RUSH UNIVERSITY MEDICAL CENTER

Biomarkers of phenotypic plasticity associate with clinical outcomes in patients with locally-advanced NSCLC treated with chemoradiotherapy with and without surgery
Marta Batus, MD; Mary J Fidler, MD; James Clark; Ellis Ziel, MD; Mark Pool, MD; Sanjib Basu, PhD; David Sher, MD; Lela Buckingham, PhD; Michael Liptay, MD; Gary Chmielewski, MD; William Warren, MD; Kelly A. Kaiser-Walters, BS; Shruthi Melinamani, MD; Brett Mahon, MD; Jeffrey A. Borgia, PhD; and Philip Bonomi, MD

RESULTS BACKGROUND
Thoracic chemoradiotherapy (CRT) with or without surgery (S) is the standard-of-care in management of stage III NSCLC. Despite the wide prevalence of the disease, there are sparse data investigating pre-treatment molecular markers in this population.
Survivin-Post
ECAD-high VIM-low

A total of 119 patients receiving chemoradiotherapy with adequate tumor specimens for analysis were enrolled in this study; 61 had definitive chemoradiation whereas 58 had pulmonary resection after chemoradiation. Patients (n=79) with low nuclear survivin immunostaining (score 6) had significantly improved PFS as compared to those patients (n=34) with higher expression levels (14.1 vs. 10.5 months, p=0.042).

OBJECTIVE
The objective of this retrospective study was to evaluate the relationships between clinical outcomes and expression for

Survivin-Pre

Cytoplasmic staining

Patients (n=72) with a cytoplasmic vimentin score 5 had

superior PFS than the with higher expression levels (n=33) (13.17 vs. 9.99 months, p=0.045).

biomarkers associated with either the epithelial-to-mesenchymal transition, or EMT (E-cadherin and vimentin), or a lung cancer stemNuclear staining

cell phenotype (CD133), DNA repair enzyme (ERCC1), and cell

survival/apoptosis (BCL-2, surviving and PTEN).
ERCC1-SQC ERCC1-ADC

High nuclear ERCC1 values (n=72) were associated with a worse OS than those patients (n=44) with low immunostaining (22.7 vs. 59.1 months; p=0.023).

METHODS
Cohorts: Stage III NSCLC pts who were treated with chest radiation (4065Gy) and concurrently with platinum doublet and had sufficient pretreatment tissue were included in this study.

Patients with low cytoplasmic significantly better OS than

E-cadherin those

(n=25) had a (n=85) with

patients

immunostaining scores (62.6 vs. 24.6 months, p=0.036).

The cytoplasmic vimentin/ E-cadherin ratio provided the most

impressive separation of cohort performance with high V/E ratios being associated with a poor PFS (12.6 vs 3.1 months; score

Surgical pts received 40-45 Gy of radiation preoperatively and those who were not surgical candidates received 60-65 Gy.
Immunohistochemistry was used to detect nuclear and cytoplasmic expression of ERCC1, bcl-2, survivin, PTEN, vimentin, E-cadherin, and CD133. Scores were calculated using the Allred scoring system. The log-rank tests used to evaluate progression free survival (PFS) and overall survival (OS) with Kaplan-Meier plots used to plot group characteristics.
Biomarker
E-Cadherin Vimentin CD133 ERCC1 BCL-2 Survivin PTEN

DEMOGRAPHICS
Age at Diagnosis (yrs) Median Mean Gender Female Male Race Caucasian African American Hispanic Other Definitive No (Surgical) Yes Side Right Left 65.79 64.96 62 57 89 26 3 1 61 58 81 38 52% 48% 75% 22% 3% 1% 51% 49% 68% 32% Smoking Status Never Former Current <NA> Stage at consent Stage III Histopathology Adenocarcinoma Squamous Cell Large Cell Other Performance Status 0 1 1.5 2 <NA> 12 73 25 9 119 50 49 3 17 13 41 2 6 57 10% 61% 21% 8% 100% 42% 41% 3% 14% 11% 34% 2% 5% 48%

ratio 10 cutoff; p=0.00073). No significant associations with cytoplasmic CD133 were observed in this cohort for either PFS or OS.

CONCLUSIONS
The association of inferior overall survival in locally-advanced NSCLC patients whose tumors express high ERCC1, high cytoplasmic Ecadherin (which is associated with mesenchymal phenotype and lower adherence of cells which are able to metastasize easier), and lower progression free survival with high survivin and high vimentin/ E-cadherin ratio suggests that combining inhibitors of survivin with those that inhibit DNA repair as well as EMT pathways might improve outcomes in this group of patients.

Clone
G-10 V9 AC133 8F1 124 N/A 28H6

Supplier
Santa Cruz Biotech. Dako Miltenyi Biotec Labvision/ Neomarkers Dako Novus Biologicals Biogenex