NOT ALL ANTIBIOTICS PULSED DOSED ERADICATE BIOFILM LIKE BORRELIA

Ceftriaxone Pulse Dosing Fails to Eradicate Biofilm-like Microcolony B. burgdorferi Persisters Which Are Sterilized by Daptomycin/Doxycycline/Cefuroxime Drug Combination without Pulse Dosing

Jie Feng1, Shuo Zhang1, Wanliang Shi1 and Ying Zhang1*

• 1Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, USA

Although the majority of Lyme disease patients can be cured, at least 10-20% of the patients continue to suffer from persisting symptoms such as fatigue, muscular and joint pain, and neurologic impairment after standard 2-4 week antibiotic treatment. While the causes for this post-treatment Lyme disease symptoms are unclear, one possibility is due to B. burgdorferi persisters that are not effectively killed by current antibiotics such as doxycycline or amoxicillin used to treat Lyme disease. A previous study showed that four rounds of ceftriaxone pulse dosing treatment eradicated B. burgdorferi persisters in vitro using a relatively young late log phase culture (5 day old). In this study, we investigated if ceftriaxone pulse dosing could also eradicate B. burgdorferi persisters in older stationary phase cultures (10 day old) enriched with more resistant microcolony form of persisters. We found that ceftriaxone pulse dosing could only eradicate planktonic log phase B. burgdorferi spirochetal forms and round body forms but not more resistant aggregated biofilm-like microcolony persisters enriched in stationary phase cultures. Moreover, we found that not all drugs are suitable for pulse dosing, with bactericidal drugs ceftriaxone and cefuroxime being more appropriate for pulse dosing than bacteriostatic drug doxycycline and persister drug daptomycin. We also showed that drug combination pulse dosing treatment was more effective than single drug pulse dosing. Importantly, we demonstrate that pulse dosing treatment impaired the activity of the persister drug daptomycin and its drug combination against B. burgdorferi persisters and that the most effective way to kill the more resistant biofilm-like microcolonies is the daptomycin/doxycycline/ceftriaxone triple drug combination without pulse dosing. Our findings indicate pulse dosing may not always work as a general principle but rather depends on the specific drugs used, with cidal drugs being more appropriate for pulse dosing than static or persister drugs, and that drug combination approach with persister drugs is more effective at killing the more resistant microcolony form of persisters than pulse dosing. These observations may have implications for more effective treatment of Lyme disease. Future studies are required to validate these findings in animal models of B. burgdorferi persistence.

http://journal.frontiersin.org/article/10.3389/fmicb.2016.01744/abstract

DRUG ACTIVITIES AGAINST BORRELIA PERSISTERS - LYME DISEASE PERSISTERS

A Drug Combination Screen Identifies Drugs Active against Amoxicillin-induced Round Bodies of Borrelia burgdorferi Persisters from an FDA Drug Library

Jie Feng1, Wanliang Shi1, Shuo Zhang1, David Sullivan1, Paul Auwaerter2 and Ying Zhang1*

• 1Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, USA
• 2Fisher Center for Environmental Infectious Diseases, School of Medicine, Johns Hopkins University, USA

Although currently recommended antibiotics for Lyme disease such as doxycycline or amoxicillin cure the majority of the patients, about 10-20% of patients treated for Lyme disease may experience lingering symptoms including fatigue, pain, or joint and muscle aches. Under stress conditions such as starvation or antibiotic exposure, Borrelia burgdorferi can develop round body forms, which are a type of persister bacteria that are not killed by current Lyme antibiotics. To identify more effective drugs that are active against the round bodies of B. burgdorferi, we established a round body persister model induced by amoxicillin and screened the Food and Drug Administration (FDA) drug library consisting of 1581 drug compounds and also 22 drug combinations using the SYBR Green I/propidium iodide (PI) viability assay. We identified 23 drug candidates that have higher activity against the round bodies of B. burgdorferi than either amoxicillin or doxycycline. Eleven of these scored better than metronidazole and tinidazole which have been previously described to be active against round bodies. While some drug candidates such as daptomycin and clofazimine overlapped with a previous screen against stationary phase B. burgdorferi persisters, additional drug candidates active against round bodies we identified include artemisinin, ciprofloxacin, nifuroxime, fosfomycin, chlortetracycline, sulfacetamide, sulfamethoxypyridazine and sulfathiozole. Two triple drug combinations had the highest activity against round bodies and stationary phase B. burgdorferi persisters: artemisinin/cefoperazone/doxycycline and sulfachlorpyridazine/daptomycin/doxycycline. These findings confirm and extend previous findings that certain drug combinations have superior activity against B. burgdorferi persisters in vitro, even if pre-treated with amoxicillin. These findings may have implications for improved treatment of Lyme disease.

