The FGF2-FGFR1 Autocrine signaling Pathway Functions as a Novel Oncogene Driver in NSCLC and approaches to inhibit FGFR.

Fred R. Hirsch, MD, PhD

Lynn Heasley, PhD

Murry Wynes, PhD

Paul A. Bunn Jr. MD

Ross Camidge, MD, PhD

University of Colorado Anschutz Medical Campus

Normal and Malignant Growth Control

Endocrine Paracrine Autocrine

LUNG CANCER GROWTH FACTORS

SCLC NSCLC

BK TGF

GRP (Bombesin) EGF

IGF-1 NGF

IGF-2 IGF-1

FGF FGF

Primarily Evaluated in: 1992-2000, 2000-2009, 2006-

Blume-Jensen and Hunter (2001) Nature 411:355-65

Receptor Tyrosine Kinases as Oncogene Drivers in NSCLC

Blume-Jensen and Hunter (2001) Nature 411:355-65

Receptor Tyrosine Kinases as Oncogene Drivers in NSCLC

FGFR structure, signaling, and dysregulation in cancer.

Brooks A N et al. Clin Cancer Res 2012;18:1855-1862

©2012 by American Association for Cancer Research

The FGFR1 gene is amplified, but not mutated, in a significant fraction of lung cancers

Frequent and Focal FGFR1 Amplification Associates with Therapeutically

Tractable FGFR1 Dependency in Squamous Cell Lung Cancer.

Weiss et al. Sci Transl Med. 2010 Dec 15;2(62):62ra93.

Inhibitor-Sensitive FGFR1 Amplification in Human Non-Small Cell Lung

Cancer.

Dutt et al. PLoS ONE. 2011 6(6): 220351.

Amplification of FGFR1 is relatively frequent in SCC and may be “the oncodriving” event.

SNP Array

~10% of SCC Amplification of 8p11-12

FGFR1 in this region

FISH Assay with BAC probe

~22% of SCC Amplification

of FGFR1 Weiss et al. Sci Transl Med. 2010 Dec 15;2(62):62ra93.

*FGFR1 amplification

FGFR inhibitors are effective in amplified cell lines

Heasley et al.

Dutt et al.

Weiss et al.

FGFR TKIs

AZD 8020, AZD4547

BDJ398

Ponatinib

Dovitinib

Brivanib

Many VEGFR TKIs

Bayer

Others

FGFR 1,2,3,4

A phase II study of ponatinib in cohorts of patients with lung cancer preselected

using FGFR1 candidate predictive biomarkers.

University of Colorado SPORE

Drs. Camidge (PI), Bunn, Heasley, Wynes

and Hirsch

Protein

FGFR1 IHC

Collaborative Enzyme

Enhanced Reactive-

ImmunoAssay

In Situ Hybridization

- FGFR1 Black &

Chromosome 8 Red

mRNA Gene Copy

Number

In Situ Hybridization

Quantitative Nuclease Protection Assay

Quantitative PCR

n=57

FGFR1 gene copy number varies in

NSCLC

5/57 (9%) FGFR1 >4 (clusters)

15/57 (26%) FGFR1 >3

5/57 (9%) FGFR1:CH8 Ratio >2

FGFR-specific TKI sensitivity is NOT significantly

associated with lung cancer cell line histology

SK

-ME

S-1

H52

0

H15

81

H17

03

H1

25

H15

7

H44

1

H46

0

HC

C15

HC

C44

HC

C95

NE

-18

Co

lo69

9

DM

S-1

14

SW

15

73

H66

1

H12

99

H17

34

H14

35

H18

69

H21

26

FGFR1

FGFR2

FGFR3

EGFR

PDGFRα

PDGFRβ

Na/K

ATPase

E-Cadherin

FGFR1 protein levels associated withTKI

sensitivity in lung cancer cell lines

SK

-ME

S-1

H52

0

H15

81

H17

03

H1

25

H15

7

H44

1

H46

0

HC

C15

HC

C44

HC

C95

NE

-18

Colo

69

9

DM

S-1

14

SW

1573

H66

1

H12

99

H17

34

H14

35

H18

69

H21

26

FGFR1

FGFR2

FGFR3

EGFR

PDGFRα

PDGFRβ

Na/K

ATPase

E-Cadherin

FGFR1 protein levels is not always associated

with gene amplification in lung cancer cell lines

O 2+ 4+

In situ hybridization to measure FGFR1 mRNA in

FFPE lung tumor specimens

FGFR1 expression is most significantly

associated with FGFR-specific TKI sensitivity

FGFR1 Gene Copy number >4

FGFR1 mRNA >3

mRNA Gene

2 2 0

Double Negative=15

mRNA Gene

3 0 3

Double Negative=8

mRNA Gene

5 2 3

Double Negative=23

ADC and SCC SCC ADC

FGFR1 gene copy number, alone, may

underestimate FGFR1-driven lung tumors

Trial

In Vitro Studies

Determine potential predictive

biomarkers based on sensitivity

or resistance to ponatinib in cell

lines.

- FGFR1 mRNA

- FGFR1 Protein

- FGFR1 Gene Copy Number

Population Studies

Determine the biomarkers

positivity frequency, overlap,

and associations to age,

smoking, gender and histology

in a lung cancer patient

population. Determine any

prognostic associations.

Phase II

Determine the objective

response rate to ponatinib in

molecularly defined stage IIIB/IV

lung cancer patients.

- FGFR1 mRNA

- FGFR1 Protein

- FGFR1 Gene Copy Number

Expanded Phase II

Choose biomarker associated

with a least a 40% response rate.

M+

vs

M-

Stratify

Ponatinib

vs

Standard Tx

Randomize

ORR

PFS

OS

Putative contribution of FGFR1 to

Oncogene-Defined lung cancer

THANKS TO THE

UCCC “DREAM TEAM"

UC AMC

•Fred R. Hirsch

•Murry Wynes

•Paul Bunn

•Barb Helfrich

•Dan Merrick

•Dara Aisner

•Leila Varella-Garcia

UC AMC

•Ross Camidge

•Robert Doebele

•Aik Choon Tan

•Jihye Kim

•James DeGregori

•Vadyym Zaberezhnyy

•Matias Casas

• Lynn Heasley, PhD

• Lindsay Marek, PRA

• Trista Hinz, PRA

• Katie Singleton, Cancer Biology

• Emily Kleczko, Cancer Biology

• Katie Ware, Cancer Biology