Safety and pharmacodynamics of an engineered E. coli Nissle for the treatment of phenylketonuria: a first-in-human phase 1/2a study

Puurunen, Marja; Vockley, Jerry; Searle, Shawn; Sacharow, Stephanie; Phillips, John A.; Denney, William S.; Goodlett, Benjamin D.; Wagner, David; Blankstein, Larry; Castillo, Mary Joan; Charbonneau, Mark R.; Isabella, Vincent M.; Sethuraman, Vasu V.; Riese, Richard J.; Kurtz, Caroline B.; Brennan, Aoife M.

doi:10.1038/s42255-021-00430-7

Cited by 106 publications

(84 citation statements)

References 23 publications

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“…Engineered strains of EcN have been successfully used to diagnose and treat bacterial infections 1 , 2 , cancers 3 – 5 , gastrointestinal bleeding 6 , inflammatory disorders 7 – 9 , and obesity 10 in a variety of animal models. EcN strains engineered to treat metabolic disorders are being evaluated in human clinical trials with promising early-phase results 11 , 12 . However, there are important safety concerns associated with organisms genetically engineered for medical applications.…”

Section: Introductionmentioning

confidence: 99%

Genetically stable CRISPR-based kill switches for engineered microbes

et al. 2022

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Microbial biocontainment is an essential goal for engineering safe, next-generation living therapeutics. However, the genetic stability of biocontainment circuits, including kill switches, is a challenge that must be addressed. Kill switches are among the most difficult circuits to maintain due to the strong selection pressure they impart, leading to high potential for evolution of escape mutant populations. Here we engineer two CRISPR-based kill switches in the probiotic Escherichia coli Nissle 1917, a single-input chemical-responsive switch and a 2-input chemical- and temperature-responsive switch. We employ parallel strategies to address kill switch stability, including functional redundancy within the circuit, modulation of the SOS response, antibiotic-independent plasmid maintenance, and provision of intra-niche competition by a closely related strain. We demonstrate that strains harboring either kill switch can be selectively and efficiently killed inside the murine gut, while strains harboring the 2-input switch are additionally killed upon excretion. Leveraging redundant strategies, we demonstrate robust biocontainment of our kill switch strains and provide a template for future kill switch development.

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Section: Introductionmentioning

confidence: 99%

Genetically stable CRISPR-based kill switches for engineered microbes

et al. 2022

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“…Since its discovery in 1957, EcN has played an important role as a Gram-negative probiotics in regulating the intestinal microbiota and suppressing enteritis ( Sassone-Corsi et al, 2016 ), due to its excellent safety profile, good tolerability, clear genetic background, availability of genetic manipulation, and excellent colonization properties ( Westendorf et al, 2005 ; Luo et al, 2020 ). Moreover, with the boom of synthetic biology, EcN has been used as a engineered strain vector for the treatment of phenylketonuria in a phase I clinical study ( Puurunen et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Engineered Bacteria EcN-MT Alleviate Liver Injury in Cadmium-Exposed Mice via its Probiotics Characteristics and Expressing of Metallothionein

Zou

Chen

et al. 2022

Front. Pharmacol.

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Cadmium (Cd) exposure is a widespread problem in many parts of the world, but effective means to treat Cd exposure is still lacking. Hence, an engineered strain expressing metallothionein (MT) named Escherichia coli Nissle 1917 (EcN)-MT was constructed, and its potential in the treatment of Cd exposure was evaluated. The in vitro studies showed that metallothionein expressed by EcN-MT could significantly bind Cd. Further, the in vivo results indicated that EcN-MT strain could reduce 26.3% Cd in the liver and increase 24.7% Cd in the feces, which greatly decreased malondialdehyde (MDA) levels and increased catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD) levels in liver, and reduced the expression of toll-like receptor4 (TLR4), nuclear factor-κB (NF-κB), the myeloid differentiation factor 88 (Myd88) andincreased B-cell lymphoma 2 (Bcl-2)/Bcl-2-Associated X (Bax). Moreover, high throughput sequencing results indicated that EcN-MT strain greatly enhanced the beneficial bacteria of Ruminococcaceae, Lactobacillaceae, Akkermansia, Muribaculaceae, Lachnospiraceae, Dubosiella and restored the disturbed microbial ecology to the normal level. Therefore, the high Cd binding capacity of the expressed metallothionein, together with the beneficial characteristics of the host bacteria EcN, makes EcN-MT a sound reagent for the treatment of subchronic Cd exposure-induced liver injury.

