OPTICAL IMAGING IN BREAST CANCER: FLUORESCENCE & ČERENKOV. Outline. Example 1: Detecting lymph node involvement. with. Exogenous molecular imaging



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OPTICAL IMAGING IN BREAST CANCER: FLUORESCENCE & ČERENKOV Lymph node imaging Whole breast RT Brian W. Pogue PhD Outline 1. Dual-probe fluorescence imaging for lymph node cancer detection 2. Cerenkov Imaging in Radiation Therapy 3. Hybrid Molecular imaging Example 1: Detecting lymph node involvement with Exogenous molecular imaging 1

Fluorescence Signal Fluorescence Signal LYMPH NODE DETECTION Lymphoscintigraphy Methylene Blue procedure 1) Procedures today remove nodes for the purpose of assay 2) Delay between surgery and lymph node analysis 3) High morbidity (axillary nodes) 4 Dual Dye injection: Targeted & Reference Methylene Blue untargeted EGFR targeted 5 EPIDERMAL GROWTH FACTOR RECEPTOR TARGETED FLUORESCENT IMAGING Methylene blue EGFR binding peptide + IRDye800 0.15 700 Signal Dorsal Node 0.1 700 Signal Ventral node 700 Signal Dorsal vessel 0.05 0 0 10 20 30 40-0.05 minutes after injection 0.4 0.3 800 Signal Dorsal Node 0.2 800 Signal Ventral node 0.1 0 0 10 20 30 40-0.1 Minutes after injection Courtesy: Alisha D Souza 6 Alisha DSouza 2

FLUORESCENCE IMAGING OF AXILLARY LYMPH NODES Injecting Rat Forepaw High Variability!! DUAL REPORTER COMPARTMENT MODEL Targeted Tracer Untargeted Tracer dc f Fl Cl Fl k3 C f k4cb dt dcb k3c f k4cb dt dc r = F l C l - F l C f dt C t BP = k 3/k 4 ( ) = C ( t) + F l C t ( )*e - F l 1+BP t Tichauer et al., Phys Med Biol 2012 8 SIZE AFFINITY TRANSPORT EFFECTS Affibody (7 kda) Shorter clearance time Lower affinity Longer clearance time Higher affinity 9 3

In Vivo Imaging Fluorescence intensity [rel.) l l t LYMPH NODE METASTASIS CANCER MODEL Control Tumor Bearing MDA-MB-231-luc-D3H2LN Tichauer et al., Nature Medicine (in press) Ken Tichauer Ph.D. KINETICS OF EGFR-TARGETED TRACER CAN PREDICT TUMOR BURDEN Tumor Bearing Erb-IRDye800 IgG-IRDye680 EGFR Concentration Map F 1 BP C t C t F C t e 0 0 2 2 Erb-IRDye800 IgG-IRDye680 EGFR Concentration Map Control node Raw Intensity shows no difference 0 0 2 2 Binding ratio has large difference!! RECEPTOR CONCENTRATION VALIDATED WITH Q-PCR Quantitative PCR of entire node Ex Vivo DNA Assay 12 Tichauer et al., Nature Medicine (in press) 4

THE FINANCIAL BARRIERS TO MOLECULAR IMAGING: Lack of existing paradigm for molecular guided surgery = NO BILLING CODE Modest financial payback for diagnostics (vs. therapeutics) http://lippincottschwartzlab.nichd.nih.gov/images/3dculture.jpg REDUCING COSTS: GMP PEPTIDE SYNTHESIS & PHASE 0 TRIAL GMP Recombinant synthesis Run $3M Peptide Production Run $1.0M Toxicity Testing Rodents + Large Animal $3M Toxicity Testing (Rodents only) $0.5M Phase 1 Trial $3M Phase 0 Trial $0.5M Total = $9M Total = $2M Recombinant production route cannot be financed by the NCI, but peptide synthesis can!! Industry/Academic Partnership for Targeted Fluorescent Receptor probe Affibody AB LI-COR Dartmouth Affibody IRDye800 -- ABY-029 0.24mg 2015 2016 2017 5

Photons per Electron PET Radionuclides Cerenkov Imaging in Radiation Therapy The Čerenkov effect GEANT4 simulations Dose deposition Čerenkov production D = 1 ρ Φ dt dx de N = 1 ρ Φ dn dx de Cherenkov radiation and its applications, Jelley Glaser, et. al., Med. Phys. (2013) 2 Čerenkov emission, energy & dose 10 6 Cerenkov Emission in Tissue 10 4 I 1 λ 3 Radiation Therapy Energies 10 0 10-2 Cerenkov intensity per unit volume Mass stopping power (J*m 2 /kg) 10 2 10-4 10 0 0 5 10 15 20 Energy (MeV) 10-2 10-1 10 0 10 1 10 2 10 3 Energy(MeV) High production of Cerenkov in EBRT relative to PET or brachytherapy Production is largely flat with energy > 1 MeV 6

position Multiple Angle Beam Imaging Images at different angles 0 O 15O 30 O 45 O ROTATE Reconstructed 3D volume with FBP Sinogram 0 O 90 O 180 O Angle Glaser et al, Optics Lett. 2013 Beam Tomography with Čerenkov sinogram FBP recon Field A sinogram FBP recon Field B Glaser, et. al., Opt. Lett. 38, 634-636 (2013) 7 3D ČERENKOGRAPHY OF LINAC BEAMS Square beam Complex shaped beam Glaser et al, Optics Lett. 2013 7

