David Corn

[(Methyl)1-11C]-Acetate Metabolism in Hepatocellular Carcinoma

PurposeStudies have established the value of [(methyl)1-11C]-acetate ([11C]Act) combined with 2-deoxy-2[18F]fluoro-d-glucose (FDG) for detecting hepatocellular carcinoma (HCC) using positron emission tomography (PET). In this study, the metabolic fate of [11C]Act in HCC was characterized.MethodsExperiments with acetic acid [1-14C] sodium salt ([14C]Act) were carried out on WCH-17 cells and freshly derived rat hepatocytes. PET scans with [11C]Act were also carried out on woodchucks with HCC before injection of [14C]Act. The radioactivity levels in different metabolites were quantified with thin-layer chromatography.ResultsIn WCH-17 cells, the predominant metabolite was phosphatidylcholine (PC). Regions of HCCs with the highest [11C]Act uptake had higher radioactivity accumulation in lipid-soluble compounds than surrounding hepatic tissues. In those regions, PC and triacylglycerol (TG) accumulated more radioactivity than in surrounding hepatic tissues.ConclusionsHigh [11C]Act uptake in HCC is associated with increased de novo lipogenesis. PC and TG are the main metabolites into which the radioactive label from [11C]Act is incorporated in HCC.

Imaging Lipid Synthesis in Hepatocellular Carcinoma with [Methyl-11C]Choline: Correlation with In Vivo Metabolic Studies

PET with [methyl-11C]-choline (11C-choline) can be useful for detecting well-differentiated hepatocellular carcinoma (HCC) that is not 18F-FDG–avid. This study was designed to examine the relationship between choline metabolism and choline tracer uptake in HCC for PET with 11C-choline. Methods: Dynamic PET scans of 11C-choline were acquired using the woodchuck models of HCC. After imaging, [methyl-14C]-choline was injected, and metabolites from both HCC tissue samples and the surrounding hepatic tissues were extracted and analyzed by radio–high-performance liquid chromatography. The enzymatic activities of choline kinase and choline-phosphate cytidylyltransferase were assayed for correlation with the imaging and metabolism data. Results: PET with 11C-choline showed an HCC detection rate of 9 of 10. The tumor-to-liver ratio for the 9 detected HCCs was 1.89 ± 0.55. Hematoxylin-eosin staining confirmed that all spots with high tracer uptake were well-differentiated HCCs. Variation of radioactivity distribution within HCCs indicated a heterogeneous uptake of choline. The activities of both choline kinase and choline-phosphate cytidylyltransferase were found to be significantly higher in HCC than in the surrounding hepatic tissues. The major metabolites of 11C-choline were phosphocholine in HCC and betaine and choline in the surrounding hepatic tissues at 12 min after injection; in HCC, phosphocholine rapidly converted to phosphatidylcholine at 30 min after injection. Conclusion: HCCs display enhanced uptake of radiolabeled choline despite a moderate degree of physiologic uptake in the surrounding hepatic tissues. Initially, increased radiolabeled choline uptake in HCCs is associated with the transport and phosphorylation of choline; as time passes, the increased uptake of radiolabeled choline reflects increased phosphatidylcholine synthesis derived from radiolabeled cytidine 5′-diphosphocholine (CDP-choline) in HCCs. In contrast, the surrounding hepatic tissues exhibit extensive oxidation of radiolabeled choline via the phosphatidylethanolamine methylation pathway, a major contributor to the observed physiologic uptake.

Transport and metabolism of radiolabeled choline in hepatocellular carcinoma.

