SLIDE 1
Disruption of the mTOR-eIF4F Axis by Selectively Targeting PI3Kd and Proteasome Potently Inhibits Cap Dependent Translation of c-Myc in Aggressive Lymphomas
Changchun Deng, M.D., Ph.D. Mark Lipstein, B.S. Luigi Scotto, Ph.D. Michael Mangone, Ph.D. Owen A. O’Connor, M.D., Ph.D. Columbia University Medical Center Department of Medicine Center for Lymphoid Malignancies
SLIDE 2 Therapeutic Strategy Targeting c-Myc in Cancer Is Urgently Needed
- C-MYC rearrangement is a risk factor for poor survival in diffuse large B cell
lymphoma (DLBCL)
Savage et al. Blood 2009; Barrans et al. JCO 2010; Copie-Bergman et al. Blood 2015
- C-MYC expression is a risk factor for poor survival in DLBCL
Green TM et al., J Clin Oncol. 2012; Johnson NA, et al., J Clin Oncol. 2012; Hu S et al., Blood. 2013
- However, no drugs specifically targeting the activity of c-Myc have been
approved for any cancer.
- C-Myc is a master transcription factor, and lacks enzymatic activity
- Structurally, c-Myc lacks globular functional domains for small molecule targeting
- The extended interaction between the c-Myc and Max offers no apparent site for
positioning a small-molecule inhibitor.
- Targeting the BET bromodomains is a promsing strategy for c-Myc driven cancer
McKeown and Bradner, CSH Perspective 2014
SLIDE 3
Potential Strategies to Silence the Translation of c-Myc through Targeting the mTOR-eIF4F Axis
mTOR c-Myc eIF4E 4EBP1 eIF4E eIF4A eIF4G P P 4EBP1 PI3Kd (eIF4F)
Translation
Proteasome Amino acids AKT
Dibble CC and Cantley LC. Trends Cell Biol, 2015 Suraweera, A., et al., Mol Cell, 2012 Quy, P.N., et al., J Biol Chem, 2013 Hutter, G., et al., Leukemia, 2012 Zhang, Y., et al., Nature, 2014
Carfilzomib TGR-1202 (Cfz) (TG) Bortezomib Idelalisib (Bz) (Cal)
? ?
SLIDE 4
Cal-101 (mM) TGR-1202 (mM) Observed Inhibition in the DLBCL cell line LY10
PI3Kd Inhibitors and Proteasome Inhibitors Synergistically Inhibit DLBCL
Carfilzomib (nM) Bortfezomib (nM)
SLIDE 5
Dual Inhibition of PI3Kd and Proteasome Is Most Synergistic with TG&Cfz Followed by Cal&Cfz > TG&Bz > Cal&Bz
Cal-101 (mM) TGR-1202 (mM) Carfilzomib (nM) Bortfezomib(nM) Excess Over BLISS (EOB) in the DLBCL cell line LY10
SLIDE 6
Dual Inhibition of PI3Kd and Proteasome Is Most Synergistic with TG&Cfz Followed by Cal&Cfz > TG&Bz > Cal&Bz
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
L y 1 0
E x p e c te d % In h ib itio n O b s e rv e d % In h ib itio n
SLIDE 7 TGR-1202&Carfilzomib Is the Most Synergistic Combination in DLBCL, Mantle Cell Lymphoma, and Multiple Myeloma
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
L y 1
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
S U D H L -2
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
J e k o -1
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
Z 1 3 8
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
S U D H L -4
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
M M .1 s
Observed Inhibition (%) Expected Inhibition (%)
SLIDE 8 TGR-1202&Carfilzomib Is the Most Synergistic Combination in T Cell Lymphoma
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
H 9
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
HH
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
P 1 2
2 5 5 0 7 5 1 0 0 2 5 5 0 7 5 1 0 0
P F 3 8 2
Observed Inhibition (%) Expected Inhibition (%)
SLIDE 9 TGR-1202 and Carfilzomib Are Synergistic in Primary CLL and MCL Cells but Not Toxic to Healthy Lymphocytes
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
C L L # 1
C e ll V ia b ility (% )
C fz T G 2 .5 m M + C fz T G 5 m M + C fz T G 7 .5 m M + C fz
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
C L L # 1 B z C a l 2 .5 m M + B z C a l 5 m M + B z C a l 7 .5 m M + B z
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
C L L # 2
C e ll V ia b ility (% )
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
C L L # 2
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
M C L
[C fz ] n M C e ll V ia b ility (% )
2 .5 5 .0 7 .5 2 5 5 0 7 5 1 0 0
M C L
[B z ] n M
a . b . c . d . e . f.
