Cedars Sinai Medical Center Gene Expression Analysis for targeted Chemotherapy in Lung Cancer Patients PF-00299804 Robert J McKenna Jr. MD Head, Thoracic Surgery Cedars Sinai Medical Center

Stage of Lung Cancer at Diagnosis- SEER Data 3 2 4 1

Adjuvant Chemotherapy No clear supportive data in Stage 1 Benefit seen in tumors greater than 4 cm Statistically significant benefit in Stage II and Stage IIIA Benefit all with cis- platin based chemo.

Chemotherapy Doublets for Lung Cancer Treatment Schiller JH. NEJM 2002; 346:92-98

Lung Cancer Chemotherapy Overall, any doublets have about a 40% response rate That means 60% chemo given has no clinical response Adjuvant treatment increases survival 5-10% That means 90% given with no change in survival

Lung Cancer Chemotherapy Future of lung cancer chemotherapy Get better drugs There is some benefit from current drugs Individualize treatment for patients

EGFR Inhibition The epidermal growth factor receptor is on the surface of cells. It transmits signals to the cell, including to grow and divide. Erlotinib has shown benefit in NSCLC. Overall magnitude of the benefit is small. Magnitude of benefit in responders is high.

Primary Tumor Work Tissue is obtained RNA is evaluated in the laboratory to ensure samples are of high quality Microarray analysis is performed using the Agilent system Molecular data is correlated with clinical data and patient outcome

AGILENT MICROARRAY PLATFORM 1. Pre treatment core vs tumor reference 2. Pre- vs post 3. Post-vs pre- Fluor reversal mRNAs Cy3 Cy5 22,000 spots each containing a 60mer representing a known gene. The two probes hybridize to the spots in proportion to concentration of the specific RNA in each probe.

RNA extraction of Lung Tumors RIN 9.0 S-06-38515 RIN 9.3 S-06-38568 RIN 9.4 S-07-00211 RIN N/A

Lung Tumors: Cluster All Probes 22,000 Genes Significantly Changed 3 7 T u m o r s Normals

Keratin Expression in 337 Lung Tumors Keratins 337 Tumors

Krt5/6, 14/16 Positive Lung Tumors (11%)

Squamous Lung Tumors on Cluster 3 7 T u m o r s Agilent 44K Chip

Neuroendocrine Tumors on Cluster 3 7 T u m o r s Agilent 44K Chip

Conclusions from primary samples High quality RNA has been obtained from nearly all of the samples Clear molecular subtypes can be found Data correlating gene expression with patient outcome will be available soon

Cell Line Collaboration We are evaluating novel compounds in a panel of 60 NSCLC cell lines Correlate cell line data with the data from primary tissue For a pre-surgical study, we selected the irreversible pan-HER inhibitor PF-299804

Clinical Trial A presurgical study to evaluate molecular changes that occur in human non-small cell lung cancer tissue after short term exposure to PF-00299804

Appropriate Additional Therapy Appropriate Follow-Up 5-11 days of drug Diagnostic Biopsy Lung Mass Surgery Appropriate Additional Therapy Assess Response to any Further Therapy Appropriate Follow-Up

Clinical Trial Evaluate molecular changes with chemo Evaluate changes in cell metabolism with chemo

Future of Lung Cancer Chemo Individualize treatment based upon a patient’s tumor markers or gene analysis