Louise M. Fanchon

Feasibility of In Situ, High-Resolution Correlation of Tracer Uptake with Histopathology by Quantitative Autoradiography of Biopsy Specimens Obtained Under 18F-FDG PET/CT Guidance

Core biopsies obtained using PET/CT guidance contain bound radiotracer and therefore provide information about tracer uptake in situ. Our goal was to develop a method for quantitative autoradiography of biopsy specimens (QABS), to use this method to correlate 18F-FDG tracer uptake in situ with histopathology findings, and to briefly discuss its potential application. Methods: Twenty-seven patients referred for a PET/CT-guided biopsy of 18F-FDG–avid primary or metastatic lesions in different locations consented to participate in this institutional review board–approved study, which complied with the Health Insurance Portability and Accountability Act. Autoradiography of biopsy specimens obtained using 5 types of needles was performed immediately after extraction. The response of autoradiography imaging plates was calibrated using dummy specimens with known activity obtained using 2 core-biopsy needle sizes. The calibration curves were used to quantify the activity along biopsy specimens obtained with these 2 needles and to calculate the standardized uptake value, SUVARG. Autoradiography images were correlated with histopathologic findings and fused with PET/CT images demonstrating the position of the biopsy needle within the lesion. Logistic regression analysis was performed to search for an SUVARG threshold distinguishing benign from malignant tissue in liver biopsy specimens. Pearson correlation between SUVARG of the whole biopsy specimen and average SUVPET over the voxels intersected by the needle in the fused PET/CT image was calculated. Results: Activity concentrations were obtained using autoradiography for 20 specimens extracted with 18- and 20-gauge needles. The probability of finding malignancy in a specimen is greater than 50% (95% confidence) if SUVARG is greater than 7.3. For core specimens with preserved shape and orientation and in the absence of motion, one can achieve autoradiography, CT, and PET image registration with spatial accuracy better than 2 mm. The correlation coefficient between the mean specimen SUVARG and SUVPET was 0.66. Conclusion: Performing QABS on core-biopsy specimens obtained using PET/CT guidance enables in situ correlation of 18F-FDG tracer uptake and histopathology on a millimeter scale. QABS promises to provide useful information for guiding interventional radiology procedures and localized therapies and for in situ high-spatial-resolution validation of radiopharmaceutical uptake.

Pathology-validated PET image data sets and their role in PET segmentation

Positron emission tomography (PET)/computed tomography has recently been finding broader application for the diagnosis, treatment and therapy assessment of malignant disease. Accurate definition of the tumor border is extremely important for the success of localized tumor therapies. PET promises to provide the metabolically active tumor volume and, at present, it is used for target definition in a variety of tumors. This process is, however, subject to uncertainties of different origin. Resolving these uncertainties is challenging, since validating PET images and segmentation contours against tumor pathology is experimentally difficult. In addition to accurate lesion contouring, this challenges validation of PET tracers and investigations of tumor functional heterogeneity. In this paper, we briefly review the present studies providing PET image data sets with pathology validation. We focus on the specimen handling techniques aimed at achieving higher geometrical accuracy of the pathology-derived “ground truth”. We also summarize the main findings obtained for the PET segmentation techniques which have been tested with the help of these data sets. Finally, we provide a critical summary of the current state of the art in pathological validation of PET images and briefly discuss future possibilities in this direction.

Ga-68 DOTATOC PET/CT-Guided Biopsy and Cryoablation with Autoradiography of Biopsy Specimen for Treatment of Tumor-Induced Osteomalacia

Abstract Tumor-induced osteomalacia (TIO) is a rare paraneoplastic syndrome caused by small benign tumors of mesenchymal origin also known as phosphaturic mesenchymal tumors mixed connective tissue variant. Excellent prognosis is expected with eradication of the culprit tumor. These small tumors are notoriously difficult to localize with conventional imaging studies; this often leads to an extensive work up and prolonged morbidity. We report a patient with clinical diagnosis of TIO whose culprit tumor was localized with Ga-68 DOTATOC PET/CT and MRI. Biopsy and cryoablation were performed under Ga-68 DOTATOC PET/CT guidance. Autoradiography of the biopsy specimen was performed and showed in situ correlation between Ga-68 DOTATOC uptake and histopathology with millimeter resolution.

