Transgenic animals capable of being induced to delete senescent cells

Patent metadata

Field	Value
Publication number	US-9968076-B2
Application number	US-201514792208-A
Country	US
Kind code	B2
Filing date	Jul 6, 2015
Priority date	Jun 21, 2011
Publication date	May 15, 2018
Grant date	May 15, 2018

Abstract

This document provides a transgenic mouse for studying the role of senescent cells in an age-related phenotype. A recombinant polypeptide is expressed in senescent cells under control of a p16INK4a promoter. The polypeptide can be triggered to directly induce cell death in senescent cells. As a result, progression of one or more age-related phenotypes is delayed in the mouse. An example is the INK-ATTAC mouse which also expresses a marker polypeptide in senescent cells.

First claim

What is claimed is:

1. A transgenic mouse for studying the role of senescent cells in an age-related phenotype, the genome of which contains a transgene, said transgene containing a p16 INK4a promoter sequence operably linked to a nucleic acid sequence encoding a polypeptide so as to cause said polypeptide to be expressed in senescent cells in said mouse,
wherein said polypeptide directly induces cell death in cells in which said polypeptide is expressed when activated by a compound, and wherein administration of said compound to said mouse activates said polypeptide, thereby directly inducing cell death in senescent cells and inducing cell death in less than 10% of non-senescent cells, whereby progression of said age-related phenotype is delayed in said mouse relative to another such transgenic mouse that is not administered said compound.

2. The transgenic mouse of claim 1 , wherein said polypeptide comprises a caspase 8 polypeptide sequence.

3. The transgenic mouse of claim 1 , wherein said polypeptide comprises an FKBP polypeptide sequence.

4. The transgenic mouse of claim 1 , wherein said polypeptide is an FKBP-caspase 8 fusion polypeptide.

5. The transgenic mouse of claim 1 , wherein said compound is AP20187.

6. The transgenic mouse of claim 1 , wherein the genetic background of said transgenic mouse is a BubR1 H/H genetic background.

7. The transgenic mouse of claim 1 , wherein said transgene comprises nucleic acid encoding a marker polypeptide.

8. The transgenic mouse of claim 7 , wherein said marker polypeptide is a GFP polypeptide.

9. A transgenic mouse, the genome of which contains a transgene, that comprises:
(a) a p16INK4a promoter sequence operably linked to a nucleic acid sequence encoding a polypeptide that directly induces cell death in cells in which said polypeptide is expressed when activated by a compound; and
(b) an internal ribosome entry site (IRES) sequence followed by a nucleic acid sequence encoding a marker polypeptide;
wherein senescent cells in said mouse express both said polypeptide and said marker polypeptide, and wherein administration of said compound to said transgenic mouse activates said polypeptide, thereby directly inducing cell death in senescent cells and inducing cell death in less than 10% of non-senescent cells; whereby progression of an age-related phenotype is delayed in said mouse relative to another such transgenic mouse that is not administered said compound.

10. The transgenic mouse of claim 9 , wherein the polypeptide comprises a caspase 8 polypeptide sequence.

11. The transgenic mouse of claim 9 , wherein the polypeptide comprises an FKBP polypeptide sequence.

12. The transgenic mouse of claim 9 , wherein the polypeptide is an FKBP-caspase 8 fusion polypeptide.

13. The transgenic mouse of claim 9 , wherein the compound is AP20817.

14. The transgenic mouse of claim 9 , wherein the genetic background of the transgenic mouse is a BubR1 H/H genetic background.

15. The transgenic mouse of claim 9 , wherein the marker polypeptide is a GFP polypeptide.

16. The transgenic mouse of claim 1 , wherein the age-related phenotype is sarcopenia.

17. The transgenic mouse of claim 1 , wherein the age-related phenotype is a cataract.

18. The transgenic mouse of claim 1 , wherein the age-related phenotype is loss of adipose tissue.

19. The transgenic mouse of claim 9 , wherein the age-related phenotype is sarcopenia.

20. The transgenic mouse of claim 9 , wherein the age-related phenotype is a cataract.

21. The transgenic mouse of claim 9 , wherein the age-related phenotype is loss of adipose tissue.

22. A method for removing senescent cells from a tissue, comprising contacting said tissue with a compound, wherein the genome of cells in said tissue contains a recombinant nucleic acid in which a p16INK4a promoter sequence controls expression of a polypeptide so as to cause said polypeptide to be expressed in senescent cells, wherein said polypeptide directly induces cell death in cells in which said polypeptide is expressed when activated by said compound, and wherein contacting said tissue with said compound directly induces cell death in senescent cells and induces cell death in less than 10% of non-senescent cells, whereby progression of an age-related phenotype is delayed in said tissue as a result of contacting said tissue with said compound.

23. The method of claim 22 , wherein said promoter in said recombinant nucleic acid controls expression of a marker polypeptide.

24. The method of claim 23 , wherein said marker polypeptide is a GFP polypeptide.

25. A method of determining whether delayed onset of an age-related phenotype coincides with a reduction in the number of senescent cells in a tissue, said method comprising:
(a) obtaining a plurality of mice according to claim 1 ;
(b) treating a proportion of the mice in said plurality with said compound, thereby directly inducing cell death in senescent cells in the treated mice; and
(c) imaging or measuring a sign of the age-related phenotype in the treated mice or in the mice that were untreated.

26. The method of claim 25 , wherein said compound is AP20187.

27. The method of claim 25 , wherein said age-related phenotype is expression of senescence-related markers in inguinal adipose tissue (IAT).

28. The method of claim 27 , wherein said sign is staining for senescence-associated beta-galactosidase (SA β Gal).

29. The method of claim 25 , wherein said age-related phenotype is replicative arrest.

30. The method of claim 29 , wherein said sign is lower incorporation of BrdU.

31. A method of determining whether delayed onset of an age-related phenotype coincides with a reduction in the number of senescent cells in a tissue, said method comprising:
(a) obtaining a plurality of mice according to claim 9 ;
(b) treating a proportion of the mice in said plurality with said compound, thereby directly inducing cell death in senescent cells in the treated mice; and
(c) imaging or measuring a sign of the age-related phenotype in the treated mice or in the mice that were untreated.

Assignees

• Mayo Found Medical Education & Res

Inventors

• Kirkland James L
• Tchkonia Tamar
• Van Deursen Jan M A
• Baker Darren J

Classifications

• A61K49/0008

  Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure · CPC title

• A01K67/0275 Primary

  Genetically modified vertebrates, e.g. transgenic · CPC title

• C12N15/62

  DNA sequences coding for fusion proteins · CPC title

• A01K2217/203

  Animal model comprising inducible/conditional expression system, e.g. hormones, tet · CPC title

• A01K2217/052

  inducing gain of function · CPC title