Compositions and methods for improving mitochondrial function

Patent metadata

Field	Value
Publication number	US-10973860-B2
Application number	US-201916293789-A
Country	US
Kind code	B2
Filing date	Mar 6, 2019
Priority date	Jan 23, 2018
Publication date	Apr 13, 2021
Grant date	Apr 13, 2021

Abstract

Provided herein are methods and compositions comprising a bacterium or a metabolite thereof for enhancing mitochondrial and/or peroxisomal function.

First claim

The invention claimed is:

1. A composition comprising a measured amount of a bacterial species that comprises and expresses nucleic acid sequences encoding membrane-bound PQQ-dependent glucose dehydrogenase (mGDH), ubiquinol-cytochrome c reductase iron-sulfur subunit, TonB-dependent receptor, carbon-nitrogen hydrolase, and ubiquinol oxidase subunit II, or an extract or fraction derived therefrom,
wherein the extract or fraction of the bacterial species comprises membrane-bound PQQ-dependent glucose dehydrogenase (mGDH), ubiquinol-cytochrome c reductase iron-sulfur subunit, TonB-dependent receptor, carbon-nitrogen hydrolase, and ubiquinol oxidase subunit II expressed from the nucleic acid sequences,
wherein the bacterial species is from the Acetobacteriaceae family, and
wherein one or more of the nucleic acid sequences are exogenous nucleic acid sequences.

2. The composition of claim 1 , wherein the bacterium is Gluconobacter spp, Acetobacter spp., Gluconoacaetobacter spp., Acidomonas spp, Ameyamaea spp., Asaia spp., Granulibacter spp., Kozakia spp., Neoasaia spp., Neokomagataea spp., Saccharibacter spp., Swaminathania spp., or Tanticharoenia spp.

3. The composition of claim 1 , wherein the measured amount of the bacteria is lyophilized.

4. The composition of claim 1 , wherein the composition further comprises one or more added bacterial metabolites selected from the group consisting of gluconic acid, 2-keto-gluconic acid, 5-keto-gluconic acid, and 2,5-diketo-D-gluconic acid.

5. The composition of claim 1 , wherein the composition is formulated as a food, a beverage, a feed composition, a probiotic, a nutritional supplement, or a pharmaceutical composition.

6. The composition of claim 1 , which further comprises a prebiotic.

7. The composition of claim 1 , further comprising a pharmaceutically acceptable carrier.

8. The composition of claim 1 , wherein the composition is formulated for oral administration.

9. The composition of claim 8 , wherein the composition is an enteric-coated formulation.

10. A method for increasing cellular ATP production in at least one cell type of a subject in need thereof, the method comprising administering to the subject the composition of claim 1 effective to increase cellular ATP production in at least one cell type compared to a subject without the administration.

11. The method of claim 10 , wherein the administered composition produces one or more of gluconic acid, 2-keto-gluconic acid, 5-keto-gluconic acid, and 2,5-diketo-D-gluconic acid.

12. The method of claim 10 , wherein the subject is human.

13. The method of claim 10 , wherein mitochondrial biogenesis is maintained or increased compared to a subject without the administration.

14. The method of claim 10 , wherein the activity of complex I and/or complex II of the mitochondrial electron transport chain is increased in the one or more cell types compared to a subject without the administration.

15. The method of claim 10 , wherein the administering increases mitochondrial membrane potential compared to a subject without the administration.

16. The method of claim 10 , wherein the administering increases one or more of:
expression of peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α), and/or mitochondrial transcription factor A (TFAM);
AMP-activated protein kinase (AMPK) phosphorylation level;
nuclear respiratory factor-2 (Nrf2) protein level;
PGC-1α mRNA level;
TFAM mRNA level;
mitochondrial DNA replication;
mitochondrial DNA copy number (mtDNA); and
expression of at least one mitochondrial β-oxidation enzyme, compared to a subject without the administration.

17. The method of claim 10 , wherein cellular ATP production is increased by at least 10% compared to the cellular ATP production prior to administration of the composition.

18. A method for making a bacterial extract, the method comprising culturing a bacterial species that comprises and expresses nucleic acid sequences encoding membrane-bound PQQ-dependent glucose dehydrogenase (mGDH), ubiquinol-cytochrome c reductase iron-sulfur subunit, TonB-dependent receptor, carbon-nitrogen hydrolase, and ubiquinol oxidase subunit II
wherein the bacterial species is from the Acetobacteriaceae family, and wherein one or more of the nucleic acid sequences are exogenous nucleic acid sequences,
in a medium comprising:
(i) standard Lysogeny broth (1% tryptone, 0.5% yeast extract, and 1% sodium chloride) containing 1% glucose,
(ii) standard Hestrin-Schramm broth (D-glucose 2%, 0.5% yeast extract, 0.5% peptone, 0.27% disodium phosphate, 0.115% citric acid) or
(iii) CaCO3 medium comprising 8% glucose, 0.5% yeast extract, 0.2% mannitol, 0.05% magnesium sulphate, and 10% calcium carbonate.

19. The method of claim 18 , wherein the bacterial species produces one or more of gluconic acid, 2-keto-gluconic acid, 5-keto-gluconic acid, and 2,5-diketo-D-gluconic acid.

20. A method of treating a mitochondrial electron transport chain disorder, the method comprising administering to a subject having a mitochondrial electron transport chain disorder, a therapeutically effective amount of the composition of claim 1
thereby reducing at least one symptom of the mitochondrial electron transport chain disorder compared to a subject without the administration.

21. The method of claim 20 , wherein the mitochondrial electron transport chain disorder comprises a disorder or impaired activity in Complex I and/or Complex II.

22. The method of claim 20 , wherein the mitochondrial electron transport chain disorder is NADH dehydrogenase (NADH-CoQ reductase) deficiency, succinate dehydrogenase deficiency, Leigh Disease, mitochondrial DNA depletion, or mitochondrial insufficiency.

23. The method of claim 20 , wherein the at least one symptom is selected from the group consisting of: myopathy, mitochondrial encephalomyopathy, failure to thrive, developmental delay, hypotonia, lethargy, respiratory failure, ataxia, myoclonus, lactic acidosis, seizures, fatigue, nystagmus, poor reflexes, difficulty eating or swallowing, breathing difficulties, ataxia, congenital myopathy, infantile myopathy and hepatopathy.

24. A method for increasing the biogenesis of cellular mitochondria or peroxisomes, the method comprising administering to a subject, the composition of claim 1
effective to increase the biogenesis of cellular mitochondria or peroxisomes compared to a subject without the administration.

25. The method of claim 24 , wherein the size and/or number of peroxisomes is increased compared to a subject without the administration.

26. The method of claim 24 , wherein mitochondrial activity is increased compared to a subject without the administration.

27. The method of claim 26 , wherein the administered composition produces one or more of gluconic acid, 2-keto-gluconic acid, 5-keto-gluconic acid, and/or 2,5-diketo-D-gluconic acid.

Assignees

• Massachusetts Gen Hospital

Inventors

• Ruvkun Gary
• Govindan J Amaranath
• Jayamani Elamparithi

Classifications

• C12Y110/0301

  Ubiquinol oxidase (H+-transporting) (1.10.3.10) · CPC title

• C12Y110/02002

  Ubiquinol-cytochrome-c reductase (1.10.2.2), i.e. electron-transport-complex-III · CPC title

• C12Y101/99

  with other acceptors (1.1.99) · CPC title

• C12P7/50

  having keto groups, e.g. 2-ketoglutaric acid · CPC title

• C12N15/74 Primary

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title