William N. Rom, MD, MPHSol and Judith Bergstein Professor of

Medicine and Environmental Medicine

NYU School of Medicine

NYU Global Institute of Public Health

Sabbatical, Environmental Protection

Agency

Biomarkers for the Early Detection of Lung Cancer

Biomass and Hut Lung• In Asia, Africa, and Latin America, cooking causes indoor air pollution

by burning wood, dung, charcoal and coal (Biomass) without

chimneys.

• Nearly half (2.8B) the people in the world use polluting inefficient

stoves to cook their food each day.

• Hut Lung: Increased COPD in women; pneumonia in children; lung

cancer especially in Chinese women.

• Global Burden of Disease Study 2010 estimates 3.2M deaths from

outdoor and 3.5M deaths from indoor pollution.

• Ranks 3rd (after high blood pressure and tobacco smoking) in Global

DALYs attributable to the 25 leading risk factors in 2010.

• IARC has declared particulate air pollution a class 1 carcinogen.

Gold JA, Jagirdar J, Hay JG Addrizzo-Harris D, Naidich DP, Rom WN. Hut Lung: A domestically acquired particulate lung disease. Medicine 2000; 79:310-317.

High-resolution chest CT-scan demonstrating numerous 2- to 3-mm nodules.

Bellevue Patient from Bangladesh with Indoor Cooking Exposure >35 Years: Alveolar Macrophages have Phagocytosed Anthracotic Particles

(X400)

A. Transbronchial biopsy specimen (40x). B. Extensive deposition of anthracotic

pigment with pneumocyte hyperplasia (400X).

Potential Use of a Serum Lung Cancer Biomarker Test

Early diagnosis

Avoid missing a cure

Avoid abortive

thoracotomies

Avoid anxiety/radiation

of prolonged follow-up

with CT-scan

Indeterminate Nodule on CT8-30mm

Serum test

PET, Bronchoscopy,

Fine Needle

Aspiration, VATS

Watchful Waiting

Repeat CT-scans

- +

NYU Lung Cancer Biomarker Center

• NYU Lung Cancer Biomarker Center recruited 1182 study

subjects (mean age 63 yr, smoking 42 p-y); studied risk factors

for presence of:

• 52% had Non-Calcified Nodules >4 mm at baseline (BENIGN).

Most were stable and 9.7% of solid and 26.2% of sub-solid resolved

• 30 lung cancers with 3 synchronous—for a total of 33 screen-

detected lung cancers (10 incident and 23 prevalent).

(MALIGNANT).

• A subgroup of 13 prevalent cancers were indolent—stable for a

prolonged period—and all stage I adenocarcinomas.

Greenberg AK, Lu F, Goldberg JD, Eylers E, Tsay JC, Yie TA, Naidich D, McGuinness GM, Pass H, Tchou-Wong KM, Addrizzo-Harris D, Chachoua A,

Crawford B, Rom WN. CT Scan screening for lung cancer: Risk factors for nodules and malignancy in a high-risk urban cohort. PLoS ONE 2012; 7(7):

e39403. Epub 2012 Jul 2. PMID: 22768300

Survival

0 20 40 60 80 100 120

405060

708090

100

Time

Su

rviv

al p

rob

ab

ility

(%

)

Number at risk

Group: I

10 4 0 0 0 0 0

Group: IP

12 11 9 4 1 0 0

Group: p

10 8 7 6 5 2 0

group

I

IP

p

Median Disease-Free Survival After Diagnosis

All Cancers 42 Months(6 – 113)

Incident (Aggressive n=10) 13 Months (6 – 33) 2 expired to

lung Ca and 1 lost to F/U

Indolent Prevalent (Non-Aggressive

n=13)

47 Months(5 – 96) 2 expired Lung

Ca and CVA; 3 local recurrences

Prevalent (Non-Aggressive n=10) 81.5 Months (11 – 113) 2 expired

lung Ca and 1 local recurrence

Plasma osteopontin in screened lung cancers (n=10) and controls (n=33).

Joseph S, Harrington R, Walter D, Beck A, Litton T, Hirsch N, Blasberg J, Rom WN, Pass H, Donington J. Osteopontin velocity differentiates lung cancers from controls in a CT screening population. Cancer Biomarkers, in press.

SOMAmers: Unique DNA Reagents for Protein Detection• Red, green, purple atoms are modified nucleotides

that provide binding energy

BOUND COMPLEXIN SOLUTION

• Single-stranded nucleic acids that bind with high affinity and specificity-currently available SOMAmers (1,043).•The limits of detection average (300 fM) and the overall dynamic range spans 7 logs using different dilutions with a ~6% coefficient of variation.

