Arend Heerschap

Magnetic resonance tracking of dendritic cells in melanoma patients for monitoring of cellular therapy

The success of cellular therapies will depend in part on accurate delivery of cells to target organs. In dendritic cell therapy, in particular, delivery and subsequent migration of cells to regional lymph nodes is essential for effective stimulation of the immune system. We show here that in vivo magnetic resonance tracking of magnetically labeled cells is feasible in humans for detecting very low numbers of dendritic cells in conjunction with detailed anatomical information. Autologous dendritic cells were labeled with a clinical superparamagnetic iron oxide formulation or 111In-oxine and were co-injected intranodally in melanoma patients under ultrasound guidance. In contrast to scintigraphic imaging, magnetic resonance imaging (MRI) allowed assessment of the accuracy of dendritic cell delivery and of inter- and intra-nodal cell migration patterns. MRI cell tracking using iron oxides appears clinically safe and well suited to monitor cellular therapy in humans.

Skeletal muscles of mice deficient in muscle creatine kinase lack burst activity

To understand the physiological role of the creatine kinase-phosphocreatine (CK-PCr) system in muscle bioenergetics, a null mutation of the muscle CK (M-CK) gene was introduced into the germline of mice. Mutant mice show no alterations in absolute muscle force, but lack the ability to perform burst activity. Their fast-twitch fibers have an increased intermyofibrillar mitochondrial volume and an increased glycogenolytic/glycolytic potential. PCr and ATP levels are normal in resting M-CK-deficient muscles, but rates of high energy phosphate exchange between PCr and ATP are at least 20-fold reduced. Strikingly, PCr levels decline normally during muscle exercise, suggesting that M-CK-mediated conversion is not the only route for PCr utilization in active muscle.

Altered Ca2+ responses in muscles with combined mitochondrial and cytosolic creatine kinase deficiencies

We have blocked creatine kinase (CK)-mediated phosphocreatine (PCr) -->/<-- ATP transphosphorylation in skeletal muscle by combining targeted mutations in the genes encoding mitochondrial and cytosolic CK in mice. Contrary to expectation, the PCr level was only marginally affected, but the compound was rendered metabolically inert. Mutant muscles in vivo showed significantly impaired tetanic force output, increased relaxation times, altered mitochondrial volume and location, and conspicuous tubular aggregates of sarcoplasmic reticulum membranes, as seen in myopathies with electrolyte disturbances. In depolarized myotubes cultured in vitro, CK absence influenced both the release and sequestration of Ca2+. Our data point to a direct link between the CK-PCr system and Ca2+-flux regulation during the excitation and relaxation phases of muscle contraction.

Antiangiogenic therapy of cerebral melanoma metastases results in sustained tumor progression via vessel co-option

Purpose: In the brain, tumors may grow without inducing angiogenesis, via co-option of the dense pre-existent capillary bed. The purpose of this study was to investigate how this phenomenon influences the outcome of antiangiogenic therapy. Experimental Design: Mice carrying brain metastases of the human, highly angiogenic melanoma cell line Mel57-VEGF-A were either or not treated with different dosages of ZD6474, a vascular endothelial growth factor (VEGF) receptor 2 tyrosine kinase inhibitor with additional activity against epidermal growth factor receptor. Effect of treatment was evaluated using contrast-enhanced magnetic resonance imaging (CE- MRI) and (immuno)morphologic analysis. Results: Placebo-treated Mel57-VEGF-A brain metastases evoked an angiogenic response and were highlighted in CE-MRI. After treatment with ZD6474 (100 mg/kg), CE-MRI failed to detect tumors in either prevention or therapeutic treatment regimens. However, (immuno)histologic analysis revealed the presence of numerous, small, nonangiogenic lesions. Treatment with 25 mg/kg ZD6474 also resulted in efficient blockade of vessel formation, but it did not fully inhibit vascular leakage, thereby still allowing visualization in CE-MRI scans. Conclusions: Our data show that, although angiogenesis can be effectively blocked by ZD6474, in vessel-dense organs this may result in sustained tumor progression via co-option, rather than in tumor dormancy. Importantly, blocking VEGF-A may result in undetectability of tumors in CE-MRI scans, leading to erroneous conclusions about therapeutic efficacy during magnetic resonance imaging follow-up. The maintenance of VEGF-A-induced vessel leakage in the absence of neovascularization at lower ZD6474 doses may be exploited to improve delivery of chemotherapeutic agents in combined treatment regimens of antiangiogenic and chemotherapeutic compounds.

Method for quantitative mapping of dynamic MRI contrast agent uptake in human tumors.

A method is presented for the acquisition and analysis of dynamic contrast‐enhanced (DCE) MRI data, focused on the characterization of tumors in humans. Gadolinium (Gd) contrast was administered by bolus injection, and its effect was monitored in time by fast T1‐weighted MRI. A simple algorithm was developed for automatic extraction of the arterial input function (AIF) from the DCE‐MRI data. This AIF was used in the pixelwise pharmacokinetic determination of physiological vascular parameters in normal and tumor tissue. Maps were reconstructed to show the spatial distribution of parameter values. To test the reproducibility of the method 11 patients with different types of tumors were measured twice, and the rate of contrast agent uptake in the tumor was calculated. The results show that normalizing the DCE‐MRI data using individual coregistered AIFs, instead of one common AIF for all patients, substantially reduces the variation between successive measurements. It is concluded that the proposed method enables the reproducible assessment of contrast agent uptake rates. J. Magn. Reson. Imaging 2001;14:457–463.

