Identification of Glucose Transport Modulators In Vitro and Method for Their Deep Learning Neural Network Behavioral Evaluation in Glucose Transporter 1–Deficient Mice

Abstract

Metabolic flux augmentation via glucose transport activation may be desirable in Glucose transporter 1 (Glut1) deficiency (G1D) and dementia, whereas suppression might prove useful in cancer. Using lung adenocarcinoma cells that predominantly express Glut1 relative to other glucose transporters, we screened 11,613 compounds for effects on the accumulation of an extracellularly-applied fluorescent glucose analog. 5 drugs currently prescribed for unrelated indications or preclinically-characterized robustly enhanced intracellular fluorescence.Additionally identified were 37 novel activating and 9 inhibitory compounds lacking previous biological characterization. Because few glucose-related mechanistic or pharmacological studies were available for these compounds, we developed a method to quantify G1D mouse behavior to infer potential therapeutic value. To this end, we designed a 5-track apparatus to record and evaluate spontaneous locomotion videos. We applied this to a G1D mouse model that replicates the ataxia and other manifestations cardinal to the human disorder. Because the first two drugs that we examined in this manner (baclofen and acetazolamide) exerted various impacts on several gait aspects, we used deep learning neural networks to more comprehensively assess drug effects. Using this method, 49 locomotor parameters differentiated G1D from control mice. Thus, we used parameter modifiability to quantify efficacy on gait. We tested this by measuring the effects of saline as control and glucose as G1D therapy. The results indicate that this in vivo approach can estimate preclinical suitability from the perspective of G1D locomotion. This justifies the use of this method to evaluate our drugs or other interventions and sort candidates for further investigation.

A concise study of acetazolamide in glucose transporter type 1 deficiency (G1D) epilepsy

“…Although ACZ may act similarly to other carbonic anhydrase inhibitors via enhancement of γ‐aminobutyric acid (GABA)–mediated inhibition, 5 it is unlikely that this mechanism will be the sole one at play in G1D. In fact, other potential actions include enhancement of cellular glucose flux 6 . In addition, the biological G1D context extends to additional manifestation‐related or therapeutically modulable unsuspected loci such as red blood cell Glut1 content, 18 which might also be susceptible to ACZ.…”

“…In fact, other potential actions include enhancement of cellular glucose flux. 6 In addition, the biological G1D context extends to additional manifestation-related or therapeutically modulable unsuspected loci such as red blood cell Glut1 content, 18 which might also be susceptible to ACZ.…”

“…However, this does not necessarily imply that ACZ exerts a similar effect on brain cell glucose transport. In lieu of this, ACZ displayed activity on G1D mouse locomotion at doses commensurate with human pharmacological use, suggesting neural impact 6 and thus potential utility worth exploring in individuals with G1D.…”

Isolation of the murine Glut1 deficient thalamocortical circuit: wavelet characterization and reverse glucose dependence of low and gamma frequency oscillations

“…This limits the value of EEG to elucidate mechanisms without resorting to source localization estimates (Rajasekaran et al, 2022b). Fifth, despite a traditional paucity of glucose transporter activators, an array of commercialized drugs and other poorly biologically characterized compounds robustly enhance glucose transport into cancer cells rich in Glut1 (Kathote et al, 2023), enabling the investigation of glucose influx-enhancing therapies. These considerations provide the motivation to develop an ex vivo circuit preparation that faithfully replicates a known neurophysiological G1D phenotype as a testbed for investigating treatments.…”

Isolation of the murine Glut1 deficient thalamocortical circuit: wavelet characterization and reverse glucose dependence of low and gamma frequency oscillations

“…This limits the value of EEG to elucidate mechanisms without resorting to source localization estimates ( Rajasekaran et al, 2022 ). Fifth, despite a traditional paucity of glucose transporter activators, an array of commercialized drugs and other poorly biologically characterized compounds robustly enhance glucose transport into cancer cells rich in Glut1 ( Kathote et al, 2023 ), enabling the investigation of glucose influx-enhancing therapies. These considerations provide the motivation to develop an ex vivo circuit preparation that faithfully replicates a known neurophysiological G1D phenotype as a testbed for investigating treatments.…”