imiquimod, an immune-response modifier, in the treatment of transitional cell carcinoma of the...
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IMIQUIMOD, AN IMMUNE-RESPONSE IMIQUIMOD, AN IMMUNE-RESPONSE MODIFIER, IN THE TREATMENT OF MODIFIER, IN THE TREATMENT OF
TRANSITIONAL CELL CARCINOMA OF TRANSITIONAL CELL CARCINOMA OF THE BLADDERTHE BLADDER
Douglas Scherr, M.D.Douglas Scherr, M.D.
Department of UrologyDepartment of Urology
New York Presbyterian Hospital-Weill Cornell New York Presbyterian Hospital-Weill Cornell Medical CenterMedical Center
IntroductionIntroduction
• Imiquimod is a potent synthetic Imiquimod is a potent synthetic immunomodulatorimmunomodulator
• Currently used as a first-line Currently used as a first-line treatment for genital condyloma (Aldara, 3M)treatment for genital condyloma (Aldara, 3M)
• Effective against several skin cancers Effective against several skin cancers including melanoma and basal cell including melanoma and basal cell carcinomacarcinoma
IntroductionIntroduction
• Induces both innate and acquired immune Induces both innate and acquired immune responsesresponses
• Toll-like receptor 7 (TLR-7) agonistToll-like receptor 7 (TLR-7) agonist
• Stimulates dendritic cells to mature and Stimulates dendritic cells to mature and secrete pro-inflammatory cytokinessecrete pro-inflammatory cytokines
• Mechanism strikingly similar to bacillus Mechanism strikingly similar to bacillus Calmette-Guerin (BCG)Calmette-Guerin (BCG)
IntroductionIntroduction
• Hypothesis: Hypothesis: – Imiquimod may have therapeutic potential Imiquimod may have therapeutic potential
against transitional cell carcinoma of the bladderagainst transitional cell carcinoma of the bladder
• Objectives:Objectives:– To determine if Imiquimod has direct effects on To determine if Imiquimod has direct effects on
bladder cancer cells bladder cancer cells in vitroin vitro– To determine if Imiquimod has anti-tumor effects To determine if Imiquimod has anti-tumor effects
in vivoin vivo in an intact immune system in an intact immune system
Cell ViabilityCell Viability(MTT Assay)(MTT Assay)
Cytokine ProductionCytokine Production (ELISA)(ELISA)
ApoptosisApoptosis(TUNEL Assay)(TUNEL Assay)
ImiquimodImiquimod
Bladder CancerBladder CancerCell LinesCell Lines
(J82, T24, TCC-SUP)(J82, T24, TCC-SUP)
24 hrs24 hrs
In vitroIn vitro Methods Methods
Figure 1. Cell Proliferation Assays
MTT Proliferation Assay after 24hrs
0
20
40
60
80
100
120
0 50 100 150 200 250
Imiquimod Concentration (mcg/ml)
Frac
tiona
l %
J82
T24
TCCSUP
Cell ViabilityCell Viability
ApoptosisApoptosis
Control (PBS) GroupControl (PBS) Group Imiquimod GroupImiquimod Group
ApoptosisApoptosis
Imiquimod GroupImiquimod GroupControl (PBS) GroupControl (PBS) Group
Apoptosis in vitro
0
4
8
12
16
20
24
% a
popt
otic
cel
ls PBS treated
Imiquimod treated
0.8%
17.1%
ApoptosisApoptosis
IL-6 Secretion
0
200
400
600
800
1000
1200
1400
1600
1800
2000
pg/m
l PBS treated
Imiquimod treated
TNF-alpha Secretion
0
200
400
600
800
1000
1200
1400
pg/m
l PBS treated
Imiquimod treated
930
425
1340
480
Cytokine ProductionCytokine Production
In vivoIn vivo Methods Methods
• Orthotopic, syngeneic, immune competent Orthotopic, syngeneic, immune competent mouse model mouse model (Bochner et al J. Urology, 2003)(Bochner et al J. Urology, 2003)
• Murine bladder tumor (MBT-2) cells instilled Murine bladder tumor (MBT-2) cells instilled intravesically into C3H miceintravesically into C3H mice
• Treatment regimen:Treatment regimen:– Day 0: Tumor cells instilledDay 0: Tumor cells instilled– Day 1:Day 1: Treatment with Imiquimod or PBS Treatment with Imiquimod or PBS– Day 8: Retreated Day 8: Retreated – Day 15: Mice sacrificed Day 15: Mice sacrificed
ImiquimodImiquimodControl (PBS)Control (PBS)
ConclusionsConclusions
• Imiquimod is a potent immune-response Imiquimod is a potent immune-response modifiermodifier
• Has direct biological effects on bladder Has direct biological effects on bladder cancer cell linescancer cell lines
• Preliminary data suggests anti-tumor Preliminary data suggests anti-tumor effects effects in vivoin vivo
• Imiquimod may be a potential Imiquimod may be a potential immunotherapy for bladder cancerimmunotherapy for bladder cancer
AcknowledgementsAcknowledgements
PI: PI: Douglas Scherr, MDDouglas Scherr, MD
Contributors:Contributors:
Hideki Kawamoto, MDHideki Kawamoto, MD
Xuecke You, MDXuecke You, MD
Mathew Albert, PhDMathew Albert, PhD
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