Compositions and methods relating to myomaker-induced muscle cell fusion

Patent metadata

Field	Value
Publication number	US-2016136240-A1
Application number	US-201414900005-A
Country	US
Kind code	A1
Filing date	Jun 27, 2014
Priority date	Jun 27, 2013
Publication date	May 19, 2016
Grant date	—

Abstract

The present disclosure describes the fusogenic activity of the Myomaker protein. This polypeptide, when expressed in non-muscle cells, is able to drive fusion of the cell with a muscle cell, but not with other non-muscle cells. The use of this protein and cell expressing it in the delivery of exogenous genetic material to muscle cells also is described.

First claim

What is claimed is:

1 . A cell transformed with an exogenous nucleic acid encoding a Myomaker polypeptide under the control of a promoter active in said cell.

2 . The cell of claim 1 , wherein said cell is a human cell.

3 . The cell of claim 1 , where said cell is a non-muscle cell.

4 . The cell of claim 1 , wherein said cell is a fibroblast, bone marrow cell or blood cell.

5 . The cell of claim 1 , wherein said exogenous nucleic acid is under the control of constitutive promoter.

6 . The cell of claim 1 , wherein said exogenous nucleic acid is under the control of an inducible promoter.

7 . The cell of claim 1 , wherein said exogenous nucleic acid is incorporated into a chromosome of said cell.

8 . The cell of claim 1 , wherein said exogenous nucleic acid is carried episomally by said cell.

9 . The cell of claim 1 , wherein said cell expresses a detectable marker.

10 . The cell of claim 1 , wherein said cell is transformed to express a gene of interest other than Myomaker.

11 . A method of preparing a non-muscle cell fusion partner comprising transferring into a non-muscle cell a nucleic acid acid encoding a Myomaker protein under the control of a promoter active in said cell.

12 . The method of claim 11 , wherein said cell is stably transformed.

13 . The method of claim 11 , wherein said cell is transiently transfected.

14 . The method of claim 11 , further comprising transferring into said cell a nucleic acid encoding or sufficient to produce a detectable marker.

15 . The method of claim 11 , wherein said exogenous nucleic acid is under the control of constitutive promoter.

16 . The method of claim 11 , wherein said exogenous nucleic acid is under the control of an inducible promoter.

17 . The method of claim 11 , wherein said cell is a human cell.

18 . The method of claim 11 , wherein said cell is a fibroblast, bone marrow cell or blood cell.

19 . The method of claim 11 , where said exogenous nucleic acid further encodes a selectable marker.

20 . The method of claim 11 , wherein said cell is transformed to express a gene of interest other than Myomaker.

21 . A method of fusing a non-muscle cell to a muscle cell comprising:
(a) providing a non-muscle cell expressing an exogenous Myomaker protein in said non-muscle cell; and (b) contacting said non-muscle cell with a muscle, wherein said non-muscle cell expressing Myomaker protein will fuse with said muscle cell.

22 . The method of claim 21 , wherein said non-muscle cell is a human cell.

23 . The method of claim 21 , wherein said non-muscle cell is a fibroblast, bone marrow cell or blood cell.

24 . The method of claim 21 , wherein step (b) is performed in vitro.

25 . The method of claim 21 , wherein step (b) is performed in vivo.

26 . The method of claim 21 , wherein said non-muscle cell expresses a detectable marker.

27 . The method of claim 21 , wherein said non-muscle cell expresses a selectable marker.

28 . The method of claim 24 , wherein said muscle cell is an isolated muscle cell.

29 . The method of claim 21 , wherein said muscle cell is located in intact muscle tissue.

30 . The method of claim 21 , wherein said muscle cell is a myoblast.

31 . A method of delivering a gene of interest to a muscle cell comprising:
(a) providing a non-muscle cell expressing an exogenous Myomaker protein, wherein said non-muscle cell further comprises a gene of interest; and (b) contacting said non-muscle cell with a muscle, wherein said non-muscle cell expressing Myomaker protein will fuse with said muscle cell and deliver said gene of interest to said muscle cell.

32 . The method of claim 31 , wherein said non-muscle cell is a human cell.

33 . The method of claim 31 , wherein said non-muscle cell is a fibroblast, bone marrow cell or blood cell.

34 . The method of claim 31 , wherein step (b) is performed in vitro.

35 . The method of claim 31 , wherein step (b) is performed in vivo.

36 . The method of claim 31 , wherein said non-muscle cell expresses a detectable marker.

37 . The method of claim 31 , wherein said non-muscle cell expresses a selectable marker.

38 . The method of claim 34 , wherein said muscle cell is an isolated muscle cell.

39 . The method of claim 31 , wherein said muscle cell is located in intact muscle tissue.

40 . The method of claim 31 , wherein said muscle cell exhibits a pathologic phenotype, and said gene of interest correct said genotype.

41 . The method of claim 40 , wherein said pathologic phenotype is the underexpression or absence of a normal gene product.

42 . The method of claim 40 , wherein said pathologic phenotype is the expression of a defective gene product.

43 . The method of claim 40 , wherein said muscle cell has a pathologic phenotype associated with congenital myopathy, sarcopenia, amyotrophic lateral sclerosis, muscular dystrophy, Pompe disease or rhabdomyosarcoma.

44 . The method of claim 40 , wherein said non-muscle cell is delivered to an affected muscle tissue comprising said muscle in a subject.

45 . The method of claim 44 , wherein delivery is via intramuscular injection.

46 . The method of claim 44 , wherein delivery is repeated at least once.

47 . The method of claim 44 , wherein a secondary therapy is administered to said subject.

48 . The method of claim 40 , wherein said non-muscle cell is delivered to said muscle cell ex vivo and subsequently implanted in to a subject.

49 . The method of claim 48 , wherein said muscle cell is comprised in intact muscle tissue.

50 . The method of claim 31 , wherein said muscle cell is a myoblast.

Assignees

• Univ Texas
• Univ Texas Sy Stem Board Of Regents

Inventors

• Olson Eric
• Millay Douglas P

Classifications

• A61K45/06

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• C12N5/16

  Animal cells · CPC title

• C07K14/4716

  Muscle proteins, e.g. myosin, actin · CPC title

• A61K38/1719 Primary

  Muscle proteins, e.g. myosin or actin · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title