Aberrant methylation of DNA-damage-inducible transcript 3 promoter is a common event in patients with myelodysplastic syndrome

Abstract Abstract 5048 Background and Objectives: Myelodysplastic syndrome (MDS) is one of the most threatening hematological malignancies. Recently, the epigenetic changes have been recognized in the MDS, and some research found that the aberrant DNA methylation had close relationship with the MDS. Meanwhile, some studies showed that the activation of the Wnt signaling pathway and abnormal methylation of the Wnt antagonists had close relationship with hematological malignancies, such as AML AALL Aand CLL, but less is known in MDS. In our research, we studied the methylation status of Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) in the patients with MDS and evaluated the role of them in the pathogenesis and progression of MDS, with providing a new theory support for clarifying the complicated pathogenesis and progression of MDS. Methods: The methylation status of Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) in pretreatment bone marrow samples from 53 patients with MDS was measured by methylation-specific polymerase chain reaction (MSP). On the other hand, we collected the clinical materials of the patients with MDS and follow up the patients. Then the correlation between methylation and clinical features as well as prognosis of MDS patients was analyzed byχ2 test and Kaplan-Meier method. Results: In 53 bone marrow samples, the methylation frequencies of the Wnt antagonists were as follows: SFRP4 for 62.3 % (33/53), DKK1 for 45.3 % (24/53), HDPR1 for 34.0 % (18/53), SFRP1 for 15.1 % (8/53), DKK3 for 9.4 % (5/53), and WIF-1 for 5.7 % (3/53). After analyzing individual tumor suppressor gene, clinical parameters and prognostic information, it was found that the patients with the percentage of BM blast above 5% had higher methylation frequency of HDPR1 than the patients with the percentage of BM blast bellow 5% (44.4%∼a13.3%, P=0.034); besides, the methylation frequency of HDPR1 exhibited significant differences in the MDS subtypes (P=0.019), and was significantly correlated with the WPSS (P=0.037); In addition, Kaplan-Meier survival curve indicated that the mean overall survival of patients with methylation was significantly shorter than that of patients without HDPR1 methylation (457.3 days vs. 919.6 days, P=0.037) (Fig.1). Conclusion: The methylation of the Wnt antagonists (DKK1, DKK3, HDPR1, WIF-1, SFRP1 and SFRP4) were observed in patients with MDS and their methylation frequencies were different. The aberrant methylation of HDPR1 may play an important role in the progression and prognosis of MDS. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Aberrant methylation of the RIZ1 gene in myelodysplastic syndrome and acute myeloid leukemia

Leukemia Research ◽

10.1016/j.leukres.2010.08.002 ◽

2011 ◽

Vol 35 (4) ◽

pp. 516-521 ◽

Cited By ~ 12

Author(s):

Naoki Mori ◽

Kentaro Yoshinaga ◽

Kaori Tomita ◽

Mari Ohwashi ◽

Toshiaki Kondoh ◽

...

Keyword(s):

Acute Myeloid Leukemia ◽

Myelodysplastic Syndrome ◽

Myeloid Leukemia ◽

Aberrant Methylation ◽

Acute Myeloid

Download Full-text

Suppression of the DNA damage response in acute myeloid leukemia versus myelodysplastic syndrome

Oncogene ◽

10.1038/onc.2009.69 ◽

2009 ◽

Vol 28 (22) ◽

pp. 2205-2218 ◽

Cited By ~ 37

Author(s):

S Boehrer ◽

L Adès ◽

N Tajeddine ◽

W K Hofmann ◽

S Kriener ◽

...

