Bacteria engineered to treat diseases that benefit from reduced gut inflammation and/or tightened gut mucosal barrier
Genetically engineered bacteria, pharmaceutical compositions thereof, and methods of treating or preventing autoimmune disorders, inhibiting inflammatory mechanisms in the gut, and/or tightening gut mucosal barrier function are disclosed.
1. A genetically engineered non-pathogenic bacterium comprising a non-native gene encoding interleukin (IL)-22, wherein the gene is operably linked to a promoter, and a non-native gene cassette for producing butyrate, wherein the gene cassette is operably linked to a promoter, wherein at least one promoter comprises any one of SEQ ID NOs: 142-157.
2. The bacterium of claim 1 , wherein the gene or the gene cassette is located on a chromosome in the bacterium.
3. The bacterium of claim 1 , wherein the gene or the gene cassette is located on a plasmid in the bacterium.
4. The bacterium of claim 1 , wherein the bacterium is a probiotic bacterium.
5. The bacterium of claim 4 , wherein the bacterium is selected from the group consisting of Bacteroides, Bifidobacterium, Clostridium, Escherichia, Lactobacillus , and Lactococcus.
6. The bacterium of claim 5 , wherein the bacterium is Escherichia coli strain Nissle.
7. The bacterium of claim 1 , wherein the bacterium is an auxotroph in a gene that is complemented when the bacterium is present in a mammalian gut.
8. The bacterium of claim 7 , wherein the bacterium is an auxotroph in diaminopimelic acid or an enzyme in the thymidine biosynthetic pathway.
9. A pharmaceutically acceptable composition comprising the bacterium of claim 1 ; and a pharmaceutically acceptable carrier.
10. The bacterium of claim 1 , wherein the gene cassette for producing butyrate comprises a butyrate kinase (buk) gene and a phosphate butyryltransferase (pbt) gene.
11. The bacterium of claim 1 , wherein the gene cassette for producing butyrate comprises an acyl-CoA thioesterase (tesB) gene.
12. The bacterium of claim 1 , wherein the promoter operably linked to the gene encoding IL-22 and the promoter operably linked to the gene cassette are different promoters.
13. The bacterium of claim 1 , wherein the promoter operably linked to the gene encoding IL-22 and the promoter operably linked to the gene cassette for producing butyrate are separate copies of the same promoter.
14. The bacterium of claim 1 , wherein the gene cassette comprises bcd2, etfB3, etfA3, thiA1, hbd, crt2, pbt, and buk.
15. The bacterium of claim 1 , wherein the gene cassette comprises ter, thiA1, hbd, crt2, pbt, and buk.
16. The bacterium of claim 1 , wherein the gene cassette comprises bcd2, etfB3, etfA3, thiA1, hbd, crt2, and tesB.
17. The bacterium of claim 1 , wherein the gene cassette comprises ter, thiA1, hbd, crt2, and tesB.
18. A genetically engineered non-pathogenic bacterium comprising a non-native gene encoding interleukin (IL)-22, and a non-native gene cassette for producing butyrate, wherein the gene and the gene cassette are operably linked to the same copy of a promoter comprising any one of SEQ ID NOs: 142-157.