This document discusses biomarkers for lung cancer diagnosis, prognosis, and prediction of treatment response. It emphasizes the importance of tumor tissue samples for biomarker analysis but also explores opportunities for using liquid biopsies of blood and other body fluids. A variety of biomarker types are considered, including mutations, gene fusions, protein expression, and microRNAs. The need for standardized biomarker testing methodologies and validation is stressed, along with organizing reference laboratories through country networks.

1. Molecular Biology: diagnostic,
prognostic and predictive biomarkers
Dra. Eloisa Jantus Lewintre
Laboratorio de Oncología Molecular
Fundación para la investigación del Hospital General Universitario de Valencia

2. Biomarkers for LUNG CANCER
BLOOD
sample
Evolve
Locoregional
Mestastatic
Modified from Wistuba et al, Nat Rev Clin Oncol 2011

3. Biomarkers for LUNG CANCER
Tissue
SAMPLE/ MATRIX ISSUES
Blood
(Serum/Plasma)
CNB
BAL, Sputum
Place of sample ?
How many ?
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive
Diagnosis
WHEN?
Response evaluation
Progression
What methodology
should be used ?

4. Biomarkers for LUNG CANCER
Fresh/ frozen
Tissue
FFPE
CNB
SAMPLE/ MATRIX ISSUES
Blood
BAL, Sputum
Place of sample ?
How many ?
HETEROGENEITY

5. Biomarkers for LUNG CANCER
SAMPLE/ MATRIX ISSUES
Tissue
Fresh/ frozen
FFPE
Biobank

6. Biomarkers for LUNG CANCER
Types of tumor specimens in lung cancer
TUMOR CELLS
ARE NEEDED !!

7. Biomarkers for LUNG CANCER
SAMPLE/ MATRIX ISSUES
Blood
Biobannk

8. Biomarkers for LUNG CANCER
MATRIX
CTCs
Cell free DNA
Serum
Plasma (EDTA)
Proteins
Metabolites

9. Biomarkers for LUNG CANCER

10. T. Mok
PLASMA
TUMOR
PLASMA C3

11. Biomarkers for LUNG CANCER
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive

12. Biomarkers for LUNG CANCER
RESPONSE
RISK
P. Massion
DIAGNOSTIC
PROGNOSIS

13. Biomarkers for LUNG CANCER
Complement system: Innate immunity
Ajona, JNCI 2013

14. Biomarkers for LUNG CANCER

15. • Caucasic
• Three discovery studies -> plasma patients
with benign and stage IA lung cancer (n=147)
• The classifier was validated on an
independent set of plasma samples (n = 104)
• The classifier proteins are modulated by
transcription regulators (NF2L2, AHR, MYC,
and FOS) associated with lung cancer, lung
inflammation, and oxidative stress networks

16. Biomarkers for LUNG CANCER
Sozzi, G
JCO in press 2013
1000 controls
85 lung cancer
miRNA signature: 24 miRNAS
(high, intermediate, low)
Blood samples
Diagnostic value: NPV: 99%
Prognostic biomarker and Predictive of
response
Superior diagnostic performance of LDCT if combined with miRNA tests

17. Biomarkers for LUNG CANCER
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive

18. Ca de pulmón
Biomarkers for LUNG CANCER
Ferté, C. et al. Nat. Rev. Clin. Oncol. 7, 367–380 (2010)

19. Biomarkers for LUNG CANCER
Table 2 . KRAS mutations: correlation with Histology
KRAS
WT
Mutated
SCC
ADC*
Others
Total
67
51
16
134
98.5%
79.7%
88.9%
89.3%
1
13
2
16
1.5%
20.3%
11.1%
10.7%
p
Table 3. KRAS mutations: correlation with Smoking Status
0.002
KRAS
Non
Smoker
Former
Smoker
Current
Smoker
Total
WT
13
54
67
134
6
4
6
16
37.5%
25%
37.5%
100%
Mutated
p
0.007

20. Biomarkers for LUNG CANCER
Table 2 . KRAS mutations: correlation with Histology
KRAS
WT
Mutated
SCC
ADC*
Others
Total
p
67
51
16
134
98.5%
79.7%
88.9%
89.3%
1
13
2
16
1.5%
20.3%
11.1%
10.7%
Table 3. KRAS mutations: correlation with Smoking Status
0.002
KRAS
Non
Smoker
Former
Smoker
Current
Smoker
Total
WT
13
54
67
134
6
4
6
16
37.5%
25%
37.5%
100%
Mutated
23
mese
s
81.2
mese
s
p
0.007

