Post on 30-Mar-2020
Targeting The Stress-Induced Cytoprotective Chaperone, Clusterin, to
Overcome Treatment Resistance in Advanced Prostate Cancer
Martin Gleave MD, FRCSC, FACS
Professor, Department of Urologic SciencesUniversity of British ColumbiaB.C. Leadership Chair in Prostate ResearchDirector, The Prostate Centre at VGH
Disclosure of Conflicts of InterestPatent - OGX-011Founder - OncoGenex TechnologiesConsultant - CSO, OncoGenex Technologies
Castration Resistance and Prostate Cancer
HormoneTherapy
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? ? Molecular mechanisms
Seru
m P
SA
Tum
or Volum
e
1. Androgen receptor (AR) related• Overamplification (hypersensitive)• Mutations (promiscuous)• cross-talk - TK, PKA, AKT, STAT3 (phosphorylation, co-regulators)
2. Adaptation• Up-regulation of survival genes (Bcl-2, clusterin, Hsp27, YB-1)• Increased alternative GF pathways (her2/neu; IGF-1/IGFBP2&5; IL-6/STAT3)
High-Throughput Bioprofiling of Hormone-Treated Prostate Cancers to Identify Stress-Induced Targets
Gene expression profiling for Target I.D.
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0 5 10 15 20 25 30 35 40 45
Days
PSA
(ng/
mL)
Cx
AD/ +DHT
AD/ -DHT
AI/ -DHT
Tumour Models
Human CaP
Untreated 1 - 3 months NHT
6 months NHT 8 months NHT HRPC
Post-Hormone-Treated TMA
Tissue Bank:-Xenografts -Post-NHT-treated human CaP-pre-surgery NHT + Taxotere RP specimens-rapid autopsy HRPC specimens
TMA profiling for Target Validation
Hypothesis:Treatment-induced changes in gene
expression after hormone therapy render cells resistant to castration- and chemo-therapy.
++ PSA- Bcl-2
- EGFR- clusterin
- IGFBP 2&5- TGFβ
++IGFBP 3 & 4-YB-1
++survivin-Hsp27
+ PKC-α
- PSA++ Bcl-2++Bclx-L- EGFR
+++ clusterin++++IGFBP 5IGFBP 3 & 4
+c-myc+YB-1
-survivin+Hsp27
+ PKC-α
++ PSA++ Bcl-2++Bclx-L+ EGFR
+++ clusterin++ IGFBP 2++ IGFBP 5
++YB-1++ survivin+++Hsp27
Changes in Gene Expression After Castration Changes in Gene Expression After Castration and During AI Progression and During AI Progression
Androgen-dependent
Tumour regression
Androgen-independent
Clonal selectionAdaptive responses
Hormone withdrawal Progression
AI
ADAI AI
Untreated CaP 3 months NHT
6 month NHT+Tax HRPC
Untreated 1 - 3 months NHT
6 months NHT 8 months NHT HRPC
Post-Hormone-Treated TMA
TMA profiling for Target Validation
Clusterin
Vinculin
Docetaxel Rx (nM) 0 10 25 50 100 500
Docetaxel Rx in PC-3
Transcriptionally activated by- Hormone withdrawal (a.k.a. TRPM-2)- Chemotherapies- Radiotherapy - Targeted therapies (Herceptin, Velcade, OGX-225, etc)
Therapeutic Stress Increases Clusterin Levels in CaP Tumor Models and Human Tissues
Androgen Ablation inShionogi Tumors
GAPDH
Clusterin
Cancer Research 60; 170, 2000
0 3 7 21Days Post-Castration
SCLU-2: Stress-induced Cytoprotective Chaperone
1. Transcriptionally activated by HSF-1, repressed by p53
a. Increased by diverse array of therapeutic triggers (HT, CT, RT, velcade, herceptin)
b. Increased by cell survival factors like androgen, IGF-1
2. Intrinsically disordered and flexible protein
3. Potent inhibitors of aggregation of client proteins under stress conditions
4. Often associated with neurodegenerative diseases and cancer
5. Interact with and inhibit activated Bax; enhances NF-kB transcriptional activity
QuickTime™ and aTIFF (LZW) decompressor
are needed to see this picture.
Clusterin: Cytoprotective MechanismsClusterin: Cytoprotective Mechanisms
QuickTime™ and aTIFF (LZW) decompressor
are needed to see this picture.