Altered choline (Cho) metabolism in cancerous cells can be used as a basis for molecular imaging with PET using radiolabeled Cho. In this study, the metabolism of tracer Cho was investigated in a woodchuck hepatocellular carcinoma (HCC) cell line (WCH17) and in freshly derived rat hepatocytes. The transporter responsible for [(11)C]-Cho uptake in HCC was also characterized in WCH17 cells. The study helped to define the specific mechanisms responsible for radio-Cho uptake seen on the PET images of primary liver cancer such as HCC. Cells were pulsed with [(14)C]-Cho for 5 min and chased for varying durations in cold media to simulate the rapid circulation and clearance of [(11)C]-Cho. Radioactive metabolites were extracted and analyzed by radio-HPLC and radio-TLC. The Cho transporter (ChoT) was characterized in WCH17 cells. WCH17 cells showed higher (14)C uptake than rat primary hepatocytes. [(14)C]-Phosphocholine (PC) was the major metabolite in WCH17. In contrast, the intracellular Cho in primary hepatocytes was found to be oxidized to betaine (partially released into media) and, to a lesser degree, phosphorylated to PC. [(14)C]-Cho uptake by WCH17 cells was found to have both facilitative transport and nonfacilitative diffusion components. The facilitative transport was characterized by Na(+) dependence and low affinity (K(m) = 28.59 ± 6.75 μM) with partial energy dependence. In contrast, ChoT in primary hepatocytes is Na(+) independent and low affinity. Our data suggest that transport and phosphorylation of Cho are responsible for the tracer accumulation during [(11)C]-Cho PET imaging of HCC. WCH17 cells incorporate [(14)C]-Cho preferentially into PC. Conversion of [(14)C]-PC into phosphatidylcholine occurred slowly in vitro. Basal oxidation and phosphorylation activities in surrounding hepatic tissue contribute to the background seen in [(11)C]-Cho PET images.

Percutaneous CT-guided Cryoablation of the Dorsal Penile Nerve for Treatment of Symptomatic Premature Ejaculation

PURPOSE To evaluate expansion of image-guided interventional cryoablation techniques usually employed for pain management to address the feasibility, safety, and efficacy of treatment for a urologic condition with otherwise limited treatment options, premature ejaculation (PE). MATERIALS AND METHODS Prospective institutional review board approval was obtained, and 24 subjects with PE were enrolled. All patients underwent unilateral percutaneous computed tomography-guided cryoablation of the dorsal penile nerve (DPN). Postprocedural intravaginal ejaculatory latency times (IELTs) and PE Profile (PEP) results served as outcome variables. In addition, subjects were asked whether they would have the procedure done again based on their experience at the 180- and 360-day marks. RESULTS The technical success rate was 100%. Baseline average IELT was 54.7 seconds ± 7.8 (n = 24), which increased to a maximum of 256 seconds ± 104 (n = 11; P = .241) by day 7 and decreased to 182.5 seconds ± 87.8 (n = 6; P = .0342) by day 90. The mean IELT remained at 182.5 seconds ± 27.6 at day 180 (n = 23; P<.0001) and decreased to 140.9 seconds ± 83.6 by 1 year (n = 22; P<.001). PEP scores improved overall, IELTs significantly improved at 180 and 360 days, and 83% of subjects reported that they would undergo the procedure again if given the same opportunity. There were no procedure-related complications. CONCLUSIONS CT-guided percutaneous unilateral cryoablation of the DPN is a feasible, safe, single-day outpatient procedure for the treatment of symptomatic PE.

Imaging early stage osteogenic differentiation of mesenchymal stem cells

Stem cells, such as mesenchymal stem cells (MSCs), contribute to bone fracture repair if they are delivered to the injury site. However, it is difficult to assess the retention and differentiation of these cells after implantation. Current options for non‐invasively tracking the transplanted stem cells are limited. Cell‐based therapies using MSCs would benefit greatly through the use of an imaging methodology that allows cells to be tracked in vivo and in a timely fashion. In this study, we implemented an in vivo imaging methodology to specifically track early events such as differentiation of implanted human MSCs (hMSCs). This system uses the collagen type 1 (Col1α1) promoter to drive expression of firefly luciferase (luc) in addition to a constitutively active promoter to drive the expression of green fluorescent protein (GFP). The resulting dual‐promoter reporter gene system provides the opportunity for osteogenic differentiation‐specific luc expression for in vivo imaging and constitutive expression of GFP for cell sorting. The function of this dual‐promoter reporter gene was validated both in vitro and in vivo. In addition, the ability of this dual‐promoter reporter system to image an early event of osteogenic differentiation of hMSCs was demonstrated in a murine segmental bone defect model in which reporter‐labeled hMSCs were seeded into an alginate hydrogel scaffold and implanted directly into the defect. Bioluminescence imaging (BLI) was performed to visualize the turn‐on of Col1α1 upon osteogenic differentiation and followed by X‐ray imaging to assess the healing process for correlation with histological analyses.