PLoS ONE 2010; 5(12): e15003. (December 2010)

SOMAmer

Target Protein

SomaLogic: Training and Verification Study Subjects

SomaLogic Aptamer Panel

PLoS One. 2010 Dec 7;5(12):e15003

Integrated Diagnostics: Scatter Plot of Nodule Size vs. Classifier Score of 247 Patients Using 13 Proteins

Li X, Hayward C, Fong P-Y, Dominguez M, Hunsucker SW, Lee LW, McLean M, Law S, Butler H, Schirm M, Gingras O, Lamontagne J, Allard R, Chelsky D,

Price ND, Lam S, Massion PP, Pass H, Rom WN, Vachani A, Fang KC, Hood L, Kearney P. A Blood-Based Proteomic Classifier for the Molecular

Characterization of Pulmonary Nodules. Science Translational Medicine 2013; 5: 207ra142.

Autoantibodies to Tumor Associated Antigens (TAAs)

136 Samples analyzed by enzyme immunoassay with recombinant

proteins of 10 TAAs in collaboration with Eng Tan (Scripps)

TAAs: p53, c-myc, IMP1, p62/IMP2, IMP3/Koc, Cyclin A, Cyclin B1, Cyclin D1,

CDK2, Survivin

ROC Curve Based on Stepwise Multiple Logistic Regression and Log Transformed Biomarkers to Classify Cancer vs. No Cancer

(including no nodules, solid nodules, and ground glass opacities groups)

Rom WN, Goldberg JD, Addrizzo-Harris D, Watson HN, Khilkin M, Greenberg AK, Naidich DP, Crawford B, Eylers E, Liu D, Tan EM. Identification of an

autoantibody panel to separate lung cancer from smokers and nonsmokers. BMC Cancer 2010; 10: 234.

Area Under Curve = 0.907

using c-myc, Cyclin A,

Cyclin B1, Cyclin D1,

CDK2, and Survivin.

Airway Genomics: A. Spira, BU

• Can profiles of gene expression in large-airway epithelial cells using

bronchial brushing RNA provide insights into how individual smokers

differ in their responses to cigarette smoke?

• Can such profiling detect smokers in whom the mutagenic effects of cigarette smoke have resulted in lung cancer presenting as a SPN?

Nat Med. 2007 Mar;13(3):361-6.

Affymmetrix Gene Microarrays of PBMCs • 29 gene classifier scores in 137 NSCLC patients and 91 high-risk smoker controls

with non-malignant lung disease. • A positive score indicates classification as cancer, a negative score as non-

malignant disease.

Gene Expression Pattern was Significantly Altered Post-Resection

Gene Classifier had Upregulated Genes in NK Cell Pathway, Ceramide and ErbB.

Wave Spectroscopic Microscopy of Cell Nanoarchitecture Determines Disorder Strength

Roy H et al. Optical detection of buccal epithelial nanoarchitectural alterations in patients harboring lung cancer: Implications for screening. Cancer Res 2010; 70: 7748.

ROC Curves for Ld Differentiating Lung Cancer vs. COPD (red) and

Lung Cancer vs. non-smokers (blue).

Breath Markers of Lung Cancer

• Lung cancer patients may have induction of cytochrome P450 enzymes accelerating catabolism of VOCs so that their abundance in 1.0 L of alveolar breath may provide an early detection biomarker.

• Chest 2003; 123: 2115, Cancer Biomarkers 2007; 3: 95.

Menssana Research’s

breath collection

apparatus collects about

200 VOCs in a breath

sample and gas

chromatography can

analyze picomolar

concentrations.

ROC Curves for Breath Markers of Lung Cancer

Breath Markers of Lung Cancer

Acknowledgements

•Katie Schliessman Research Administrator. Shanni Subryan Research Coord.

•EDRN: NYU Lung Cancer Biomarker Center: Study Nurse:

Siham.Akleh@nyumc.org or 263-6126 to enroll smokers >50 yr >20 pack-yrs;

Jim Messina Data manager, and Jackie Polenco,Scheduler.

•Pulmonary Faculty: Drs. James Tsay, John Munger, Michael Weiden, Leo Segal,

Eric Leibert, Doreen Addrizzo-Harris, Eric Tang PhD.

•Pulmonary Fellow: Vikram Mukherjee MD

•Research Laboratory Technician: Ting-An Yie MS.

•NYU SoM Varick GlycoLab: Ingrid Gils, Jennifer Thomson, Jordan Preiss

•Bioinformatics: Marko I. Vuskovic, D. Sci.

•Radiologists: David Naidich MD, Jane Ko MD, Maria Schiao MD, GeorgeAnn

McGuinness MD.

•Thoracic Surgery: Harvey Pass MD, Margaret Huflejt PhD, Jessica Donington

MD, Bernard Crawford MD.

•NCI Early Detection Research Network UO1 CA086137 Sudhir Srivastava PhD,

MPH, Karl Krueger PhD, Lynn Sorbara PhD, T32 NIEHS 007267. Con Edison.