a sub-network of signaling protein complexes in aa may provide new molecular candidates for...
TRANSCRIPT
A sub-network of signaling protein complexes in
Alopecia Areata may provide new molecular
candidates for pharmacologic targeting
Adam G. Schrum, PhD
Assistant Professor of Immunology
Mayo Clinic College of Medicine
CD28 a b
CD8
SLAP 130
TCR
a b CD3
e g
z z
CD4
CBP
CSK
CD45
LCK
FYN Zap70
Calcineurin NFATc
Calmodulin Ca2+
Bcl-10
CD3
de
IKKa
IKKb
IKKg TBK1
IKKe
TANK
IkB
NFkB
MALT1
GRB2
SOS1
RAS
RAF1
MEK1
ERK1/ERK2
DOK1
DOK2
SHIP1
GADS
SLP-76
Nck
PAK
WASP
WIP
F-actin
ARP2/3
ITK
CDC42 RAC1
MEKK1
JNKK1
JNK1
SHP1/2
CARMA1 LAT PLCg CpA
PKCq VAV
PI3K
AKT
PKCa
PKCb
Cbl-b
B7.1/2 MHC
1 2
APC MHC
1
T cell
THEMIS CD6
Do different signals through TCR affect
immunologic tolerance and AA?
Qualitative
Quantitative
Kinetic
What could be
“different” ?
Responders: Jurkat
APCs: Raji
+/- SEE, 5 min
PiSCES: Proteins in Shared Complexes detected by Exposed Surface epitopes:
a multiplex matrix analysis to assess PPI network signatures
Smith SEP, Neier SC, et al., Aug 2, 2016 Science Signaling (AAAS)
PiSCES network analysis &
visualization
Responders: Jurkat
APCs: Raji
+/- SEE, 5 min
SEE / null, 5 min SEE CD28+ / SEE CD28–, 5 min
Subject T cells
(millions) %CD4
SP %CD8
SP %CLA+CCR4+ Age Sex
aa1 2 20.9 56.9 12.9 29 m
aa2 3 65.4 28 74.2 50 f
aa3 8.4 14.7 76.3 17.4 50 f
aa4 3.87 88.9 8.7 44.5 81 f
aa5 1.32 21.6 59.3 21.7 39 f
aa6 1.58 85.1 7.7 71.5 41 f
aa7 1.58 69.3 25 16.6 59 m
cntl1 1.5 82.9 6.1 24.4 51 m
cntl2 1.6 78 17.9 50.5 77 m
cntl3 6.8 91.8 5.1 84.2 51 m
cntl4 6.2 98.1 0.3 79.1 76 m
cntl5 2.8 85.9 12.3 59 89 f
T cells from volunteer patient punch-biopsies
Overall similar response
pattern between Control
and AA patient T cells to
anti-CD3/anti-CD28
stimulation
PiSCES data distinguishes AA from Control patient T cells,
and suggests increased basal TCR signaling in AA
AA > Ctrl
Ctrl > AA
Difference in basal levels
ITK
Thy1
Fyb
TCR
a b
z z
LCK Zap70
CD3
de
GADS
Nck
SLP-76
LAT VAV
plasma membrane
GRB2
SOS1
LAT
PKCq
PLCg Fyn
Zap70
CD3
e g
SKAP55 Integrin activation
Cell adhesion
Ras-Raf-ERK Differentiation
Clonal expansion
AA > CtrlCtrl > AA
Upon exogenous stimulation, PiSCES network activity
suggests disease-associated signaling profile in
Alopecia Areata patients
Acknowledgements
Schrum Lab
• Stephen E.P. Smith, PhD
(currently Assistant
Professor, U. WA,
Seattle)
• Steven Neier
• Brendan Reed
• Tessa Davis
Diana Gil, PhD
• Alejandro Ferrer, PhD
• Robert Stiles
Collaborators
• Claudia Neuhauser, PhD (U. MN, Comput. Biol.)
• Jeanette Eckel-Passow, PhD (Mayo Biostatisitics)
• Jason P. Sinnwell (Mayo Biostatistics)
• Gabriel Sciallis, MD (Mayo Dermatology)
• Carrie Wieland, MD (Mayo Dermatology)
• Shelly Torgerson, MD/PhD (Mayo Dermatology)
Funding partners
• R01 NIGMS
• Mr. Bernie Fineman (Mayo Benefactor)