Keyword(s):

Dna Damage ◽

Acute Myeloid Leukemia ◽

Myelodysplastic Syndrome ◽

Dna Damage Response ◽

Myeloid Leukemia ◽

Damage Response ◽

Acute Myeloid

Download Full-text

p53 Co-Activator Aspp1 Induces Apoptosis in Damaged Hematopoietic Stem Cells and Prevents Malignant Transformation

Blood ◽

10.1182/blood.v124.21.603.603 ◽

2014 ◽

Vol 124 (21) ◽

pp. 603-603 ◽

Cited By ~ 1

Author(s):

Masayuki Yamashita ◽

Eriko Nitta ◽

Toshio Suda

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Bone Marrow ◽

Dna Damage ◽

Hematopoietic Stem Cells ◽

Hematological Malignancies ◽

Aberrant Methylation ◽

Self Renewal ◽

Hematopoietic Stem ◽

Apoptotic Genes

Abstract Accumulation of DNA damage in hematopoietic stem cells (HSCs) is associated with aging, bone marrow failure and development of hematological malignancies. Mutation accumulation in HSCs precedes the development of leukemia and lymphoma, and these “pre-leukemic HSCs” can survive after chemotherapy, contributing to the relapse of the disease. Thus, understanding for the DNA damage response at a HSC level is a matter of critical importance for lifelong hematopoiesis, yet the protection mechanism for HSCs from DNA damage accumulation remains to be elucidated. During our study on the response of HSCs to ionizing radiation (IR), we have detected higher responsiveness of HSCs to DNA damage compared with committed progenitor cells: higher p53 activation was observed in HSC-enriched LSK (Lin-Sca1+cKit+) cells and LT-HSCs (CD150+CD41-CD48-LSK) than in myeloid progenitor-enriched LKS- cells. Of note, when treated with 4 Gy IR, LSK cells exhibited stronger upregulation of pro-apoptotic genes Bax, Noxa and Puma compared with LKS- cells, whereas upregulation of survival-contributing p21 and Mdm2 genes was comparable between the two populations. Corresponding to such characteristic behavior, we have identified apoptosis-stimulating protein of p53 1 (Aspp1) as a novel specific regulator of HSCs that provides HSCs with high sensitivity to apoptosis. We found that mRNA and protein of Aspp1 were specifically detected in LSK cells and LT-HSCs. To uncover the roles of Aspp1 in the regulation of HSCs, we evaluated HSCs of adult Aspp1 knockout (KO) mice. These mutant mice exhibited a major increase in the absolute number of LSK cells (1.5-fold; P<0.05) and LT-HSCs (2-fold; P<0.0005). Furthermore, self-renewal capacity of Aspp1-null HSCs was significantly enhanced as measured by serial competitive bone marrow (BM) transplantation assays (P<0.01). To assess the cause of enhanced self-renewal of Aspp1-null HSCs, we examined gene expression profile of Aspp1-null LSK cells before and after BM transplantation using multiplex quantitative RT-PCR array. Aspp1-null LSK cells showed higher expression of multiple quiescence-related genes including Tek, Mpl and Ndn. In line with this, Ki67 staining revealed that Aspp1-null LSK cells showed resistance to the loss of quiescence after serial BM transplantation (P<0.01), and Aspp1 KO mice showed accelerated recovery of peripheral blood and BM when treated with a single dose of 5-FU (P<0.05). Moreover, when serially transplanted or subjected to 4 Gy IR in vivo, Aspp1-null LSK cells exhibited higher resistance to apoptosis which was detected as decreased proportion of Annexin V-positive cells (P<0.05). Gene expression analysis consistently revealed that the induction of pro-apoptotic genes Bax, Noxa and Puma was impaired in irradiated Aspp1-null LSK cells. As a result of the reduced apoptosis, Aspp1-null LSK cells exhibited the tendency to retain persistent DNA damage after genotoxic stress as assessed by γH2AX and 53BP1 foci (chi-square test, P<0.05). Importantly, by breeding Aspp1 KO mice with Mx1-Cre mice and p53flox/flox mice, we verified that Aspp1 synergized with p53 to regulate self-renewal and genomic integrity of HSCs beyond its canonical p53-dependent function. Aspp1 loss further enhanced self-renewal capacity of HSCs in a p53-null background when assayed by serial BM transplantation (P<0.05). Likewise, Aspp1 deficiency further accentuated the accumulation of DNA damage after IR exposure in the absence of p53 (P<0.05). Consequently, whereas approximately half of the recipients receiving p53-null LSK cells died of thymic lymphoma, the recipient mice transplanted with LSK cells deficient for both Aspp1 and p53 were 100% lethal within 6 months after BM transplantation (log-rank test, P<0.01). These mice succumbed to hematological malignancies, mostly T-cell acute lymphoblastic lymphoma and leukemia (ALL) (88%) but also B-cell (6%) and myeloid (6%) malignancies. Taken together, our study demonstrates that Aspp1 attenuates HSC quiescence and induces apoptosis in damaged HSCs, in both p53-dependent and -independent manners, thereby inhibiting the development of leukemia and lymphoma in conjunction with p53 in HSCs. As loss of Aspp1 expression due to aberrant methylation of its promoter has already been proven to be an independent poor prognosis factor in ALL patients, Aspp1 may be a potential target for stem cell-directed therapy of leukemia and lymphoma. Disclosures No relevant conflicts of interest to declare.

