Bacteria engineered to treat metabolic diseases
Genetically engineered bacteria, pharmaceutical compositions thereof, and methods of attenuating metabolic diseases are disclosed.
1. A bacterium comprising a gene cassette that encodes a biosynthetic pathway for producing a short chain fatty acid, wherein the short chain fatty acid is butyrate, propionate, or acetate, and wherein the gene cassette is operably linked to a promoter that is induced by exogenous environmental conditions found in the mammalian gut and that is not associated with the gene cassette in nature, and wherein the bacterium further comprises a gene or gene cassette for producing indole-3-acetic acid or tryptamine.
2. The bacterium of claim 1 , wherein the bacterium further comprises a gene or gene cassette for the production of GLP1.
3. The bacterium of claim 1 , wherein the promoter is
induced by low-oxygen or anaerobic conditions;
selected from a FNR-inducible promoter, an ANR-inducible promoter, and a DNR-inducible promoter;
regulated by a reactive nitrogen species (RNS); or
regulated by a reactive oxygen species (ROS).
4. The bacterium of claim 3 , wherein the promoter is a FNR-inducible promoter.
5. The bacterium of claim 3 , wherein the gene cassette and operatively linked promoter are present on a plasmid in the bacterium.
6. The bacterium of claim 3 , wherein the gene cassette and operatively linked promoter are present on a chromosome in the bacterium.
7. The bacterium of claim 1 , wherein the bacterium is an auxotroph comprising a deletion or mutation in a gene required for cell survival and/or growth.
8. The genetically engineered bacterium of claim 7 , wherein the bacterium is an auxotroph in diaminopimelic acid or an enzyme in the thymidine biosynthetic pathway.
9. The bacterium of claim 1 , wherein the bacterium comprises a kill switch.
10. The bacterium of claim 1 , wherein the bacterium further comprises a second gene encoding a substance toxic to the bacterium, wherein the second gene is under the control of a second promoter that is directly or indirectly induced by an exogenous environmental condition not naturally present in a mammalian gut.
11. The bacterium of claim 1 , wherein the bacterium is a non-pathogenic bacterium.
12. The bacterium of claim 1 , wherein the bacterium is a probiotic or a commensal bacterium.
13. The bacterium of claim 12 , wherein the bacterium is selected from the group consisting of Bacteroides, Bifidobacterium, Clostridium, Escherichia, Lactobacillus , and Lactococcus.
14. The bacterium of claim 13 , wherein the bacterium is Escherichia coli strain Nissle.
15. A pharmaceutically acceptable composition comprising the bacterium of claim 1 ; and a pharmaceutically acceptable carrier.
16. The pharmaceutically acceptable composition of claim 15 , wherein the composition is formulated for oral administration.
17. The pharmaceutically acceptable composition of claim 15 or claim 16 , wherein the composition is for use in treating a metabolic disease or disorder.
18. The pharmaceutically acceptable composition of claim 17 , wherein the metabolic disease or disorder is selected for the group consisting of type 1 diabetes; type 2 diabetes; metabolic syndrome; Bardet-Biedel syndrome; Prader-Willi syndrome; non-alcoholic fatty liver disease; tuberous sclerosis; Albright hereditary osteodystrophy; brain-derived neurotrophic factor (BDNF) deficiency; Single-minded 1 (SIM1) deficiency; leptin deficiency; leptin receptor deficiency; pro-opiomelanocortin (POMC) defects; proprotein convertase subtilisin/kexin type 1 (PCSK1) deficiency; Src homology 2B1 (SH2B1) deficiency; pro-hormone convertase 1/3 deficiency; melanocortin-4-receptor (MC4R) deficiency; Wilms tumor, aniridia, genitourinary anomalies, and mental retardation (WAGR) syndrome; pseudohypoparathyroidism type 1A; Fragile X syndrome; Borjeson-Forsmann-Lehmann syndrome; Alstrom syndrome; Cohen syndrome; and ulnar-mammary syndrome.
19. The pharmaceutically acceptable composition of claim 18 , wherein the metabolic disease or disorder is type 1 diabetes or type 2 diabetes.