Effects of crenolanib, a nonselective inhibitor of PDGFR, in a mouse model of transient middle cerebral artery occlusion

Abstract

Neurogenesis in the subventricular zone (SVZ) plays a vital role in neurologic recovery after stroke. However, only a small fraction of newly generated neuroblasts from the SVZ will survive long-term. Successful migration and survival of neuroblasts requires angiogenesis, lesion-derived chemo-attractants, and appropriate local microenvironments, which are partly regulated by the platelet-derived growth factor receptor (PDGFR) signaling pathway. In this study, we investigated the effects of PDGFR inhibition in a mouse model of transient middle cerebral artery occlusion (MCAO). We blocked the pathway using a selective pan-PDGFR inhibitor, crenolanib, during the acute post-MCAO phase (days 1–3) or during the sub-acute phase (days 7–9). Downregulating the PDGFR signaling pathway with crenolanib from day 1 to day 3 after MCAO significantly decreased the migration of neuroblasts from the SVZ to the peri-infarct region, decreased angiogenesis, and lowered expression of vascular endothelial growth factor, stromal cell-derived factor-1, and monocyte chemotactic protein-1. Downregulation of the PDGFR signaling pathway on days 7 to 9 with crenolanib significantly increased apoptosis of the neuroblasts that had migrated to the peri-infarct region, increased the number of activated microglia, and decreased the expression of brain-derived neurotrophic factor, neurotrophin-3, and interleukin-10. Crenolanib treatment increased the apoptosis of pericytes and decreased the pericyte/vascular coverage, but had no effects on apoptosis of astrocytes. We conclude that PDGFR signaling pathway plays a vital role in the SVZ neurogenesis after stroke, it can also affect angiogenesis, lesion-derived chemo-attractants and local microenvironment, which are of importance in the stroke-induced neurogenesis.

Carbon monoxide-releasing molecule-3 protects against ischemic stroke by suppressing neuroinflammation and alleviating blood-brain barrier disruption

“…We subjected mice to the tMCAO model of ischemic stroke as previously described [ 29 – 31 ]. We briefly anesthetized each mouse with an intraperitoneal injection of 5% chloral hydrate, made a midline neck incision, and carefully separated the arteries from adjacent tissues.…”

“…We sacrificed mice on day 3 after surgery for immunofluorescence analysis as previously described [ 30 , 31 , 39 ]. Briefly, mice were anesthetized with an intraperitoneal injection of 5% chloral hydrate and perfused with PBS and 4% paraformaldehyde.…”

“…We sacrificed the mice on day 3 after surgery for Western blot analysis as previously described [ 31 , 40 ]. Protein was extracted from the whole right hemisphere, separated on 6–12% glycine gels, and transferred onto polyvinylidene difluoride membranes (Millipore, Bedford, MA).…”

“…We sacrificed the mice on day 3 post-surgery and prepared the brain homogenates for ELISA as previously described [ 31 ]. We homogenized the right hemisphere using a protein extraction kit (Sangon Biotech, Shanghai, China) and measured protein concentration with a bicinchoninic acid protein assay kit (Sangon Biotech).…”

A Human Multi-Lineage Hepatic Organoid Model for Liver Fibrosis

“…To determine if the pathways identified by our transcriptomic analyses were essential to the pathogenesis of ARPKD liver fibrosis, we examined the effect that three clinically used inhibitors of the PDGFR signaling (Crenolanib 68,69 , Sunitinib 70,71 and Imatinib 72,73 ) and a γ-secretase inhibitor that is commonly used to block NOTCH signaling (DAPT 74 ) had on the extent of ARPKD organoid fibrosis. Three different methods were used to assess collagen abundance in ARPKD organoids: quantitative collagen immunostaining in whole organoids, and measurement of hydroxyproline and collagen mRNA levels.…”