Compositions and method for manipulating pim-1 activity in circulatory system cells

Abstract

The invention provides compositions (e.g., pharmaceutical compositions) comprising nucleic acids encoding the serine/threonine kinase PIM-1, and methods for making and using them; including methods for inducing cellular proliferation, and protecting cardiac cells from hypoxia and cellular apoptosis. The invention provides compositions (e.g., pharmaceutical compositions) comprising nucleic acids encoding PIM-1, and methods for enhancing the regenerative potential of stem cells in the heart.

What is claimed is: 
     
         1 . A method, comprising identifying a patient in need of enhanced PIM activity in a vascular system cell or tissue, and enhancing levels of PIM in the vascular system cell or tissue of the patient to alter a functional characteristic of cells in that tissue. 
     
     
         2 . The method of  claim 1 , wherein:
 (a) the patient has experienced cardiac injury and the enhanced PIM levels facilitate cardiac regeneration to repair that injury;   (b) the enhancing step comprises enhancing endogenous production of PIM in the vascular system tissue;   (c) the enhancing step comprises administering to the patient an exogenous PIM;   (d) the exogenous PIM comprises a PIM enzyme in association with a cellular delivery moiety;   (e) the cellular delivery moiety is a translocation domain that is attached to the PIM enzyme;   (f) the enhancing step comprises administering cells to the patient that produce enhanced levels of PIM;   (g) the administered cells are stem cells or vascular system progenitor cells;   (h) the administered cells comprise a PIM-encoding polynucleotide operatively linked to a non-PIM promoter, a cardiac-specific promoter, a promoter active in a cardiac cell or a stem cell, or a PIM promoter;   (i) the promoter is or comprises a α-myosin heavy chain promoter; a cardiac troponin-T promoter or a MLC-2v promoter; or   (j) the enhancing step comprises: administering cells to a vascular tissue of the patient, and expressing enhanced levels of PIM from the administered cells.   
     
     
         3 . A PIM enzyme-delivering or enhancing material for treatment of vascular system disease or injury. 
     
     
         4 . The PIM enzyme-delivering or enhancing material of  claim 3 , comprising:
 (a) a PIM enzyme linked to a cellular delivery agent;   (b) a cell for introduction into a human or animal, wherein the cell has been altered to permit enhanced production of a PIM; or   (c) comprising an inducer of endogenous PIM expression.   
     
     
         5 . The PIM enzyme-delivering or enhancing material of  claim 3 , wherein:
 (a) the cell is a progenitor cell or a stem cell, and wherein the alteration comprises a PIM-encoding polynucleotide under control of a non-PIM promoter, a cardiac-specific promoter, a promoter active in a cardiac cell or a stem cell, or a PIM promoter; or   (b) the promoter is or comprises a α-myosin heavy chain promoter; a cardiac troponin-T promoter or a MLC-2v promoter.   
     
     
         6 . A composition, comprising a vascular system cell or a cell that is differentiable or can differentiate into a vascular system cell, wherein the cell comprises a PIM-encoding polynucleotide sequence operably linked to a non-PIM promoter, a cardiac-specific promoter, a promoter active in a cardiac cell or a stem cell, or a PIM promoter. 
     
     
         7 . The composition of  claim 6 , wherein:
 (a) the promoter is or comprises a α-myosin heavy chain promoter; a cardiac troponin-T promoter or a MLC-2v promoter;   (b) the cell is a stem cell or cardiac progenitor cell, or the is differentiable (can differentiate) into a myocyte or a heart cell;   (c) the cell is a mesenchymal stem cell; or   (d) the promoter is an inducible promoter.   
     
     
         8 . A method for treating, preventing or ameliorating a cardiac disease or an injury, comprising enhancing levels of PIM within diseased or injured cardiac tissue. 
     
     
         9 . The method of  claim 8 , wherein the cardiac disease or injury comprises: ischemic injury, hypoxic injury, myocardial infarction, traumatic cardiac injury, cardiac hypertrophy, overpressure injury, congestive heart failure, apoptosis-inducing injury or disease, bacterial infection, viral infection, and conditions that create an enhanced risk of any of the foregoing. 
     
     
         10 . A pharmaceutical composition formulated for administration to a heart muscle comprising:
 (i) (a) a Pim-1 encoding nucleic acid;   (b) a Pim-1 encoding nucleic acid inserted in an expression construct or expression vehicle, or a naked Pim-1 encoding nucleic acid operably linked to a promoter;   (c) the pharmaceutical composition of (b), wherein the expression construct or expression vehicle comprises or consists of a vector, a plasmid, a recombinant virus or an artificial chromosome;   (d) the pharmaceutical composition of (c), wherein the expression construct or expression vehicle comprises or consists of a recombinant adeno-associated viral vector; an adenovirus vector, a retroviral vector; or a lentiviral vector;   (e) the pharmaceutical composition of (d), wherein the expression construct or expression vehicle comprises or consists of an immunodeficiency virus derived vector;   (f) the pharmaceutical composition of (e), wherein the immunodeficiency virus derived vector comprises or consists of a human immunodeficiency virus (HIV) derived vector; or   (g) the pharmaceutical composition of (f), wherein the human immunodeficiency virus (HIV) derived vector comprises or consists of a human immunodeficiency virus-1 (HIV-1) derived vector;   (h) the pharmaceutical composition of any of (a) to (g), wherein the Pim-1 encoding nucleic acid is operably linked to a promoter;   (i) the pharmaceutical composition of (h), wherein the promoter is a constitutive or an inducible promoter; or   (j) the pharmaceutical composition of (i), wherein the promoter is constitutively or inducibly active in a heart cell (a myocyte); and,   (ii) a pharmaceutically acceptable excipient.   wherein the pharmaceutical composition formulated for administration to a heart muscle.   
     
