et ai., 1981; Duffy et ai., 1982; Kennedy et ai., 1982;. Reivich et ..... curse. J Nucl Med 29: 1603-1604. Di Chiro G, Brooks RA, Patronas NJ, Bairamian D, Kornblith.
Journal of Cerebral Blood Flow and Metabolism
10:190-198 © 1990 Raven Press, Ltd., New York
Deoxyglucose Lumped Constant Estimated in a Transplanted Rat Astrocytic Glioma by the Hexose Utilization Index Alexander M. Spence, Michael M. Graham, Mark Muzi, Gregory L. Abbott, Kenneth A. Krohn, Rajesh Kapoor, and Steven D. Woods Division of Neurology and Imaging Research Laboratory, Department of Radiology, University of Washington School of Medicine, Seattle, Washington, U.S.A.
Summary: The lumped constant (LC) for calculating the regional glucose (glc) metabolic rate by the deoxyglucose (DG) method was estimated in a transplanted rat glioma and normal rat brain. First, the hexose utilization index (HUI) was measured at 1.5, 3.0, and 4.5 min in right hemisphere glioma implants and uninvolved contralateral hemisphere following bolus intravenous injections of eH]DG and P4C]glucose. At these times, the glioma HUI values were 0.639, 0.732, and 0.712, respectively, and the coordinate left hemisphere values were 0.432, 0.449, and 0.418. Second, the volumes of distribution of DG and glucose were determined to be 0.436 and 0.235 in
glioma implants and 0.402 and 0.237 in left hemisphere, respectively. Third, following simultaneous intracarotid injections of eH]DG and [14C]glucose, the ratio KTIKI was 1.1 in glioma grafts and 1.3 in left hemisphere. With these values for HUI, volume of distribution, and KJ ra tio, the LC in this rat glioma was estimated to be 2.1 times higher than the left hemisphere LC (p < 0.02). These results suggest that measurement of brain tumor CMRglc using a normal brain LC may significantly overestimate the true tumor CMRg!c. Key Words: Lumped constant Deoxyglucose-Rat glioma-Glucose metabolism Hexose utilization index-CMR g!c.
e4C]deoxyglucose (DG) with quantitative autora diography and e8F]fluorodeoxyglucose (FDG) with
constants for the exchange of DG from the plasma to precursor pool across the blood-brain barrier and back, and the phosphorylation and dephosphoryla tion (Phelps et aI., 1979) of DG, and (b) a lumped constant (LC) that is the ratio of the utilization of DG to that of glucose (Sokoloff et aI., 1977; Phelps et aI., 1979; Crane et aI., 1981). Measurements of the kinetic rate constants (Sokoloff et aI., 1977; Phelps et aI., 1979; Lammertsma et ai., 1987) and LC (Sokoloff et ai., 1977; Phelps et aI., 1979; Crane et ai., 1981; Duffy et ai., 1982; Kennedy et ai., 1982; Reivich et ai., 1984; Gjedde et aI., 1985; Takei et al., 1986) in normal animal and human brain tissue have been reported. For pathological tissues such as ma lignant brain tumors, the correct values of the ki netic constants and LC may differ enough from nor mal brain to influence the accurate measurement of CMRglc with DG (Kato et aI., 1985; Koeppe et ai., 1987; Nakai et aI., 1987a,b; Wienhard et ai., 1987). In a companion paper, we reported measurements of the kinetic rate constants in a transplanted rat astrocytic glioma model (Graham et ai., 1989). The
positron emission tomography (PET) are used to measure the regional utilization of glucose (CMRglc) in animal (Sokoloff et aI., 1977) and hu man brain (Phelps et aI., 1979). This method is based on a three-compartment model of DG (or FDG) and glucose consumption and an operational equation for the glucose metabolic rate calculated from the amount of accumulated isotopic DG and DG-6-phosphate (DG-6-P) in the tissue over time and the arterial plasma isotope time-activity curve. The operational equation contains (a) kinetic rate
Received March 28, 1989; revised August 25, 1989; accepted August 29, 1989. Address correspondence and reprint requests to Dr. A. M. Spence at Neurology, RG-27, University of Washington, Seattle, WA 98195, U.S.A. Abbreviations used: CMRglc' cerebral metabolic rate of glu cose; DG, deoxyglucose; PDG, eSPlfluorodeoxyglucose; HUI, hexose utilization index; LC, lumped constant; TLC, thin-layer chromatography.
