Maximizing Protein Production Efficiency in Cancer Therapeutics Katherine Thomas

Pancreatic Cancer

445,220

PANCAN Pancreatic Cancer Fact Sheet 2013

TRAIL •  TNF-related apoptosis-inducing ligand •  Induces apoptosis by binding to an

extrinsic death receptor •  In order to be active, TRAIL must form a

homotrimer •  TRAIL therapies are more effective in

cancer cells than human cells A Genetically Encoded Multifunctional TRAIL Trimer Facilitates Cell-Specific Targeting and Tumor Cell Killing;

2010

Shepard B et al. Anti-Infective Agents in Medicinal Chemistry 2010;8:87-101

TRAIL •  Non-covalently bonded

TRAIL is sensitive to degredation

•  Must form a homotrimer in order to become active

TR3 •  Covalently-linked trimer

developed through genetic fusion

•  Can be further genetically modified

TR3 is a powerful inducer of apoptosis.

Spitzer D et al. Mol Cancer Ther 2010;9:2142-2151

Target Proteins

•  Surface proteins •  Over-expressed in pancreatic and ovarian

cancer cells

Mesothelin MUC16

http://www.epitomics.com/images/IVD/AC-0130_ihc_17343_large.jpg http://www.mybiosource.com/images/tds/protocol_images/MBS245325_IH.jpg

FhMeso1-TR3

Targeting

Targeting

Meso-TR3 TR3

Garg et al

Need for Mass Production

Signal Peptides

•  Leader sequences •  Direct protein to

secretory pathway •  Destination: Golgi

http://www.nobelprize.org/nobel_prizes/medicine/laureates/1999/illpres/protein.html

Kozak Sequence •  Initiation of translation •  Contains start codon

http://www.biomedcentral.com/content/supplementary/1471-2164-13-21-s9.png

Results

pTR3

cd59 signal peptide cd59 signal peptide

Kozak sequence

pTR3-mod59

cd59 signal peptide Kozak sequence

pTR3-mod59

Honeybee melittin signal peptide

Kozak sequence

pTR3-Mel

LEX signal peptide Kozak sequence

pTR3-LEX

Future Research •  Mass Drug Production

–  In-vivo – Clinical Trials

•  Genetic Modification – Target Proteins – Cancer Stem Cells – Additional Drugs

Acknowledgements

•  Dr. Hawkins •  Dr. Spitzer •  Jesse Gibbs •  Christy Hornick

Thank You