http://journal.frontiersin.org/article/10.3389/fmicb.2016.00743/full 

Links into earlier posts on Prof Zhang research can be found:- 

http://lookingatlyme.blogspot.co.uk/2015/10/lyme-disease-persister-drugs-dr-ying.html

FDA APPROVED DRUGS FOR PERSISTER CELLS - LYME DISEASE- FROM JOHNS HOPKINS

Subject Category: Antimicrobials and drug resistance

Citation: Emerging Microbes & Infections (2014) 3, e49; doi:10.1038/emi.2014.53
Published online 2 July 2014

Identification of novel activity against Borrelia burgdorferi persisters using an FDA approved drug library

OPEN

Jie Feng¹, Ting Wang¹, Wanliang Shi¹, Shuo Zhang¹, David Sullivan¹, Paul G Auwaerter² and Ying Zhang¹

¹Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA

1. ²Fisher Center for Environmental Infectious Diseases, School of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA

Correspondence: Y Zhang, E-mail: yzhang@jhsph.edu

Received 17 April 2014; Revised 30 May 2014; Accepted 3 June 2014

Link to the article here and interesting table of activity against Borrelia burgdorferi persisters here

ABSTRACT

Although antibiotic treatment for Lyme disease is effective in the majority of cases, especially during the early phase of the disease, a minority of patients suffer from post-treatment Lyme disease syndrome (PTLDS). It is unclear what mechanisms drive this problem, and although slow or ineffective killing of Borrelia burgdorferi has been suggested as an explanation, there is a lack of evidence that viable organisms are present in PTLDS. Although not a clinical surrogate, insight may be gained by examining stationary-phase in vitro Borrelia burgdorferi persisters that survive treatment with the antibiotics doxycycline and amoxicillin. To identify drug candidates that can eliminate B. burgdorferi persisters more effectively, we screened an Food and Drug Administration (FDA)-approved drug library consisting of 1524 compounds against stationary-phase B. burgdorferi by using a newly developed high throughput SYBR Green I/propidium iodide (PI) assay. We identified 165 agents approved for use in other disease conditions that had more activity than doxycycline and amoxicillin against B. burgdorferi persisters. The top 27 drug candidates from the 165 hits were confirmed to have higher anti-persister activity than the current frontline antibiotics. Among the top 27 confirmed drug candidates from the 165 hits, daptomycin, clofazimine, carbomycin, sulfa drugs (e.g., sulfamethoxazole), and certain cephalosporins (e.g. cefoperazone) had the highest anti-persister activity. In addition, some drug candidates, such as daptomycin and clofazimine (which had the highest activity against non-growing persisters), had relatively poor activity or a high minimal inhibitory concentration (MIC) against growing B. burgdorferi. Our findings may have implications for the development of a more effective treatment for Lyme disease and for the relief of long-term symptoms that afflict some Lyme disease patients.

Keywords: 

Borrelia burgdorferi; drug discovery; FDA approved drug library; persisters; SYBR Green I

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Interesting discussion on this important paper on Lymenet Europe forum thanks to Camp Other 

'Lyme Research Alliance has been funding Dr. Kim Lewis' team to do research on Borrelia burgdorferi persister cells and to find a treatment that will kill persisters (either a new antibiotic or antibiotic and an additional drug/metabolite which awakens dormant cells).

Today, an open access paper has been published by Nature by both Auwaerter (who has been adamant that people cannot get persistent infections with Lyme disease) and Zhang, who has worked on persister cells and published in Nature prior to this about them. Zhang and Lewis have done persister research together.

In an interesting turn of events, Auwaerter and Zhang and co. have been doing research on FDA-approved drugs already in their database which may work on Borrelia persister cells. While Auwaerter issues his usual caveats that there isn't established evidence of persistent infection in people with post treatment Lyme disease/chronic Lyme disease, it is notable that he would be working on this kind of research and write about the uncertainties concerning Lyme disease.' 

http://www.lymeneteurope.org/forum/viewtopic.php?f=5&t=5419

An excellent explanation about the difference between antibiotic resistance and persister cells found on Camp Other blog here