QuickTime™ and aTIFF (LZW) decompressor
are needed to see this picture.QuickTime™ and a
TIFF (LZW) decompressorare needed to see this picture.
sCLU-2 is a COMMD1 and ubiquitin binding partner in cancer cells
Zoubeidi et al, 2007
Input IgG LNCLU2 LNmock
IP: CLU
COMMD1
Input LNmock LNCLU2
IP: COMMD1
CLU
Clu COMMD1 Merge
1. sClu-2 and COMMD1 co-localize in cytoplasm with a juxtanuclear aggregation
0 50 100 250 500 1000 (ng)
Clusterin
Clu transient transfection
Clusterin
COMMD1
Vinculin
COMMD1
sCLU-2
Vinculin
siRNA (25 nM) Scr Clu
Clu Knockdown increasesLevels of COMMD1 & total Iκ-Bα
I-κBα
2. sCLU-2 Levels Negatively Correlate with COMMD1 & Ik-B Levels
sCLU-2 Enhances TNF-a induced NF-kB Nuclear Translocation and Transcriptional Activity
Zoubeidi et al, 2007
10min (TNF-α) 0min (TNF-α)
LNCLU2
LNmock
p65-NF-κB
0100200300400500600700800900
1000
no treatment TNF-a
NF-
κB-lu
c ac
tivity
(fo
ld in
duct
ion/
cont
rol) LN-V
LN-CluLNCLU2
LNmock
1. sClu-2 Overexpression ↑↑ NF-kB Activity
0
0.2
0.4
0.6
0.8
1
1.2
siRNA Scr siRNA Clu
NF-
kB-lu
c ac
tivity
(fo
ld in
duct
ion/
cont
rol)
PC3
siRNA Scr
siRNA Clu
PC3
p65-NF-κB
10min (TNF-α) 0min (TNF-α)
2. sClu-2 Knockdown ↓↓ NF-kB Activity
Clusterin Expression Levels Positively Correlate with NF-κB - regulated Genes
B Western Analysis
P T T-scr T-siRNA
66kDacIAP2
MCP-1 12kDa
7.5kDMCP-2
Perc
ent c
hang
e In
pro
tein
exp
ress
ion
20
100
80
60
40
cIAP2
MCP-1MCP-2
A
NGAL 845bp
sPLA2-IIa
2255bpSema3C
P T T-scr T-siRNA
Perc
ent c
hang
e In
mR
NA
exp
ress
ion
20
100
80
60
40
Northern Analysis
MIP3α 3000bp
Sema 3c
NGALsPLA2
MIP3α
854bp
* *
Ettinger et al, 2006
All Genes filtered1.5 Fold up in OE (red) 1.5 fold down in KD (blue)
OE KDAll Genes
NGAL
Clusterin
OE KD OE KD
Filtered Genes
sClu-2 Enhances COMMD1 and I-κBαDegradation by the Proteasome
Zoubeidi et al, 2007
I-κBα
p65
p50
I-κBα
p50
ubububP P
p65
nucleus
cytoplasm
20S
19S
19S
ububub
CLU
NF-ΚB
Stressp50
p65
NF-ΚB
COMMD1
P P
I-κBα
Functional Significance of sCLUFunctional Significance of sCLU--2 Over2 Over--expression expression in Prostate Cancerin Prostate Cancer
Cancer Research 60;2547, 2000; Clin Can Res 8:3276-84, 2002Cancer Research 60;170, 2000;
LNCaP over-expressing CLU
LNCaP-CluControls
0102030405060708090
100
0.0001 0.005 0.001 0.005 0.01 0.05 0.1 0.5 1
LNCaP/T1
LNCaP/C
% v
iabl
e ce
lls
Taxol concentration (nM)
IC50
LNCaPsCLU-2
• sCLU overexpression is antiapoptotic: confers broad spectrum treatment resistance including hormone, radiation, and chemo-therapy
CLU ASO (OGX-011) Suppress Clu Levels in PC-3 CellsMM Oligo OGX-011
Cel
l pro
lifer
atio
n co
mpa
red
to c
ontr
ol
Docetaxel concentration (nM)
0
20
40
60
80
100
120
0 15 31 63 125 250 500 1000
PC3-R SiRNA Scr
PC3-R SiRNA Clu
OGX-011 Chemosensitizes PC-3 Cells to Docetaxel
Inhibition of Clusterin Expression Enhances Activity of Chemotherapy in Prostate Cancer Cells
OGX-011 Enhances Taxol Activity in PC3 Tumours in vivo
PC3 Tumor Volume (cm3)
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1
1.2
1.4
1.6
1.8
2
0 1 2 3 4 5 6 7 8
Weeks
ASO Clusterin+ Taxol
MM Clusterin + Taxol
Clin Cancer Res 6:1655, 2000
