Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Dec 15;128(24):2774-2784.
doi: 10.1182/blood-2015-11-683490. Epub 2016 Oct 18.

Metformin improves defective hematopoiesis and delays tumor formation in Fanconi anemia mice

Qing-Shuo Zhang  1 , Weiliang Tang  2 , Matthew Deater  1 , Ngoc Phan  1 , Andrea N Marcogliese  3 , Hui Li  2 , Muhsen Al-Dhalimy  4 , Angela Major  5 , Susan Olson  4 , Raymond J Monnat Jr  2   6 , Markus Grompe  1
Affiliations

Metformin improves defective hematopoiesis and delays tumor formation in Fanconi anemia mice

Qing-Shuo Zhang et al. Blood. .

Abstract

Fanconi anemia (FA) is an inherited bone marrow failure disorder associated with a high incidence of leukemia and solid tumors. Bone marrow transplantation is currently the only curative therapy for the hematopoietic complications of this disorder. However, long-term morbidity and mortality remain very high, and new therapeutics are badly needed. Here we show that the widely used diabetes drug metformin improves hematopoiesis and delays tumor formation in Fancd2-/- mice. Metformin is the first compound reported to improve both of these FA phenotypes. Importantly, the beneficial effects are specific to FA mice and are not seen in the wild-type controls. In this preclinical model of FA, metformin outperformed the current standard of care, oxymetholone, by improving peripheral blood counts in Fancd2-/- mice significantly faster. Metformin increased the size of the hematopoietic stem cell compartment and enhanced quiescence in hematopoietic stem and progenitor cells. In tumor-prone Fancd2-/-Trp53+/- mice, metformin delayed the onset of tumors and significantly extended the tumor-free survival time. In addition, we found that metformin and the structurally related compound aminoguanidine reduced DNA damage and ameliorated spontaneous chromosome breakage and radials in human FA patient-derived cells. Our results also indicate that aldehyde detoxification might be one of the mechanisms by which metformin reduces DNA damage in FA cells.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Metformin administration enhances hematopoiesis. (A) CBCs after 6 months of treatment with metformin. D2, Fancd2; HGB, hemoglobin; NS, not significant. The data are pooled results from 17 to 19 individual mice in each group. (B) Representative flow cytometry profiles for placebo and metformin-treated Fancd2−/− mice. The percentages on the profiles indicate the mean value for each group. PI, propidium iodide. (C) Quantification of CD34−KSL frequency in bone marrow. The data represent the percentage of CD34−KSL cells in all nucleated bone marrow cells from 15 mice in each group.
Figure 2.
Figure 2.
Metformin administration helps FA HSPCs maintain quiescence. (A) Representative flow cytometry profiles of the cell cycle analysis for KSL cells: placebo-treated Fancd2+/+ KSL cells (i), MET-treated Fancd2+/+ KSL cells (ii), placebo-treated Fancd2−/− KSL cells (iii), and MET-treated Fancd2−/− KSL cells (iv). The percentages on the profiles indicate the mean value for each group. (B) Statistical analysis of the cell cycle status. Data are poled results from 10 to 15 mice.
Figure 3.
Figure 3.
Metformin administration improves the function of FA bone marrow cells. (A) Representative pictures of spleens analyzed in the CFU-S assay. (B) Statistical analysis of CFU-S assays. Forty thousand donor bone marrow cells were injected intravenously into each recipient mouse. Data represent 3 or 4 donors in each group of mice, with 2 to 4 recipients for each donor. (C) Schematic chart to show the procedures of poly(I:C) experiments. Three-month-old mice were injected intraperitoneally with either poly(I:C) or saline at 8 mg/kg body weight. The mice were harvested either 2 weeks (for CFU-S assay) or 3 weeks (for CBC analysis) after the completion of poly(I:C) treatment. (D) Statistical analysis of CFU-S assays after poly(I:C) administration. Data represent 8 or 9 donors in each group of mice, with 2 to 4 recipients for each donor. Total recipients in each group ranged from 23 to 28 mice. (E) Statistical analysis of CBC tests after poly(I:C) administration. Data are pooled results from 11 to 17 mice each group for wild-type mice and 18 to 19 mice each group for Fancd2−/− mice.
