Metronomic gemcitabine suppresses tumour growth, improves perfusion, and reduces hypoxia in human pancreatic ductal adenocarcinoma.

BACKGROUND: The current standard of care for pancreatic cancer is weekly gemcitabine administered for 3 of 4 weeks with a 1-week break between treatment cycles. Maximum tolerated dose (MTD)-driven regimens as such are often associated with toxicities. Recent studies demonstrated that frequent dosing of chemotherapeutic drugs at relatively lower doses in metronomic regimens also confers anti-tumour activity but with fewer side effects. METHODS: Herein, we evaluated the anti-tumour efficacy of metronomic vs MTD gemcitabine, and investigated their effects on the tumour microenvironment in two human pancreatic cancer xenografts established from two different patients. RESULTS: Metronomic and MTD gemcitabine significantly reduced tumour volume in both xenografts. However, K(trans) values were higher in metronomic gemcitabine-treated tumours than in their MTD-treated counterparts, suggesting better tissue perfusion in the former. These data were further supported by tumour-mapping studies showing prominent decreases in hypoxia after metronomic gemcitabine treatment. Metronomic gemcitabine also significantly increased apoptosis in cancer-associated fibroblasts and induced greater reductions in the tumour levels of multiple pro-angiogenic factors, including EGF, IL-1alpha, IL-8, ICAM-1, and VCAM-1. CONCLUSION: Metronomic dosing of gemcitabine is active in pancreatic cancer and is accompanied by pronounced changes in the tumour microenvironment.

Morphological and parametric estimation of fetal neural stem cell migratory capacity in the rat brain.

Magnetic resonance imaging (MRI) can non-invasively monitor the migratory behavior of magnetically labeled stem cells after transplantation. Signal changes associated with the clearance of the contrast agent due to cell death and leaked tracer in the interstitial space must be better understood in order to accurately interpret imaging results. In this study, fetal neural stem cells were labeled with superparamagnetic iron oxide (SPIO) particles and transplanted into the corpus callosum of the adult rat. MRI was performed on the day of transplantation and at one week. Control subjects received injections of either non-viable, labeled cells or loose SPIO particles. Two quantitative image analysis algorithms were developed to evaluate imaging results: 1) signal intensity drop-out areas were segmented and compared on a pixel-wise basis between initial and one week images; and 2) signal intensity profiles of transplanted materials at one week were parametrically modeled to estimate migration speed. Segmentation results showed that the number of pixels segmented at one week was significantly greater than the initial number of segmented pixels for subjects receiving injections of viable cells as compared to controls (p<0.05). The average speed of migration of viable cells along the corpus callosum was 69.2+/-41.1 microm/d and was significantly higher than controls (p<0.05). This study demonstrates an in vivo assay to quantitatively evaluate stem cell migration that can be used in different experimental paradigms.

Biomedical engineering and society: policy and ethics.

Biomedical engineering impacts health care and contributes to fundamental knowledge in medicine and biology. Policy, such as through regulation and research funding, has the potential to dramatically affect biomedical engineering research and commercialization. New developments, in turn, may affect society in new ways. The intersection of biomedical engineering and society and related policy issues must be discussed between scientists and engineers, policy-makers and the public. As a student, there are many ways to become engaged in the issues surrounding science and technology policy. At the University of Washington in Seattle, the Forum on Science Ethics and Policy (FOSEP, www.fosep.org) was started by graduate students and post-doctoral fellows interested in improving the dialogue between scientists, policymakers and the public and has received support from upper-level administration. This is just one example of how students can start thinking about science policy and ethics early in their careers.

In vivo manganese MR imaging of calcium influx in spontaneous rat pituitary adenoma.

BACKGROUND AND PURPOSE: Rapid uptake of the calcium analog manganese (Mn2+) into spontaneous pituitary adenoma during MR imaging of aged rats generated the hypothesis that neuroendocrine tumors may have a corresponding increase in calcium influx required to trigger hormonal release. A goal of this study was to investigate the potential for clinical evaluation of pituitary adenoma by MR imaging combined with administration of Mn2+ (Mn-MR imaging). MATERIALS AND METHODS: Mn-MR imaging was used to characterize the dynamic calcium influx in normal aged rat pituitary gland as well as spontaneous pituitary adenoma. To confirm the validity of Mn2+ as a calcium analog, we inhibited Mn2+ uptake into the olfactory bulb and pituitary gland of normal rats by using the calcium channel blocker verapamil. Rats with adenomas received fluorodeoxyglucose-positron-emission tomography (FDG-PET) scanning for characterization of tumor metabolism. Mn2+ influx was characterized in cultured pituitary adenoma cells. RESULTS: Volume of interest analysis of the normal aged pituitary gland versus adenoma indicated faster and increased calcium influx in adenoma at 1, 3, 11, and 48 hours. Mn2+ uptake into the olfactory bulb and pituitary gland of normal rats was inhibited by calcium channel blockers and showed dose-dependent inhibition on dynamic MR imaging. FDG-PET indicated correlation between tumor energy metabolism and Mn2+ influx as well as tumor size. CONCLUSION: These results indicate that adenomas have increased activity-dependent calcium influx compared with normal aged pituitary glands, suggesting a potential for exploitation in the clinical work-up of pituitary and other neuroendocrine tumors by developing Mn-MR imaging for humans.

