Pe u ba ion o Cell-Sub ype Speci ic Ac i e Kinome
Ne wo ks in Schizoph enia
Ali Imami1,, Khaled Alganem1, Nicholas Henkel1, Alex Joyce1, Emily De ine3, Jessica Ji on1, Elizabe h Shedd o 1,
Abdul Hamoud1, Ja ek Melle 3, Robe E. McCullumsmi h1, 2
1Uni e si y o Toledo Depa men o Neu osciences and Psychia y, Toledo, OH, 2P omedica, Toledo, OH; 3Cincinna i Child en’s Hospi al
Medical Cen e , Cincinna i, OH; Uni e si y o Cincinna i, OH
DEPARTMENT OF
NEUROSCIENCES
AND PSYCHIATRY
UNIVERSITY OF TOLEDO
STE
CK1
AGC
CAMK
CMGC
TKL
STE
CK1
AGC
CAMK
CMGC
TKL
STE
CK1
AGC
CAMK
CMGC
TKL
A B
STE
CK1
AGC
CAMK
CMGC
TKL
C D
Py amidal Neu ons
SCZ s CTL
DLPFC
SCZ s CTL
Ra PFC
HLD s CTL
hiPSC Neu ons
DISC1 s CTL
Schizoph enia is a se ious illness wi h signi ican
e ec s on pa ien s and hei amilies. Because
he disease impac s execu i e unc ion, he Do -
sola e al P e on al Co ex (DLPFC) has been a
ocus o s udy. We p e iously epo ed al e ed
p o ein kinase ac i i y, including AKT, in pos mo -
em b ain samples om pa ien s wi h schizoph e-
nia. Now, we ex end his esea ch o he cellula
le el, concen a ing on on al co ical py amidal
neu ons. Using lase cap u e mic odissec ion,
we isola ed DLPFC py amidal neu ons om
ma ched pai s o schizoph enia and con ol pos -
mo em b ain samples (n = 20 pe g oup). We
hen used he PamChip STK kinome a ay assay
o high- h oughpu analysis o kinase ac i i y.
Wi h well-es ablished bioin o ma ics me hods,
we iden i ied ups eam kinases in ol ed in
schizoph enia. Se e al kinases o in e es
eme ged, such as c-Jun N- e minal kinases
(JNK), ex acellula signal- egula ed kinases
(ERK), and p38 mi ogen-ac i a ed p o ein kinas-
es (P38). O hese, he P38 kinases we e o pa -
icula impo ance as hey a e in ol ed in he in-
lamma ion cascade and immune unc ion. Nex ,
we applied a new echnique o iden i y kinase in-
e ac ion ne wo ks wi hin high- h oughpu kinase
ac i i y da a. This modeling enabled us o de ec
ne wo k-le el changes in py amidal neu ons in
schizoph enia. This is he i s s udy o analyze
he subkinome a he cellula le el in schizoph e-
nia and o e eal ac i e kinome ne wo k changes
in his o en-de as a ing illness. P e ailing hy-
po heses ega ding he e iology o schizoph enia
include abe an synapse u no e , which is me-
dia ed by mic oglia and immune unc ion pa h-
ways. Ou indings o e a c ucial new s a ing
poin o unde s anding how dis up ed signaling
ne wo ks may media e he pa hophysiology o
his o en de as a ing se e e men al illness.
Backg ound
Me hods
Pai wise SCZ s CTL
Conclusions
• The CAMK Family, and MAP Kinases in pa icula , appea o ha e an in acellula ole in
exci a o y neu ons he pa hogenesis o schizoph enia.
• The indi idual MAP Kinase membe s, JNK, ERK and P38, show di e en ial ac i i y in schizoph enia
subjec s.
• JNK and P38 ac i i y was inc eased, while ERK was dec eased, wi h he kinome a ay in SCZ.
• We con i med dec eased ERK ac i i y using a highly selec i e indi idual kinase ac i i y assay.
