Optical Bio-im aging w ith Polym er Nanoparticles I ck Chan Kw on, - - PDF document

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Optical Bio-im aging w ith Polym er Nanoparticles

I ck Chan Kw on, Ph.D Biom edical Research Center Korea I nstitute of Sci. & Tech.

2 0 0 8 . 4 . 1 7 . US-Korea Nano Forum

OA Normal

+ Probe

• I nhibitor

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+ Probe + Probe + Probe

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• I nhibitor

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+ Probe + Probe + Probe

Wednesday, Jan. 16, 2008

Judah Folkman, Cancer Pioneer

By Alice Park

• M. Judah Folkman, biomedical pioneer, dies at 74

Surgeon, research scientist, teacher and mentor, created field of angiogenesis

January 16, 2008

Judah Folkman, Researcher, Dies at 74

Children's m ourns the death of

• Dr. Judah Folkm an

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“On January 14, Dr. Judah Folkman, founder of the field of angiogenesis, died unexpectedly in Denver, Colo., while en route to Vancouver for one of the thousands of lectures that he gave to scientists around the world. A visionary and scientific pioneer, Dr. Folkman was founder and director of the Vascular Biology Program at Children's Hospital Boston, and a professor of Pediatric Surgery and Cell Biology at Harvard Medical School.” from Bess Andrews, Childrens Hospital “When Dr. Folkman first proposed, in the 1970s, that a cancer could be kept in check by cutting off its blood supply, he faced skepticism from a scientific com m unity that simply wasn't ready for his ideas. But he persevered, even when there were setbacks, and today, more than 1 ,0 0 0 laboratories w orldw ide are engaged in the study of angiogenesis, the field he founded. As a result of Dr. Folkman's vision and resilience, more than 1 0 new cancer drugs are currently on the m arket, and more than 1 .2 m illion patients w orldw ide are now receiving anti-angiogenic therapy.” from Robert Cook, Harvard Science Correspondent

W hat is Tum oral Angiogenesis

Carm eliet et al. Nature 2 0 0 0 ;4 0 7 :2 4 9 -2 5 7

Pathological angiogenesis is a hallm ark of cancer. W ithout blood vessels, tum ors can not grow beyond a critical size ( ~ 5 m m in diam eter) due to the lack of oxygen and nutrients for their survival.

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W hen Tum oral Angiogenesis Starts

http:/ / w w w .angiow orld.com

Beyond the critical volum e of 2 cubic m illim eters, oxygen and nutrients have difficulty diffusing to the cells in the center of the tumor, causing a state of cellular hypoxia that marks the onset

• f tum oral angiogenesis.

VEGF and bFGF are first synthesized inside tumor cells and then secreted into the surrounding tissue. When they encounter endothelial cells, they bind to specific proteins, called receptors, sitting on the outer surface of the cells. The binding of either VEGF or bFGF to its appropriate receptor activates a series of relay proteins that transmits a signal into the nucleus of the endothelial cells. The nuclear signal ultimately prompts a group of genes to m ake products needed for new endothelial cell grow th.

The Angiogenesis Signaling Cascade

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The activation of endothelial cells by VEGF or bFGF sets in motion a series of steps toward the creation of new blood vessels. First, the activated endothelial cells produce matrix m etalloproteinases (MMPs). The MMPs break down the extracellular matrix—support material that fills the spaces between cells and is made of proteins and polysaccharides. Breakdown of this matrix permits the migration of endothelial cells. As they migrate into the surrounding tissues, activated endothelial cells begin to divide.

Endothelial Cell Activation

• A cell activated by a lack of
• xygen releases angiogenic

molecules that attract inflam m atory and endothelial cells and promote their proliferation.

• During their migration,

inflammatory cells also secrete molecules that intensify the angiogenic stimuli.

The Angiogenic Sequence

• The endothelial cells that form the blood vessels respond to the

angiogenic call by differentiating and by secreting matrix m etalloproteases (MMP), which digest the blood-vessel walls to enable them to escape and migrate toward the site of the angiogenic stimuli.

http:/ / w w w .angiow orld.com

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Hobbs SK et. al. Proc Natl Acad Sci USA 1 9 9 8 ;9 5 :4 6 0 7 - 4 6 1 2 .