MO-G-17A-09: Quantitative Autoradiography of Biopsy Specimens Extracted Under PET/CT Guidance

PURPOSE To develop a procedure for accurate determination of PET tracer co ncentration with high spatial accuracy in situ by performing Quantitative Autoradiography of Biopsy Specimens (QABS) extracted under PET/CT guidance. METHODS Autoradiography (ARG) standards were produced from a gel loaded with a known co ncentration of FDG biopsied with 18G and 20G biopsy needles. Specimens obtained with these needles are generally cylindrical: up to 18 mm in length and about 0.8 and 0.6 mm in diameter respectively. These standards, with similar shape and density as biopsy specimens were used to generate ARG calibration curves.Quantitative ARG was performed to measure the activity co ncentration in biopsy specimens extracted from ten patients. The biopsy sites were determined acco rding to PET/CTs obtained in the operating room. Additional CT scans were acquired with the needles in place to co nfirm co rrect needle placements. The ARG images were aligned with the needle tip in the PET/CT images using the open source CERR software. The mean SUV calculated from the specimen activities (SUVarg) were co mpared to that from PET (SUVpet) at the needle locations. RESULTS Calibration curves show that the relation between ARG signal and activity co ncentration in those standards is linear for the investigated range (up to 150 kBq/ml). The co rrelation co efficient of SUVarg with SUVpet is 0.74. Discrepancies between SUVarg and SUVpet can be attributed to the small size of the biopsy specimens compared to PET resolution. CONCLUSION The calibration procedure using surrogate biopsy specimens provided a method for quantifying the activity within the biopsy cores obtained under FDG-PET guidance. QABS allows mapping the activity concentration in such biopsy specimens with a resolution of about 1mm. QABS is a promising tool for verification of biopsy adequacy by comparing specimen activity to that expected from the PET image. A portion of this research was funded by a research grant from Biospace Lab, 13 rue Georges Auric 75019 Paris, FRANCE.

Technical Note: Scintillation well counters and particle counting digital autoradiography devices can be used to detect activities associated with genomic profiling adequacy of biopsy specimens obtained after a low activity 18F‐FDG injection

PURPOSE Genomic profiling of biopsied tissue is the basis for precision cancer therapy. However, biopsied materials may not contain sufficient amounts of tumor deoxyribonucleonic acid needed for the analysis. We propose a method to determine the adequacy of specimens for performing genomic profiling by quantifying their metabolic activity. METHODS We estimated the average density of tumor cells in biopsy specimens needed to successfully perform genomic analysis following the Memorial Sloan Kettering Integrated Mutation Profiling of Actionable Cancer Targets (MSK-IMPACT) protocol from the minimum amount of deoxyribonucleonic acid needed and the volume of tissue typically used for analysis. The average 18 F-FDG uptake per cell was assessed by incubating HT-29 adenocarcinoma tumor cells in 18 F-FDG containing solution and then measuring their activity with a scintillation well counter. Consequently, we evaluated the response of two devices around the minimum expected activities which would indicate genomic profiling adequacy of biopsy specimens obtained under 18 F-FDG PET/CT guidance. Surrogate samples obtained using 18G core needle biopsies of gels containing either 18 F-FDG-loaded cells in the expected concentrations or the corresponding activity were measured using autoradiography and a scintillation well counter. Autoradiography was performed using a CCD-based device with real-time image display as well as with digital autoradiography imaging plates following a 30-min off-line protocol for specimen activity determination against previously established calibration. RESULTS Cell incubation experiments and estimates obtained from quantitative autoradiography of biopsy specimens (QABS) indicate that specimens acquired under 18 F-FDG PET/CT guidance that contained the minimum amount of cells needed for genomic profiling would have an average activity concentration in the range of about 3 to about 9 kBq/mL. When exposed to specimens with similar activity concentration, both a CCD-based autoradiography device and a scintillation well counter produced signals with sufficient signal-to-background ratio for specimen genomic adequacy identification in less than 10 min, which is short enough to allow procedure guidance. CONCLUSION Scintillation well counter measurements and CCD-based autoradiography have adequate sensitivity to detect the tumor burden needed for genomic profiling during 18 F-FDG PET/CT-guided 18G core needle biopsies of liver adenocarcinoma metastases.