Figure 4.
Figure 4.
Metformin prevents FA patient–derived cells from developing radials and chromosomal breaks. (A) Representative pictures of spontaneous radials and breaks in PD259i human FA-A fibroblasts. The arrows indicate a chromosomal break (left) or a radial (right). (B-C) Statistical quantitation of radials and breaks in PD259i human FA-A fibroblasts after aminoguanidine or metformin treatment. PD259i cells were maintained in DMEM supplemented with 10% fetal bovine serum and penicillin/streptomycin. Cells were cultured with metformin or aminoguanidine for 48 hours before metaphase spreads were made. Fifty metaphases for each sample were scored for radial contents and chromosomal breakage. Data are combined results from 6 independent experiments. AG, aminoguanidine.
Figure 5.
Figure 5.
Aldehyde sensitivity of human FA cells and the detoxification of aldehydes by aminoguanidine. (A) Formaldehyde dose-dependent survival of EUFA316 human FA-G mutant lymphoblastoid cells compared with an isogenic, FANCG-complemented EUFA316 control. Complementation of patient cells was performed by stably transducing FANCG-mutant EUFA316 cells with a retrovirus expressing a wild-type human FANCG complementary DNA. EUFA316 and EUFA316+FANCG cells were cultivated in RPMI 1640 medium supplemented with 10% fetal bovine serum and penicillin/streptomycin. (B) Aminoguanidine shows dose-dependent rescue of EUFA316 cells from formaldehyde-induced cell death. (C) MMC dose-dependent survival of EUFA316 and wild-type controls. (D) Aminoguanidine provided a mild protection on EUFA316 cells from MMC-induced cell death. (E-F) Statistical quantitation of radials and chromosomal breaks in 259i human FA-A fibroblasts treated with C3, the ADH5 inhibitor. Metformin was added to the cell culture at 10 µM and maintained at the same concentration throughout the experiment. One hour later, C3 was added at 100 µM. Forty-eight hours later, cells were harvested for breakage and radial analysis. Data are combined results from 4 independent experiments.
Figure 6.
Figure 6.
Metformin protects Fancd2−/− mice from tumor development. Kaplan-Meier survival curves of the Fancd2−/−Trp53+/− mice and Fancd2+/+Trp53+/− mice. For Fancd2−/−Trp53+/− mice, the data represent 31 mice for metformin treatment and 60 mice for placebo treatment. For Fancd2+/+Trp53+/− mice, the data represent 30 mice for metformin treatment and 60 mice for placebo treatment. Tumor samples and selected tissues were fixed in 10% phosphate-buffered formalin, stained with hematoxylin and eosin, and examined under a microscope. The Kaplan-Meier survival curves were generated by Prism 6.0c Software (GraphPad Software, Inc.), and P values were calculated using the log-rank (Mantel-Cox) test.
See this image and copyright information in PMC

Comment in

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Shimamura A, Alter BP. Pathophysiology and management of inherited bone marrow failure syndromes. Blood Rev. 2010;24(3):101-122. - PMC - PubMed
    1. Kim H, D’Andrea AD. Regulation of DNA cross-link repair by the Fanconi anemia/BRCA pathway. Genes Dev. 2012;26(13):1393-1408. - PMC - PubMed
    1. Kottemann MC, Smogorzewska A. Fanconi anaemia and the repair of Watson and Crick DNA crosslinks. Nature. 2013;493(7432):356-363. - PMC - PubMed
    1. Kutler DI, Singh B, Satagopan J, et al. . A 20-year perspective on the International Fanconi Anemia Registry (IFAR). Blood. 2003;101(4):1249-1256. - PubMed
    1. Ceccaldi R, Parmar K, Mouly E, et al. . Bone marrow failure in Fanconi anemia is triggered by an exacerbated p53/p21 DNA damage response that impairs hematopoietic stem and progenitor cells. Cell Stem Cell. 2012;11(1):36-49. - PMC - PubMed

Publication types

MeSH terms