Perm eablility of Angiogenic Vessel

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EPR effect

( Enhanced Perm eability And Retention)

Accumulation of Evans blue-albumin complex in tumor tissue and normal skin in tumor-bearing mice. Tumor S-180 was injected into the skin.

1day 6 h 12 h 2day 5 day 6day 3day 0h 1day 6 h 12 h 2day 5 day 6day 3day 0h 1day 6h 12h 2day 3day 0h 1day 6h 12h 2day 3day 0h

NI F channel NI F + W hite light channel

3 day 4 day 5 day

Cy 5 .5 GC-Cy 5 .5 HGC-Cy 5 .5

Norm al

1 0 μg/ m l 5 0 μg/ m l 1 m g/ m l 1 0 0 μg/ m l

7 Activatable Nanoparticle I m aging Angiogenesis

AuNP AuNP Probe Cy5.5 Sub. Wavenumber (nm)

400 500 600 700 800

Absorbance (a.u.) AuNP AuNP Probe Cy5.5 Sub.

• DTT

+ DTT

Gold Nanoparticle for MMP Activation

I n press, Angew Chem I ntern Ed ( 2 0 0 8 )

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Wavenumber (nm)

700 720 740 760 780 800

Fluorescence Intensity (a.u.)

200 400 600 800 1000 1200

Time (min)

20 40 60 80 100 120

Fluorescence Intensity (a.u.)

200 400 600 800 1000 1200

w/o MMP w/ MMP-2 (M2) w/ MMP-3 (M3) w/ MMP-7 (M7) w/ MMP-13 (M13) w/ MMP-2/Scramble (M2S) w/ MMP-2/Inhibitor (M2I) 1 3 7 14 MMP-2 conc. (nmol/L) Buffer M2 M3 M7 M13 M2S M2I

I n press, Angew Chem I ntern Ed ( 2 0 0 8 )

Gold Nanoparticle for MMP Activation

I n press, Angew Chem I ntern Ed ( 2 0 0 8 )

Gold NP for MMP-2 positive Tum or I m aging

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1H 6H 12H 1day 3day 5day 7day 9day

Long Circulating Self-assem bled Glycol Chitosan Nanoparticle

2day 4day 6day 8day 10day

• SCC-7cell (3x106 cell), tail vein injection of HGC-Cy5.5 (5mg/ kg)

`

Self-assem bled Glycol Chitosan Nanoparticle

O HO O O HO NH2 O CH2OCH2CH2OH CH2OCH2CH2OH NH S n

PBS (pH 7.4) C h l

• r
• f
• r

m S

• n

i c a t i

• n

Decrease in polymer concentraion

5 0 0 nm 3 3 3 nm 1 5 0 nm

Published in Colloid Polym Sci ( 2 0 0 6 ) 2 8 4 : 5 0 6 – 5 1 2

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Probe Probe + inh. No treat MMP-2

1 2 3

1: MMP-2 probe 2: MMP-2 probe + inh. 3: No treat

Polym er NP for MMP-2 positive Tum or I m aging

Molecular I m aging

Visualization of biological process Molecular events at m olecular and celluar level I n living system s Using rem ote im aging detectors “The characterization and m easurem ent of biological processes in living anim als, m odel system s, and hum ans at the cellular and m olecular level by using rem ote im aging detectors”

Lucker GD and Piw nica-W orm s D Acad. Radiol. 2 0 0 1 ;8 ;4

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Reduction in Tum or Size MMP-1 3 Specific Nanoprobe for OA

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2 6 4 8 12 10 weeks

Histology vs. MMP-13 Detection Histology vs. MMP-13 Detection

Histology Grading MMP-13 Expression

Normal Grade 1 Grade 2 Grade 3 Grade 4

OA: Right Normal: Left

Theragnosis

Diagnosis, Molecular I m aging Targeted Therapy Evaluation of Therapeutic Effect

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Biom arkers

Abnorm al cell DNA Protein RNA Metabolite

Lazer Optics Photosensitizer Photonics DNA probes Protein probes RNA probes Metabolite probes

Molecular Probes Tailored Pharm aceuticals Personalized Medicine Bioinform atics

Future of Medicine

Theranostic im aging