Inhibition of Src kinase activity attenuates amyloid associated microgliosis in a murine model of Alzheimer's disease.
BACKGROUND
Microglial activation is an important histologic characteristic of the pathology of Alzheimer's disease (AD). One hypothesis is that amyloid beta (Aβ) peptide serves as a specific stimulus for tyrosine kinase-based microglial activation leading to pro-inflammatory changes that contribute to disease. Therefore, inhibiting Aβ stimulation of microglia may prove to be an important therapeutic strategy for AD.


METHODS
Primary murine microglia cultures and the murine microglia cell line, BV2, were used for stimulation with fibrillar Aβ1-42. The non-receptor tyrosine kinase inhibitor, dasatinib, was used to treat the cells to determine whether Src family kinase activity was required for the Aβ stimulated signaling response and subsequent increase in TNFα secretion using Western blot analysis and enzyme-linked immunosorbent assay (ELISA), respectively. A histologic longitudinal analysis was performed using an AD transgenic mouse model, APP/PS1, to determine an age at which microglial protein tyrosine kinase levels increased in order to administer dasatinib via mini osmotic pump diffusion. Effects of dasatinib administration on microglial and astroglial activation, protein phosphotyrosine levels, active Src kinase levels, Aβ plaque deposition, and spatial working memory were assessed via immunohistochemistry, Western blot, and T maze analysis.


RESULTS
Aβ fibrils stimulated primary murine microglia via a tyrosine kinase pathway involving Src kinase that was attenuated by dasatinib. Dasatinib administration to APP/PS1 mice decreased protein phosphotyrosine, active Src, reactive microglia, and TNFα levels in the hippocampus and temporal cortex. The drug had no effect on GFAP levels, Aβ plaque load, or the related tyrosine kinase, Lyn. These anti-inflammatory changes correlated with improved performance on the T maze test in dasatinib infused animals compared to control animals.


CONCLUSIONS
These data suggest that amyloid dependent microgliosis may be Src kinase dependent in vitro and in vivo. This study defines a role for Src kinase in the microgliosis characteristic of diseased brains and suggests that particular tyrosine kinase inhibition may be a valid anti-inflammatory approach to disease. Dasatinib is an FDA-approved drug for treating chronic myeloid leukemia cancer with a reported ability to cross the blood-brain barrier. Therefore, this suggests a novel use for this drug as well as similar acting molecules.
School of Medicine and Health Sciences, 504 Hamline St, Room 118, Grand Forks, ND 58203, USA.
Gunjan
G
Dhawan
Colin K
CK
Combs
D000544:Alzheimer Disease / Q000188:drug therapy / Q000201:enzymology*; D016229:Amyloid beta-Peptides / Q000008:administration & dosage* / Q000633:toxicity; D000818:Animals; D000831:Animals, Newborn; D002460:Cell Line; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D000069439:Dasatinib; D004195:Disease Models, Animal; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D008137:Longitudinal Studies; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D017628:Microglia / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology*; D010446:Peptide Fragments / Q000008:administration & dosage* / Q000633:toxicity; D047428:Protein Kinase Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D011743:Pyrimidines / Q000494:pharmacology / Q000627:therapeutic use; D013844:Thiazoles / Q000494:pharmacology / Q000627:therapeutic use; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000502:physiology*
D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D013844:Thiazoles; C075222:amyloid beta-protein (1-42); D019061:src-Family Kinases; D000069439:Dasatinib