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DEOXYGLUCOSE LUMPED CONSTANT IN RAT GLIOMA present paper reports an estimate of the ratio of the LC in this glioma relative to normal brain based on the hexose utilization index (HOI) approach of Crane et al. (1981) and Pardridge et al. (1982a,b). Our chief finding is that the LC in intracerebral grafts of this rat glioma is significantly higher than in contralateral cerebral tissue. MATERIALS AND METHODS Isotopes, enzymes, and biochemicals
D-[6-14C]glucose (specific activity, 55 mCi/mmol) and 2-deoxy-D-[l,2-3H]glucose (specific activity, 50 Cilmmol) were obtained from American Radiolabeled Chemicals (St. Louis, MO, U.S.A.) and ICN (Irvine, CA, U.S.A.), respectively. Chemical and radiochemical purity were greater than 99% as analyzed by thin-layer chromatogra phy (TLC) methods suggested by the manufacturer and MacGregor et al. (1981), high-performance liquid chro matography (Brownlee Polypore H column eluted with 0.01 N H2S0 at 0.3 mllmin), and by enzymatic degra�a 4 tion. Anion exchange resin, Sephadex-A25 (PharmacIa, Piscataway, NJ, U.S.A.) was regenerated in the chlori�e form and fully equilibrated with Tris buffer A (10 mM Tns HCI, pH 7.4, 0.01% sodium azide) prior to use. Hexoki nase (n-glucose:ATP phosphotransferase, E.C. 2.7.1.2), glucose-6-phosphate dehydrogenase (n-glucose-6phosphate:NADP+ I-oxidoreductase, E.C. 1.1.1.49), ATP, and NAD+ were purchased from Sigma Chemical Company (St. Louis, MO, U.S.A.). Instagel and Soluene350 were acquired from Packard Instrument Company (Downers Grove, IL, U.S.A.). Rat brain tumor model
The rat brain tumor model used in this work has been reported in detail elsewhere ( � pence and 0
Assuming = 1 (Sokoloff et aI., 1977), this ap proach yielded a LC of 0.517 ± 0.051 in normal rat brain and 1.168 ± 0.171 in intracerebral 36B-1O im plants. From these data, the ratio LCT/LCN com putes to be 2.26 ± 0.73 (SD), which agrees very well with the median LCT/LCN of 2.1 determined in the present paper. The present results and our enzyme kinetic stud ies cited above (Kapoor et aI., 1989) indicate that the LC in the pathological tissue, 36B-I0 rat glioma, is significantly higher than normal brain tissue. Therefore, application of the DG approach for mea suring accurately the glucose metabolic rate in this glioma requires using the LC determined for this pathological tissue. Since the calculated CMRglc in the DG approach is inversely proportional to the LC, determination of glucose utilization in 36B-I0 glioma using the normal brain rather than the true tumor LC overestimates the glucose metabolic rate by roughly a factor of 2. PET studies in humans with gliomas have been reported to show that grade of malignancy corre lates with higher CMRglc assessed with FDG (Di Chiro et aI., 1982, 1984; Kornblith et aI., 1984; Pa tronas et aI., 1985; Di Chiro, 1987; Alavi et aI., 1988; Di Chiro and Brooks, 1988a; Ogawa et aI., 1988) and that recurrence can be distinguished from radionecrosis (Patronas et aI., 1982; Doyle et aI., 1987). The former point has not been corroborated in all laboratories (Tyler et aI., 1987, 1988). In all of J Cereb Blood Flow Metab, Vol. 10, No.2, 1990
these human investigations, either the normal brain tissue LC was used to calculate the glioma CMRglc or images were analyzed nonquantitatively for "hot spots" indicative of foci of suspected malignant ac tivity (Patronas et aI., 1985; Doyle et aI., 1987; Di Chiro and Brooks, 1988a,b). Our rat data do not resolve the important questions raised by these hu man studies on quantitative FDG imaging of glio mas based on the normal brain LC or nonquantita tive "hot spot" imaging. However, they sharpen the questions and suggest the need for assessments of the LC in human gliomas for accurate quantita tion of CMRglc . Acknowledgment: The authors wish to thank Drs. James Bassingthwaighte and Finbar O'Sullivan for their critical review and suggestions. This work was supported by NIH grant, number CA 42045.
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