Jacques Delforge

Positron emission tomography with 11C CGP-12177 to assess beta-adrenergic receptor concentration in idiopathic dilated cardiomyopathy.

BACKGROUND Positron emission tomography (PET) with 11C-labeled CGP-12177 (CGP) has been shown to have the potential to noninvasively measure beta-adrenergic receptor concentration in dog heart. The present study was undertaken to evaluate the clinical value of this technique. METHODS AND RESULTS Eight normal subjects and 10 patients with heart failure related to an idiopathic cardiomyopathy were studied. Estimation of beta-receptor concentration was based on a graphic method applied on myocardial PET time-concentration curves obtained after an intravenous injection of 11C-CGP followed 30 minutes later by a coinjection of labeled and unlabeled CGP. The clinical tolerance of these injections was good. Left ventricular concentration of beta-receptors was decreased in patients compared with controls (3.12 +/- 0.51 versus 6.60 +/- 1.18 pmol/mL, respectively; p < 0.001). This 53% decrease agrees with previous in vitro data. In eight of the 10 patients, the beta-receptor concentration obtained from PET was compared with the beta-receptor density determined on left ventricular endomyocardial biopsy samples by in vitro binding technique using 3H-CGP-12177. Results obtained with both techniques were correlated (r = 0.79, p = 0.019). Moreover, decreased beta-receptor concentration correlated with the beta-contractile responsiveness to intracoronary dobutamine infusion (r = 0.83, p = 0.003), indicating a direct link between changes in the receptor number and its biological function. CONCLUSIONS PET appears to be a safe and reliable method of assessing in vivo changes in the number of left ventricular beta-adrenergic receptor sites of patients with idiopathic cardiomyopathy.

Quantification of benzodiazepine receptors in human brain using PET, [11C]flumazenil, and a single-experiment protocol

A kinetic method using a multiinjection protocol, positron emission tomography (PET), and [11C]flumazenil as a specific ligand was used to study in vivo the flumazenil-benzodiazepine receptor interactions in the human brain. The model structure is composed of three compartments (plasma, free, and bound ligand) and five parameters (including the benzodiazepine receptor concentration). The arterial plasma concentration, after correction for metabolites, was used as the input function. The experimental protocol, which consisted of three injections of labeled and/or unlabeled ligand, allowed the evaluation of the five model parameters in various brain regions from a single experiment. In particular, the concentration of receptor sites available for binding (B′max) and the equilibrium dissociation constant (KDVK, VR being the volume of reaction) were estimated in five brain regions, including the pons, in which these parameters are identified for the first time (B′max = 4.7 ± 1.7 pmol/ml and KDVR = 4.4 ± 1.3 pmol/ml). Due to the large range of measured receptor concentrations, a linear correlation between B′max and KDVR was pointed out (r = 0.88, p < 0.0005) and was interpreted as a linear relationship between B′max and VR, the parameter KD being assumed constant. This result and its concordance with the published data are discussed. Simulation of the usual two-experiment Scatchard analysis, using the pons as a reference region, showed that the bias on the receptor concentration estimates introduced by this method is significant (from 20 to 40%) but can be corrected using an estimate of the receptor concentration in the pons. Furthermore, we propose a new experimental protocol, based on a Scatchard analysis of the PET data obtained with a partial-saturation experiment. This single-injection protocol is entirely noninvasive, and thus the estimation of the benzodiazepine receptor concentration and of the flumazenil affinity is now possible in human patients using a single 1-h experiment without blood sampling.

Identifiability analysis and parameter identification of an in vivo ligand-receptor model from PET data

The identification of the model parameters from data obtained with a single tracer injection leads to disappointing numerical results, since most of the parameters have to be considered as unidentifiable. A protocol including two injections, a first injection of the labeled ligand and a second injection of the cold ligand (displacement experiment), leads to two very different numerical solutions, which is surprising, since such multiplicity of solutions was not indicated by a preliminary theoretical identifiability study. It is shown that a three-injections protocol, including both a displacement and a coinjection experiment, makes it possible to determine which of these two solutions is biologically valid.<<ETX>>

Modeling Analysis of [11C]Flumazenil Kinetics Studied by PET: Application to a Critical Study of the Equilibrium Approaches