Short echo time 1H‐MRSI of the human brain at 3T with minimal chemical shift displacement errors using adiabatic refocusing pulses

The chemical shift displacement error (CSDE) is an often‐underestimated problem in slice selection for localized proton spectroscopy at higher fields. With the proposed semi‐localized by adiabatic selective refocusing (LASER) pulse sequence, this problem is dealt with by using RF pulses with bandwidths in the order of 5 kHz. A combination of conventional nonadiabatic slice‐selective excitation of proton spins, together with double slice‐selective refocusing of the spins by two pairs of adiabatic full‐passage (APF) pulses, produces a spin echo in a volume of interest (VOI) at an echo time down to 30 ms. An illustration of the CSDE of conventional point‐resolved spectroscopy (PRESS) and the semi‐LASER sequence is shown with a measurement of the brain of a volunteer at 3T. With one application of the technique to a patient with a glioblastoma multiforme (GBM), its clinical functionality is demonstrated. With sharp selection profiles and a small CSDE, voxels close to the edge of the VOI can also be used for evaluation. With the additional advantage of being relatively insensitive for B1 inhomogeneities, the semi‐LASER technique can be viewed as a superior substitute for conventional PRESS MR spectroscopic imaging (MRSI) at 3T and beyond. Magn Reson Med, 2007.

Combined quantitative dynamic contrast-enhanced MR imaging and (1)H MR spectroscopic imaging of human prostate cancer.

To differentiate prostate carcinoma from healthy peripheral zone and central gland using quantitative dynamic contrast‐enhanced (DCE) magnetic resonance (MR) imaging and two‐dimensional 1H MR spectroscopic imaging (MRSI) combined into one clinical protocol.

Creatine kinase B-driven energy transfer in the brain is important for habituation and spatial learning behaviour, mossy fibre field size and determination of seizure susceptibility

Creatine kinases are important in maintaining cellular‐energy homeostasis, and neuroprotective effects have been attributed to the administration of creatine and creatine‐like compounds. Herein we examine whether ablation of the cytosolic brain‐type creatine kinase (B‐CK) in mice has detrimental effects on brain development, physiological integrity or task performance. Mice deficient in B‐CK (B‐CK–/–) showed no gross abnormalities in brain anatomy or mitochondrial ultrastructure, but had a larger intra‐ and infrapyramidal mossy fibre area. Nuclear magnetic resonance spectroscopy revealed that adenosine triphosphate (ATP) and phosphocreatine (PCr) levels were unaffected, but demonstrated an apparent reduction of the PCr ⇆ ATP phosphorus exchange capacity in these mice. When assessing behavioural characteristics B‐CK–/– animals showed diminished open‐field habituation. In the water maze, adult B‐CK–/– mice were slower to learn, but acquired the spatial task. This task performance deficit persisted in 24‐month‐old, aged B‐CK–/– mice, on top of the age‐related memory decline normally seen in old animals. Finally, a delayed development of pentylenetetrazole‐induced seizures (creating a high‐energy demand) was observed in B‐CK–/– mice. It is suggested that the persistent expression of the mitochondrial isoform ubiquitous mitochondrial CK (UbCKmit) in the creatine/phospho‐creatine shuttle provides compensation for the loss of B‐CK in the brain. Our studies indicate a role for the creatine–phosphocreatine/CK circuit in the formation or maintenance of hippocampal mossy fibre connections, and processes that involve habituation, spatial learning and seizure susceptibility. However, for fuelling of basic physiological activities the role of B‐CK can be compensated for by other systems in the versatile and robust metabolic‐energy network of the brain.

Fast dynamic gadolinium‐enhanced MR imaging of urinary bladder and prostate cancer

Among the noninvasive imaging modalities, contrast enhanced magnetic resonance (MR) imaging is the most powerful tool with which to visualize vascularity. Common pathology only shows microvessel density, whereas dynamic MR imaging is sensitive to the total endothelial surface area of perfused vessels. Therefore, dynamic MR imaging may be of additional value in tumor staging and in evaluating therapies that affect the perfused microvessel density or surface area, such as chemo‐, radiation, or anti‐angiogenic therapy. In urinary bladder cancer, this technique results in improved local and nodal staging, in improved separation of transurethral granulation tissue and edema from malignant tumor, and in improved evaluation of the effect of chemotherapy. In prostate cancer, dynamic MR imaging may be of help in problematic cases. This technique can assist in determining seminal vesicle infiltration, in depicting of minimal capsular penetration, and in recognizing tumors within the transitional zone. Also, based on very rapid enhancement, very poorly differentiated tumors can be recognized. Evaluation of the effects of therapy is another promising area, however a lot of research remain to be done. This article reviews some basics of fast enhancement techniques, provides practical information, and shows recent developments, in using these fast techniques for staging and grading of bladder and prostate cancer, and for evaluating the effect of therapy. J. Magn. Reson. Imaging 1999;10:295–304.

Tumor vascular architecture and function evaluated by non-invasive susceptibility MRI methods and immunohistochemistry.

To investigate the physiological origins responsible for the varying blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) responses to carbogen (95% O2/5% CO2) breathing observed with different tumor types.