21. Biomarkers for LUNG CANCER
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive

22. Ca de pulmón
Biomarkers for LUNG CANCER
De-selection
Ferté, C. et al. Nat. Rev. Clin. Oncol. 7, 367–380 (2010)

23. Biomarkers for LUNG CANCER
ADC
SCC
Li et al, J Clin Oncol 2013

24. Biomarkers for LUNG CANCER
ADC
CUALES DEBEN SER
EVALUADOS COMO
BIOMARCADORES
PREDICTIVOS????

25. Biomarkers for LUNG CANCER
•EGFR
•ALK
•KRAS
•HER2
•BRAF
•ROS1
•RET
mut and amplif
BRAF
RET
Sensitive to Vemurafenib / Dabrafenib/ MEK inh
Sensitive to RET kinase inh

26. Biomarkers for LUNG CANCER

27. Biomarkers for LUNG CANCER
Drilon, 2013
Stahel, 2013

28. Biomarkers for LUNG CANCER

29. NGS, Foundation Medicine
In total, 3 of 91 tumor samples which had
no other sign of cancer-causing genetic
abnormalities, had fusions involving NTRK1.

30. Asian population!!

31. Biomarkers for LUNG CANCER
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive
What methodology
should be used ?

32. Biomarkers for LUNG CANCER
EGFR mutation testing methodology
Test sensitivity
Mutations detected

33. Biomarkers for LUNG CANCER
ALK
ALK fusion variants in NSCLC
Peters et al., 2013 Lung Cancer

34. Biomarkers for LUNG CANCER
FISH
IHC
RNAseq

35. Biomarkers for LUNG CANCER
Tissue should be priorized for EGFR and ALK testing.
EGFR and ALK results should be available within 2
weeks (10 working days)
•In resection specimens, testing is not recommended
in lung cancer that lack morphological or IHC evidence
of adenocarcinoma differentiation
•In limited specimens (biopsies, cytology), testing may
be performed in SCC or small cell histology + clinical
criteria
Lindenman, JTO 2013

36. Biomarkers for LUNG CANCER
BUT…..
Biomarkers can be
determined in BLOOD
samples
MULTIPLEX ANALYSIS (NGS)
are more informative ….
Same amount of sample

37. Biomarkers for LUNG CANCER

38. Biomarkers for LUNG CANCER
Diagnostic
TYPE OF BIOMARKER
Prognostic
Predictive
REBIOPSIES
What we should do
after resistance to
targeted therapies?

39. Biomarkers for LUNG CANCER
Consider liquid biopsies!!

40. RESISTANCE TO EGFR TKI
P. Giannikopoulos, WCLC 2013

41. N=16
sample
sample
P. Giannikopoulos, WCLC 2013
AXL overexpression in 2

42. R. Thomas, WCLC 2013

43. HGF (circulating
levels) in SCLC is
prognostic 
may be
predictive?
E. Arriola,2013

44. Biomarkers for LUNG CANCER
Rebiopsies … why?

45. Biomarkers for LUNG CANCER
Squamous cell carcinoma

46. Biomarkers for LUNG CANCER
Squamous cell carcinoma : FGFR

47. Squamous cell carcinoma: FGFR

48. Biomarkers for LUNG CANCER
Squamous cell carcinoma

49. Biomarkers for LUNG CANCER
Squamous cell carcinoma : HLA-A

50. Biomarkers for LUNG CANCER
BIOMARKERS TESTS

51. Li T, et al, J Clin Oncol 2013

52. Biomarkers for LUNG CANCER

53. Li T, et al, J Clin Oncol 2013

54. Biomarkers for LUNG CANCER

56. Biomarkers for LUNG CANCER

57. Take a Biomarkers for LUNG CANCER
home message ...

58. Take a Biomarkers for LUNG CANCER
home message ...

59. 3rd and 4th generation sequencing
for mutation analysis
Transcriptomic
miRNAs
Epigenetics
Proteomics and Metabolomics
Validation of the new methodologies:
standardization, CLIA
References laboratories: networks
organized by contries