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Pseudomonas aeruginosa infection causes endothelial cell Tau aggregation: evidence for an endothelial tauopathy (1176.5)
Author(s) -
Morrow Kyle,
Ochoa Cristhiaan,
Alexeyev Mikhail,
Frank Dara,
Balczon Ronald,
Stevens Troy
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1176.5
Subject(s) - tauopathy , microbiology and biotechnology , matrigel , tau protein , biology , mutant , extracellular , chemistry , angiogenesis , cancer research , neurodegeneration , biochemistry , alzheimer's disease , medicine , disease , pathology , gene
Our laboratory recently demonstrated that Pseudomonas aeruginosa exoenzyme Y (ExoY) stimulates Tau hyperphosphorylation and insolubility ‐ hallmarks of a “Tauopathy”. Unlike neurodegenerative Tauopathies, we did not observe formation of Tau aggregates indicating that other pathogenic factors may be required to elicit a full spectrum Tauopathy. Thus, we tested whether P. aeruginosa ExoU induces Tau aggregation, causing an acute infectious Tauopathy. Pulmonary microvascular endothelial cells (PMVECs) were engineered to conditionally (Tet‐On) express codon‐optimized activity‐attenuated mutant ExoU encoding both myc‐ and protein degradation domain‐epitopes. Multiple strains of P. aeruginosa have been generated that allow systematic investigation of exoenzyme function. The background bacterial strain is PA103, a clinically relevant strain that possesses ExoU. We also used a strain that introduces only ExoY (ExoY+) to infect PMVECs. PMVEC gap formation was induced in a time‐dependent manner following ExoY+ infection as well as by doxycycline‐induced ExoU expression or PA103 infection. Cell extracts and supernatants were collected and immunoblotted with pan‐Tau and Tau oligomer‐specific antibodies. Gap formation was paralleled by a loss in intracellular Tau expression and hyperphosphorylation, with an increase in extracellular Tau oligomers. ExoU induces endothelial cell Tau release and aggregation, suggesting P. aeruginosa causes an endothelial Tauopathy. Grant Funding Source : HL66299, HL60024