Murine leukemia virus envelope gp70 is a shared biomarker for the high-sensitivity quantification of murine tumor burden

The preclinical development of anticancer drugs including immunotherapeutics and targeted agents relies on the ability to detect minimal residual tumor burden as a measure of therapeutic efficacy. Real-time quantitative (qPCR) represents an exquisitely sensitive method to perform such an assessment. However, qPCR-based applications are limited by the availability of a genetic defect associated with each tumor model under investigation. Here, we describe an off-the-shelf qPCR-based approach to detect a broad array of commonly used preclinical murine tumor models. In particular, we report that the mRNA coding for the envelope glycoprotein 70 (gp70) encoded by the endogenous murine leukemia virus (MuLV) is universally expressed in 22 murine cancer cell lines of disparate histological origin but is silent in 20 out of 22 normal mouse tissues. Further, we detected the presence of as few as 100 tumor cells in whole lung extracts using qPCR specific for gp70, supporting the notion that this detection approach has a higher sensitivity as compared with traditional tissue histology methods. Although gp70 is expressed in a wide variety of tumor cell lines, it was absent in inflamed tissues, non-transformed cell lines, or pre-cancerous lesions. Having a high-sensitivity biomarker for the detection of a wide range of murine tumor cells that does not require additional genetic manipulations or the knowledge of specific genetic alterations present in a given neoplasm represents a unique experimental tool for investigating metastasis, assessing antitumor therapeutic interventions, and further determining tumor recurrence or minimal residual disease.

Incidence of hypercoagulable events after image-guided percutaneous cryoablation of renal tumors: a single-center experience.

PURPOSE To identify retrospectively hypercoagulable events that occurred over time in patients who underwent image-guided percutaneous renal cryoablation and compare the incidence with a cohort of patients who underwent surgical partial nephrectomy (PN) during the same time period. MATERIALS AND METHODS An electronic medical record database was queried for patients who underwent percutaneous image-guided renal mass cryoablation or PN between September 2006 and June 2012. Records were examined for thrombotic events during the year following the procedure in each group. Incidence rates, Kaplan-Meier estimates, and patient demographic variables were compared using the stratified log-rank test and t test for independent samples. RESULTS The study comprised 114 cryoablation cases. The cumulative incidence of thrombotic events after 1 year was 4.39%. The incidence per 100 person-years was 4.84. There were 105 PN cases. The cumulative incidence of thrombotic events after 1 year was 1.0%. The incidence per 100 person-years was 1.14. The person-time incidence rate difference for these two groups did not reach statistical significance (P = .0894). CONCLUSIONS The incidence of thrombotic events in patients who underwent percutaneous renal cryoablation in this study was not significantly different than a comparable cohort who underwent surgical PN during the same time period.

Effect of the time to intervention on the outcome of thrombosed dialysis access grafts managed percutaneously.

PURPOSE We aimed to investigate the effect of the time interval from the clinical presentation of a thrombosed dialysis access graft to intervention on procedure success. MATERIALS AND METHODS Records from two academic institutions for patients who underwent percutaneous thrombectomy of occluded surgical hemodialysis graft access sites in interventional radiology from 2006 to 2011 were reviewed retrospectively. The following data were recorded: gender, age, time and date of the initial request for a thrombectomy and the procedure, age of the surgical access, angiographic outcome, and clinical outcome (successful or unsuccessful postinterventional dialysis). Univariate and multivariate logistic regression were used to evaluate whether the time to intervention significantly affected the study endpoint. RESULTS In total, 268 percutaneous thrombectomies were performed in 139 patients. Of these 224 (83.5%) were categorized as successful and 44 (16.4%) as unsuccessful. The time to intervention was 19.9±30.1 vs. 22±35 hours for successful and unsuccessful procedures, respectively. The difference between the two was not significant, and there were also no significant differences in covariate distributions between successful and unsuccessful outcomes. CONCLUSION During the first 72 hours following clinical presentation of a thrombosed dialysis access graft, time to intervention may be considered independent of procedure outcome.

Radio-deoxynucleoside Analogs used for Imaging tk Expression in a Transgenic Mouse Model of Induced Hepatocellular Carcinoma.

