The document describes a laser-induced choroidal neovascularization model in pigs for studying retinal diseases. It discusses the Micron X imaging system for obtaining high-quality images of the retina and fluorescein angiograms in large animals like pigs. The model involves using laser shots to induce CNV lesions and then treating them with drugs like aflibercept to evaluate effectiveness. Quantitative analysis of the lesions is done using OCT, fluorescein angiography, and flat mounts, showing that aflibercept significantly reduces CNV development. The pig model is said to be more clinically relevant and translatable than rodent models for testing new therapies.

1. Studying Retinal Function in Large Animals:
Laser-Induced Choroidal Neovascularization in Pigs
Dr. David Culp and Dr. Christine van Hover present novel
technology designed to overcome the challenges of large
animal eye and eye-brain research.

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3. Christine van Hover, PhD
Chief Scientist,
Phoenix Research Labs
W. David Culp, PhD
Director of Research,
Powered Research, LLC
Studying Retinal Function in Large Animals:
Laser-Induced Choroidal Neovascularization in Pigs

4. Copyright 2017 Phoenix Research Labs and InsideScientific. All Rights Reserved.
Christine van Hover, PhD
Chief Scientist,
Phoenix Research Labs
Micron X: Optimized Image System
for Large Animal Eye Research

5. Micron X
Imaging
Large animal camera

6. And: Anterior segment, Ganzfeld ERG, Laser-induced CNV
image
examine
measure
Geography
Micron IV
Structure
OCT2
Function
ERG
Phoenix Research Labs

7. The Micron X is optimized for large animals
✓ Wide angle 100°
field-of-view
✓ Resolution of 8 um
✓ Bright field and
fluorescein angiography
Images courtesy of Powered Research

8. Why large animals?
• Critical step between
rodents and clinical trials
• Similar eye size to
humans
• Allergy testing

9. Four Step Process
1. Sedate and dilate the animal
2. Position and apply gel
3. Contact the Micron X lens onto
the eye. Adjust light level, focus,
and gain
4. Take images and video as the
Micron X scans the retina

10. Imaging Versatility in a Range of Species
Cat retina Pig retina Dog retina

11. Albino rabbit retina
Imaging
Versatility
Crisp images of
both light albino
retinas and darkly
pigmented retinas
Pigmented macaque primate
retina

12. Stunning Fluorescein Angiography Ability
Bright field macaque primate retina Fluorescein angiography
Images courtesy of Ora Clinical

13. Fluorescein Angiography Video Ability
Video of fluorescein infiltration in a macaque primate
Images courtesy of Ora Clinical
Artery
Vein

14. Fluorescein Angiography Video Ability
Slowed down video of fluorescein infiltration in a macaque primate
Images courtesy of Ora Clinical

15. Time lapse in a macaque primate:
Clearly see the arteries, capillaries, and then veins filling with fluorescein
Fluorescein Angiography Time Lapse
Images courtesy of Ora Clinical

16. Bright field pig retina
with laser shots
Fluorescein angiography of pig
retina showing CNV induction
Images courtesy
of Powered Research
CNV Induction in Pigs

17. Eylea® treatment
significantly
decreases CNV
development
Pig retina with
laser shots and no
treatment shows
CNV development
Saline
Eylea®
Treating CNV Induction in Pigs with Eylea®
Images courtesy of Powered Research

18. Micron X: A unique retinal imaging
system optimized for eye research using
large animals. Micron X delivers bright
field images of pinpoint resolution and
high contrast. Angiograms using
fluorescein can also be obtained utilizing
the Phoenix Research Labs Micron X.
Learn More >>

19. Copyright 2017 D. Culp, Phoenix Research Labs and InsideScientific. All Rights Reserved.
W. David Culp, PhD
Director of Research,
Powered Research, LLC
Evaluation of Aflibercept in a Laser-Induced
Model of Choroidal Neovascularization in Swine

20. • Preclinical contract research
organization based in Research
Triangle Park, North Carolina.
• Provide high quality and cost
effective, non-GLP studies.
• More than 80% of our business
is in the ophthalmic space.
Learn More >>

21. Preclinical Models of Choroidal
Neovascularization (CNV) are Needed to
Assess New Therapies…
Limitations of current models:
• Pharmacokinetically (e.g. rodent models)
• Pathophysiologically (e.g. VEGF induced models)
• Economically (e.g. non-human primates)

22. Swine Model of Laser-Induced
Choroidal Neovascularization (CNV)
• Weanling Swine, Sus scrofa domesticus
• Female and male
• 6-12 week, 12 – 20 kilograms
• Approximately 6 lesions per swine eye,
equidistant from the optic nerve were
created using an 810 nm diode laser
and an indirect ophthalmoscope

23. Prior to laser Post laser 2 weeks 3 weeks 4 weeks
BSS
Aflibercept
Swine Model of Laser-Induced CNV
Clinical images

25. Prior to laser Post laser 2 weeks 4 weeks
Swine Model of Laser-Induced CNV
OCT images

26. 0
2
3
4
0
50
100
150
200
250
Adjacent-lesion retinal thickness
Weeks after injection
mean+/-SDuM
BSS
Aflibercept
0
2
3
4
0
50
100
150
200
250
Central-lesion retinal thickness
Weeks after injection
mean+/-SDuM
BSS
Aflibe
150
200
250
Central-lesion retinal thickness
-SDuM
BSS
Aflibercept
Swine Model of Laser-Induced CNV
OCT measurements

27. Day 7 BSS
Day 7 Eylea®
Micron X, Phoenix Research Labs

28. Day 14 BSS
Day 14 Eylea®
Micron X, Phoenix Research Labs

29. CTLF:
Corrected Total Lesion
Fluorescence
Integrated Density:
Area of lesion X Mean
fluorescence of
background readings

30. Swine Model of Laser-Induced CNV
FA measurements

31. BSS
Aflibercept
W
eek
2
W
eek
4
0
100000
200000
300000
400000
Weeks after injection
µm
2
IsolectinIB4Signal
BSS
Aflibercept
Swine Model of Laser-Induced CNV
Flat mount measurements
Blue : DAPI (nucleus); Green : AF-488 Phalloidin (F-actin); Red : AF-594 Isolectin IB4 (blood vessels and microglia)

32. Aflibercept
200 X magnification;
hematoxylin and
eosin staining
Swine Model
of Laser-Induced
CNV:
Histopathology
Day 28
BSS

33. ✓ The larger size of the swine eye is more relevant to human eyes.
✓ The model provides quantitative assessment by evaluating
neovascularization by fluorescein angiography and flat mount analysis.
✓ The model is easy to reproduce and is economical.
✓ Characterization of this sensitive and translatable model allows
effective evaluation and proof of concept of new treatments or delivery.
Conclusions

34. Acknowledgments
• Dr. Brian Gilger, Professor of Ophthalmology,
North Carolina State University
• Kristie Powell, Powered Research
• Dr. Grazia Spiga, Powered Research
• Justin Prater, Powered Research
info@poweredresearch.com

35. Christine van Hover, PhD
Chief Scientist,
Phoenix Research Labs
Send Email >>
W. David Culp, PhD
Director of Research,
Powered Research, LLC
Send Email >>
Thank You:
For additional information on the products and applications presented
during this webinar please visit www.phoenixreslabs.com