Quantification of lactate metabolism with [11C]lactate

The available repertoire of PET tracers enables quantification of glucose, fatty acid, and amino-acid metabolism, but the measurement of lactate/lactic acid (2-hydroxypropanoate) metabolism in humans has not been performed directly. Lactate however is an important substrate for most tissues, including heart and brain, and altered lactate metabolism has been associated with insulin resistance and cancer. Furthermore, monocarboxylate transporters (Halestrap, 2013), which allow lactate to pass cell membranes, may be potential targets for diagnostics and drugs.

Biochemistry of lactate

lactic acid

Lactate exists in the body as two stereoisomers, L-lactate and D-lactate. L-lactate dehydrogenase (LDH) catalyses reactions where L-lactate is formed from pyruvate while NADH is oxidized to NAD+, and vice versa. Glucose (via glycolysis) and L-alanine are the main sources of pyruvate. L-lactate can be concerted back to glucose via gluconeogenesis. Oxidative decarboxylation of pyruvate to acetyl-CoA and CO2 is irreversible reaction, catalysed by pyruvate dehydrogenase (PDH) complex in the mitochondria. Acetate (from acetyl-CoA) is then consumed in the tricarboxylic acid (TCA) cycle forming CO2.

D-lactate is endogenously formed from methylglyoxal through glyoxylase system and as byproduct of glycolysis. Some foods (sour milk products, apples, tomatoes) are also sources of D-lactate. D-lactate is efficiently removed from the body via oxidation to pyruvate, and the concentrations of D-lactate are small compared to L-lactate.

Lactate transport across plasma membranes requires either proton-coupled (MCTs, SLC16 family) or sodium-coupled monocarboxylate transporters (SMCTs, SLC5 family). Nonionic diffusion accounts for only about 5% of total lactate transport across erythrocyte membranes (De Bruijne et al, 1983). These transporter families also transport other substrates than lactate such as pyruvate, acetate, acetoacetate, and β-hydroxybutyrate. Transporter subtypes show somewhat different affinities towards the substrates, and their expression is organ-specific.

Lactate metabolism

The brain

The brain shows normally a small net release of L-lactate. During intense physical exercise and increased blood lactate levels the brain starts to extract more L-lactate than it releases. Most of the extracted L-lactate is oxidised; at rest L-lactate accounts for about 8% of cerebral energy requirements, but during intense exercise it may increase to 60%. In the brain, astrocytes mainly consume glucose, releasing lactate, which is the main fuel for the neurons (Magistretti and Allaman, 2015).

In rat cerebral cortex using L-1-[11C]lactate and [18F]FDG, it was shown that lactate is readily oxidized in brain activity dependent manner. Increasing plasma lactate concentration resulted in the reduction of cerebral glucose utilization (Wyss et al, 2011).

Skeletal muscle

Muscle can switch quickly from a net lactate producer to a net lactate consumer, depending on the plasma lactate concentration. Resting skeletal muscle in postabsorbtive state release more L-lactate than is taken up, providing about 40% of the total L-lactate released into circulation. L-lactate concentration is higher in the muscle tissue than in the blood (3 mM vs 1.4 mM, respectively). During exercise, L-lactate release and extraction are both increased. Low and moderate exercise does not increase L-lactate concentration in the plasma; the work rate where lactate concentration starts to rise is referred to as the lactate threshold. As plasma lactate levels increase, the non-exercising muscles take up more lactate than they release.

Myocardial muscle

During rest, heart muscle extracts more L-lactate than it releases. Most but not all of the extracted L-lactate is oxidized. L-lactate uptake correlates positively with plasma lactate concentration, and negatively with concentration of free fatty acids.

Liver and kidney

In postabsorbtive state liver and kidneys consume L-lactate, converting it mainly to glucose. Insulin decreases lactate uptake and glucose production. Lactate uptake is increased during exercise in the liver.

The gut may be a substantial L-lactate net producer, but the liver extracts most of it before it can reach the main circulation.

