Phenotypic Screening for Small Molecules that Protect β-Cells from Glucolipotoxicity

Abstract

Type 2 diabetes is marked by progressive β-cell failure, leading to loss of β-cell mass. Increased levels of circulating glucose and free fatty acids associated with obesity lead to β-cell glucolipotoxicity. There are currently no therapeutic options to address this facet of β-cell loss in obese type 2 diabetes patients. To identify small molecules capable of protecting β-cells, we performed a high-throughput screen of 20,876 compounds in the rat insulinoma cell line INS-1E in the presence of elevated glucose and palmitate. We found 312 glucolipotoxicity-protective small molecules (1.49% hit rate) capable of restoring INS-1E viability, and we focused on 17 with known biological targets. 16 of the 17 compounds were kinase inhibitors with activity against specific families including but not limited to cyclin-dependent kinases (CDK), PI-3 kinase (PI3K), Janus kinase (JAK), and Rho-associated kinase 2 (ROCK2). 7 of the 16 kinase inhibitors were PI3K inhibitors. Validation studies in dissociated human islets identified 10 of the 17 compounds, namely, KD025, ETP-45658, BMS-536924, AT-9283, PF-03814735, torin-2, AZD5438, CP-640186, ETP-46464, and GSK2126458 that reduced glucolipotoxicity-induced β-cell death. These 10 compounds decreased markers of glucolipotoxicity including caspase activation, mitochondrial depolarization, and increased calcium flux. Together, these results provide a path forward toward identifying novel treatments to preserve β-cell viability in the face of glucolipotoxicity.

Pancreatic Beta-cell Dysfunction in Type 2 Diabetes

“…Among them, KD025, ETP-45,658, BMS-536,924, AT-9283, PF-03,814,735, torin-2, AZD5438, CP-640,186, ETP-46,464, and GSK2126458 were observed to decrease the markers of glucolipotoxicity including caspase activation and mitochondrial depolarization. 68 Thus, in future, use of phenotypic screening could become a time saving method to identify the novel compounds and molecules to study changes in β-cells.…”

PSL Chemical Biology Symposia Third Edition: A Branch of Science in its Explosive Phase

“…To identify small molecules able to protect beta cells, they adapted this assay to high‐throughput screening of 384 K compounds. This led to the discovery of a molecule named BRD5653 which resulted in high luciferase activity in T2D human islets even when glucose concentrations are high [53] . However, studies on the mechanism of action did not reveal the target of BRD5653.…”

“…This led to the discovery of a molecule named BRD5653 which resulted in high luciferase activity in T2D human islets even when glucose concentrations are high. [53] However, studies on the mechanism of action did not reveal the target of BRD5653. To identify its target, a genome-wide CRISPR screen revealed several receptors as being involved in the regulation of glucose blood concentrations.…”

Búsqueda de nuevos fármacos antihelmínticos basada en el análisis estructura-función de proteínas que unen ácidos grasos (FABPs) de cestodos mediante técnicas bioinformáticas y quimioinformáticas

“…Estos métodos han evolucionado desde el tamizaje de baja capacidad (que examina sólo un pequeño conjunto de compuestos) hasta las estrategias de HTS ya mencionadas, que permiten evaluar más de 100.000 compuestos en solo un día (Hertzberg & Pope, 2000). Estos métodos representan una forma interesante de buscar nuevas oportunidades de reposicionamiento, ya que, además de evaluar el impacto en un objetivo molecular, hoy en día es posible adaptar los métodos al tamizaje fenotípico y, de esta manera, evaluar los medicamentos en sistemas complejos (Blay et al, 2020;Brito et al, 2020;Small et al, 2022).…”