Global histone modification fingerprinting in human cells using epigenetic
reverse phase protein array
#MMPMID28326191
Partolina M
; Thoms HC
; MacLeod KG
; Rodriguez-Blanco G
; Clarke MN
; Venkatasubramani AV
; Beesoo R
; Larionov V
; Neergheen-Bhujun VS
; Serrels B
; Kimura H
; Carragher NO
; Kagansky A
Cell Death Discov
2017[]; 3
(?): 16077
PMID28326191
show ga
The balance between acetylation and deacetylation of histone proteins plays a
critical role in the regulation of genomic functions. Aberrations in global
levels of histone modifications are linked to carcinogenesis and are currently
the focus of intense scrutiny and translational research investments to develop
new therapies, which can modify complex disease pathophysiology through
epigenetic control. However, despite significant progress in our understanding of
the molecular mechanisms of epigenetic machinery in various genomic contexts and
cell types, the links between epigenetic modifications and cellular phenotypes
are far from being clear. For example, enzymes controlling histone modifications
utilize key cellular metabolites associated with intra- and extracellular
feedback loops, adding a further layer of complexity to this process. Meanwhile,
it has become increasingly evident that new assay technologies which provide
robust and precise measurement of global histone modifications are required, for
at least two pressing reasons: firstly, many approved drugs are known to
influence histone modifications and new cancer therapies are increasingly being
developed towards targeting histone deacetylases (HDACs) and other epigenetic
readers and writers. Therefore, robust assays for fingerprinting the global
effects of such drugs on preclinical cell, organoid and in vivo models is
required; and secondly, robust histone-fingerprinting assays applicable to
patient samples may afford the development of next-generation diagnostic and
prognostic tools. In our study, we have used a panel of monoclonal antibodies to
determine the relative changes in the global abundance of post-translational
modifications on histones purified from cancer cell lines treated with HDAC
inhibitors using a novel technique, called epigenetic reverse phase protein
array. We observed a robust increase in acetylation levels within 2-24?h after
inhibition of HDACs in different cancer cell lines. Moreover, when these cells
were treated with N-acetylated amino acids in addition to HDACs, we detected a
further increase in histone acetylation, demonstrating that these molecules could
be utilized as donors of the acetyl moiety for protein acetylation. Consequently,
this study not only offers a novel assay for diagnostics and drug screening but
also warrants further research of the novel class of inexpensive, non-toxic
natural compounds that could potentiate the effects of HDAC inhibitors and is
therefore of interest for cancer therapeutics.
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