Therapeutic engineered microbial cell system and methods for treating hyperuricemia and gout
The present disclosure relates to engineered microbial cells that have been engineered to include a uricase, a uric acid transporter, or both a uricase and a uric acid transporter. The engineered microbial cells of the present disclosure are useful in degrading uric acid inside the engineered microbial cell. The engineered microbial cells of the present disclosure are useful in methods of treating hyperuricemia. The engineered microbial cells of the present disclosure are also useful in methods of treating gout, and in particular chronic refractory gout.
1 . An engineered bacterial cell, the cell comprising:
a first nucleic acid encoding an intracellular uric acid degrading polypeptide operatively linked to a constitutive heterologous promoter; and
a second nucleic acid encoding a uric acid transporter operatively linked to a heterologous promoter, wherein the uric acid transporter transports uric acid into the engineered bacterial cell;
wherein the engineered bacterial cell has increased intracellular uric acid degrading activity and has increased uric acid uptake activity as compared to an otherwise identical non-engineered bacterial cell that does not comprise the first and second nucleic acids.
2 . The engineered bacterial cell of claim 1 , wherein the uric acid transporter comprises an amino acid sequence having at least 95% sequence identity with the full length amino acid sequence of any one of SEQ ID NOs: 9-17.
3 . The engineered bacterial cell of claim 1 , wherein the uric acid degrading polypeptide is a uricase.
4 . The engineered bacterial cell of claim 3 , wherein the uricase is a fungal uricase.
5 . The engineered bacterial cell of claim 3 , wherein the uricase comprises an amino acid sequence having at least 95% sequence identity to the full length amino acid sequence of one of SEQ ID NOs: 1-7.
6 . The engineered bacterial cell of claim 3 , wherein the uricase is a bacterial uricase.
7 . A method of lowering uric acid in a gastrointestinal tract of a recipient, the method comprising:
administering to the gastrointestinal tract of the recipient the engineered bacterial cell of claim 1 .
8 . The method of claim 7 , wherein the recipient suffers from a condition selected from the group consisting of: gout, rheumatoid arthritis, cerebral stroke, heart disease, arrhythmia, chronic renal disease, and chronic refractory gout.
9 . The method of claim 7 , wherein the recipient suffers from chronic refractory gout.
10 . The method of claim 7 , wherein the engineered bacterial cell is administered to the gastrointestinal tract of the recipient by oral administration.
11 . The method of claim 7 , wherein the recipient is administered 10 6 to 10 12 of the engineered bacterial cells.
12 . The method of claim 7 , further comprising determining that the recipient has a serum uric acid level of ≥6.8 mg/dl prior to administration.
13 . The engineered bacterial cell of claim 1 , wherein the first and second nucleic acids are integrated into the chromosome of the engineered bacterial cell.
14 . The engineered bacterial cell of claim 1 , wherein the uric acid degrading polypeptide and the uric acid transporter encoded by the first and second nucleic acids are endogenous to the engineered bacterial cell.
15 . The engineered bacterial cell of claim 1 , wherein the first and second nucleic acids are located in a single operon wherein mRNA transcribed from the first and second nucleic acids is independently translated to produce distinct protein products.