Adipose tissue

Adipose tissue is a net producer of L-lactate, especially after glucose or insulin challenge. In diabetic and obese subjects the lactate production by adipose tissue is higher than in lean subjects with normal insulin sensitivity in the basal state, but the insulin-induced increase in lactate production is attenuated.

Blood cells

White blood cells, platelets, and erythrocytes are net producers of L-lactate. Erythrocytes lack the mitochondria and can produce ATP only by the non-oxidative glycolytic pathway, with L-lactate as the end product. Lactate transport of erythrocytes varies between species, being high in humans and dogs.

Lactate in blood plasma

L-Lactate concentration is healthy subjects is generally less than 2 mM, but during physical exercise can increase to 10 mM. Fasting plasma lactate concentration is higher in patients with type 1 and type 2 diabetes and in obese subjects (hyperlactatemia) compared to subjects who are lean or have normal insulin sensitivity, mainly caused by increased lactate production in adipose tissue. Glucose challenge and insulin increase plasma lactate concentration, but less so in insulin resistant subjects.

Lactic acidosis is diagnosed when plasma L-lactate concentration is > 5 mM and blood pH is < 7.35.

D-lactate concentrations in plasma and urine are higher in diabetic than in normal subjects because of increased production of methylglyoxal.

PET tracers

Lactate has been labelled in the 1- (carboxylic) and 3-position. If L-lactate is labelled in the C-1 position (L-1-[11C]lactate), the 11C label is released as [11C]CO2 already when pyruvate is converted to acetyl-CoA by PDH complex in mitochondria (Wyss et al., 2011). But if L-lactate is labeled in the C-3 position (L-3-[11C]lactate), the 11C label follows acetyl-CoA into the TCA cycle, where it finally is released as [11C]CO2 (Herrero et al., 2007), but during PET studies most of the 11C label will be attached to other metabolites of the TCA cycle.

Lactate, labeled into C-1 and C-3 position

Figure 1. L-1-[11C]lactate and L-3-[11C]lactate, labeled into C-1 and C-3 position, respectively.

L-1-[11C]lactate

Wyss et al (2011) used L-1-[11C]lactate to study L-lactate oxidation in the brain of rats, using beta-probe system instead of PET. Data was analyzed using a one-tissue compartmental model including parameter for vascular volume. Uptake rate of L-lactate can then be calculated as the product of K1 and plasma lactate concentration. Rate constant k2 represents the back-diffusion of labeled lactate to blood and the release of [11C]CO2. It can be assumed that LDH, not PDH, is the rate-limiting step for the release of [11C]CO2 in the rat brain (Wyss et al., 2011).

Labeled metabolites in plasma

The main label-carrying metabolite in rat blood is [11C]CO2, when L-lactate is labeled in C-1 position (van Hall et al, 2009; Wyss et al, 2011).

L-3-[11C]lactate

Herrero et al (2007) showed in a dog study that under conditions of net lactate extraction, L-3-[11C]lactate depicts myocardial metabolism of exogenous lactate and that measurements of lactate metabolism are feasible with PET using monoexponential clearance analysis (kmono representing L-lactate oxidation), or applying compartmental model with two compartments (vascular and extravascular/cytosolic) with four rate constants and vascular volume fixed to 0.08, providing estimates of L-lactate extraction, back-diffusion, and oxidation.

Labeled metabolites in plasma

In the dog study (Herrero et al, 2007), the fraction of parent tracer (including some labeled pyruvate which could not be separated from lactate in the applied method) stayed on a relatively high level (about 40%) because of lactate back-diffusion. The fractions of neutral metabolites (mainly glucose, possibly glycerol) and [11C]CO2 increased by time, while the fraction of basic metabolites (mainly alanine, possibly other amino acids) was relatively stable (usually less than 10%) during the 60 min study. Interventions had high impact on the plasma fractions.



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Created at: 2015-05-28
Updated at: 2017-06-28
Written by: Vesa Oikonen