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Transcript of Malignant Melanoma and CDKN2A Kate Coffin .
Malignant Melanoma and CDKN2A
Kate Coffin
http://www.liuding.com/blog/uploaded_images/fig8-794407.jpg
MelanomaGenetics and Environment
Skin cancer of melanocytes
Pre-existing nevus or new nevus
Different areas of skin
ABCDE assessment system of skin
http://www.aad.org/skin-conditions/dermatology-a-to-z/melanoma/signs-symptoms/melanoma-signs-and-symptoms
Dangers of Melanoma
Leading cause of death due to skin disease
2012 estimates in US76,250 new cases9,180 deaths
National Cancer Institute http://www.cancer.gov/cancertopics/types/melanomahttp://www.cancer.gov/aboutnci/servingpeople/cancer-statistics/snapshots
Metastasis
Spreading to other areas of skin and other tissues
14% 5 year survival rate
Miller, A et al. (2006). Melanoma. The New England Journal of Medicine. 355(1), 51-65. PMID:16822996
CDKN2A Cyclin-dependent kinase inhibitor 2A
Inhibits CDK4
Tumor Suppressor
>30 germline mutations
http://en.wikipedia.org/wiki/File:Protein_CDKN2A_PDB_1a5e.png
CDKN2A Domains – Ankyrin repeats
Pfam SMART
Found in archaea, bacteria, eukarya, and poxviruesIn other proteins associated with human disease
http://smart.embl-heidelberg.de/http://pfam.sanger.ac.uk/
CDKN2A Gene OntologyBiological Processes
- cell cycle arrest - cell cycle checkpoint- induction of apoptosis - negative regulation of cell growth
Cellular Components- nucleus - cytoplasm
Molecular Functions- cyclin-dependent kinase inhibitor - protein kinase binding
http://amigo.geneontology.org/cgi-bin/amigo/go.cgi
CDKN2A Protein Phylogeny
http://www.ebi.ac.uk/Tools/msa/clustalw2/
Human CDKN2A Interaction Network
http://string-db.org/
Human CDKN2A Interaction Network
http://string-db.org/
Human CDKN2A Interaction Network
http://string-db.org/
Human CDK4 Interaction Network
http://string-db.org/
CDK4 Domains – Protein Kinase
http://pfam.sanger.ac.uk/
Pfam
Conserved from E. coli to humansCatalytic component of Serine/Threonine kinases
CDK4 Gene OntologyBiological Processes
- regulation of cell cycle - regulation of cell proliferation- signal transduction
Cellular Components- cyclin-dependent protein kinase holoenzyme complex- transcription factor complex
Molecular Functions- cyclin-dependent protein kinase activity - ATP bindinghttp://amigo.geneontology.org/cgi-bin/amigo/go.cgi
Model of CDKN2A and CDK4
CDK4CDKN2A
Model of CDKN2A and CDK4
CDK4CDKN2A
http://chungkitblog.files.wordpress.com/2012/01/ellen-family-visual-medium.jpg
Regulated Growth
Model of CDKN2A and CDK4
CDK4CDKN2A
Model of CDKN2A and CDK4
CDK4CDKN2A
http://upload.wikimedia.org/wikipedia/commons/thumb/6/6c/Melanoma.jpg/250px-Melanoma.jpg
UnregulatedGrowth
Model of CDKN2A and CDK4
CDK4CDKN2A
?
Model of CDKN2A and CDK4
CDK4CDKN2A
?
http://chungkitblog.files.wordpress.com/2012/01/ellen-family-visual-medium.jpg
Regulated Growth
QuestionCan I find a compound that mimics
CDKN2A by inhibiting CDK4?
QuestionCan I find a compound that mimics
CDKN2A by inhibiting CDK4?
HypothesisIf a compound were known to interact with and
inhibit CDK4, it could be used as a drug for melanoma
Experiment – Identify Compound
http://pubchem.ncbi.nlm.nih.gov/
Results – Identify Compound
2-Methyl-5-[(4-methylphenyl)amino]benzothiazole-4,7-dione
http://pubchem.ncbi.nlm.nih.gov/
Cdk4 Inhibitor III
Results – Identify Compound
http://pubchem.ncbi.nlm.nih.gov/
QuestionCan I find a compound that mimics
CDKN2A by inhibiting CDK4?