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“…To estimate the effect of daily SYNB8802 administration on UOx excretion in healthy subjects and EH patients, dosing simulations were performed for three‐times daily doses (TID) of 0, 1 × 10 11 , 2 × 10 11 , or 5 × 10 11 SYNB8802 cells with meals over 10 days on a diet containing 200 mg/day oxalate. SYNB8802 dose levels were selected based on well‐tolerated doses of related synthetic biotics (Kurtz et al , 2019 ; Puurunen et al , 2021 ). Compared to simulated healthy subjects receiving no therapy, subjects receiving SYNB8802 exhibited lowered UOx excretion after 10 days by 14 ± 3, 25 ± 5, and 38 ± 7% for 1 ×10 11 , 2 × 10 11 , and 5 × 10 11 cell dose levels, respectively (Fig 5B ).…”

Section: Resultsmentioning

confidence: 99%

“…In contrast, Escherichia coli Nissle 1917 (EcN) is a non‐colonizing, probiotic strain (Kurtz et al , 2018 ), providing a flexible platform for the development of engineered bacterial therapeutics (synthetic biotics). EcN strains that consume toxic metabolites within the GI tract have been developed (Isabella et al , 2018 ) and are safe and well‐tolerated in healthy volunteers (Kurtz et al , 2019 ; Puurunen et al , 2021 ). The non‐colonizing nature of EcN enables predictable pharmacodynamics, reduced risk of genetic drift in vivo , limited interactions with host microbiota and the complete clearance after dosing is stopped (Charbonneau et al , 2020 ).…”

Section: Introductionmentioning

confidence: 99%

An engineered bacterial therapeutic lowers urinary oxalate in preclinical models and in silico simulations of enteric hyperoxaluria

Lubkowicz

James

et al. 2022

Molecular Systems Biology

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Enteric hyperoxaluria (EH) is a metabolic disease caused by excessive absorption of dietary oxalate leading to the formation of chronic kidney stones and kidney failure. There are no approved pharmaceutical treatments for EH. SYNB8802 is an engineered bacterial therapeutic designed to consume oxalate in the gut and lower urinary oxalate as a potential treatment for EH. Oral administration of SYNB8802 leads to significantly decreased urinary oxalate excretion in healthy mice and non‐human primates, demonstrating the strain's ability to consume oxalate in vivo . A mathematical modeling framework was constructed that combines in vitro and in vivo preclinical data to predict the effects of SYNB8802 administration on urinary oxalate excretion in humans. Simulations of SYNB8802 administration predict a clinically meaningful lowering of urinary oxalate excretion in healthy volunteers and EH patients. Together, these findings suggest that SYNB8802 is a promising treatment for EH.

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Safety and pharmacodynamics of an engineered E. coli Nissle for the treatment of phenylketonuria: a first-in-human phase 1/2a study

Cited by 106 publications

References 23 publications

Genetically stable CRISPR-based kill switches for engineered microbes

Genetically stable CRISPR-based kill switches for engineered microbes

Engineered Bacteria EcN-MT Alleviate Liver Injury in Cadmium-Exposed Mice via its Probiotics Characteristics and Expressing of Metallothionein

An engineered bacterial therapeutic lowers urinary oxalate in preclinical models and in silico simulations of enteric hyperoxaluria

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