Inactivation of HOXA genes by hypermethylation in myeloid and lymphoid malignancy is frequent and associated with poor prognosis.
OBJECTIVE
The HOX genes comprise a large family of homeodomain-containing transcription factors, present in four separate clusters, which are key regulators of embryonic development, hematopoietic differentiation, and leukemogenesis. We aimed to study the role of DNA methylation as an inducer of HOX gene silencing in leukemia.


METHODS
Three hundred and seventy-eight samples of myeloid and lymphoid leukemia were quantitatively analyzed (by COBRA analysis and pyrosequencing of bisulfite-modified DNA) for methylation of eight HOXA and HOXB cluster genes. The biological significance of the methylation identified was studied by expression analysis and through re-expression of HOXA5 in a chronic myeloid leukemia (CML) blast crisis cell line model.


RESULTS
Here, we identify frequent hypermethylation and gene inactivation of HOXA and HOXB cluster genes in leukemia. In particular, hypermethylation of HOXA4 and HOXA5 was frequently observed (26-79%) in all types of leukemias studied. HOXA6 hypermethylation was predominantly restricted to lymphoid malignancies, whereas hypermethylation of other HOXA and HOXB genes was only observed in childhood leukemia. HOX gene methylation exhibited clear correlations with important clinical variables, most notably in CML, in which hypermethylation of both HOXA5 (P = 0.00002) and HOXA4 (P = 0.006) was strongly correlated with progression to blast crisis. Furthermore, re-expression of HOXA5 in CML blast crisis cells resulted in the induction of markers of granulocytic differentiation.


CONCLUSIONS
We propose that in addition to the oncogenic role of some HOX family members, other HOX genes are frequent targets for gene inactivation and normally play suppressor roles in leukemia development.
Strathdee
Gordon
G
Centre for Oncology and Applied Pharmacology, Cancer Research UK. G.R.Strathdee@newcastle.ac.uk
Holyoake
Tessa L
TL
Sim
Alyson
A
Parker
Anton
A
Oscier
David G
DG
Melo
Junia V
JV
Meyer
Stefan
S
Eden
Tim
T
Dickinson
Anne M
AM
Mountford
Joanne C
JC
Jorgensen
Heather G
HG
Soutar
Richard
R
Brown
Robert
R
D001752:Blast Crisis; D018899:CpG Islands; D019175:DNA Methylation; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D007938:Leukemia / Q000235:genetics* / Q000401:mortality / Q000473:pathology; D007945:Leukemia, Lymphoid / Q000235:genetics / Q000401:mortality / Q000473:pathology; D007951:Leukemia, Myeloid / Q000235:genetics / Q000401:mortality / Q000473:pathology; D011379:Prognosis; D014157:Transcription Factors