OGX-011
OGX-225 100 nM+
Control ODN
OGX-225 100 nM+
Olig
ofec
tam
ine
OG
X-2
25 1
00 n
M
10 n
M
100
nM
500
nM
10 n
M
100
nM
500
nM
PC-3
Vinculin
CLU
60 kD
40 kD
Clusterin Clusterin is expressed in 65%is expressed in 65% of of Primary BreastPrimary Breast Cancers Cancers
So et al, Mol Cancer Ther, 2005
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0 0.7 1 1.4 2 2.5 2.7 3Log Paclitaxel concentration (nM)
Cel
l pro
lifer
atio
n co
mpa
red
to c
ontro
l
s iRNA Clusterin
siRNA ScramOGX-011
ASO -Scram
Vinculin
+ O
GX
-011
+ Pa
clita
xel
+ Pa
clita
xel
+
OG
X-0
11
60 kDa
40 kDa
55 kDa
CLU ASO (OGXCLU ASO (OGX--011) Suppresses 011) Suppresses sCLU sCLU Levels andLevels andChemosensitizes Chemosensitizes MCFMCF--77 Xenografts Xenografts to to Paclitaxel Paclitaxel in vivoin vivo
In vitro In vitro
Mea
n T
umor
Siz
e (m
m3 )
0
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1000
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3000
1 2 3 4 5S
()
OGX-011SCR
Weeks
* *
*
*
Paclitaxel
Paclitaxel
*
**
60 kDa
55 kDa
s-CLU
n-CLU
Vinculin
ASO SCR OGX-011In vivo In vivo
From Bench to Bedside: Translational Research in Action
sCLU sCLU as a Therapeutic Target:as a Therapeutic Target:Preclinical Studies For Proof of PrinciplePreclinical Studies For Proof of Principle
Of Mice and MenClusterin:•Stress-induced survival response•confers resistance•knockdown enhances chemo & HT in many tumor models
Antisense ClusterinAntisense Clusterin: OGX: OGX--011 Product Description011 Product Description
MOEMOE P-SLicensed from UBC for development by OncoGenex in collaboration with Isis
2nd generation antisense molecule4-13-4 21-mer MOE gapmer oligonucleotide
Advantages of 2'MOE analoguesIncreased potency and resistance to degradationFacilitates more convenient dosing regimen
once-weekly infusionJ Pharmacol Exp Ther. 298(3):934-40, 2001 O C
O
O
G
O
O
T
OO
O
O
A
OO
PO
OO
PO
A
OO
P
OO
P
OO
P
-S
-S
-S
-S
-S
OCH2CH2OCH3
OCH2CH2OCH3
H
H
OCH2CH2OCH3
CLU Isoform 1sCLU-1
CLU Isoform 2sCLU-2
Nuclear splice variant 1nCLU-v1ΔII
siRNA CLU ex3 siRNA CLU ex1
siRNA CLU ex3 OGX-011 (CLU ex2 ASO)
siRNA CLU ex3 OGX-011 (CLU ex2 ASO)
siRNA CLU ex1
I II III IV
IISpliced out
5’-UTR, v1ATG ATGATG
5’-UTR, v2
V VI VII VIII
II III IVATGATG
I III IV5’-UTR, v1
ATG ATG
siRNA CLU ex2
Nuclear localization signalER-targeting signal
Nuclear localization signalER-targeting signal
Nuclear localization signal
IX3’-UTR
V VI VII VIII IX3’-UTR
V VI VII VIII IX3’-UTR
siRNA CLU ex2
NCIC IND.153: Phase I PreNCIC IND.153: Phase I Pre--Surgery Surgery pkpk/pd /pd Trial of OGXTrial of OGX--011 011 -- Tissue Tissue PkPk datadata
Chi et al, JNCI. 97:1287-96, 2005
OGX-011 Dose (mg)6404803201600
Con
cent
ratio
n of
OG
X-0
11 in
Pro
stat
e Ti
ssue
(µ
g of
OG
X-0
11 p
er g
ram
of p
rost
ate
tissu
e) 7
6
5
4
3
2
1
08040
Ptrend <.001
•25 men with localized CaP treated with 5 weeks of NHT + escalating doses of OGX-011
Chi et al, JNCI. 97:1287-96, 2005
IND.153: Target Regulation Data:Dose-dependent suppression of clusterin in Regional Lymph Nodes
Untreated
Controls
6 weeks NHT+ OGX-011(dose escalation) 160 mg
