Identification and use of the putative Bacteroides ovatus xylanase promoter for the inducible production of recombinant human proteins

Scott

Farrar

et al. 2009

Gut

100

Section: Methodsmentioning

confidence: 99%

Xylan-regulated delivery of human keratinocyte growth factor-2 to the inflamed colon by the human anaerobic commensal bacterium Bacteroides ovatus

Scott

Farrar

et al. 2009

Gut

100

“…Regulation of heterologous protein production by recombinant B ovatus using xylan is a novel and important safety feature of this drug delivery system. The recent identification of the B ovatus xylan operon promoter 11 enabled human cytokine producing strains to be generated. The Bacteroides are dominant among commensal anaerobes and have been identified in the mucin layer coating the colonic mucosa, 14 making them ideal for therapeutic protein delivery directly to the injured epithelium.…”

Section: Discussionmentioning

confidence: 99%

“…The resultant sequence was cloned into B ovatus genomic deoxyribonucleic acid (DNA) by conjugation using a suitable vector as demonstrated previously. [11][12][13] Transconjugates were detected by performing colony PCR using recombinant genes. Reverse transcription PCR was performed to detect recombinant and native gene transcripts figure 1 Diagrammatic representation of the proposed Bacteroides ovatus xylan-controlled protein delivery system.…”

Section: Methodsmentioning

confidence: 99%

Novel xylan-controlled delivery of therapeutic proteins to inflamed colon by the human anaerobic commensal bacterium

2013

annals

INTRODUCTION Growth factors such as keratinocyte growth factor-2 (KGF-2) and transforming growth factor-beta (TGF-β) are important immunoregulatory and epithelial growth factors. They are also potential therapeutic proteins for inflammatory bowel disease. However, owing to protein instability in the upper gastrointestinal tract, it is difficult to achieve therapeutic levels of these proteins in the injured colon when given orally. Furthermore, the short half-life necessitates repeated dosage with large amounts of the growth factor, which may have dangerous side effects, hence the importance of temporal and spatial control of growth factor delivery. METHODS The human commensal gut bacterium, Bacteroides ovatus, was genetically engineered to produce human KGF-2 or TGF-β 1 (BO-KGF or BO-TGF) in a regulated manner in response to the dietary polysaccharide, xylan. The successful application of BO-KGF or BO-TGF in the prevention of dextran sodium sulphate induced murine colitis is presented here. RESULTS This novel drug delivery system had a significant prophylactic effect, limiting the development of intestinal inflammation both clinically and histopathologically. The ability to regulate heterologous protein production by B ovatus using xylan is both unique and an important safety feature of this drug delivery system. CONCLUSIONS The use of genetically engineered B ovatus for the controlled and localised delivery of epithelial growth promoting and immunomodulatory proteins has potential clinical applications for the treatment of various diseases targeting the colon.

“…No. EU334491) was then cloned upstream of the tgf-b and the construct introduced into chromosomal DNA of B. ovatus (BOV975) by conjugation 28,29 to generate the strain BO-TGF.…”

Section: Generation Of Tgf-b1 Expressing Strains Of B Ovatusmentioning

confidence: 99%

“…27 A potential advantage of this method of delivery is that the bacterium is administered less frequently. Furthermore, with this system the protein is delivered in a controlled manner 28 and in smaller doses, reducing the likelihood of side effects. Here we tested the efficacy of B. ovatus-delivered TGF-b 1 in an experimental mouse model of acute colitis.…”

mentioning

confidence: 99%

Treatment of colitis with a commensal gut bacterium engineered to secrete human tgf-β1 under the control of dietary xylan

Inflammatory Bowel Diseases

Scott

Farrar

et al. 2011

Self Cite