dna repair and intracellular communication

DNA Repair and Intracellular Communication

Daniel B. Yarosh, Ph.D.SVP, Basic Science Research

Estee Lauder Co.

1953: Birth of Molecular Genetics Watson and Crick

Heredity / Control of Cell FunctionDNA is a sensor for cell damage

UVINDIRECT

Free Radicals

DIRECT

DNA-AGC^T-

DNA-AoGCT-

CellularOxidationProducts

ANTIOXIDANTS

UV DAMAGE TO DNA

Sunscreens

Thymine CytosineCyclobutane pyrimidine dimer

Direct DNA Damage

Can Antioxidants Prevent CPD?

Murray et al. (Pinnell) JAAD 59:418, 2008

15% Vitamin C, 1% Vitamin E and 0.5% Ferulic Acid (CEFer)

Indirect DNA Damage

8-oxo-Guanine

ROS

UV DAMAGE TO DNAType of Damage DNA Repair Enzyme

Direct:Cyclobutane pyrimidine T4 endonuclease V

dimer UV EndonucleasePhotolyase

Indirect:ROS → 8-oxo-Gua OGG1

T4 Endonuclease V

Bacteriophage T4 recombinant protein

Recognition and incision in one polypeptide

Liposome Delivery

DNA repair enzymes

LIPOSOME

DNA Repair in Human Skin

Untreated

Single UVB dose (1,500J/m2)6h treatment with the DNA Repair Lotion (2mg/cm2)

DNA Repair Lotion

Zastrow and Olivier, 2004 ICP, Jeju Korea

DNA Repair in Human Skin

UV

UV+Photosomes

Human skin: UVB

CPD (green)

Photosomes (1h) reduce CPD by 45%

(Stege et al., PNAS 97:1790, 2000)

OGG1 Repair of 8-oxo-Gua

SKH‐1 mice: UV followed by OGG1 liposomes

Wulff et al., Photochem. Photobiol. 84:317, 2008

8oxoGua

Organ Transplant Patientsand Skin Cancer

180,000 living transplant patients

Nickel sulfate:wheal and flareimmune response

UV erythema and suppressionof immune response

Photosomes prevent UV-B erythemaand restore immune response

Stege et al., PNAS 97:1790, 2000

DNA Damage and Immunity

Local Contact Hypersensitivity

FITC

FITCDNA

RepairUV-B

3 days 5 days 1 day

measureearswelling

DNA Repair Prevents Immunosuppression at Remote Sites

FITC +UV +Placebo +T4N55

6

7

8

9

10

CH

S R

espo

nse

Mea

n Ea

r Sw

ellin

g

p<0.05

Kripke et al. Exp Derm 5:173, 1996

Langerhans Cells Flee Skin

Kuchel et al. PPS 4:577, 2005

Dendritic cells in human skin (red)After 4 days of solar simulating light

Treated with Placebo liposomes (Empty)

orT4N5 Liposome Lotion

T-cells in Lymph Node

Phase contrast FITC (antigen) DiI (Liposome marker)

Yarosh et al, JID 103:461, 1994

Cell to Cell Communication

2 MED (UVA,B) 6 hours

anti-IL-10 antibody staining

T4N5 LiposomeLotion

Placebo

Wolf et al. JID 114:149-156, 2000

DNA Damage induces IL-10

DNA repair reduces expression of IL-10

Human Skin

Wound Healing Protease: MMP-1

Direct UV damageto collagen

UV to fibroblast

UV to keratinocytereleases cytokine

MMP-1 release

collagen

Induction of MMP-1 in Skin

Modified from: Fisher et al., New England Journal of Medicine, 337:1419, 1997

Cell to Cell Communication

UVB 500 J/m2

Keratinocytes+ DNA Repair

24h media

Fibroblasts

48 h media

MMP-1Collagen

MMP-1 Gene Expression

Normal human fibroblasts treated with media from NHEKNHEK: 500 J/m2 UV-B + T4N5 Liposomes (1 µg/ml) for 1 hr post-UV.MMP-1 mRNA expression measured by Real-Time PCR (RT-PCR)

1

1.5

2

2.5

3

3.5

4

0 10 20 30

UV-BUV-B+T4N5None

Hours Media Exposure

Rel

ativ

e In

crea

se M

MP-

1Human Fibroblasts Receiving NHEK Media

50

100

MM

P-1

/GA

PD

H (%

Con

trol)

Significance: * p<0.05

0.0 0.1 1.0 0.1 1.0

150

200

(%)

- UVB

+ UVB

0.0 0.1 1.0

MMP-1

GAPDH

0.1 1.0Blank

LiposomeUV

Endonuclease(liposomal M. luteus

extract)

(%)

- UVB

+ UVB BlankLiposome

UVEndonuclease

(liposomal M. luteus extract)

UV endonuclease significantly suppressed the Kc-induced MMP-1 expression in Fb

OGG1 Inhibition of MMP-1MMP1 in Fb

48h after UV-Kc media

0.0 1.0 2.50.00

0.25

0.50

0.75

1.00

1.25 Empty LiposomesOGG1 Liposomes

Liposomes (µg/ml)

MM

P-1

(ng/

ml)

DNA Damage and Cell Communication

Kinase CascadeSelective transcript activation

CYTOKINES:IMMUNE SUPPRESSIONWOUND HEALINGVIRAL ACTIVATION

DNA PK (PI3K family)ATM, ATR, mTOR

RNA-Pol

The New Synthesis• DNA damage injures a cell• Injured cells communicate

– Cytokines release– Immune suppression– Wound healing response– Viral activation

• Undamaged cells respond to DNA damage• Extracellular signals contribute to skin

cancer

Acknowledgements

AGI Dermatics Cosmos ResearchKen Smiles, PhD Hitoshi Masaki, PhDMatt Canning Yasunobu OchiaiKelly Dong Kuri ObayashiStephanie Nay Yuri OkanoNoah DamaghiStephaney Picart THE XP STUDY GROUPNelli Markova, PhD Margaret Kripke labNeveena Juruvoska, PhD Jean Krutmann lab