ns
2
4
6
8
10
CTL
SCZ
Pai 01
ns
2
4
6
8
10
CTL
SCZ
Pai 02
ns
2
4
6
8
10
CTL
SCZ
Pai 03
ns
2
4
6
8
10
CTL
SCZ
Pai 04
ns
2
4
6
8
10
CTL
SCZ
Pai 05
ns
2
4
6
8
10
CTL
SCZ
Pai 06
ns
2
4
6
8
10
CTL
SCZ
Pai 07
ns
2
4
6
8
10
CTL
SCZ
Pai 08
*
2
4
6
8
10
CTL
SCZ
Pai 09
ns
2
4
6
8
10
CTL
SCZ
Pai 11
ns
2
4
6
8
10
CTL
SCZ
Pai 12
ns
2
4
6
8
10
CTL
SCZ
Pai 13
ns
2
4
6
8
10
CTL
SCZ
Pai 14
ns
2
4
6
8
10
CTL
SCZ
Pai 15
ns
2
4
6
8
10
CTL
SCZ
Pai 16
ns
2
4
6
8
10
CTL
SCZ
Pai 17
ns
2
4
6
8
10
SCZ
CTL
Pai 18
ns
2
4
6
8
10
SCZ
CTL
Pai 19
CTL
SCZ
Pai 01
CTL
SCZ
Pai 02
CTL
SCZ
Pai 03
CTL
SCZ
Pai 04
CTL
SCZ
Pai 05
CTL
SCZ
Pai 06
CTL
SCZ
Pai 07
CTL
SCZ
Pai 08
CTL
SCZ
Pai 09
CTL
SCZ
Pai 11
CTL
SCZ
Pai 12
CTL
SCZ
Pai 13
CTL
SCZ
Pai 14
CTL
SCZ
Pai 15
CTL
SCZ
Pai 16
CTL
SCZ
Pai 17
CTL
SCZ
Pai 18
CTL
SCZ
Pai 19
Legend. Phylogene ic ees o kinome a ay ac i i y p o iles in (A) Schizoph enia (SCZ) s
Con ol (CTL) do sola e al p e on al co ex (DLPFC) py amidal neu ons, (B) egion-le el,
(C) human iPSC-de i ed on oco ical neu ons wi h DISC1 mu a ion and (D)
halope idol- ea ed (HDL) a p e on al co ex (PFC) subs a es. (E) Qua ile- anked kinase
ac i i y compa ison ac oss all pai s. (F) MAP Kinase Family-Speci ic (P38, JNK and ERK)
ac i i y ac oss all pai s. Red colo means s a is ically signi ican esul (p < 0.1) and la ge
ci cles ep esen bigge e ec sizes. (G) AssayQuan a ge ed ERK ac i i y assay o SCZ
s CTL showing a dec ease in ac i i y be ween g oups
Abou Me
DAPK
AMPK
SGK
PRKDC
ATR
MLCK
PKN
CK2
CAMK2
QIK
ATM
PKCA
COT
IRAK
SLK
MTOR
GSK
ULK
TTK
MST
VRK1
NIK
RAF
MLK
CLK
CAMKK
PEK
CAMK1
LKB
NEK
NUAK
MELK
BUD32
KHS
TAO
MAPKAPK
STE11
RIPK
PDHK
RAD53
MARK
WNK
MOS
PDK1
PAKB
BARK2
PIM
IKK
MSN
HAL
PHK
CHK1
BRSK
PKD
NDR
RSK
PASK
TLK
GRK
FRAY
PKG
AKT
NAK
PKCH
PKCI
PAKA
PKA
DMPK
AUR
PLK
NMO
CK1
STE7
DYRK
PKCD
STKR
P38
CDK
ERK
JNK
P01
P02
P03
P04
P05
P06
P07
P08
P09
P10
P11
P12
P13
P14
P15
P16
P17
P18
P19
-1.6
-1.4
-1.2
-1.0
-0.8
-0.6
-0.4
-0.2
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
P10
P12
P11
P15
P18
P04
P19
P09
P05
P16
P08
P14
P17
P13
P02
P06
P07
P03
P01
o
g
2
F
o
l
d
C
h
a
n
g
e
P10
P11
P12
P02
P18
P16
P19
P04
P15
P05
P08
P14
P09
P01
P13
P06
P17
P03
P07
P10
P12
P18
P15
P11
P14
P19
P16
P02
P01
P04
P03
P06
P09
P17
P07
P05
P08
P13
Subjec Pai
E ec Size
0.0
0.2
0.4
0.6
0.8
1.0
JNK
ERK
P38
Kinase Family
P38
Co e Func ion
JNK
ERK
Change in SCZ Impac ed p ocesses Clinical Rele anceMembe s
• P38α
• P38β
• P38γ
• P38δ
• JNK1
• JNK2
• JNK3
• ERK1
• ERK2
• ERK3/4
• ERK5
• ERK7
• In lamma ion
• Cy okine Signaling
•Mic oglia Ac i a ion
• S ess esponse
• Apop osis
• Synap ic P uning
• Cell G ow h
• Cell Di e en ia ion
• Synap ic Plas ici y
• Neu oin lamma ion
• Immune Response
• Neu o-Glial Communica ion
• Neu onal s ess signaling
• Dend i ic emodeling
• Long- e m po en ia ion
• Lea ning and Memo y
ci cui s
Links SCZ o immune
dys unc ion; candida e o
an i-in lamma o y he apy
D i es excessi e synapse
u no e ; po en ial s ess
pa hway modula ion a ge
Reduced plas ici y may
explain cogni i e de ici s;
con i med by in- i o assay
F
E
G
CTRL SCZ
0
100
200
300
400
ERK Ac i i y
slope
p=0.0319
