Organoid Imaging Strategies for Accelerating Translational …

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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