     
         11 . A liposome comprising the pharmaceutical compound of  claim 10 . 
     
     
         12 . A nanoparticle comprising the pharmaceutical compound of  claim 10 . 
     
     
         13 . A method for inducing, upregulating or inserting a PIM-1 nucleic acid or a Pim-1 kinase activity in a cardiac or vascular cell, tissue or organ, comprising:
 (a) (i) providing a Pim-1 encoding nucleic acid; and inserting the Pim-1 encoding nucleic acid into the cardiac or vascular cell, tissue or organ; (ii) providing a cell expressing and/or secreting a Pim-1 kinase; (iii) administering Pim-1 kinase or a Pim-1 expressing nucleic acid to the cardiac or vascular cell, tissue or organ; or, (iv) providing a compound that induces or upregulates PIM-1 nucleic acid or a Pim-1 kinase activity in a cardiac or vascular cell, tissue or organ;   (b) the method of (a), wherein the Pim-1 encoding nucleic acid comprises or consists of a Pim-1 encoding message (a Pim-1 encoding mRNA), or a PIM-1 gene;   (c) the method of (a) or (b), wherein the Pim-1 encoding nucleic acid comprises or consists of a human Pim-1 encoding nucleic acid, or a human Pim-1 encoding message (mRNA), or a human PIM-1 gene, or a human PIM-1 gene locus;   (d) the method of any of (a) to (c), wherein the cell is a human cell, a stem cell, an adult stem cell, a hematopoietic stem cell, an adipose-derived stem cell, a mesenchymal stem cell, a c-kit +  stem cell, a human stem cell, an autologous or allogeneic stem cell, an embryonic cell, a tissue-specific resident stem cell, an allogeneic or autologous cell, a progenitor cell, a placental and/or cord blood cell, a Sca-1+ cell, or a CD34+ cell;   (e) the method of any of (a) to (d), wherein the Pim-1 encoding nucleic acid is inserted into a cardiac or vascular cell, tissue or organ ex vivo or in vivo;   (f) the method of any of (a) to (e), wherein a Pim-1 encoding nucleic acid is inserted in an expression construct or expression vehicle;   (g) the method of any of (f), wherein the expression construct or expression vehicle comprises or consists of a vector, a plasmid, a recombinant virus or an artificial chromosome;   (h) the method of any of (g), wherein the expression construct or expression vehicle comprises or consists of a recombinant adeno-associated viral vector; an adenovirus vector, a retroviral vector; or a lentiviral vector;   (i) the method of any of (h), wherein the expression construct or expression vehicle comprises or consists of an immunodeficiency virus derived vector;   (j) the method of any of (i), wherein the immunodeficiency virus derived vector comprises or consists of a human immunodeficiency virus (HIV) derived vector;   (k) the method of any of (j), wherein the human immunodeficiency virus (HIV) derived vector comprises or consists of a human immunodeficiency virus-1 (HIV-1) derived vector;   (l) the method of any of (a) to (k), wherein the Pim-1 encoding nucleic acid is inserted into a cell that does not express wild type (normal) levels of PIM-1 protein;   (m) the method of (l), wherein the Pim-1 encoding nucleic acid is inserted into a cell that does not express wild type (normal) levels of PIM-1 protein-encoding message (mRNA);   (n) the method of (m), wherein the Pim-1 encoding nucleic acid is inserted into a cell that does not comprise a wild type (normal) PIM-1 gene or genomic Pim-1 encoding nucleic acid;   (o) the method of any of (a) to (n), wherein the Pim-1 encoding nucleic acid is inserted into a cardiac or vascular cell, tissue or organ ex vivo and the cardiac or vascular cell, tissue or organ is implanted into an individual in need thereof;   (p) the method of any of (a) to (o), wherein the Pim-1 encoding nucleic acid is inserted into a heart cell, cardiac or vascular tissue or cardiac or vascular organ or a myocyte cell ex vivo and the cell is implanted into a cardiac or vascular cell, tissue or organ or a myocardium (a heart) in need thereof;   (q) the method of any of (a) to (n), wherein the Pim-1 encoding nucleic acid is in vivo inserted into a cardiac or vascular cell, tissue or organ in an individual in need thereof;   (r) the method of (q), wherein the Pim-1 encoding nucleic acid is inserted into a cardiac or vascular cell, tissue or organ or a heart cell or a myocyte cell or a heart in vivo;   (s) the method of (r), wherein the individual has congestive heart failure, or has had a myocardial infarction, or heart muscle damage;   (t) the method of any of (a) to (s), wherein the cardiac or vascular cell, tissue or organ is or is contained in: a heart cell (a myocyte), a heart tissue or a heart; or, a kidney, a liver, a spleen, a pancreas, skin, mucosa, an eye, a brain, a lung, a nerve, a sensory nerve, a peripheral nerve, a spinal cord, a blood vessel, an endothelium, a muscle, a smooth muscle, a skeletal muscle, a heart muscle or a bone or bone marrow;   (t) the method of (a), wherein the compound that induces or upregulates PIM-1 nucleic acid or a Pim-1 kinase activity in a cardiac or vascular cell, tissue or organ comprises: an interleukin, a cytokine and/or a paracrine factor involved in survival and/or proliferative signaling; an up-regulator of AKT serine/threonine kinase; insulin-like growth factor-1 (IGF-1); insulin; leukemia inhibitory factor (LIF); granulocyte-macrophage colony-stimulating factor (GM-CSF); or epidermal growth factor (EGF);   (u) the method of any of (a) to (t), wherein the wherein Pim-1 activity in the cardiac or vascular cell, tissue or organ is increased by administering an exogenous Pim-1 kinase to the population of cells;   (v) the method of (u), wherein Pim-1 activity is increased by contacting a population of cells with a transfected cell that expresses an exogenous PIM-1 gene; or   (w) the method of (u), wherein the population of cells comprises stem cells.   
     