320 mg
480 mg
640 mg
80 mg
0
50
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150
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250
300
350
400
450
No treatment >2 m NHT OGX011-40mg
OGX011-80mg
OGX011-160mg
OGX011-320mg
OGX011-480mg
OGX011-640mg
Treatment Grou
Clinical Proof-of-Concept:Dose-dependent Decreases in Clusterin Levels in RP Specimens using LCM and Real-Time PCR
Chi et al, J Nat.Canc.Inst. 97:1287-96, 2005
Clinical Trial Development with Clinical Trial Development with ClusterinClusterin ASO (OGXASO (OGX--011)011)
Established 640 mg once weeklyas phase II dose (OBD) based on
tissue pk and pd criteria
Completed:2 regimens found well tolerated -• OGX-011 + weekly Taxotere• OGX-011 + q 3 weekly Taxotere
• 30-60% decrease in serum CLU levels
• 10 pts Phase I• 70 pts Phase II• ASCO May 2007
NCIC IND.165:First Line mHRPC
Phase II RCT -Taxotere +/- OGX-011
NCIC IND.164: Phase II Metastatic Breast- Taxotere +
OGX-011
• 82/80 pts enrolled• ASCO May 2007
• 16 pts enrolled
Phase II stage III/IV NSCLC
Gem/cis + OGX-011
CUOG P-06a:2nd line mHRPC
Phase RCT Taxotere or mitoxantrone + OGX-
011
• 42/40 pts enrolled
NHTPre - surgery
Solid Tumors +docetaxel n = 35
Results as of May 24, 2007
Median Follow-up 12.7 months
Estimated Median Survival 14.1 months
Number of Patients Surviving ≥ 1 year 25/46 = 54% *
Number of Patients Surviving ≥ 18 months 8/22 = 36%
Number of Deaths 37/81 (46%)Median Progression-Free Survival (range) 4.6 months (0.06-15.6+)
Historical Controls* Phase 1 and 2 (n=81)
Median Survival 8.0 – 10.8 months 14.1 Months (estimated)
•Data from five randomized clinical trials using gemcitabine plus platinum-based chemoin 1st line NSCLC (1260 patients)
Phase 2 Study in 1st Line NSCLC:Treatment Schema
•81 pts with stage IIIB/IV NSCLC treated with gem/cis plus OGX-011
Laskins et al, ASCO, 2007
Randomized Open label, multicentre trial comparing docetaxel +/- OGX-011 in men with mHRPC (PI - K. Chi).
PSA Response Rates
BEST RESPONSE CRITERIA
Arm A(OGX-011 + Docetaxel)
N=40
Arm B(Docetaxel)
N=41
% Change in favor of OGX-
011
≥ 50% PSA Decline at 12 weeks 45% 34%
PSA Response (50% decline -confirmed)
50% 51% NA
22%
10%
32%
2%
32%
≥ 80% PSA Decline 38% 73%
PSA Progression (PSAWG Criteria)
0% 100%
PSA Non-Progression/Non-Response
45% 41%
Inevaluable 3% NA
NCIC IND.165: Taxotere +/- OGX-011 in First-Line mHRPC
Chi et al, ASCO, 2007
RECISTCRITERIA
Arm A(OGX-011 +
Docetaxel) n=26
Arm B(Docetaxel)
n=23
% Change in favor
of OGX-011Disease Control(CR+PR+SD)
92% 74% 24%
Inevaluable 4% 4% N/A
Complete Response 0% 0% N/APartial Response 19% 22% -14%Stable Disease 73%
9.7 months52%
7.6 months40% 28%
Progressive Disease 4% 22% 82%
Median PFS 7.3 months 5.9 months 24%
NCIC IND.165: Taxotere +/- OGX-011 in First-Line mHRPC
Chi et al, ASCO, 2007
NCIC IND.165: Taxotere +/- OGX-011 in First-Line mHRPC
Median for Arm A (OGX-011 + Docetaxel): 7.26 months (95%CI 5.22-9.33)Median for Arm B (Docetaxel): 5.85 months (95% CI 3.61-10.74)
At Risk0
20
40
60
80
100
Arm A: OGX-011 + DocetaxelArm B: Docetaxel
0.04041
5.02118
10.024
15.000Arm B
