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GRP78/BiP/HSPA5/Dna K is a universal therapeutic target for human disease.

Booth L, Roberts JL, Cash DR, Tavallai S, Jean S, Fidanza A, Cruz-Luna T, Siembiba P, Cycon KA, Cornelissen CN, Dent P - J. Cell. Physiol. (2015)

Bottom Line: The GRP78 inhibitor OSU-03012 (AR-12) interacted with sildenafil (Viagra) or tadalafil (Cialis) to rapidly reduce GRP78 levels in eukaryotes and as a single agent reduce Dna K levels in prokaryotes.Pre-treatment with OSU-03012/sildenafil reduced expression of the coxsakie and adenovirus receptor in parallel with it also reducing the ability of a serotype 5 adenovirus or coxsakie virus B4 to infect and to reproduce.OSU-03012 as a single agent at clinically relevant concentrations killed laboratory generated antibiotic resistant E. coli and clinical isolate multi-drug resistant N. gonorrhoeae and MRSE which was in bacteria associated with reduced Dna K and Rec A expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298.

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OSU-03012 kills Neisseria gonorrhoeae bacteria. (A) Neisseria gonorrhoeae (FA19) were grown in liquid culture and treated with vehicle or with OSU-03012 (0–8 μM). The optical density of each treatment was determined 0–4h after treatment as indicated in each graphical presentation (n = 3 +/ − SEM) # P < 0.05 less than CDM value; $ P < 0.05 less than corresponding time = 0 density value. (B) From bacteria isolated during Panel A: Upper images: the morphology of the coccoid bactreia did not alter following OSU-03012 exposure. Lower western blot: OSU-03012 reduces expression of Dna K and Rec A but not GrpE in FA19 bacteria. (C) Neisseria gonorrhoeae (FA19) were grown in liquid culture and bacteria were treated with vehicle or with OSU-03012 (1.5–3.0 μM) as indicated in the presence or absence of sildenafil (4 μM) or tadalafil (4 μM). The optical density of each treatment and the bacterial protein content was determined after 0–240 min (n = 3 +/− SEM) # P < 0.05 less than value in CDM treated cells; $ P < 0.05 less than time = 0 value.
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fig08: OSU-03012 kills Neisseria gonorrhoeae bacteria. (A) Neisseria gonorrhoeae (FA19) were grown in liquid culture and treated with vehicle or with OSU-03012 (0–8 μM). The optical density of each treatment was determined 0–4h after treatment as indicated in each graphical presentation (n = 3 +/ − SEM) # P < 0.05 less than CDM value; $ P < 0.05 less than corresponding time = 0 density value. (B) From bacteria isolated during Panel A: Upper images: the morphology of the coccoid bactreia did not alter following OSU-03012 exposure. Lower western blot: OSU-03012 reduces expression of Dna K and Rec A but not GrpE in FA19 bacteria. (C) Neisseria gonorrhoeae (FA19) were grown in liquid culture and bacteria were treated with vehicle or with OSU-03012 (1.5–3.0 μM) as indicated in the presence or absence of sildenafil (4 μM) or tadalafil (4 μM). The optical density of each treatment and the bacterial protein content was determined after 0–240 min (n = 3 +/− SEM) # P < 0.05 less than value in CDM treated cells; $ P < 0.05 less than time = 0 value.

Mentions: Similar data to that obtained in E. coli with OSU-03012 were also found Neisseria gonorrhoeae strains FA19, FA1090, MS11 and 1291 (Fig. 8A, data not shown). OSU-03012 treatment did not appear to alter the morphology of these coccoid bacteria, though as before, we noted that OSU-03012 reduced expression of Dna K and Rec A (Fig. 8B). In FA19, OSU-03012 and PDE5 inhibitors could interact to cause a greater than additive amount of bacteria killing (Fig. 8C). Data in prior panels had shown that unlike the growth inhibitory antibiotic chloramphenicol, OSU-03012 exhibited true bacteriocidal effects on cells that had yet to enter log-phase growth. Finally, using F89 and HO41 drug resistant N. gonorrhoeae isolates we determined whether OSU-03012 exhibited any antibiotic effect and whether it could enhance/restore the bacteriocidal properties of approved standard of care antibiotics. Treatment of F89 and HO41 bacteria with OSU-03012 caused a dose-dependent reduction in bacterial growth (Fig. 9A). Treatment of F89 and HO41 bacteria with OSU-03012 variably enhanced the lethality of Ceftriaxone, Ciprofloxin, or Azithromycin over a 5 h time course (Figures 9B and 9 C). Methicillin-resistant Staphylococcus epidermidis (MRSE) bacteria were noted to be at least as sensitive to OSU-03012 as were N. gonorrhoeae (F89, HO41), possibly moreso i.e. 2 μM OSU-03012 was not growth inhibitory in F89 and HO41 but reduced MRSA growth by ∼50%, and co-exposure of bacteria to OSU-03012 and sildenafil modestly enhanced the cyto-toxic effect beyond that of OSU-03012 alone (Fig. 9D).


