Targeted cell death

Abstract

The present invention provides compositions and methods for studying neuropathy. The compositions and methods provided herein are particularly useful for screening agents of therapeutic and/or diagnostic potential.

What is claimed is: 
     
         1 . A transgenic animal comprising:
 a nucleotide sequence encoding a cell death mediator protein (CDMP) operably linked to a cell type-specific expression regulatory element;   wherein said animal exhibits a greater degree of neuropathy relative to an animal without said nucleotide sequence.   
     
     
         2 . The animal of  claim 1 , wherein said animal is a mammal, primate, or rodent. 
     
     
         3 . The animal of  claim 1 , wherein said animal is a mouse, rat, guinea pig, dog, cat, rabbit, pig, chimpanzee or monkey. 
     
     
         4 . The animal of  claim 1 , wherein said neuropathy comprises neuronal demyelination and/or defect in the blood brain barrier. 
     
     
         5 . The animal of  claim 1 , wherein said animal exhibits an increase in apoptotic oligodendrocytes or pericytes relative to that of a control animal. 
     
     
         6 . The animal of  claim 1 , wherein said CDMP is caspase 2, caspase 5, caspase 8, caspase 9, caspase 10 or caspase 11. 
     
     
         7 . The animal of  claim 1 , wherein said CDMP is a chimeric protein comprising a binding domain for a FK506-type ligand, a FKBP12-type ligand, cyclosporin A-type ligand, tetracycline or steroid ligand. 
     
     
         8 . The animal of  claim 1 , wherein expression of said CDMP is inducible. 
     
     
         9 . The animal of  claim 1 , wherein expression of said CDMP is ectopically confined to the central nervous system. 
     
     
         10 . The animal of  claim 1 , wherein the apoptosis promoting activity of said CDMP is inducible. 
     
     
         11 . The animal of  claim 10 , wherein said apoptosis promoting activity is specifically in the neural cells of said animal. 
     
     
         12 . The animal of  claim 10 , wherein said activity is induced by a chemical inducer of dimerization (CID). 
     
     
         13 . The animal of  claim 10 , wherein said CDMP is an inducible caspase 9 (iCP9). 
     
     
         14 . The animal of  claim 12 , wherein said CID is AP20187. 
     
     
         15 . The animal of  claim 1 , wherein said cell type-specific expression regulatory element is a neural or mural cell specific regulatory element. 
     
     
         16 . The animal of  claim 15 , wherein said neural cell specific regulatory element is a glial cell specific regulatory element. 
     
     
         17 . The animal of  claim 16 , wherein said glial cell is an oligodendrocyte, astrocyte, microglial cell, or Schwann cell. 
     
     
         18 . The animal of  claim 16 , wherein said glial cell specific regulatory element is from a gene selected from CC1, myelin basic protein (MBP), ceramide galactosyltransferase (CGT), proteolipid protein (PLP). oligodendrocyte-myelin glycoprotein (OMG), cyclic nucleotide phosphodiesterase (CNP), NOGO, myelin protein zero (MPZ), peripheral myelin protein 22 (PMP22), protein 2 (P2), GFAP, AQP4, PDGFα, RG5, pGlycoprotein, neurturin (NRTN), artemin (ARTN), persephin (PSPN), PDGFR-α, PDGFR-β, or sulfatide. 
     
     
         19 . The animal of  claim 1 , wherein said nucleic acid sequence is operably linked to a second nucleic acid sequence encoding a marker protein. 
     
     
         20 . A method of screening for a biologically active agent that modulates a phenomenon associated with a demyelination or blood brain barrier disorder comprising:
 a) contacting a candidate agent with a cell comprising a nucleic acid encoding a cell death mediator protein (CDMP), wherein said nucleic acid is operably linked to a glial cell specific expression regulatory element;   b) detecting an effect on said phenomenon; and,   c) selecting said agent as effective to modulate said phenomenon if the level of activity of said CDMP is modulated relative to a control cell.   
     
     
         21 . The method of  claim 20 , wherein said cell is a neuronal, glial or mural cell. 
     
     
         22 . The method of  claim 20 , wherein said cell is an oligodendrocyte, astrocyte, microglial cell, pericyte, or Schwann cell. 
     
     
         23 . A method of screening for a biologically active agent that modulates a phenomenon associated with a demyelination or blood brain barrier disorder comprising:
 a) administering a candidate agent to a non-human transgenic animal, wherein said phenomenon occurs in said animal upon expression of a nucleic acid sequence encoding a cell death mediator protein (CDMP); wherein said nucleic acid sequence expression is regulated by a cell-specific expression regulatory element;   b) activating said CDMP to effect apoptosis in at least one cell in said animal, wherein said cell is associated with a demyelination blood brain barrier disorder; and,   c) detecting an effect of said agent upon said phenomenon.   
     
