Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Methods and compositions for treating mitochondrial disease or disorders and heteroplasmy

a mitochondrial disease or disorder and composition technology, applied in the field of compositions, can solve the problems of high morbidity and mortality, difficult management and treatment of patients with mitochondrial disease or disorders, and considerable morbidity, so as to reduce endogenous mtdna copy number, reduce mitochondrial mass, and reduce endogenous mitochondrial function

Pending Publication Date: 2020-02-20
IMEL BIOTHERAPEUTICS INC
View PDF0 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes methods for reducing the number of copies of a specific gene called mtDNA in cells and using exogenous mitochondria (mtDNA from a non-senescent cell) to treat senescence and extend lifespan in cells. The methods involve using an agent that reduces the function of mitochondria or the number of copies of mtDNA in cells. This can lead to a reduction in mitochondrial mass and the exogenous mitochondria can have enhanced function relative to the endogenous mitochondria. The technical effects of the patent include the ability to treat age-related diseases and extend lifespan in cells.

Problems solved by technology

Mitochondrial disease or disorders can involve any organ or tissue, characteristically involve multiple systems, typically affecting organs that are highly dependent on aerobic metabolism, and are often relentlessly progressive with high morbidity and mortality.
The management and treatment of patients with mitochondrial disease or disorders remains challenging.
For the vast majority of patients, the condition is relentlessly progressive leading to considerable morbidity and, in those most severely affected, death.
The classical method to remove endogenous mtDNA involves long term treatment of cells with low concentrations of ethidium bromide (EtBr), a known carcinogen and teratogen, limiting its application for therapeutic purposes.
In addition to the potential for unwanted side effects, the EtBr protocol can take several months, which further limits its clinical use.
This complete depletion of mtDNA severely hinders the ability of a cell to ingest exogenous mitochondria.
Other mitochondrial transfer protocols have attempted to add mitochondria without depletion of endogenous mtDNA, but this approach has been found to be inefficient or harmful to a cell.
For example, mitochondrial transfer using simple coincubation has been reported to be ineffective and not equally efficient among different cell types.
Additional techniques to transfer have involved injection using invasive instruments, which caused harm to the recipient cell, or other invasive instruments, such as nanoblades, but all were less efficient than coincubation (Caicedo et al, Stem Cells International, (2017), vol.
Accordingly, current methods of mitochondrial transfer are not only impractical for the clinical setting, but they are also inefficient, harmful to recipient cells and / or time intensive.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Methods and compositions for treating mitochondrial disease or disorders and heteroplasmy
  • Methods and compositions for treating mitochondrial disease or disorders and heteroplasmy
  • Methods and compositions for treating mitochondrial disease or disorders and heteroplasmy

Examples

Experimental program
Comparison scheme
Effect test

examples

[0348]The examples in this section are offered by way of illustration, and not by way of limitation. The following examples are presented as exemplary embodiments of the invention. They should not be construed as limiting the broad scope of the invention.

example i

ion of the MirC Protocol Revealed that XbaI Degraded mtDNA In Vitro and the MTS Expression Vector Targeted Mitochondria

[0349]A scheme of the method used to generate a mitochondria replaced cell (MirC) is provided in FIG. 1A. First, the mammalian expression vector used to express the XbaI restriction enzyme fused to a mitochondrial-targeted sequence (MTS) was engineered by cloning the MTS-XbaI sequence into the pCAGGS vector using standard techniques known in the art (FIG. 1B). Among mitochondrial transfer signals (MTS) being reported we utilized the ND4 signal sequence in this study. The resultant expression vector also contained the puromycin resistance gene to allow for selection (FIG. 1B).

[0350]XbaIR is one of the most powerful endonucleases and a standard sequence of mtDNA named under Cambridge reference sequence (CRS) in human mitochondria genome has as many as five recognition sites targeted by the particular endonuclease (FIG. 1D). It was verified by an in vitro endonuclease ...

example ii

se MTS-XbaIR Treatment Exhibits Improved Degradation of mtDNA Relative to the Conventional Method of EtBr

[0354]The efficiency and efficacy of the MTS-XbaIR expression vector relative to the conventional method that employs ethidium bromide (EtBr) was evaluated according to the scheme illustrated in FIG. 2A. The placental venous endothelium-derived cell line EPC100 with DsRed labeled mitochondria were cultured in pyruvate-free DMEM (Wako cat #044-29765) with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin (P / S), and at day 0 the cells were either untreated (“normal”), transfected with the MTS-XbaIR expression vector (“MTS-XbaIR”), or treated with 50 ng / mL of EtBr. At day 1, the cells were cultured in DMEM with 10% FBS, and 1% P / S supplemented with 100 μg / mL pyruvate and 50 μg / mL uridine. Quantitative polymerase chain reaction (qPCR) was performed according to methods known in the art at days 3 and 5 to measure the mtDNA relative to the housekeeping gene, (3-actin (Actb). ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Fractionaaaaaaaaaa
Fractionaaaaaaaaaa
Fractionaaaaaaaaaa
Login to View More

Abstract

The present invention provides methods and compositions for generation of mitochondria replaced cells (MirC), and therapeutic methods for using such compositions for treating a subject having an age-related disease or syndrome, mitochondrial disease or disorder, or otherwise in need of mitochondrial replacement. Also provided are methods and compositions for producing a recipient cell having a mitochondrial disease or disorder, as well as methods and compositions for producing or enhancing production of an inducible pluripotent stem cell (iPSC). In addition, methods and compositions to enhance mitochondrial transfer are also included.

Description

[0001]This application claims the benefit of U.S. Provisional Application No. 62 / 718,891, filed Aug. 14, 2018, U.S. Provisional Application No. 62 / 731,731 filed Sep. 14, 2018, and U.S. Provisional Application No. 62 / 817,987 filed Mar. 13, 2019, which are incorporated herein by reference in their entirety.SEQUENCE LISTING[0002]The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Aug. 13, 2019, is named 14595-001-999_SL.txt and is 12,905 bytes in size.Field of the Invention[0003]The present invention provides a composition of cells with reduced mitochondrial DNA and / or replacement of mitochondrial DNA, methods for their production, and methods for treating various diseases associated with genetic or age-related mitochondrial dysfunctions.Background of the Invention[0004]Mitochondria play a major and critical role in cellular homeostasis, and are i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K35/28C12N5/077
CPCA61K31/436C12N5/0669A61K45/06A61K35/28C12N5/0647C12N2510/00C12N5/0636C12N5/0656A61K2239/31A61K39/4644A61K2239/46A61K39/461A61K39/4611A61K39/4621A61K2239/38A61K39/46433A61P3/10A61P25/28
Inventor GOJO, SATOSHIKAMI, DAISUKEMAEDA, HIDEKI
Owner IMEL BIOTHERAPEUTICS INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com