Kathote Gauri, Ma Qian, Angulo Gustavo, Chen Hong, Jakkamsetti Vikram, Dobariya Aksharkumar, Good Levi B, Posner Bruce, Park Jason Y, Pascual Juan M
Rare Brain Disorders Program, Department of Neurology (G.K., Q.M., G.A., V.J., A.D., L.B.G., J.M.P.), Department of Biochemistry (H.C., B.P.), Department of Pathology (J.Y.P.), Department of Physiology (J.M.P.), Department of Pediatrics (J.M.P.), and Eugene McDermott Center for Human Growth & Development/Center for Human Genetics (J.Y.P., J.M.P.), University of Texas Southwestern Medical Center, Dallas, Texas.
J Pharmacol Exp Ther. 2023 Mar;384(3):393-405. doi: 10.1124/jpet.122.001428. Epub 2023 Jan 12.
Metabolic flux augmentation via glucose transport activation may be desirable in glucose transporter 1 (Glut1) deficiency syndrome (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 9,646 compounds for effects on the accumulation of an extracellularly applied fluorescent glucose analog. Five drugs currently prescribed for unrelated indications or preclinically characterized robustly enhanced intracellular fluorescence. Additionally identified were 37 novel activating and nine inhibitory compounds lacking previous biologic 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 five-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. SIGNIFICANCE STATEMENT: There are few or no activators and few clinical inhibitors of glucose transport. Using Glut1-rich cells exposed to a glucose analog, we identified, in highthroughput fashion, a series of novel modulators. Some were drugs used to modify unrelated processes and some represented large but little studied chemical compound families. To facilitate their preclinical efficacy characterization regardless of potential mechanism of action, we developed a gait testing platform for deep learning neural network analysis of drug impact on Glut1-deficient mouse locomotion.
通过激活葡萄糖转运来增强代谢通量在葡萄糖转运蛋白1（Glut1）缺乏综合征（G1D）和痴呆症中可能是有益的，而在癌症中抑制代谢通量可能被证明是有用的。我们使用相对于其他葡萄糖转运蛋白主要表达Glut1的肺腺癌细胞，筛选了9646种化合物，以研究其对细胞外应用的荧光葡萄糖类似物积累的影响。目前用于其他无关适应症或临床前已充分表征的五种药物显著增强了细胞内荧光。此外，还鉴定出37种新型激活化合物和9种缺乏先前生物学表征的抑制化合物。由于针对这些化合物的葡萄糖相关机制或药理学研究很少，我们开发了一种量化G1D小鼠行为的方法，以推断其潜在治疗价值。为此，我们设计了一种五轨道装置来记录和评估自发运动视频。我们将此应用于一个G1D小鼠模型，该模型复制了人类疾病的共济失调和其他主要表现。由于我们以这种方式研究的前两种药物（巴氯芬和乙酰唑胺）对几个步态方面产生了不同影响，我们使用深度学习神经网络更全面地评估药物效果。使用这种方法，49个运动参数区分了G1D小鼠和对照小鼠。因此，我们使用参数可变性来量化对步态的疗效。我们通过测量生理盐水作为对照和葡萄糖作为G1D治疗的效果来对此进行测试。结果表明，这种体内方法可以从G1D运动的角度估计临床前适用性。这证明了使用这种方法来评估我们的药物或其他干预措施，并对候选药物进行进一步研究的合理性。意义声明：葡萄糖转运的激活剂很少或没有，临床抑制剂也很少。使用暴露于葡萄糖类似物的富含Glut1的细胞，我们以高通量方式鉴定了一系列新型调节剂。其中一些是用于改变无关过程的药物，一些代表了庞大但研究较少的化合物家族。为了便于对它们进行临床前疗效表征，而不考虑其潜在作用机制，我们开发了一个步态测试平台，用于对药物对Glut1缺陷小鼠运动的影响进行深度学习神经网络分析。
