Byoung Chul Cho, M.D., Ph.D.

Yonsei Cancer Center

Yonsei University College of Medicine

JE-UK Laboratory of Molecular Cancer Therapeutics

FALCON (Fight Against Lung Cancer Oncology Network)

Fibroblast Growth Factor Receptor 1 Gene Amplification Is Associated with

Poor Survival and Cigarette Smoking Dosage in Resected Squamous Cell Lung Cancer

Lung Cancer Mutation ConsortiumIncidence of Driver Mutations in Adenocarcinoma

Mutation found in 54% (280/516) oftumors completely tested (CI 50-59%)

Kris et al ASCO 2011

ROS1

Squamous Cell Carcinoma of Lung

Lung squamous cell carcinoma (SqCC) accounts for ~30% of non-small cell lung cancer

Currently, lung SqCC lacks any druggable target

~90% are male smokers (Korean Cancer Registry)

Despite advances in personalized treatment of adenocarcinoma, effective targeted therapies for SqCC has remained elusive

Frequencies of Potential Driver Mutations inLung Squamous Cell Carcinoma

Lancet oncol 2011;12:175

Mutation Frequency (%)

EGFR <5

ALK <5

HER2 0

BRAF 0

KRAS <5

PIK3CA <5

AKT1 <5

MAP2K1 0

MET <5

~70% un-known

FGFR1 is Amplified in Lung Squamous Cell Carcinoma

Weiss J. Sci Transl Med 2010

8p.12

By FISH, high FGFR1 amplification (≥ 9 copies): 22% (34/153)

FGFR1 amplifications are associated with FGFR inhibitor activity

Weiss J. Sci Transl Med 2010

To investigate the frequency and the prognostic impact of FGFR1 amplification in surgically resected lung SqCC

To evaluate the association between smoking does and FGFR1-amplification

Study Purpose

Patient and Method

SqCC patients that underwent radical resection of a primary lung cancer at Severance Hospital between 1998 and 2009.

Selection criteria (n= 262): availability of tumor tis-sue from the primary lung cancer, smoking-data, and survival data

Construct a tissue-microarray with 2-mm diameter cores (3 cores per tumor)

FGFR1 FISH assay was performed on the tissue-mi-croarrays using FGFR1-probe that hybridizes to the band 8p12-8p11.23 with Spectrum Orange (red) and CEP 8 with Spectrum Green (Abbott Molecular®)

Prespecified Criteria1

“high-amplification” FGFR1 copy number ≥ 9 “low-amplification” FGFR1 copy number >2 or <9 “disomy” FGFR1 copy number = 2

Gene Copy Number

1Weiss J et al. Sci Transl Med 2010

FGFR1 protein & mRNA Expression

IHC analysis was performed using FGFR1 Ab (Epitomics, Burlingame, CA) Only clear membranous staining of the tumor cells was

considered positive and cytoplasmic or granular staining was considered negative or trace

Scoring system (0-400): % of positive tumor cells (0% to 100%) X dominant staining pattern (1: negative or trace, 2: weak, 3: moderate, 4: intense)

mRNA expression analysis was performed by QuantiGene Reagent Systems in FFPE tissue samples

Patient characteristics according to FGFR1 gene amplification by FISH

FGFR1 IHC staining & Gene Copy Number by FISH

34 (13%) 105 (40.1%) 123 (46.9%)

Association between FGFR1 GCN and FGFR1 protein & mRNA Expression

FGFR1 Amplification Is Associated with Poor Sur-vival in Resected Lung SqCC Patients

FGFR1 high ampFGFR1 high amp

Kim HR, Soo RA, Cho BC. J Clin Oncol 2012

Variable Category Disease-free survival Overall survival

HR 95% CI P HR 95% CI P

Sex Female vs.

male

0.68 0.26-1.74 0.42 0.70 0.27-1.79 0.46

Pathologic stage III vs. I+II 2.24 1.45-3.45 <0.0001 2.78 1.76-4.38 <0.0001

Smoking Smoker vs.

never smoker

1.60 0.84-3.05 0.15 1.35 0.70-2.58 0.35

Adjuvant therapy Yes vs. no 1.13 0.74-1.73 0.56 1.08 0.68-1.72 0.71

FGFR1 FISH FGFR1 amp+ vs

FGFR1 amp-

2.24 1.45-3.45 <0.001 1.83 1.15-2.89 0.01

Multivariate Survival Analyses

Treatment outcome to EGFR-TKI according to FGFR1 Amplification

Independent dataset of unresectable, previously treated lung SqCC (N= 47)

FGFR1 Amplification is a Smoking-as-sociated Oncogenic Mutation

P value was tested by χ2 test for linear trend

Kim HR, Soo RA, Cho BC. J Clin Oncol 2012

Intratumoral Heterogeneity? Whole tissue section FISH in 51 randomly selected tumors

(31 high FGFR1-amp, 10 low FGFR1-amp and 10 disomy tumors)

Homogenous FGFR1 distribution in high amplified tumors - at least 80% of nuclei per area exhibited ≥ 9 copies of FGFR1

Majority of areas displayed low amplified FGFR1 signals In

low amplified tumors Two FGFR1 signals were homogenously distributed in

disomy cases No FGFR1 amplification in peritumoral normal tissue High correlation of FGFR1 GCN between primary & metastatic

lesion1

Summary of Whole Tissue Section FISH

1Friederike Goeke et al. Chest Apr 12, 2012

FGFR1 amplification may be involved not in early tumorigenesis, but in early disease progression

Conclusion: FGFR1 Amplification- A New “Druggable” Target in

SqCC The first high-frequency (13%) therapeutic target of

smoking-associated SqCC FGFR1 amplification induced a strong FGFR1 de-

pendency in FGFR1 amplified SqCC FGFR1 amplification is a negative prognostic factor

in resected lung SqCC FGFR1-amplification is associated with cigarette-

smoking in a dose-dependent manner Strongly implies that FGFR1-amplification is an onco-

genic-aberration caused by tobacco-carcinogen