18 F-fluoromisonidazole predicts evofosfamide uptake in pancreatic tumor model

BackgroundQuantitative imaging can facilitate patient stratification in clinical trials. The hypoxia-activated prodrug evofosfamide recently failed a phase III trial in pancreatic cancer. However, the study did not attempt to select for patients with hypoxic tumors. We tested the ability of 18F-fluoromisonidazole to predict evofosfamide uptake in an orthotopic xenograft model (BxPC3).MethodsTwo forms of evofosfamide were used: (1) labeled on the active moiety (3H) and (2) on the hypoxia targeting nitroimidazole group (14C). Tumor uptake of evofosfamide and 18F-fluoromisonidazole was counted ex vivo. Autoradiography of 14C and 18F coupled with pimonidazole immunohistochemistry revealed the spatial distributions of prodrug, radiotracer, and hypoxia.ResultsThere was significant individual variation in 18F-fluoromisonidazole uptake, and a significant correlation between normalized 18F-fluoromisonidazole and both 3H-labeled and 14C-labeled evofosfamide. 18F-fluoromisonidazole and 14C-evofosfamide both localized in hypoxic regions as identified by pimonidazole.Conclusion18F-fluoromisonidazole predicts evofosfamide uptake in a preclinical pancreatic tumor model.

Evaluation of the tumor registration error in biopsy procedures performed under real-time PET/CT guidance

Purpose: The purpose of this study is to quantify tumor displacement during real‐time PET/CT guided biopsy and to investigate correlations between tumor displacement and false‐negative results. Methods: 19 patients who underwent real‐time 18F‐FDG PET‐guided biopsy and were found positive for malignancy were included in this study under IRB approval. PET/CT images were acquired for all patients within minutes prior to biopsy to visualize the FDG‐avid region and plan the needle insertion. The biopsy needle was inserted and a post‐insertion CT scan was acquired. The two CT scans acquired before and after needle insertion were registered using a deformable image registration (DIR) algorithm. The DIR deformation vector field (DVF) was used to calculate the mean displacement between the pre‐insertion and post‐insertion CT scans for a region around the tip of the biopsy needle. For 12 patients one biopsy core from each was tracked during histopathological testing to investigate correlations of the mean displacement between the two CT scans and false‐negative or true‐positive biopsy results. For 11 patients, two PET scans were acquired; one at the beginning of the procedure, pre‐needle insertion, and an additional one with the needle in place. The pre‐insertion PET scan was corrected for intraprocedural motion by applying the DVF. The corrected PET was compared with the post‐needle insertion PET to validate the correction method. Results: The mean displacement of tissue around the needle between the pre‐biopsy CT and the postneedle insertion CT was 5.1 mm (min = 1.1 mm, max = 10.9 mm and SD = 3.0 mm). For mean displacements larger than 7.2 mm, the biopsy cores gave false‐negative results. Correcting pre‐biopsy PET using the DVF improved the PET/CT registration in 8 of 11 cases. Conclusions: The DVF obtained from DIR of the CT scans can be used for evaluation and correction of the error in needle placement with respect to the FDG‐avid area. Misregistration between the pre‐biopsy PET and the CT acquired with the needle in place was shown to correlate with false negative biopsy results.

SU-F-J-07: Evaluating the Adequacy of Biopsy Specimens for Genetic Signature Assessment by Measuring the Metabolic Activity in Specimens Obtained Under 18F-FDG PET/CT Guidance

PURPOSE Genetic profiling of biopsied tissue is the basis for personalized cancer therapy. However biopsied materials may not contain sufficient amounts of DNA needed for analysis. We propose a method to determine the adequacy of specimens for performing genetic profiling by quantifying metabolic activity. METHODS We measured the response of two radiation detectors to the activity contained in the minimum amount of tumor cells needed for genetic profiling in biopsy specimens obtained under 2-deoxy-2-(18 F)fluoro-D-glucose (18 F-FDG) PET/CT guidance. The expected tumor cell concentration in biopsy specimens was evaluated from the amount of DNA needed (∼100 µg) and the number of pathology sections typically used for the analysis. The average 18 F-FDG uptake per cell was measured by incubating KPC-4662 pancreatic tumor cells and HT-29 colorectal adenocarcinoma tumor cells in 18 F-FDG containing solution (activity concentrations between 0.0122 and 1.51 MBq/mL and glucose concentrations of 3.1 and 1 g/L) for 1 to 1.75 hours and then measuring the activity of a known number of cells. Measurements of surrogate specimens obtained using 18G needle biopsies of gels containing these cells in expected concentrations (∼104 µL-1 ) were performed using an autoradiography CCD based device (up to 20 min exposure) and a scintillation well counter (∼1 min measurements) about 3 and 5 hours after the end of incubation respectively. RESULTS At start of autoradiography there were between 0.16 and 1.5 18 F-FDG molecules/cell and between 1.14 and 5.43×10718 F-FDG molecules/mL. For the scintillation well counter, sample to minimum-detectable-count rate ratios were greater than 7 and the counting error was less than 25% for ≤80 s measurement times. Images of the samples were identifiable on the autoradiograph for ∼10 min and longer exposure times. CONCLUSION Scintillation well counter measurements and CCD based autoradiography have adequate sensitivity to detect the tumor burden needed for genetic profiling in 18G core needle biopsies. Supported in part through the NIH/NCI Cancer Center Support Grant P30 CA008748 and by a sponsored research agreement with Biospace Lab S.A.