HypothesisIf a compound were known to interact with and
inhibit CDK4, it could be used as a drug for melanoma
HypothesisIf Cdk4 were inhibited by Cdk4 Inhibitor III in a Cdkn2a CKO mouse, the tumor cells would not proliferate and
metastasize
Why choose a mouse?
Skin and vascular systems
Previous cancer model
Cdkn2a – Cdk4 interaction
http://info.criver.com/flex_content_area/images/nude_mouse-2.jpg
Why choose a mouse?
Skin and vascular systems
Previous cancer model
Cdkn2a – Cdk4 interaction
https://www.aalaslearninglibrary.org/fileupload_content/Course245/14_Nude_tumor.jpg
Why choose a mouse?
Skin and vascular systems
Previous cancer model
Cdkn2a – Cdk4 interaction
http://string-db.org/
Experiment – Treatment GroupsCdkn2a +/+
http://info.criver.com/flex_content_area/images/nude_mouse-2.jpghttp://www.miketiffee.com/uploaded_images/chemoIV-739375.jpg
1
Experiment – Treatment GroupsCdkn2a +/+ Cdkn2a -/- (CKO)
http://info.criver.com/flex_content_area/images/nude_mouse-2.jpghttp://www.miketiffee.com/uploaded_images/chemoIV-739375.jpg
1 2
Experiment – Treatment GroupsCdkn2a +/+ Cdkn2a -/- (CKO)
Cdkn2a +/+& Cdk4 Inhibitor III
http://info.criver.com/flex_content_area/images/nude_mouse-2.jpghttp://www.miketiffee.com/uploaded_images/chemoIV-739375.jpg
1 2
3
Experiment – Treatment GroupsCdkn2a +/+ Cdkn2a -/- (CKO)
Cdkn2a +/+& Cdk4 Inhibitor III
Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
http://info.criver.com/flex_content_area/images/nude_mouse-2.jpghttp://www.miketiffee.com/uploaded_images/chemoIV-739375.jpg
1
43
2
Experiment – Graft Human MelanomaCdkn2a +/+ Cdkn2a -/- (CKO)
Cdkn2a +/+& Cdk4 Inhibitor III
Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
https://www.aalaslearninglibrary.org/fileupload_content/Course245/14_Nude_tumor.jpghttp://www.miketiffee.com/uploaded_images/chemoIV-739375.jpg
1
3 4
2
Data Collection
Microarray analysis ofCdkn2a and Cdk4expression in cells of mice
https://commons.wikimedia.org/wiki/File:DNA_microarray.svg
Expected Microarray Results1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Left Well: Cdkn2a Right Well: Cdk4
Expected Microarray Results1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Left Well: Cdkn2a Right Well: Cdk4
Expected Microarray Results1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Left Well: Cdkn2a Right Well: Cdk4
Expected Microarray Results1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Left Well: Cdkn2a Right Well: Cdk4
Expected Microarray Results1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Left Well: Cdkn2a Right Well: Cdk4
Expected Metastasis Results
1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
Expected Metastasis Results
1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
LEAST METASTASIS
Expected Metastasis Results
1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
MOST METASTASIS
LEAST METASTASIS
Expected Metastasis Results
1) Cdkn2a +/+ 2) Cdkn2a -/- (CKO)
3) Cdkn2a +/+& Cdk4 Inhibitor III
4) Cdkn2a -/- (CKO)& Cdk4 Inhibitor III
MOST METASTASIS
LEAST METASTASIS
MODERATE METASTASIS
MODERATE METASTASIS
Conclusion
CDK4 Inhibitor III could be used as an antimelanoma drug by mimicking the tumor suppressor function of CDKN2A in patients with CDKN2A loss of function mutations
Future Directions
Test other modes of drug administrationTopical cream vs Intravenous solution
Different dosages/times of administration
Study other proteins in interaction networkLook for other tumor suppressors
Study effects of UV exposure on CDKN2AHow does SPF affect this interaction?
Look for compounds in other mutated melanoma pathways
Questions?
http://www.liuding.com/blog/uploaded_images/fig8-794407.jpg