1991; 264(3):109-14. DOI: 10.1016/0165-7992(91)90125-n. View

Heddle J, Cimino M, Hayashi M, ROMAGNA F, Shelby M, Tucker J . Micronuclei as an index of cytogenetic damage: past, present, and future. Environ Mol Mutagen. 1991; 18(4):277-91. DOI: 10.1002/em.2850180414. View

Yajima N, Kurata Y, Imai E, Sawai T, Takeshita Y . Genotoxicity of genetic recombinant human erythropoietin in a novel test system. Mutagenesis. 1993; 8(3):231-6. DOI: 10.1093/mutage/8.3.231. View

Kaina B, Fritz G, Coquerelle T . Contribution of O6-alkylguanine and N-alkylpurines to the formation of sister chromatid exchanges, chromosomal aberrations, and gene mutations: new insights gained from studies of genetically engineered mammalian cell lines. Environ Mol Mutagen. 1993; 22(4):283-92. DOI: 10.1002/em.2850220418. View

Galloway S . Chromosome aberrations induced in vitro: mechanisms, delayed expression, and intriguing questions. Environ Mol Mutagen. 1994; 23 Suppl 24:44-53. DOI: 10.1002/em.2850230612. View

10.

Dertinger S, Torous D, Tometsko K . Simple and reliable enumeration of micronucleated reticulocytes with a single-laser flow cytometer. Mutat Res. 1996; 371(3-4):283-92. DOI: 10.1016/s0165-1218(96)90117-2. View

11.

Morita T, Asano N, Awogi T, Sasaki Y, Sato S, Shimada H . Evaluation of the rodent micronucleus assay in the screening of IARC carcinogens (groups 1, 2A and 2B) the summary report of the 6th collaborative study by CSGMT/JEMS MMS. Collaborative Study of the Micronucleus Group Test. Mammalian Mutagenicity.... Mutat Res. 1997; 389(1):3-122. DOI: 10.1016/s1383-5718(96)00070-8. View

12.

Dertinger S, Torous D, Tometsko K . Flow cytometric analysis of micronucleated reticulocytes in mouse bone marrow. Mutat Res. 1997; 390(3):257-62. DOI: 10.1016/s1383-5718(97)00022-3. View

13.

Glassner B, Weeda G, Allan J, Broekhof J, Carls N, Donker I . DNA repair methyltransferase (Mgmt) knockout mice are sensitive to the lethal effects of chemotherapeutic alkylating agents. Mutagenesis. 1999; 14(3):339-47. DOI: 10.1093/mutage/14.3.339. View

14.

Bodell W . Effect of cations on the formation of DNA alkylation products in DNA reacted with 1-(2-Chloroethyl)-1-nitrosourea. Chem Res Toxicol. 1999; 12(10):965-70. DOI: 10.1021/tx980200c. View

15.

ROTH R, Samson L . Gene transfer to suppress bone marrow alkylation sensitivity. Mutat Res. 2000; 462(2-3):107-20. DOI: 10.1016/s1383-5742(00)00021-1. View

16.

Kina T, Ikuta K, Takayama E, Wada K, Majumdar A, Weissman I . The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage. Br J Haematol. 2000; 109(2):280-7. DOI: 10.1046/j.1365-2141.2000.02037.x. View

17.

Fenech M . The in vitro micronucleus technique. Mutat Res. 2000; 455(1-2):81-95. DOI: 10.1016/s0027-5107(00)00065-8. View

18.

Styles J, Clark H, Festing M, Rew D . Automation of mouse micronucleus genotoxicity assay by laser scanning cytometry. Cytometry. 2001; 44(2):153-5. DOI: 10.1002/1097-0320(20010601)44:2<153::aid-cyto1095>3.0.co;2-h. View

19.

Socolovsky M, Nam H, Fleming M, Haase V, Brugnara C, Lodish H . Ineffective erythropoiesis in Stat5a(-/-)5b(-/-) mice due to decreased survival of early erythroblasts. Blood. 2001; 98(12):3261-73. DOI: 10.1182/blood.v98.12.3261. View

20.

Roth R, Samson L . 3-Methyladenine DNA glycosylase-deficient Aag null mice display unexpected bone marrow alkylation resistance. Cancer Res. 2002; 62(3):656-60. View