Multiple myeloma is characterized by widespread
epii
igenomicalter ti
a ons with pr
ti
ognos c il
imp ili t
ca i
tions
Christoph J. Heuck1, Jayesh Mehta2, Natalie Pulliam2, Richard Meagher2, Li Zhou1, Eric Vickrey2, Elizabeth
Voight2, John Greally1, Amit Verma1, Seema Singhal2
1Albert Einstein

College

of Medicine, Bronx

, NY

2Robert H. Lurie Comprehensive Cancer Center at Northwestern University, Chicago, MI
2010 ASCO
ASC Annual

Mee

t
Mee ing
t
June 48
Chicago, IL

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Epigenetics
· change
chang s
e in phenotype or gen
ge e
n ex
e pression
pr

caused by mechanisms other than changes in
the underlying DNA sequence
· Post tr
ii
anscriptional gene il
s
i
enc ng by RNA
interference
· Histone modifications
· DNA methyla
y tion

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HpaII tiny fragment Enrichment by Ligation
mediat
media ed
t
PCR
P
(HELP assay)
assa

---

The HELP assay can separate normal from malignant
plasma cells


·29 MM patients
·4 nl cont
co r
nt o
r l
o s
Group 2nl PC
Group 1

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The HELP assay identifies two subgroups of
myel
mye o
l m
o a cells
Normal PC vs. Group 1
Normal PC vs. Group 2
575
249
612
153
P-value
-log(10)
Difference of Means

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Ingenuity Pathway Analysis of differentially
meth
me yla
th
t
yla ed
t
gen
ge e
n s
e
The majority of

dif

f
dif er
f en
er tially
en
methyla
meth
t
yla ed
t
gen
ge e
n s
e
involve cancer associated pathways, with
· TNF
· P16
· PI3 Kinase
· IFN alpha
· NF kappa B
as the central nodes

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Clinical Correlation
Predictors for Group 2Pvalue
nonIgG isotype
p=0.0006
plasma cell

labeling index
inde 0.3

p=0.018
abnormal classic cytogenetics
p=0.009
complex cytogenetic
p=0.000007
abnormalities on

FISH
Mean Gr. 1Mean Gr. 2Pvalue
Hb
12.2
10.3
p=0.0023
B2MG
2.91
6.55
p=0.030

---

Conclusions
· This is the first genome wide analysis of DNA
methyla
meth
t
yla ion
t
in multiple

my

e
my loma
m
· The HELP assay clearly separates normal controls
from
fr
MM

cells

in
unsuper

vised clus

t
clus e
t ring
· The HELP assay identifies 2 distinct clusters of
myeloma cells
· Group 2 MM samples identifies patients w/ poor
prognostic markers
· The mos

t
mos significan
signific t
an dif

f
dif er
f en
er tially
en
meth

yla
meth
t
yla ed
t
genes involve the cancer pathways around P16,
PI3 Kinase, TNF, NF kappa B and Interferon alpha

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Where do we want to go from here?
· Analysis more MM samples and more normal
controls
· Gene expression studies
· Identific
Iden
a
tific tion
a
of

ta
rget
rge s
t fo
r further

functional s
t
s udies
t

---

Thank You
Verma Lab
Northwestern University
Amit Verma (PI)
Seema Singhal
Yon
Yo ka
k i Mo
Jay
Ja e
y sh
e
Meht
Meh a
t
Tushar Baghat
Richard Meagher
Amy Sharma
Natalie Pulliam
Li Zhou
Caroline Hu
Center for Epigenomics
John Greally
Gr
Shahina Maqbool

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