1. MCW Regional Cancer Therapy Program
IHC-Microarray Analysis of Patients With Peritoneal Metastases (PM) of Appendiceal or Colorectal Origin
Green D.E., Hwang M., Jayakrishnan T.T., Pappas S.G., Gamblin T.C., Turaga K.K.
MCW Regional Cancer Therapy Program

2. We have no relevant financial disclosures

3. Background Appendiceal vs. Colorectal Cancer
Genomically different via microarray, phenotypic clusters predict prognosis
-High Risk Colorectal, no 5 years
-Low Risk Appendiceal, 70% 116 mo
-High Risk Appendiceal, 25% 116 mo
Levine 2012

4. Background Colorectal Cancer KRAS mutated in 34.5%
Median survival 27.7 months
BRAF mutated in 6.5%
Median survival 11 months
Yokota, 2011

5. Background
Mutational analysis of patients with appendiceal adenocarcinoma (n=80)
BRAF 8%
KRAS 45%
EGFR 24%
PIK3CA 19%
Lilleberg 2009

6. Background: IHC and Clinical Benefit
Predictive Biomarker
Associated Treatments *
BRAF
vemurafenib (cetuximab, panitumumab)
KRAS (wt)
cetuximab, panitumumab, sorafenib (erlotinib, gefitinib)
ERCC 1
cisplatin, carboplatin, oxaliplatin
PTEN
erlotinib, gefitinib, cetuximab, panitumumab, trastuzumumab
SPARC
nab-paclitaxel
TOP2A
doxorubicin, liposomal-doxorubicin, epirubicin
TOPO1
irinotecan, topotecan TS
capecitabine, pemetrexed, fluorouracil
* Expression of biomarker determines if chemotherapy is associated with response

7. Hypothesis
We hypothesized that it is feasible to assess the immunohistochemistry and mutational analysis of patients with peritoneal carcinomatosis of appendiceal and colorectal primaries using a commercially available molecular profiling service.

8. Methods Retrospective chart review from clinical data
April June 2012
Inclusion: Appendiceal or colorectal primary, malignant PM, sufficient tissue biopsy
16 patients
Exclusion: Insufficient tissue
Molecular Profiling
Caris Target Now: Phoenix, Arizona/ Irving, Texas
Stratified data
PCI score, histology, treatment

9. Methods 23 Biomarkers examined with immunohistochemistry
ERCC1, RRM1, TS, COX-2, and TOP2A
88 Biomarkers examined with microarray analysis [Illumina Method]

10. Demographics n=16 Age (years) 51 (42-60) Female Gender 11 (69%) T4
Neoadjuvant Chemotherapy
PCI Score
17 (13-26)
CRS + HIPEC
8 (50%)

11. Comparison of General Characteristics of Colon and Appendix Cancer Patients
Colon (n=11)
Appendix (n=5)
P-value
Age (years)
51 (38-72)
51 (40-56)
0.42
Female Gender
8 (73%)
3 (60%)
1.00 T4
Neoadjuvant Chemotherapy
9 (82%)
2 (40%)
0.24
PCI Score
17 (8-27)
17 (12-20)
0.82
CRS + HIPEC
4 (36%)
4 (80%)
0.28
Median Survival (months) 30 60
0.50
9 (82%)
2 (40%)

12. Results Immunohistochemistry performed on 16 patients
Sufficient tissue for microarray analysis on 7 patients (44%)

13. Immunohistochemistry
Biomarker (IHC) *
Colon (N=11)
Appendix (N=5)
P-value
COX-2
80%
100%
0.52
ERCC1
1.00
PTEN
SPARC
36%
20%
TOP2A
0.31
TOPO1 TS
82%
*Significance as reported by the commercial test based on percent staining and staining intensity threshold

14. Mutational Analysis
10%
20%
20%
10%
40%
20%
40%
40%

15. Survival Analysis Biomarker Hazards Ratio (95% CI) P-value COX-2
0.06 ( )
0.02 *
RRM1
0.26 ( )
0.14
SPARC (Poly)
0.15 ( )
0.08 TS
1.00
* Survival not significant when adjusted for multiple comparisons

16. Targeted Therapy & Survival
Biomarker
Chemotherapy
Number
Median Survival (months)
TS +
5FU/LV* 9 30
KRAS WT
Cetuximab/panitumimab 4
ERCC1 +
Oxaliplatin 11 60
TOP2A +
Irinotecan 10
* 5 FU/LV = Fluorouracil/Leucovorin

17. Conclusions
IHC and mutational analysis using commercially available molecular profiling testing is feasible for patients undergoing surgery in setting of PM
40% of patients with colorectal primaries and PM have BRAF mutations (KRAS wt)
Possible survival benefit with COX-2 expression

18. Limitations Small sample size (n=16)
Limited microarray analyses: done on 44% patients (n=7)

19. Future Directions
Prospective evaluation of impact of microarray analysis on response to systemic chemotherapy using multi-institutional data.

20. References
Levine E.A., et al. Gene expression profiling of peritoneal metastases from appendiceal and colon cancer demonstrates unique biologic signatures and predicts patient outcomes. J Am Coll Surg. 2012;214(4): ; discussion
Lilleberg SL, et al (2009, January). Use of mutation analysis of Pseudomyxoma peritonei and appendiceal adenocarcinomas to identify somatic variants within MAPK, AKT and WNT signaling pathways: Potential markers for focused treatment strategies. Poster session presented at Gastrointestinal Cancers Symposium in San Francisco, CA.
Yokota T, et al. BRAF mutation is a powerful prognostic factor in advanced and recurrent colorectal cancer. Br J Cancer ;104(5):

21. Thank You