The multi-injection modeling approach was used for the in vivo quantitation of benzodiazepine receptors in baboon brain using positron emission tomography (PET) and [11C]flumazenil (RO 15-1788) as a specific ligand. The model included three compartments (plasma, free, and bound ligand) and five parameters (including the benzodiazepine receptor concentration). The plasma concentration after correction for the metabolites was used as the input function. The experimental protocol consisted of four injections of labeled and/or unlabeled ligand. This protocol allows the evaluation, from a single experiment, of the five model parameters in various regions of interest. For example, in the temporal cortex, the concentration of receptor sites available for binding (B′max) and the equilibrium dissociation constant (Kd) were estimated to be 70 ± 15 pmol/ml and 15.8 ± 2.2 nM, respectively. The validity of the equilibrium approach, which is the most often used quantitation method, has been studied from simulated data calculated using these model parameters. The equilibrium approaches consist of reproducing in PET studies the experimental conditions that permit the use of the usual in vitro methods such as Scatchard analysis. These approaches are often open to criticism because of the difficulty of defining the notion of equilibrium in in vivo studies. However, it appears that the basic relation of Scatchard analysis is valid over a broader range of conditions than those normally used, such as the requirement of a constant bound/free ratio. Simulations showed that the values of the receptor concentration (B′max) and the equilibrium dissociation constant (Kd) found using Scatchard analysis are always underestimated. These simulations also suggest an explanation concerning the dependency of B′max and Kd on the time point employed for the Scatchard analysis, a phenomenon found by several authors. To conclude, we propose new protocols that allow the estimation of the B′max and Kd parameters using a Scatchard analysis but based on a protocol including only one or two injections. These protocols being entirely noninvasive, it thus becomes possible to investigate possible changes in receptor density and/or affinity in patients.

The problem of structural identifiability of a linear compartmental system solved or not

Abstract In a recent paper, C. Cobelli and G. Romanin-Jacur have given a criterion for the structural identifiability of a system of biological compartments. Three comments are made here which show, in particular, that the proposed criterion is a necessary, but not a sufficient, condition.

In vivo quantification of myocardial muscarinic receptors in heart transplant patients.

Decreased myocardial adenylate cyclase activity in response to guanine nucleotide stimulation has been recently demonstrated in denervated myocardium of transplant patients, suggesting that changes in left ventricular muscarinic receptors may occur. Methods and ResultsThe concentration and affinity constants of myocardial muscarinic receptors were determined by positron emission tomography with 11C-labeled methylquinuclidinyl benzilate (MQNB), a specific hydrophilic antagonist, in six transplant patients 4.7±2.3 months after surgery and in six normal subjects. Patients had no sign of cardiac rejection at endomyocardial biopsy. After intravenous injections of MQNB, time-activity curves were obtained over different regions of interest and were fitted to a nonlinear mathematical model. No difference in the concentration of muscarinic receptors was found in transplant patients compared with control subjects: 24±4 versus 26±7 pmol/mL tissue, respectively (P=NS). The association rate constant k−1, the dissociation rate constant k−1 and thus the equilibrium-dissociation constant Kd were the same in transplant patients compared with control subjects. ConclusionsDespite known decreased GTP-stimulated adenylate cyclase activity in transplant patients, the density and affinity constants of myocardial muscarinic receptors are not altered. This suggests abnormalities of the signal-transduction function, such as a change in the guanine nucleotide binding proteins.

Kinetic Analysis of Central [76Br]Bromolisuride Binding to Dopamine D2 Receptors Studied by PET

The in vivo kinetic analysis of dopamine D2 receptors was obtained in baboon brain using positron emission tomography (PET) and [76Br]bromolisuride ([76Br]BLIS) as radioligand. An injection of a trace amount of [76Br]BLIS was followed 3 h later by an injection of a mixture of [76Br]BLIS and BLIS in the same syringe (coinjection experiment). A third injection performed at 6 h was either an excess of unlabeled ligand (displacement experiment) or a second coinjection. This protocol allowed us to evaluate in the striatum of each animal and after a single experiment the quantity of available receptors (B′max) and the kinetic parameters including the association and dissociation rate constants (k+1 VR and k–1, respectively, where VR is the volume of reaction). The cerebellum data were fitted using a model without specific binding. All the parameters were estimated using nonlinear mathematical models of the ligand-receptor interactions including or not including nonspecific binding. The plasma time-concentration curve was used as an input function after correction for the metabolites. An estimate of standard errors was obtained for each PET study and for each identified parameter using the covariance matrix. The average values of B′max and KdVR were 73 ± 11 pmol/ml tissue and 1.9 ± 0.9 pmol/ml, respectively. The nonspecific binding was identifiable in the experiment where the last injection corresponded to a second coinjection. We found that ∼6% of the striatal binding was nonspecific after a tracer injection of [76Br]-BLIS. The nonspecific binding appeared to be reversible in the striatum but irreversible in the cerebellum.