GRP78/BiP/HSPA5/Dna K is a universal therapeutic target for human disease.

Booth L, Roberts JL, Cash DR, Tavallai S, Jean S, Fidanza A, Cruz-Luna T, Siembiba P, Cycon KA, Cornelissen CN, Dent P - J. Cell. Physiol. (2015)

OSU-03012 kills Neisseria gonorrhoeae bacteria. (A) Neisseria gonorrhoeae (FA19) were grown in liquid culture and treated with vehicle or with OSU-03012 (0–8 μM). The optical density of each treatment was determined 0–4h after treatment as indicated in each graphical presentation (n = 3 +/ − SEM) # P < 0.05 less than CDM value; $ P < 0.05 less than corresponding time = 0 density value. (B) From bacteria isolated during Panel A: Upper images: the morphology of the coccoid bactreia did not alter following OSU-03012 exposure. Lower western blot: OSU-03012 reduces expression of Dna K and Rec A but not GrpE in FA19 bacteria. (C) Neisseria gonorrhoeae (FA19) were grown in liquid culture and bacteria were treated with vehicle or with OSU-03012 (1.5–3.0 μM) as indicated in the presence or absence of sildenafil (4 μM) or tadalafil (4 μM). The optical density of each treatment and the bacterial protein content was determined after 0–240 min (n = 3 +/− SEM) # P < 0.05 less than value in CDM treated cells; $ P < 0.05 less than time = 0 value.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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fig08: OSU-03012 kills Neisseria gonorrhoeae bacteria. (A) Neisseria gonorrhoeae (FA19) were grown in liquid culture and treated with vehicle or with OSU-03012 (0–8 μM). The optical density of each treatment was determined 0–4h after treatment as indicated in each graphical presentation (n = 3 +/ − SEM) # P < 0.05 less than CDM value; $ P < 0.05 less than corresponding time = 0 density value. (B) From bacteria isolated during Panel A: Upper images: the morphology of the coccoid bactreia did not alter following OSU-03012 exposure. Lower western blot: OSU-03012 reduces expression of Dna K and Rec A but not GrpE in FA19 bacteria. (C) Neisseria gonorrhoeae (FA19) were grown in liquid culture and bacteria were treated with vehicle or with OSU-03012 (1.5–3.0 μM) as indicated in the presence or absence of sildenafil (4 μM) or tadalafil (4 μM). The optical density of each treatment and the bacterial protein content was determined after 0–240 min (n = 3 +/− SEM) # P < 0.05 less than value in CDM treated cells; $ P < 0.05 less than time = 0 value.
Mentions: Similar data to that obtained in E. coli with OSU-03012 were also found Neisseria gonorrhoeae strains FA19, FA1090, MS11 and 1291 (Fig. 8A, data not shown). OSU-03012 treatment did not appear to alter the morphology of these coccoid bacteria, though as before, we noted that OSU-03012 reduced expression of Dna K and Rec A (Fig. 8B). In FA19, OSU-03012 and PDE5 inhibitors could interact to cause a greater than additive amount of bacteria killing (Fig. 8C). Data in prior panels had shown that unlike the growth inhibitory antibiotic chloramphenicol, OSU-03012 exhibited true bacteriocidal effects on cells that had yet to enter log-phase growth. Finally, using F89 and HO41 drug resistant N. gonorrhoeae isolates we determined whether OSU-03012 exhibited any antibiotic effect and whether it could enhance/restore the bacteriocidal properties of approved standard of care antibiotics. Treatment of F89 and HO41 bacteria with OSU-03012 caused a dose-dependent reduction in bacterial growth (Fig. 9A). Treatment of F89 and HO41 bacteria with OSU-03012 variably enhanced the lethality of Ceftriaxone, Ciprofloxin, or Azithromycin over a 5 h time course (Figures 9B and 9 C). Methicillin-resistant Staphylococcus epidermidis (MRSE) bacteria were noted to be at least as sensitive to OSU-03012 as were N. gonorrhoeae (F89, HO41), possibly moreso i.e. 2 μM OSU-03012 was not growth inhibitory in F89 and HO41 but reduced MRSA growth by ∼50%, and co-exposure of bacteria to OSU-03012 and sildenafil modestly enhanced the cyto-toxic effect beyond that of OSU-03012 alone (Fig. 9D).

Bottom Line: The GRP78 inhibitor OSU-03012 (AR-12) interacted with sildenafil (Viagra) or tadalafil (Cialis) to rapidly reduce GRP78 levels in eukaryotes and as a single agent reduce Dna K levels in prokaryotes.Pre-treatment with OSU-03012/sildenafil reduced expression of the coxsakie and adenovirus receptor in parallel with it also reducing the ability of a serotype 5 adenovirus or coxsakie virus B4 to infect and to reproduce.OSU-03012 as a single agent at clinically relevant concentrations killed laboratory generated antibiotic resistant E. coli and clinical isolate multi-drug resistant N. gonorrhoeae and MRSE which was in bacteria associated with reduced Dna K and Rec A expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA 23298.

Show MeSH
Related in: MedlinePlus