Transcript Control of T cell development by cellular stress response

New effector cytokine networks in IBD
Scripps Florida
- Kelly McKevitt
- Wei Cao, M.D., Ph.D.
Funding
- GlaxoSmithKline
- Scripps – Florida
Tempero Pharmaceuticals
- Radha Ramesh
- Alex Pellerin
- Thaddeus Carlson, Ph.D.
- Ivana Djuretic, Ph.D.
University of Miami
- Maria Abreu, M.D.
- Jacob McCauley, Ph.D.
- Maria Quintero
New York University
- Derya Unutmaz, M.D.
- Lina Kozaya
Mark Sundrud, Ph.D.
The Scripps Research Institute
[email protected]
Unexpected features of pathogenic
T cells associated with IBD
Scripps Florida
- Kelly McKevitt
- Wei Cao, M.D., Ph.D.
Funding
- GlaxoSmithKline
- Scripps – Florida
Tempero Pharmaceuticals
- Radha Ramesh
- Alex Pellerin
- Thaddeus Carlson, Ph.D.
- Ivana Djuretic, Ph.D.
University of Miami
- Maria Abreu, M.D.
- Jacob McCauley, Ph.D.
- Maria Quintero
New York University
- Derya Unutmaz, M.D.
- Lina Kozaya
Mark Sundrud, Ph.D.
The Scripps Research Institute
[email protected]
Disclosures
• Former employee of Tempero pharmaceuticals, Inc.
• No current relationship with Tempero or any other
company
Mechanisms of T cell-driven inflammation
2) T cell differentiation into
specialized “effector” subsets
1) T cell activation in
regional lymph nodes
5) Resolution of
4) T cell effector function in
inflammation/memory tissue/pathogen clearance 3) Tissue infiltration
Mechanisms of T cell-driven inflammation
Disease biology
Models of T cell development
Naïve CD4+
nTreg
Foxp3
IL-4
IL-12
IL-6
IL-21
TNFa
TFH
Th2
Gata3
CCR4
CRTH2
Bcl6
Th22 iTreg
AhR
CXCR5 CCR10
IL-4
IL-5
IL-13
IL-21
TGFb IL-1b
IL-12
IL-6
IL-2
IL-23
IL-22
Foxp3
Th17
Th1
RORgt
T-bet
CCR6
IL-17A
T cell
activation IL-17F
CXCR3
IFNg
“I go home today. They cured me using this new
miracle drug. But I’m afraid it doesn’t work in
humans.”
Mechanisms of T cell-driven inflammation
Human
What T cell subsets are in
autoimmune target organs?
Mouse
What regulates their
inflammatory function?
CD4+ T cell diversity
Naïve CD4+
IL-4
IL-12
IL-6
IL-21
TNFa
Th2
TFH
Gata3
CCR4
CRTH2
Bcl6
Th22 iTreg
AhR
CXCR5 CCR10
IL-4
IL-5
IL-13
IL-21
TGFb IL-1b
IL-12
IL-2 IL-6IL-23
IL-22
Foxp3
Th17
Th1
RORgt
T-bet
CCR6
T cell
IL-17A
activation IL-17F
CXCR3
IFNg
“Th17 cells are defined by and
function through IL-17”
IL-17-independent functions of Th17 cells
1. IL-17A expression is instable; “Th17 cells are a
transitional phenotype”
2. Th17 cells promote inflammation independent of IL17A
3. T cells that lack Il23r, Stat3, Rorc fail to induce
experimental colitis upon transfer into lymphopenic
mice
Doodes et al., J Immunol 2008; Codarri et al., Nat Immunol 2010; Huber et al., Gut 2010; Huber et a.,
IL-17A
Th17
What is a “Th17”
cell in humans?
Stable features of Th17 cells
Human memory T cells
• CCR6 expression defines
RORgt
effector/memory T cells with “Th17”
CCR6
features:
RORC
500
400
300
200
100
0
Chemokine receptor: R4 R4 R4 R4 X3 X3 X3 X3 R4 R4 R4 X3 X3 X3 -Cytokine: -- g 22 17 -- g 22 17 -- g 22 -- g 22 --
CCR6+
CCR6-
Naive
IL-17A
IL-17F
– Express RORgt
– Capable of producing IL-17 cytokines
mRNA (AU)
Th17
Wan et al., J Exp Med 2011
Stable features of Th17 cells
Human memory T cells
• CCR6 expression defines effector/memory
T cells with “Th17” features:
– Express RORgt
– Capable of producing IL-17 cytokines
• IL-17A is transient; Th17 is stable
• IL-17A expression is maintained by IL-2
family cytokines (e.g., IL-2, IL-7, IL-15)
- PI-3K/AKT signaling antagonizes FOXO1-mediated repression of
IL17A
Wan et al., J Exp Med 2011
CCR6
Stable features of Th17 cells
IL-17A
Wan et al., J Exp Med 2011
Pathogenic/non-pathogenic Th17 cells
Th17 “Non-pathogenic”
Naïve T cells
TGFb1
IL-6
IL-17A
IL-10
Homeostasis
IL-6 Th17 “Pathogenic”
IL-1b
TGFb3
Goreschi et al., Nature 2010
Hirota et al., NI 2011
Lee et al., NI 2012
IL-23
IL-17A
IFNg
Inflammation
“Pathogenic” Th17 cells
1.
2.
3.