Magneto-optical labeling of fetal neural stem cells for in vivo MRI tracking.

Neural stem cell therapy for neurological pathologies, such as Alzheimer's and Parkinson's disease, may delay the onset of symptoms, replace damaged neurons and/or support the survival of endogenous cells. Magnetic resonance imaging (MRI) can be used to track magnetically labeled cells in vivo to observe migration. Prior to transplantation, labeled cells must be characterized to show that they retain their intrinsic properties, such as cell proliferation into neurospheres in a supplemented environment. In vivo images must also be correlated to sensitive, histological markers. In this study, we show that fetus-derived neural stem cells can be co-labeled with superparamagnetic iron oxide and PKH26, a fluorescent dye. Labeled cells retain the ability to proliferate into neurospheres in culture, but labeling prevents neurospheres from merging in a non-adherent culture environment. After labeled NSCs were transplantation into the rat brain, their location and subsequent migration along the corpus callosum was detected using MRI. This study demonstrates an imaging paradigm with which to develop an in vivo assay for quantitatively evaluating fetal neural stem cell migration.

A viral envelope as a vehicle for tracer, drug, and gene delivery: initial biodistribution study using PET imaging.

Viral envelopes can be used as an effective vehicle to deliver imaging tracers as well as therapeutic drugs and genes. However, the current methods for in vivo tracking of viral envelopes are limited. This purpose of this study is to investigate dynamically the in vivo biodistribution of viral envelopes using positron emission tomography (PET) imaging. The hemagglutinating virus of Japan envelope (HVJ-E) was labeled with radioactive fluorine (F-18) for tracking with PET imaging. Due to the low molecular weight of F-18, the encapsulation process by HVJ-E was optimized using the cationic agent poly-L-lysine (PLL, MW 66.7 kDa) and Feridex, a magnetic resonance imaging tracer. After labeling, HVJ-Es were injected intravenously into the normal rat and followed for 2 h using high resolution PET imaging. Region of interest analysis showed a significant increase in average liver accumulation based on radioactivity as compared to all control subjects. Average brain uptake showed a significant increase in radioactivity as compared to control subjects receiving F-18-PLL complexes or F-18 alone. Control subjects showed F-18 uptake primarily in the bones. These results demonstrate a molecular imaging technique that can be used to monitor drug and gene delivery and evaluate potential targeting mechanisms.

A Viral Envelope as a Vehicle for Tracer, Drug, and Gene Delivery: Initial Biodistribution Study Using PET Imaging.

Viral envelopes can be used as an effective vehicle to deliver imaging tracers as well as therapeutic drugs and genes. However, the current methods for in vivo tracking of viral envelopes are limited. This purpose of this study is to investigate dynamically the in vivo biodistribution of viral envelopes using positron emission tomography (PET) imaging. The hemagglutinating virus of Japan envelope (HVJ-E) was labeled with radioactive fluorine (F-18) for tracking with PET imaging. Due to the low molecular weight of F-18, the encapsulation process by HVJ-E was optimized using the cationic agent poly-L-lysine (PLL, MW 66.7 kDa) and Feridex, a magnetic resonance imaging tracer. After labeling, HVJ-Es were injected intravenously into the normal rat and followed for 2 h using high resolution PET imaging. Region of interest analysis showed a significant increase in average liver accumulation based on radioactivity as compared to all control subjects. Average brain uptake showed a significant increase in radioactivity as compared to control subjects receiving F-18-PLL complexes or F-18 alone. Control subjects showed F-18 uptake primarily in the bones. These results demonstrate a molecular imaging technique that can be used to monitor drug and gene delivery and evaluate potential targeting mechanisms.

Efficiency of transfection and localization of superparamagnetic iron oxide particles in neural progenitor cells using two methods.

Stem cells represent a potentially revolutionary therapy for neurological pathologies but for which a thorough investigation of cell behavior in the living nervous system has yet to be performed. Contrast-enhanced cell tracking with magnetic resonance imaging can enable this investigation by introducing superparagmagnetic iron oxide (SPIO) particles within the cell membrane. Before magnetically labeled cells can be observed in vivo, it is essential to maximize SPIO transfer into the cell and to fully understand the localization of the contrast agent in mature neural cells. For practical applications, a quantitative evaluation of labeled cells before implantation will allow in vivo assertions. In this study, we present a comparison between two methods for magnetic transfection of neural progenitor cells: the hemmaglutinating virus of Japan envelope (HVJ-E) as a viral vector and a liposomal reagent. We show that HVJ-E is a more efficient vehicle of cell transfection using quantitative evaluation and that the iron content per cell can be predicted using a simple, automated image analysis of stained, labeled cells. Image analysis is also used in this study to show that the contrast agent is distributed in the axon after differentiation, an important aspect of understanding cell tracking in vivo.