Arm A
Time (months)
Mature Data
Progression Free Survival
Chi et al, ASCO, 2007
Indicators of Anti-cancer Activity
Consistent trend in favor of OGX-011/docetaxel arm:More patients with a 50% decline in PSA within the first 12 weeks More pts (38% vs 22%) with >80% decline in PSA; less pts (0 vs 10%) with primary PSA progression as best response Longer time on treatment and a greater median # of treatment cycles.Higher frequency and longer duration of stable measurable disease.Lower frequency of progressive disease as “best response”. Longer time to progression
NCIC IND.165: Taxotere +/- OGX-011 in First-Line mHRPC
Cel
l pro
lifer
atio
n
0
20
40
60
80
100
120
0 15 31 63 125 250 500 1000
PC3-R MM
PC3-R ASO
PC3 MM
PC3 ASO
OGX-011 in docetaxel-refractory HRPC:CLU knockdown chemosensitizes taxane-resistant PC3-dR cells to docetaxel
docetaxel +/- OGX-011
0
20
40
60
80
100
120
0 15 31 63 125 250 500 1000
PC3-R MM
PC3-R ASO
PC3 MM
PC3 ASO
CLUVinculin
PC-3
PC-3R
MM OGX01150 200 500 50 200 500 O
onl
y
CLUVinculin
In vitro Docetaxel concentration (nM)
-0.5
0
0.5
1
1.5
2
2.5
3
3.5
4
1 2 3 4 5 6 7 8 9
weeks
PC-3R, MM
PC-3R, OGX-011
PC-3, MM
PC-3, OGX-011
* *
In vivo
* - chemotherapy treatment
Tum
or V
olum
e
Phase II Feasibility Trial of OGX-011 in 2nd Line Therapy in HRPC:
RANDOMIZE
Arm ADocetaxel (75 mg/M2 IV) q 21 days and OGX-011 (640 mg IV) weekly plus Prednisone (5 mg po bid) daily
Arm BMitoxantrone (12 mg/M2 IV) day 1 and OGX-011 (640 mg IV) weekly plus Prednisone (5 mg po bid) daily.
Continue treatment until disease progression or unacceptable toxicity. If removed from treatment for any reason, follow until death
PR
D OI G
S RE EA SS SE I
ON
S FU OR LV LI OV WA L U
P
n = 41
HRPC Patients Who Had Disease Progression
During or Within 6
Months of 1st
Line Docetaxel Treatment
n = 19
n = 22
Study treatment ongoing in 11 (26%) of patients
OGX-011 in 2nd Line Therapy in HRPC:Chemosensitizes taxane-resistant patients to docetaxel
0
500
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1500
2000
2500
3000
3500
4000
4500
PSA
Valu
e
Start 1st line therapy
End 1st line therapy6 cycles
Start 2nd line therapy
Off Study Treatment for disease progression
5 Cycles
Time (2 Weeks per mark)
Patient: 009-002
0
20
40
60
80
100
120
140
160
180
200
Time (2 Weeks per mark)
PSA
Valu
e
Off Study Treatment for
disease progression
6 cycles
Start 1st line therapy
End 1st line therapy5 cycles
Bone Scan, PSA Progression
Start 2nd line therapy
Patient: 025-002
Summary: Clusterin as a Therapeutic Target in HRPC
sCLU is a stress-activated cytoprotective chaperone that is highly expressed in HRPC
Over-expression of sCLU-2 confers broad spectrum treatment resistanceInhibits protein aggregation, facilitates proteasome degration of ubiquitinated proteinsInteracts and and inhibits activated Bax, preventing cytochrome C releaseIncreases NF-kB transcriptional activity
CLU knockdown using OGX-011Enhances treatment-induced apoptosis in vitro and in vivoPre-clinical proof-of-principle in prostate, breast, lung, urothelial, melanoma, renal cell
