Organoid Imaging Strategies for Accelerating Translational Research Charles Deckard III , Joseph Daniele, Dmitriy Loza, Vandhana Ramamoorthy, Kuo-Shun Hsu, Mikhila Mahendra, Benjamin Bivona, Ningping Feng, Henry Lyu, Jing Han, Joe Marszalek T ranslational R esearch to A dvan C e T herapeutics and I nnovation in ON cology (TRACTION)
Model Development:
Mission : •
Our mission in the Technology & Innovation group at TRACTION is to develop tools that support translational research & drug development.
TRACTION maintains >300 PDX models across various cancers.
•
Research Objectives : • Implement imaging techniques to evaluate drug response in organoids (PDXO/PDO). • Build imaging workflows to enable biomarker discovery and evaluation. • Develop better understanding of the relationship between organoid and invivo response.
Organoids:
PDX
Organoids
Patient
PDX
Organoid
Organoid Imaging Strategies for Accelerating Translational Research Charles Deckard III , Joseph Daniele, Dmitriy Loza, Vandhana Ramamoorthy, Kuo-Shun Hsu, Mikhila Mahendra, Benjamin Bivona, Ningping Feng, Henry Lyu, Jing Han, Joe Marszalek T ranslational R esearch to A dvan C e T herapeutics and I nnovation in ON cology (TRACTION)
Cancer Model PDAC
Live Dead Imaging for Evaluating Drug Efficacy
PDX
Organoids
Strategy : Leverage Live/Dead Selective Dyes to quantify organoid viability in real-time.
in vivo Growth Curve
Organoid Growth Curve
4
Cell Titer Glo
100
z
4
DRAQ7 (Dead cells)
3
y
80
3
2
60
x
DEAD
2
40
1
LIVE
20
1
0 3 6 9 12151821 0
Organoid
0
0 1 2 3 4 5 6 7
Days
Days
Day 0
Day 7
(+)Apoptosis
Vehicle MAPKi
Chemo
Vehicle MAPKi
(+)Apoptosis
DRAQ7
Validated Imaging Toolbox : Hoechst Stains All Calcein Stains live
mCherry Stains live
100 µm
- co-culture/cell tracking - Drug combination testing - Detecting resistant clones
Organoid Imaging Strategies for Accelerating Translational Research Charles Deckard III , Joseph Daniele, Dmitriy Loza, Vandhana Ramamoorthy, Kuo-Shun Hsu, Mikhila Mahendra, Benjamin Bivona, Ningping Feng, Henry Lyu, Jing Han, Joe Marszalek T ranslational R esearch to A dvan C e T herapeutics and I nnovation in ON cology (TRACTION)
Cancer Model PDAC
Biomarker Imaging in Organoid and PDX Tissue: mIF & Histology
PDX
Organoids
Organoids
PDX
3D Imaging
2D Imaging
DAPI HLA-A CC3
DAPI HLA-A CC8
DAPI HLA-A CC8
200 µm
100 µm
100 µm
50 µm
Current work : Applying multi-faceted approach to more models to better understand how we can apply organoid data to in vivo studies and the clinic.
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