0% 20% 40% 60% 80% 100% 120% PBMC - 24HR PBMC - 48HR PBMC - 72HR
Viability Healthy lymphocytes
SLIDE 10 PARP Cleavage
2 4 6 8 10
Cal-Bort - Cal+Bortb TG-Cfz - TG+Cfz
TGR-1202 and Carfilzomib Synergistically Induce Apoptosis in Lymphoma Cell Lines and Primary Lymphoma Cells
Fold Change / Control
Caspase 3/7 Activity
PARP b-Actin (B: LY10) (A:LY7) PARP b-Actin (C: PF382) PARP b-actin PARP b-actin PARP b-actin (D: CLL-1) (E: CLL-2) (F: LY10)
SLIDE 11 (A: DLBCL) P-Akt P-4EBP1 4EBP1 HIF1a c-Myc Actin
P-4EBP1 c-Myc B-Actin 4EBP1
(C:CLL-1) (D: CLL-2) (B: T-ALL) GAPDH
TGR-1202 and Carfilzomib Synergistically Inhibit Phosphorylation of 4EBP1 and Expression of c-Myc
B-Actin C-Myc P-4EBP1 4EBP1
(E: MM)
SLIDE 12 C-Myc B-Actin
TGR-1202 and Carfilzomib in Combination Inhibit Cap Dependent Translation of c-Myc in DLBCL
(E) Cap dependent translation of Myc
LucR LucF
PolioV IRES MYC UTR
0% 50% 100%
R/F Luc (%control)
p=0.0013 (C) (D)
Actin c-Myc (A)
0% 50% 100% 150%
(B)
mRNA level of MYC % control
0% 50% 100% 150%
mRNA level of MYC % control
(F)
SLIDE 13 Running Enrichment Score (RES)
Gene Set 52 : SCHUHMACHER
Gene List Index
Gene Set 70 : DANG Gene Set 32 : KIM
RES RES Gene List Index
Gene Set 29 : SCHLOSSER
TGR-1202 and Carfilzomib in Combination Inhibit the c-Myc Transcription Program in DLBCL
SLIDE 14
Overexpression of eIF4E Suppresses the Synergistic Cytotoxicity of TGR-1202 and Carfilzomib and Increases the Protein Level of c-Myc
Control TG5mM & Cfz5nM B-Actin c-Myc eIF4E TG5mM & Cfz6nM TG5mM & Cfz7nM 0% 20% 40% 60% 80% 100% No Transduction GFP-EV GFP-eIF4E TG5mM & Cfz5nM TG5mM & Cfz6nM TG5mM & Cfz7nM Viability
SLIDE 15 Silence c-Myc through Simultaneous Targeting of the PI3Kd and Proteasome Pathways
eIF4F c-Myc
Translation
mTOR PI3Kd Proteasome pp-4EBP1 TGR-1202 Carfilzomib
- Optimize c-Myc-silencing therapy by targeting phosphorylation of
4EBP1
- Phase I/II clinical trial of TGR-1202 and carfilzomib in relapsed and
refractory lymphoma
SLIDE 16
Owen A. O’Connor, M.D., Ph.D. Mark Lipstein, B.S. Luigi Scotto, Ph.D. Michael Mangone, Ph.D. Xavier O. Jirau Serrano, B.S. Center for Lymphoid Malignancies Columbia University Medical Center Shirong Li, Ph.D. & Suzanne Lentzsch, M.D., Ph.D. Hematology & Oncology, CUMC Nicholas Tatonetti, Ph.D. Biomedical informatics, CUMC Charles Karan, Ph.D. Sulzberger Columbia Genome Center, CUMC
Thank you!
Funding from: Lymphoma Research Fund Irving Institute (CUMC) Pilot Award TG Therapeutics Amgen/Onyx