OGX-011, a 2nd generation ASO potently suppresses target CLU levels >90% in human CaP tissues
Anti-cancer activity observed in multi-centre Phase II trials in breast, HRPC, lungPhase III registration trial in second-line HRPC set to begin in 2008
++ PSA- Bcl-2
- EGFR- clusterin
- IGFBP 2&5- TGFβ
++IGFBP 3 & 4-YB-1
++survivin-Hsp27
+ PKC-α
- PSA++ Bcl-2++Bclx-L- EGFR
+++ clusterin++++IGFBP 5IGFBP 3 & 4
+c-myc+YB-1
-survivin+Hsp27
+ PKC-α
++ PSA++ Bcl-2++Bclx-L+ EGFR
+++ clusterin++ IGFBP 2++ IGFBP 5
++YB-1++ survivin+++Hsp27
Changes in Gene Expression After Castration Changes in Gene Expression After Castration and During AI Progression and During AI Progression
Androgen-dependent
Tumour regression
Androgen-independent
Clonal selectionAdaptive responses
Hormone withdrawal Progression
AI
ADAI AI
Post-docs
Hide MiyakeMoto Muramaki
Isis PharmaceuticalsBrett Monia
PI’sKim ChiLarry GoldenbergColleen Nelson Paul RennieAlan So
Sue EttingerAmina Zoubeidi Eliana BeraldiRichard SoweryPathology
Antonio HurtadoLadan Fazli David HuntsmanTorsten NeilsenTed Jones
Grant FundingNCICNIH SPOREDoDPCF
CollaborationsUW - B. VessellaP. Lange, P Nelson, Tia HiganoU Montreal - F. Saad
OncoGenex Tech. Inc.Scott Cormack
NCIC IND GroupElizabeth EisenhauerLesley Seymour
OGX-011: Safety Profile in >270 Patients
Well tolerated in all Phase 1 and Phase 2 studies to date Safety profile of OGX-011 in combination with docetaxel vsdocetaxel alone
Increase in Grade 1 or 2 AE’s events (fever, rigors/chills and sweating during the loading-dose week and sensory neuropathy during therapy)lymphopenia was more prevalent in the OGX-011 + docetaxel arm (no clinical sequelae)No increase in SAEs in the OGX-011 + docetaxel arm
OGX-011 in combination with gemcitabine/platinum-based or mitoxantrone regimens
Safety profile similar to that expected for regimen (no increase in expected rate of Grade 3 or higher AEs)
OGX-011 Mechanism of Action
OGX-011
↑ nCLU
⇑ Cytochrome C ↓ NF-κB ⇑ ER Stress ↓ ProteasomeActivity
⇑ Bax ⇑ COMMD1⇑ Protein Aggregation ↓ UBP
↓ sCLU ↑ Apoptosis
↑ Apoptosis
Alternative splicing of Clusterin mRNA
Clusterin: Isoforms and Splice Variants
CLU Isoform 1sCLU-1
CLU Isoform 2sCLU-2
Nuclear splice variant 2nCLU-v2ΔII
Nuclear splice variant 1nCLU-v1ΔII
siRNA CLU ex3
siRNA CLU ex3 siRNA CLU ex1
siRNA CLU ex3 OGX-011 (CLU ex2 ASO)
siRNA CLU ex3 OGX-011 (CLU ex2 ASO)
siRNA CLU ex1
I II III IV
IISpliced out
5’-UTR, v1ATG ATGATG
5’-UTR, v2
V VI VII VIII
II III IVATGATG
I III IV5’-UTR, v1
ATG ATG
5’-UTR, v2III IVATG
siRNA CLU ex2
Nuclear localization signalER-targeting signal
Nuclear localization signalER-targeting signal
Nuclear localization signal
Nuclear localization signal
IISpliced out
IX3’-UTR
V VI VII VIII IX3’-UTR
V VI VII VIII IX3’-UTR
V VI VII VIII IX 3’-UTR
siRNA CLU ex2
Fold
Cha
nge
CLU Isoform 1
EtOH R1881
Fold
Cha
nge
0
0.5
1
1.5
2
2.5
CLU Isoform 2
EtOH R1881
clusterin
tubulin
Vehi
cle0.
01 n
M
0.1
nM
1 nM
10 n
M
60 kDa
40 kDa
1 nM R1881
1 μM Casodex
48 ho
urs
60 ho
urs
72 ho
urs
ClusterinmRNA
- + + - + + - + +
- - + - - + - - +
TotalRNA
CLU is an androgen-regulated gene
Cochrane et al, JBC, 2006