     
         14 . A method for treating, preventing or ameliorating a disease or condition comprising administering to an individual in need thereof the pharmaceutical compound of  claim 10 ,
 wherein the treatment, prevention and/or amelioration of the disease or condition comprises:   the amelioration, treatment or prevention of cellular apoptosis and/or damage in a cardiac or vascular cell, tissue or organ subsequent to cellular, tissue and/or organ hypoxia, hypoxaemia or anoxia, or subsequent to pressure-overload induced hypertrophy or heart failure; or   the amelioration, treatment or prevention of cellular apoptosis and/or damage in a cardiac or vascular cell, tissue or organ subsequent to or because of a hypertrophic myocardium, an aged myocardium, a failing myocardium, an ischemic myocardium, a remodeled myocardium, a myocardium damaged by inflammation, infection, chronic stress, disease, diabetes or alcoholism; and/or oxidative damage,   by increasing or upregulating Pim-1 kinase activity in the cardiac or vascular cell, tissue or organ.   
     
     
         15 . The method of  claim 14 , wherein the cellular apoptosis and/or damage, or the hypoxia, hypoxaemia or anoxia, is caused by an infarction, trauma, surgery, reimplantation, transplantation or a toxin, or by inflammation, infection, chronic stress, diabetes or alcoholism;
 and/or oxidative damage.   
     
     
         16 . The method of  claim 14 , comprising inducing cellular dedifferentiation and/or re-expression of a stem cell marker in a cardiac or vascular cell, tissue or organ. 
     
     
         17 . The method of  claim 14 , comprising enhancing the retention of engrafted or transplanted cells, tissues or organs by overexpressing or expressing Pim-1 in the cells, tissues or organs. 
     
     
         18 . The method of  claim 14 , comprising increasing the expression of bcl-2, bcl-XL and/or phosphorylation of Bad protein in a cardiac or vascular cell, tissue or organ. 
     
     
         19 . The method of  claim 14 , comprising the amelioration, treatment or prevention of ischemia reperfusion injury in a cardiac or vascular cell, tissue or organ. 
     
     
         20 . The method of  claim 14 :
 (a) wherein the cardiac or vascular cell, tissue or organ is or is contained in: a heart cell (a myocyte), a heart tissue or a heart; or, a kidney, a liver, a spleen, a pancreas, skin, mucosa, an eye, a brain, a lung, a nerve, a sensory nerve, a peripheral nerve, a spinal cord, a blood vessel, an endothelium, a muscle, a smooth muscle, a skeletal muscle, a heart muscle or a bone or bone marrow;   (b) comprising overexpressing or expressing Pim-1 in a stem cell or a pluripotent cell to enhance the regenerative potential and/or induce proliferation of the stem cell or pluripotent cell;   (c) comprising overexpressing or expressing Pim-1 in a heart cell (a myocyte) or heart tissue to increase Bcl-XL expression in the heart cell (myocyte) or heart tissue to induce cardioprotective anti-apoptotic signaling and/or to increase myocardial survival signaling; or   (d) wherein the cell is a stem cell, an adult stem cell, a hematopoietic stem cell, an adipose-derived stem cell, a mesenchymal stem cell, a c-kit+ stem cell, a human stem cell, an autologous or allogeneic stem cell, an embryonic cell, a tissue-specific resident stem cell, an allogeneic or autologous cell, a progenitor cell, a placental and/or cord blood cell, a Sca-1+ cell, or a CD34+ cell.