     
         24 . The method of  claim 23 , wherein after step b), said animal is allowed to recover from said phenomenon. 
     
     
         25 . The method of  claim 23 , wherein said phenomenon is characterized by a loss of oligodendrocytes, astrocytes, pericytes, or Schwann cells in said animal. 
     
     
         26 . The method of  claim 23 , wherein said phenomenon is characterized by a decrease in myelinated axons or an increase in blood brain barrier permeability. 
     
     
         27 . The method of  claim 20  or  23 , wherein said regulatory element is from a gene selected from the CC1, myelin basic protein (MBP), ceramide galactosyltransferase (CGT), oligodendrocyte-myelin glycoprotein (OMG), cyclic nucleotide phosphodiesterase (CNP), NOGO, myelin protein zero (MPZ), peripheral myelin protein 22 (PMP22), protein 2 (P2), GFAP, AQP4, PDGFα, RG5, pGlycoprotein, neurturin (NRTN), artemin (ARTN), persephin (PSPN), sulfatide, PDGFR-β, PDGFR-α, or proteolipid protein (PLP) gene. 
     
     
         28 . The method of  claim 23 , wherein said demyelination disorder is multiple sclerosis. 
     
     
         29 . The method of  claim 20  or  23 , wherein said determining said effect involves a PCR, immunoassay, hybridization assay or a combination thereof. 
     
     
         30 . The method of  claim 20  or  23 , wherein said candidate agent is an antisense oligonucleotide, a peptide, an antibody, a liposome, a small interfering RNA, a small organic compound, or an inorganic compound. 
     
     
         31 . The method of  claim 23 , wherein said at least one cell is a neuronal, glial or mural cell. 
     
     
         32 . The method of  claim 31 , wherein said at least one cell is an oligodendrocyte, astrocyte, microglial cell, pericyte, or Schwann cell. 
     
     
         33 . The method of  claim 20  or  23 , wherein said CDMP is caspase 2, caspase 5, caspase 8, caspase 9, caspase 10, or caspase 11. 
     
     
         34 . The method of  claim 20  or  23 , wherein said CDMP is a chimeric protein comprising a binding domain for a FK506-type ligand, a FKBP12-type ligand, cyclosporin A-type ligand, tetracycline or steroid ligand. 
     
     
         35 . The method of  claim 20  or  23 , wherein expression of said CDMP is inducible. 
     
     
         36 . The method of  claim 20  or  23 , wherein the apoptosis promoting activity of said CDMP is inducible. 
     
     
         37 . The method of  claim 36 , wherein said activity is induced by a chemical inducer of dimerization (CID). 
     
     
         38 . The method of  claim 20  or  23 , wherein said CDMP is an inducible caspase 9 (iCP9). 
     
     
         39 . The method of  claim 38 , wherein said CID is AP20187. 
     
     
         40 . A method for compiling a profile data set for characterizing a phenomenon associated with multiple sclerosis (MS) or MS-associated condition related to multiple sclerosis comprising:
 a) providing a transgenic animal or cell comprising a nucleic acid encoding a cell death mediator protein (CDMP), wherein said nucleic acid is operably linked to a neuronal- or glial-specific expression regulatory element;   b) activating said CDMP thereby inducing apoptosis;   c) obtaining at least one surviving neuronal or glial cell following said activation; and   d) profiling RNA transcripts and/or encoded products in said surviving glial or neuronal cell; thereby compiling a profile data set characterizing a phenomenon associated with multiple sclerosis or MS-associated condition related to multiple sclerosis.   
     
     
         41 . The method of  claim 40 , wherein said CDMP is caspase 9 or caspase 11. 
     
     
         42 . The method of  claim 40 , wherein the apoptosis inducing activity of said CDMP is inducible. 
     
     
         43 . The method of  claim 40 , wherein said glial-specific expression regulatory element is from a CC1, myelin basic protein (MBP), ceramide galactosyltransferase (CGT), and proteolipid protein (PLP). the group consisting of CC1, myelin basic protein (MBP), ceramide galactosyltransferase (CGT), oligodendrocyte-myelin glycoprotein (OMG), cyclic nucleotide phosphodiesterase (CNP), NOGO, myelin protein zero (MPZ), peripheral myelin protein 22 (PMP22), protein 2 (P2), GFAP, AQP4, PDGFα, PDGFR-β, PDGFR-α, RG5, pGlycoprotein, neurturin (NRTN), artemin (ARTN), persephin (PSPN), sulfatide, or proteolipid protein (PLP) gene.