Express Il23r and induce EAE in an IL-23-dependent manner
Produce both Th17/Th1 cytokines (IL-17A + IFNg)
Express a unique (“pathogenic”) transcriptional signature
Genetics of Th17 cells in autoimmune disease
163 independent loci
IL-23R
JAK2
RORC
STAT3
Pathogenic Th17signature genes
IFNG
IL22
CSF2 (GM-CSF)
STAT4
IL1R1
IL17A
IBD-associated loci
Locus not associated with IBD
Jostens et al., Nature 2012
What distinguishes pathogenic
Th17 cells in humans?
Th1
Th17.1
Th17
Th2
MDR1 is selectively expressed
by a subset of Th17.1 cells
MDR1
Th17.1selective
ABCB1
RORC
CCR6
CTSH
“Core Th17”
signature
Th17selective
IL17A
IL17F
TEM
Ramesh et al., J Exp Med 2014; Aller et al. Science 2009
Multi-drug resistance type 1 (MDR1)
1. 1 of 50 human ATP-binding cassette (ABC) transporter genes
1. Expressed in chemoresistant tumors; transports
chemotherapeutic agents (e.g., vinblastine, doxorubicin)
1. Expressed on epithelial cells (gut, lung, kidney, BBB)
1. Transports numerous structurally-unrelated molecules
1. Pharmaceutical drugs
2. xenobiotic compounds
1. Little is known about what it does in T cells
Assessing MDR1 expression/function by FACS
Rh123
CD4+ T cells
CCR6
DMSO
10
5
10
4
15
CsA
36
10 3
10
5
10
4
.08
Elacridar
50
10 3
2
0
10
10
2
0
47
10
2
10
3
Rh123
10
4
10
5
10
5
10
4
.2
51
10 3
2
0
37oC
10
.1
0
10
49
2
10
3
10
4
10
5
2
0
.1
0
10
49
2
10
3
10
4
10
5
Memory CD4+ T cells
mRNA (fold-change)
40C
MDR1MDR1+
25
ABCB1 (MDR1)
ABCC1 (MRP1)
20
ABCC3 (MRP3)
15
ABCG2 (BCRP)
10
5
0
Rh123:
hi
lo
Blood (HC)
hi
lo
Gut (CD)
Ramesh et al., J Exp Med 2014
MDR1+ Th17 cells
1. Restricted to the CCR6+CXCR3hiCCR4lo (Th17.1) compartment
1. Enriched within CCR7lo (TEM) cells
1. Display “stem cell-like” characteristics (c-Kit, TCF7, LEF1, etc.)
Ramesh et al., J Exp Med 2014
MDR1+ Th17 cells
TEM phenotype
Subset
IL-17/ IL23R
P Th17- NP Th17IL-23
IFNg+ (mRNA) signature signature response
R6-X3hiR4loRh123hi MDR1-Th1
--
+
--
R6+X3loR4hiRh123hi MDR1-Th17
R6+X3hiR4loRh123hi MDR1-Th17.1
--
+
--
--
+
--
+
--
+++
+
--
+
R6+X3hiR4loRh123lo MDR1+Th17.1 +
Ramesh et al., J Exp Med 2014
MDR1+ Th17 cells are enriched and activated in CD
Crohn’s Disease lesion
***
80
MDR1+ 3
*
***
60
2
Pathogenic Th17
Non-pathogenic Th17
Other
*
0
on
C
C AT
i
M
M
s
N
B
B
Le
P
P
Normal
Crohn’s
0
MDR1- -1
IFNG
IL23R
TNF
CSF2
CCL20
IL22
IL2
CCL5
1
20
SOCS3
IL6ST
IL1RN
MAF
TNFRSF9
FOXP3
IL10
RORC
IL17A
IL17F
40
PB
MC
(H
P
C)
No BMC
!
rm
(C
a
l
In
gu D)!
fl a
t(
me
C
d
D)
gu
!
t(
C
D)
!
% MDR1+
Frequency (%)
(of CD4+ CD45RO+)
MDR1+ T cells
Ramesh et al., J Exp Med 2014
Conclusions: MDR1 distinguishes pathogenic Th17 cells
• MDR1 is selectively expressed
on IL-23-responsive proinflammatory Th17 cells
“Spotting the troublemakers”
• MDR1+ Th17 cells are enriched
and activated in involved areas
of the gut in Crohn’s disease
•
MDR1+
Th17 cells are refractory
to glucocorticoids used to treat
IBD
Y. Bordon, Nat Rev Immunol 2014
Ramesh et al., J Exp Med 2014; Y. Bordon, Nat. Rev. Immunol. 2014
Mechanisms of T cell-driven inflammation
Human
What T cell subsets are in
autoimmune target organs?
Mouse
What regulates their
inflammatory function?
Thank you!
Scripps Florida
- Kelly McKevitt
- Wei Cao, M.D., Ph.D.
Funding
- GlaxoSmithKline
- Scripps – Florida
Tempero Pharmaceuticals
- Radha Ramesh
- Alex Pellerin
- Thaddeus Carlson, Ph.D.
- Ivana Djuretic, Ph.D.
University of Miami
- Maria Abreu, M.D.
- Jacob McCauley, Ph.D.
- Maria Quintero
New York University
- Derya Unutmaz, M.D.
- Lina Kozaya
Mark Sundrud, Ph.D.
The Scripps Research Institute
[email protected]