WE-AB-BRA-04: Evaluation of the Tumor Registration Error in Biopsy Procedures Performed Under Real Time PET/CT Guidance

Purpose: PET/CT guidance is used for biopsies of metabolically active lesions, which are not well seen on CT alone or to target the metabolically active tissue in tumor ablations. It has also been shown that PET/CT guided biopsies provide an opportunity to verify the location of the lesion border at the place of needle insertion. However the error in needle placement with respect to the metabolically active region may be affected by motion between the PET/CT scan performed at the start of the procedure and the CT scan performed with the needle in place and this error has not been previously quantified. Methods: Specimens from 31 PET/CT guided biopsies were investigated and correlated to the intraoperative PET scan under an IRB approved HIPAA compliant protocol. For 4 of the cases in which larger motion was suspected a second PET scan was obtained with the needle in place. The CT and the PET images obtained before and after the needle insertion were used to calculate the displacement of the voxels along the needle path. CTpost was registered to CTpre using a free form deformable registration and then fused with PETpre. The shifts between the PET image contours (42% of SUVmax) for PETpre and PETpost were obtained at the needle position. Results: For these extreme cases the displacement of the CT voxels along the needle path ranged from 2.9 to 8 mm with a mean of 5 mm. The shift of the PET image segmentation contours (42% of SUVmax) at the needle position ranged from 2.3 to 7 mm between the two scans. Conclusion: Evaluation of the mis-registration between the CT with the needle in place and the pre-biopsy PET can be obtained using deformable registration of the respective CT scans and can be used to indicate the need of a second PET in real-time. This work is supported in part by a grant from Biospace Lab, S.A.

SU‐E‐T‐262: Treatment Room Activation after 15 MV Single Fraction Radiation Treatments Delivered Using Varian's TrueBeam and Trilogy Linear Accelerators

PURPOSE To measure radiation levels in treatment room due to activation after 15MV single fraction radiation treatment (SFRT) delivered to a solid water phantom. METHODS We performed radiation surveys of two LINAC treatment rooms immediately after 15 MV SFRT. We delivered a sequence of two 15 MV single fraction IMRT treatments to a phantom at the end of a typical treatment day. The first treatment delivered was 6201MU (about 12 Gy) and the second one, 15 minutes later was 12711 MU (24 Gy). Both were delivered to the pelvic region of a solid water anthropomorphic phantom. In a second technique, a 15 MV VMAT SFRT (4326 MU) was delivered using the Varian TrueBeam LINAC. Radiation measurements were recorded repetitively at four locations using a thin windowed Geiger Muller detector, a sodium iodide photon spectrometer and a pressurized ionization chamber. The four locations surveyed were: the top of the collimator head, the collimator window surface, the isocenter, and the inferior end of the patient support assembly. RESULTS Radiation levels at the isocenter at the end of the treatment day and before the two IMRT SFRTs varied from 0.06 to 0.1 mR/h. Within 2-3 minutes after finishing the second IMRT SFRT the radiation levels were approximately 10 and 1.4 mR/h at isocenter for the TrueBeam and Trilogy rooms respectively and around 0.6 mR/h for the TrueBeam VMAT SFRT. Closing the MLC and the jaws significantly reduces the radiation level at isocenter. The average half life of the mixture of radionuclides produced is about 10 minutes. CONCLUSIONS High dose single fraction IMRT treatments with 15 MV photons produce elevated treatment room activation as compared to conventional IMRT. In addition, activation levels varied between the TrueBeam and Trilogy for similar SFRT schemes. There is no funding support, disclosures, or conflict of interest.