Modula'on of the F1F0-­‐ATPase Induces Apoptosis of Mouse and Human Lamina Propria T Cells and is Efficacious in Models of IBD!
Luigi Franchi, Lycera Corp
Acknowledgements
Lycera Corp.
University of Rome “Tor Vergata”
Anthony W. Opipari
Giovanni Monteleone
Rod Morgan
Ivan Monteleone
Mark Spahr
Davide di Fusco
Brian Sanchez
Francesca Zorzi
Charles Lesch
Irene Marafini
Corrin Hepburn
Francesco Pallone
Alexander Hurd
Clarke Taylor
University of Michigan
Chad Van Huis
Peter D.R. Higgins
Don Skalitzky
Kelli Porzondek
Peter Toogood
Northwestern University
Laura Carter
Kelan Hlavaty
Gary Glick
2!
Background: Targeting the F1F0-ATPase
• 
• 
Large, 16-subunit complex embedded in mitochondrial inner membrane
Final enzyme in the electron transport chain, (aka respiratory complex V)
• 
During oxidative phosphorylation, proton flow drives rotating motor function
catalyzing the generation of ATP
• 
Lycera has discovered small molecules modulators of the F1F0 ATPase
which induce apoptosis of susceptible cells by increasing intracellular
superoxide levels
F1F0-ATPase
Sundberg et al. (2009)
3!
Induction of Apoptosis by ATPase Modulators (LYC-51194)
Apoptosis is triggered by indirect effect on the mitochondrial respiratory chain
Modulation of the F1F0-ATPase
leads to hyperpolarization of
the mitochondrial membrane
potential and an increase in the
production of superoxide
by complex III
Sundberg et al. (2009)
4!
ATPase Modulators Act Selectively on
Chronically Activated Lymphocytes
Bioenergetic and redox abnormalities sensitize chronically-activated
lymphocytes to F1Fo-ATPase modulation resulting in selective apoptosis
Normal Lymphocytes
Chronically Activated Lymphocytes
Oxidative
Phosphorylation
OXPHOS
Glycolysis
Glycolysis
• Generate ATP by aerobic glycolysis
• High anti-oxidant stores (glutathione) and
low superoxide levels
• Generate ATP by oxidative phosphorylation
• Depleted anti-oxidant stores and elevated
basal superoxide level
• ATPase Modulator induced increases in
superoxide not detoxified à apoptosis
Science Translational Medicine, 2011, 3: 67ra8
5!
ATPase Modulators are Effective
in Different Murine Model of Autoimmune Disease
•  Selective apoptosis of chronically activated lymphocytes translates to
broad efficacy profile!
•  No effect on T-dependent antibody production!
Arthritis
Lupus
GVHD
J. Pharm. Exp.
Thera. (2008)
Arthritis Rheum.
(2003)
J. Clin. Invest.
(2002)
Sci Transl Med
(2011)
ATPase
Modulators
Control
Psoriasis
6!
The ATPase Modulator LYC-51194 Reduces Severity of
TNBS-Induced Chronic Colitis
!
day 1!
Body Weight Change
(% of Day 0)
105
day 21!
day 7!
110
day 28!
!
Control! (n=6)!
100
!
95
!
90
!
85
!
80
!
LYC-51194 !(n=8)!
*!p< 0.001!
Vehicle! (n=8)!
1
!
2
!
3
!
4
!
Weeks!
LYC-51194 Dosing! (30 (mg/kg)!
Control!
LYC-51194!
Vehicle!
7!
The ATPase Modulator LYC-51194 Reduces Severity of
Adoptive Transfer-Induced Colitis!
Body Weight Change
(% of Day 0)
102
100
98
LYC-51194 !(n=6)!
96
*!p< 0.05!
Vehicle! (n=6)!
94
92
1!
2!
3!
4!
5!
Weeks!
LYC-51194 Dosing! (30 (mg/kg)!
LYC-51194!
Vehicle!
8!
The ATPase Modulator LYC-51194 Dosed Therapeutically in vivo
Improves Inflammatory Bowel Disease (IBD) in Murine Models
Histology scores following therapeutic administration of LYC-51194
5
4
*
3
p< 0.005
2
1
Histology score
Histology score
5
4
* p< 0.005
3
2
1
0
0
EtOH
TNBS
+ vehicle
TNBS
+ LYC-51194
Chronic TNBS
vehicle
LYC-51194
Adoptive Transfer
9
p< 0.005!
*!
80!
70!
60!
vehicle!
LYC-51194!
Chronic TNBS!
lamina propria T lymphocytes!
90!
% survival!
lamina propria T lymphocytes!
% survival!
The ATPase Modulator LYC-51194 Induces Death of Lamina Propria T Cells!
90
80
70
60
50
40
30
!
!
!
!
!
!
!
*!p< 0.005!
vehicle!
LYC-51194!
Adoptive transfer!
10!
lamina propria activated T lymphocytes!
% survival!
ATPase Modulator LYC-51194 Induces !
Death of LP T Cells from CD Patient Biopsies!
100!
80!
60!
40!
20!
0!
vehicle! LYC-51194! Fas!
10μM!
vehicle! LYC-51194! Fas!
10μM!
non!
inflamed!
CD!
inflamed!
Isolated lamina propria T cells from CD biopsies treated with LYC-51194 ex vivo
11!
ATPase Modulator LYC-51194 Selectively Induces !
Death of CCR9+ Gut-tropic T cells in PBMC from CD Patients!
% survival!
80!
60!
40!
20!
0!
peripheral blood T lymphocytes!
lamina propria T lymphocytes!
% survival!
100!
100!
80!
60!
40!
20!
0!
vehicle! LYC-51194!
10μM!
lamina propria!
CD!
vehicle! LYC-51194! vehicle! LYC-51194!
10μM!
10μM!
PBMC!
PBMC!
CD non-gut tropic! CD gut tropic!
12!
Conclusions!
•  ATPase Modulators are efficacious in several rodent models of
IBD
•  ATPase Modulators induce cell death in lamina propria T cells
from CD subjects
•  Taken together these data suggests that ATPase Modulator
compounds may be a promising approach for treating IBD!
!
13!