Absolute Quantification by Positron Emission Tomography of the Endogenous Ligand

The results of several recent papers have shown a significant influence of the endogenous neurotransmitters on the exogenous ligand kinetics measured by positron emission tomography. For example, several groups found that the percentage of D2 receptor sites occupied by the endogenous dopamine ranged from 25% to 40% at basal level. An obvious consequence of this significant occupancy is that the ligand-receptor model parameters, usually estimated by a model that does not take into account the endogenous ligand (EL) kinetics, can be significantly biased. In the current work, the authors studied the biases obtained by using the multiinjection approach. The results showed that in the classical ligand-receptor model, the receptor concentration is correctly estimated and that only the apparent affinity is biased by not taking the EL into account. At present, all absolute quantifications of the EL have been obtained through pharmacologic manipulation of the endogenous transmitter concentration, which is often too invasive a method to be used in patients. A theoretical reasoning showed that a noninvasive approach is necessarily based on both the apparent affinity measurement and on a multiregion approach. The correlation between the receptor concentration and the apparent affinity, previously observed with some ligands, verifies these two conditions; thus, the authors suggest that this correlation could be the result of the EL effect. To test this assumption experimentally, the effect of reserpine-induced dopamine depletion on the interactions between the D2 receptor sites and the FLB 457 is studied. With untreated baboons, the apparent FLB 457 affinity was smaller in the receptor-rich regions (striatum) than in the receptor-poor regions. This discrepancy disappeared after dopamine depletion, strongly suggesting that this affinity difference was related to the EL effect. Therefore, the purpose of the current study was to test the ability to quantify the EL based on the observed correlation between the receptor concentration and the apparent affinity. This approach offers a method for estimating the percentage of receptor sites occupied by the EL and, if its affinity is known, the free EL concentration. From the data obtained using FLB 457 with baboons, the authors found that approximately 53% of the D2 receptor sites are occupied by dopamine in the striatum and that the free dopamine concentration is approximately 120 nmol/L at basal level. This approach is transferable to patients, because the experimental data are obtained without pharmacologically induced modification of the EL.

New results on the problem of identifiability of a linear system

Abstract We suggest that the identifiability of a linear system can be studied on the basis of the structural characteristics of the matrices A , B , and C . The initial results—in particular the necessary and the sufficient conditions for identifiability, the upper limits to the number of solutions, and the demonstration of the possibilities arising from the study of constraints —are most encouraging, and should lead to further investigation of the method.

Parametric Images of Benzodiazepine Receptor Concentration Using a Partial-Saturation Injection

The in vivo quantification of the benzodiazepine receptor concentration in human brain using positron emission tomography (PET) and 11C-flumazenil (11C-FMZ), is usually based on a three-compartment model and on PET curves measured in a small number of large regions of interest; however, it should be interesting to estimate the receptor concentration for each pixel and to build quantified images of the receptor concentration. The main advantage is to allow screening of the receptor site localization and visual observation of the possible abnormalities. Up to now, all the methods described include complex experimental protocols, difficult to use in routine examinations. In this paper, we propose the partial-saturation approach to obtain parametric images of benzodiazepine receptor concentration and FMZ affinity. It consists of a single FMZ injection with a low specific activity, followed by Scatchard analysis. Like other parametric imaging methods, this partial-saturation approach can lead to a small percentage (<1%) of unrealistic values in receptor-poor regions; however, it is the only method that allows receptor concentration and affinity images to be obtained from a single-injection 40-min experiment without blood sampling. We also propose a second method in which the receptor concentration map is directly deduced from the PET image acquired 5 to 10 min after a partial-saturation injection. This method assumes a known and constant FMZ affinity value but requires only very simple corrections of this PET image. It is robust (negative values are never found) and quite simple to use in routine examination of patients (no blood sampling, single injection, only 10-min experiment).