Methods of Cell Renewal

Abstract

The present invention encompasses compositions and methods to rejuvenate cells by, expanding the replicative life span of the cells for, e.g., use in regenerative therapies. Specifically, the methods and compositions of the present invention increase the proliferation capacity and differentiation capacity and plasticity of cells.

We claim: 
     
         1 . A mammalian synthetic chromosome comprising at least one stably-integrated expression cassette expressing at least two genes selected from the group consisting of:
 a) a gene that controls telomere length;   b) a gene involved in an anti-oxidant response pathway; and   c) a gene involved in an anti-inflammatory response.   
     
     
         2 . The mammalian synthetic chromosome of  claim 1 , wherein the mammalian synthetic chromosome is a human synthetic chromosome. 
     
     
         3 . A mammalian synthetic chromosome comprising at least one of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         4 . The mammalian synthetic chromosome of  claim 3 , comprising at least two of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         5 . The mammalian synthetic chromosome of  claim 4 , comprising all three of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         6 . The mammalian synthetic chromosome of  claim 3 , wherein the TERT expression cassette comprises TERT clone RP11-117B23, the SIRT1 expression cassette comprises SIRT1 clone RP11-22C9, and the NFE2L2 expression cassette comprises NFE2L2 clone RP11-844A8. 
     
     
         7 . The mammalian synthetic chromosome of  claim 3 , wherein the mammalian synthetic chromosome is a human synthetic chromosome. 
     
     
         8 . A mammalian cell comprising a mammalian synthetic chromosome comprising at least one of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         9 . The mammalian cell of  claim 8 , wherein the mammalian synthetic chromosome comprises at least two of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         10 . The mammalian cell of  claim 8 , wherein the mammalian synthetic chromosome comprises all three of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         11 . The mammalian cell of  claim 8 , wherein the TERT expression cassette comprises TERT clone RP11-117B23, the SIRT1 expression cassette comprises SIRT1 clone RP11-22C9, and the NFE2L2 expression cassette comprises NFE2L2 clone RP11-844A8. 
     
     
         12 . The mammalian cell of  claim 8 , wherein the mammalian cell comprising the synthetic chromosome has increased replicative lifespan, differentiation capacity and/or plasticity as compared to the mammalian cell before transfection of the human synthetic chromosome. 
     
     
         13 . The mammalian cell of  claim 8 , wherein the synthetic chromosome comprises an expression cassette cDNAs of the SIRT, NFE2L2 and TERT genes engineered as a single transcript containing self-cleaving peptides between the genes. 
     
     
         14 . A stem cell comprising a synthetic chromosome comprising all three of a stably-integrated expression cassette expressing TERT, a stably-integrated expression cassette expressing SIRT1, and a stably-integrated expression cassette expressing NFE2L2. 
     
     
         15 . The stem cell of  claim 14 , wherein the stem cell is a mammalian adult-derived mesenchymal stem cell (MSC) originating from adipose tissue. 
     
     
         16 . The stem cell of  claim 14 , wherein the stem cell is a mammalian adult-derived mesenchymal stem cell (MSC) originating from bone marrow. 
     
     
         17 . The stem cell of  claim 14 , wherein the stem cell is an autologous cell from a subject to be treated. 
     
     
         18 . The mammalian synthetic chromosome of  claim 5 , wherein the expression cassette comprises an endogenous TERT promoter driving expression of TERT, an endogenous SIRT1 promoter driving expression of SIRT1, and an endogenous NFE2L2 promoter driving expression of NFE2L2. 
     
     
         19 . The mammalian cell of  claim 13 , wherein the expression cassette comprises an inducible promoter driving expression of all of TERT, SIRT1, and NFE2L2.