Engineered Escherichia coli Nissle 1917 with urate oxidase and an oxygen-recycling system for hyperuricemia treatment

Abstract

Hyperuricemia is the second most prevalent metabolic disease to human health after diabetes. Only a few clinical drugs are available, and most of them have serious side effects. The human body does not have urate oxidase, and uric acid is secreted via the kidney or the intestine. Reduction through kidney secretion is often the cause of hyperuricemia. We hypothesized that the intestine secretion could be enhanced when a recombinant urate-degrading bacterium was introduced into the gut. We engineered an Escherichia coli Nissle 1917 strain with a plasmid containing a gene cassette that encoded two proteins PucL and PucM for urate metabolism from Bacillus subtilis , the urate importer YgfU and catalase KatG from E. coli , and the bacterial hemoglobin Vhb from Vitreoscilla sp. The recombinant E. coli strain effectively degraded uric acid under hypoxic conditions. A new method to induce hyperuricemia in mice was developed by intravenously injecting uric acid. The engineered Escherichia coli strain significantly lowered the serum uric acid when introduced into the gut or directly injected into the blood vessel. The results support the use of urate-degrading bacteria in the gut to treat hyperuricemia. Direct injecting bacteria into blood vessels to treat metabolic diseases is proof of concept, and it has been tried to treat solid tumors.

RETRACTED: Effect and Potential Mechanism of Lactobacillus plantarum Q7 on Hyperuricemia in vitro and in vivo

“…Compared with the CON group, the composition of gut microbiota at the phylum level showed that the relative abundance of Firmicutes was significantly decreased, whereas Bacteroidetes and Proteobacteria were markedly increased in the MOD group ( Figure 8F ). This imbalance of gut microbiota was recovered in the Q7 group, and the corresponding symptoms of HUA were further relieved, which was consistent with the reports of Zhao et al ( 32 ). The difference of gut microbiota in each group at the genus level suggested that the relative abundance of Muribaculaceae was increased after HUA modeling, the relative abundance of Lachnospiraceae _NK4A136_group, Prevotellaceae _UCG-001 and Prevotellaceae _NK3B31_group were decreased, while those flora in the Q7 group were recovered to the level of CON group ( Figure 8G ).…”

Rational design of a genome‐based insulated system in Escherichia coli facilitates heterologous uricase expression for hyperuricemia treatment

“…In addition, there are 10 nucleobase‐ascorbate transporter (NAT) family‐related proteins in E. coli that are responsible for transporting different forms of base metabolites. Further, the ygfU gene located at the inner membrane was hypothesized to import urate 61 and overexpression of YgfU could improve the urate degradation efficiency in E. coli 62 . However, our data showed that cytoplasmic expression of uricase in the EcN strain only slightly decreased the urate level in vitro (Figure S1b), suggesting that the efficiency of urate import may be limited by the bacterial cell membrane or the affinity of urate transporters.…”

Intestinal Engineered Probiotics as Living Therapeutics: Chassis Selection, Colonization Enhancement, Gene Circuit Design, and Biocontainment

“…Traditional probiotics can be easily engineered and are the chassis of choice. For example, engineered EcN has been developed into various biopharmaceuticals, − and its genomic, transcriptomic, and metabolic properties have been intensively studied . In fact, lactic acid bacteria now support various CRISPR/Cas systems − and have long been developed as mature heterologous production platforms, biocontrol systems, and biosensors .…”

“…For instance, recombinant L. lactis heterologously expressing IL-10 has been used to treat DSS-induced model mice in situ since 2000 . This effective and simple method was immediately extended to different probiotic chassis and applied to different treatment scenarios, ,, resulting in numerous practical research findings. For example, the engineered probiotic SYNB1618, developed by Synlogic to treat phenylketonuria, has successfully completed phase II trials, and represents a milestone for living therapeutics. , …”

“…With the development of synthetic biology, various genetic tools can be used to carry out targeted modifications of traditional probiotics and convert them into smart drugs, to overcome the shortcomings of traditional probiotic treatment and achieve personalized treatment. , These engineered probiotics are predicted to colonize the afflicted region, specifically detect abnormal signals from the host, respond promptly, produce the desired drugs in situ within a preset period, and die after completing the treatment; these engineered probiotics are expected to be living therapeutics. While retaining the advantages of previous probiotic therapies, this approach can make treatment less expensive, and more patient-friendly, while maintaining the stability of specific therapeutic molecules. , Therefore, engineered probiotics are expected to elicit preventive or therapeutic effects and participate in the treatment process for various diseases, including metabolic disorders, − tumors, ,− and chronic inflammatory infections. − …”