Mitochondrial augmentation therapy for primary mitochondrial diseases
AssigneeMinovia Therapeutics Ltd.
InventorsNatalie Yivgi Ohana, Uriel Halavee, Shmuel Bukshpan, Noa Sher, Moriya Blumkin
Life of this patent
Estimating the term…
Abstract
The present invention provides human stem cells enriched with functional mitochondria, wherein the healthy functional exogenous mitochondria constitute at least 3% and less than 33% of the total mitochondria in the mitochondrially-enriched human stem cells. Further disclosed are methods for the production of such cells, and uses thereof for the treatment of primary mitochondrial diseases.
Claims29
-3. (canceled)
The method of claim 37, wherein the human exogenous mitochondria are syngeneic or allogeneic.
-6. (canceled)
The method of claim 37, wherein the mitochondrial disease or disorder is associated with a mutation in mitochondrial DNA selected from the group consisting of Pearson syndrome;
- Kearns-Sayre syndrome;
- a Mitochondrial encephalopathy lactic acidosis and stroke-like episodes (MELAS) syndrome;
- Leber's hereditary optic neuropathy (LHON);
- Neuropathy, ataxia, and retinitis pigmentosa (NARP) syndrome;
- myoclonic epilepsy with ragged red fibers (MERRF) syndrome;
- maternally inherited diabetes and deafness (MIDD);
- Alpers-like syndrome;
- Chronic progressive external ophthalmoplegia (CPEO);
- mitochondrial DNA-associated forms of Congenital lactic acidosis (CLA);
- Mitochondrial DNA depletion Syndrome (MDDS);
- mitochondrial DNA-associated forms of Leigh syndrome, Mitochondrial neurogastrointestinal encephalopathy (MNGIE) syndrome;
- Alpers syndrome;
- Friedreich's ataxia (FA);
- progressive external ophthalmoplegia (PEO);
- Sideroblastic anemia;
- an ataxia neuropathy syndrome (ANS);
- a Mendelian neurodegenerative mitochondriopathy;
- 3-methylglutaconic aciduria (MEG) deafness (D), encephalopathy (E) and Leigh-like disease (L) syndrome (MEGDEL);
- Sengers syndrome;
- Minimal change nephrotic syndrome (MCNS);
- nuclear DNA-associated forms of Congenital lactic acidosis (CLA);
- and nuclear DNA-associated forms of Leigh syndrome.
-9. (canceled)
The method of claim 37, wherein the mitochondrial disease or disorder is associated with an organ selected from the group consisting of kidney, liver, brain, muscle, pancreas, eye and any combination thereof.
The method of claim 37, wherein the symptom is selected from the group consisting of impaired walking capability, impaired motor skills, impaired language skills, impaired memory, impaired weight gain, failure to thrive, low blood alkaline phosphatase levels, low blood magnesium levels, high blood creatinine levels, low blood bicarbonate levels, low blood base excess levels, high urine glucose/creatinine ratios, high urine chloride/creatinine ratios, high urine sodium/creatinine ratios, high blood lactate levels, high urine magnesium/creatinine ratios, high urine potassium/creatinine ratios, high urine calcium/creatinine ratios, glucosuria, magnesuria, high blood urea levels, low C-Peptide level, high HbA1C level, hypoparathyroidism, ptosis, hearing loss, cardiac conduction disorder, epileptic seizures, Stroke-like episodes, impaired EEG, high blood aspartate aminotransferase (AST) levels, high blood alanine aminotransferase (ALT) levels, low ATP content and oxygen consumption in lymphocytes.
The method of claim 37, wherein said pharmaceutical composition is administered to a specific tissue or organ or systemically.
-15. (canceled)
The method of claim 37, wherein the mitochondrially-enriched human stem cells have at least one of:
- an increased mitochondrial DNA content;
- (ii) an increased level of citrate synthase (CS) activity;
- (iii) an increased content of at least one mitochondrial protein selected from SDHA and COX1;
- (iv) an increased rate of O2 consumption;
- (v) an increased rate of ATP production;
- or
- (vi) any combination thereof,
- relative to the corresponding level in the stem cells prior to mitochondrial enrichment.
-19. (canceled)
The method of claim 37, wherein the human stem cells are hematopoietic stem cells, mesenchymal stem cells, pluripotent stem cells (PSCs), induced pluripotent stem cells (iPSCs), or CD34+ cells.
-23. (canceled)
The method of claim 37, wherein the human stem cells are isolated, derived or obtained from cells of the bone marrow, adipose tissue, oral mucosa, skin fibroblasts, blood or umbilical cord blood.
The method of claim 37, wherein the human exogenous mitochondria are isolated or obtained from placenta, placental cells grown in culture or blood cells.
(canceled)
The method of claim 37, wherein the exogenous mitochondria constitute between 1% and 30% of the total mitochondria.
An ex-vivo method for enriching human stem cells with exogenous mitochondria comprising:
- providing a first composition, comprising a plurality of isolated or partially purified human stem cells from a patient afflicted with a mitochondrial disease, disorder or a symptom thereof, or from a donor;
- (ii) providing a second composition, comprising a plurality of isolated human exogenous mitochondria obtained from a donor without a pathogenic mutation in mitochondrial DNA;
- (iii) contacting the human stem cells of the first composition with the human exogenous mitochondria of the second composition, at a ratio of 0.088-176 mU CS activity per 106 stem cells thus providing a third composition;
- and
- (iv) incubating the third composition under conditions allowing the human exogenous mitochondria to enter the human stem cells thereby enriching said human stem cells with said human exogenous mitochondria, thus providing a fourth composition comprising mitochondrially-enriched human stem cells;
- wherein the human exogenous mitochondria constitute at least 1% and less than 33% of the total mitochondria in the fourth composition.
-32. (canceled)
-36. (canceled)
A method of treating a mitochondrial disease, disorder or a symptom thereof in a human patient in need of such treatment comprising administering a pharmaceutical composition to the patient, the pharmaceutical composition comprising at least about 5×105 to 5×109 human stem cells, wherein the human stem cells are enriched with human exogenous mitochondria without a pathogenic mutation in mitochondrial DNA, wherein the human exogenous mitochondria constitute at least 1% and less than 33% of the total mitochondria in the mitochondrially-enriched human stem cells.
(canceled)
A pharmaceutical composition comprising at least 105 to 2×107 human stem cells per kilogram bodyweight of a patient, in a pharmaceutically acceptable carrier capable of supporting the viability of the cells, wherein the human stem cells are enriched with human exogenous mitochondria without a pathogenic mutation in mitochondrial DNA, and wherein the human exogenous mitochondria constitute at least 1% and less than 33% of the total mitochondria in the mitochondrially-enriched human stem cells.
Dependencies are inferred from the claim text.