Purpose: A group of radiolabeled thymidine analogs were developed as radio-tracers for imaging herpes viral thymidine kinase (HSV1-tk) or its variants used as reporter gene. A transgenic mouse model was created to express tk upon liver injury or naturally occurring hepatocellular carcinoma (HCC). The purpose of this study was to use this unique animal model for initial testing with radio-labeled thymidine analogs, mainly a pair of newly emerging nucleoside analogs, D-FMAU and L-FMAU. Methods: A transgeneic mouse model was created by putting a fused reporter gene system, firefly luciferase (luc) and HSV1-tk, under the control of mouse alpha fetoprotein (Afp) promoter. Initial multimodal imaging, which was consisted of bioluminescent imaging (BLI) and planar gamma scintigraphy with [125I]-FIAU, was used for examining the model creation in the new born and liver injury in the adult mice. Carcinogen diethylnitrosamine (DEN) was then administrated to induce HCC in these knock-in mice such that microPET imaging could be used to track the activity of Afp promoter during tumor development and progression by imaging tk expression first with [18F]-FHBG. Dynamic PET scans with D-[18F]-FMAU and L-[18F]-FMAU were then performed to evaluate this pair of relatively new tracers. Cells were derived from these liver tumors for uptake assays using H-3 labeled version of PET tracers. Results: The mouse model with dual reporters: HSV1-tk and luc placed under the transcriptional control of an endogenous Afp promoter was used for imaging studies. The expression of the Afp gene was highly specific in proliferative hepatocytes, in regenerative liver, and in developing fetal liver, and thus provided an excellent indicator for liver injury and cancer development in adult mice. Both D-FMAU and L-FMAU showed stable liver tumor uptake where the tk gene was expressed under the Afp promoter. The performance of this pair of tracers was slightly different in terms of signal-to-background ratio as well as tracer clearance. Conclusion: The newly created knock-in mouse model was used to demonstrate the use of the dual-reporter genes driven by well-characterized cancer-specific transcriptional units in conjunction with in vivo imaging as a paradigm in studying naturally occurring cancer in live animals. While BLI is suitable for small animal imaging with luc expression, PET with L-FMAU seemed be the choice for liver injury or liver cancer imaging with this animal model for future investigations.

Abstract 5297: Dual-modality imaging of hepatocellular carcinoma

Choline compounds are linked to malignancy since elevation of the intensity of choline peak reflects increased biosynthesis of membrane phospholipids and can be a marker for cellular proliferation. Positron Emission Tomography (PET) images an injected tracer dose (nM∼pM) of [11C]-choline while Proton MR Spectroscopy (1H MRS) can characterize endogenous choline-containing compounds usually in the mM range. There have been studies on brain and prostate cancers with both imaging modalities although correlation between the results from PET and MRS is inconsistent. The transient radiolabeled choline metabolism measured by PET is different from the relative steady total choline contents measured by MRS. However, it begs the question of whether the two imaging modalities are measuring the same metabolic pathways of choline in cancer. For primary Hepatocellular Carcinoma (HCC), there have been [11C]-choline PET studies, both pre-clinical and clinical, showing focal uptake. Likewise, there have been 1H-MRS studies of choline with animals and humans showing significantly high choline levels in HCC. So far, there is no comparison study reported to link the two measurements in HCC. This study was conducted with a woodchuck model of hepatitis viral infection-induced HCC, which is regarded as a naturally occurring animal model of human HCC with similar pathology and clinical behavior. The original objective of the study was to examine the effects of animals being fed and fasted on PET scans using [11C]-choline. MRI and MRS were added and performed immediately before PET imaging. Experiments were first performed with animals at the fed state using a 4T MR scanner with a human head coil. Dynamic PET images were then generated from a nearby clinical PET/CT scanner right after, spanning 50 minutes starting from [11C]-choline injection. Time-activity curves and tracer contrast uptake between focal uptake in HCC and that in the surrounding hepatic tissues were calculated in liver regions. The same animals were scanned again four days later following the exact same procedure, except that the animals were fasted over night. The results from PET imaging with [11C]-choline showed that the dietary state of fed or fast had little impact on radio-choline uptake in HCC while the results from 1H MRS of choline found similar choline/lipids ratios in HCC regardless of whether the animal was fed or fasted. These results are very promising for correlating the two measurements although further experiments are needed to confirm that the two imaging modalities are measuring related lipid metabolic pathways in HCC involving choline. This work is supported in part by NCI grants R01 CA095307 and U24 CA110943. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5297. doi:10.1158/1538-7445.AM2011-5297