Download Full-text

Aberrant methylation of GTPase regulator associated with the focal adhesion kinase (GRAF) promoter is an adverse prognostic factor in myelodysplastic syndrome

European Journal Of Haematology ◽

10.1111/j.1600-0609.2010.01453.x ◽

2010 ◽

pp. no-no ◽

Cited By ~ 1

Author(s):

Jun Qian ◽

Zhen Qian ◽

Jiang Lin ◽

Dong-Ming Yao ◽

Zhao-Hui Zhu ◽

...

Keyword(s):

Prognostic Factor ◽

Myelodysplastic Syndrome ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Aberrant Methylation ◽

Adverse Prognostic Factor

Download Full-text

DNA damage check points prevent leukemic transformation in myelodysplastic syndrome

Leukemia ◽

10.1038/sj.leu.2404748 ◽

2007 ◽

Vol 21 (10) ◽

pp. 2195-2198 ◽

Cited By ~ 15

Author(s):

S Horibe ◽

M Takagi ◽

J Unno ◽

M Nagasawa ◽

T Morio ◽

...

Keyword(s):

Dna Damage ◽

Myelodysplastic Syndrome ◽

Leukemic Transformation ◽

Check Points

Download Full-text

Innovative Analyses Support a Role for DNA Damage and an Aberrant Cell Cycle in Myelodysplastic Syndrome Pathogenesis

Bone Marrow Research ◽

10.1155/2011/950934 ◽

2011 ◽

Vol 2011 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

David R. Head ◽

James W. Jacobberger ◽

Claudio Mosse ◽

Madan Jagasia ◽

William Dupont ◽

...

Keyword(s):

Cell Cycle ◽

Flow Cytometry ◽

Dna Damage ◽

Myelodysplastic Syndrome ◽

Dna Content ◽

Annexin V ◽

Aberrant Cell ◽

Cd34 Cells ◽

G2 Cells ◽

Myeloid Progenitors

We used flow cytometry to analyze the cell cycle, DNA damage, and apoptosis in hematopoietic subsets in MDS marrow. Subsets were assigned using CD45, side scatter, CD34, and CD71. Cell cycle fractions were analyzed using DRAQ 5 (DNA content) and MPM-2 (mitoses). DNA damage was assessed using p-H2A.X, and apoptosis using Annexin V. Compared to controls, MDS patients demonstrated no increased mitoses in erythroid, myeloid, or CD34+ cells. Myeloid progenitors demonstrated increased G2 cells, which with no increased mitoses suggested delayed passage through G2. Myeloid progenitors demonstrated increased p-H2A.X, consistent with DNA damage causing this delay. Annexin V reactivity was equivalent in MDS and controls. Results for each parameter varied among hematopoietic compartments, demonstrating the need to analyze compartments separately. Our results suggest that peripheral cytopenias in MDS are due to delayed cell cycle passage of marrow progenitors and that this delayed passage and leukemic progression derive from excessive DNA damage.

Download Full-text