Imaging Complex Treatment plans in real time AAPM TG Report # 119 IMRT C-shape Plan in real time Čerenkov Video Time-integrated Glaser et al, Med. Phys. (2014) VMAT Plan in real time Čerenkov Video Time-integrated David Gladstone, Thursday presentation AAPM 8

Optical dosimetry system v3.0 Low Cost option ($250 in parts) Nightvision Web cam monocular filter $50 $150 $50 $150 $5000 $2000 MONTE CARLO MODELING GEANT4 & TISSUE OPTICS PLUG-IN Glaser et al, Biomed. Opt. Express (2013) Glaser et al, Phys Med Biol 2014 Čerenkov in Humans 9

Čerenkov dose imaging tracking skin dose with room lights on! Direct Cerenkov Filtered video Jarvis et al, IJROBP (2014) W/cm 2 1 10-1 10-2 10-3 10-4 Sunlight Room light dimmed lights IMPROVEMENTS IN SNR TO ALLOW ČERENKOV IMAGING Gated Acquisition - 10 5 x gain (3 msec pulses @ 200 Hz) 10-5 10-6 10-7 10-8 Čerenkov GATED TO LINAC PULSING 10-9 Glaser et al, Optics Lett. 2012 First imaging of Čerenkov emission from human tissue Entrance Cerenkov Exit Cerenkov 6MV Entrance 6MV Exit Whole breast radiotherapy with dynamic field. Real time monitoring possible. Jarvis et al, IJROBP (2014) 10

Dynamic beam field monitoring Čerenkov emission identifies beam field dynamics due to MLC motion in real time (fps ~= 2.5). Jarvis et al, IJROBP (2014) Options for Gated Cameras Jacqueline Andreozzi, AAPM talk (Thursday AM) Video rate imaging (30 frames per second) with EM-ICCD 11

12 patient pilot trial completed Hitchcock et al, (submitted, 2014) Respiration tracking: - edge extraction - temporal correlation Breath hold Free breathing Rongxiao Zhang, AAPM talk (Thursday AM) Total Skin irradiation Jacqueline Andreozzi, AAPM talk (Thursday AM) 12

(M -1 cm -1 ) Emission intensity(normalized) Molecular Imaging In Vivo? Use blue Čerenkov to excite NIR emitting molecular reporters in tissue Oxygen sensitive phosphor x 10 4 435nm excitation 1 0.8 772nm emission 10 623nm 0.6 5 0.4 0.2 0 400 500 600 700 800 Wavelength(nm) 700 750 800 850 900 950 Wavelength(nm) Sergei Vinogradov, UPenn Zhang et al, J. Biomed Optics. 2013 Oxygen imaging with luminescence lifetime tomography Lifetime recovery t = 44us po2 = 140mmHg t = 22us po2 = 5mmHg Could be done with dose CT scan Holt et al, Phys Med Biol (in press) 13

SUMMARY: 1. Real-time water tank imaging 2. Real-time in vivo imaging 3. Molecular Sensing with g-čerenkov-luminscence Čerenkography & Čerenkoscopy Team Adam Glaser Rongxiao Zhang Jacqueline Andreozzi Lesley Jarvis, MD PhD David Gladstone, DSc Chad Kanick, PhD Scott Davis, PhD Shudong Jiang, PhD Johan Axelsson, PhD Sergei Vinogradov, PhD www.dartmouth.edu/optmed/ R01 CA109558 R21 EB017559 Audrey Prouty Development Fund QUESTIONS? 14

GEANT4 Monte Carlo: Dose vs. Cherenkov 6 MV beam 6 MV no filter 18 MV beam 18 MV no filter The relative error along the central axis between the Cherenkov light emission and dose for 0.5, 1.0, 2.0, 4.0, and 10.0 cm diameter beams for the 6X, 6FFF, 18X, and 18FFF beams GEANT4 Monte Carlo: Dose vs. Cherenkov (umbra) 6 MV umbra 6 MV no filter 18 MV umbra 18 MV no filter The relative error along the central axis between the Cherenkov light emission and dose for 0.5, 1.0, 2.0, 4.0, and 10.0 cm diameter beams for the 6X, 6FFF, 18X, and 18FFF beams GEANT4 Monte Carlo: Dose vs. Cherenkov two parallel beams 6 MV umbra 6 MV no filter 18 MV umbra 18 MV no filter (a) The central axis curve for the AP-PA treatment is plotted. (b) The corresponding lateral curves at d max, 50 cm, and at the isocenter (100 mm) are shown. 15

Čerenkoscopy of dog oral tumor Radiation Treatment Upper palate Lower jaw Treatment area Cerenkov video Cerenkov video (room lights on) LINAC Beam commissioning & Quality Assurance Treatment plan Targeted dose map Beam delivery Quality assurance plays a fundamental role in radiation treatment of cancer: while modern techniques offer the ability to deliver precise doses of radiation to tumour tissue, this advantage is lost if the equipment is not stable and accurate. Regular and precise calibration of radiotherapy apparatus is thus an essential procedure for hospitals. Treatment plan: lung nodule Beam delivery physicsworld.com New accelerator enhances radiotherapy accuracy, Nov. 19, 2008 16