Systemic delivery and regulated expression of paracrine genes for cardiovascular diseases and other conditions

Abstract

In alternative embodiments, the invention provides methods for treating, ameliorating or protecting (preventing) an individual or a patient against a disease, an infection or a condition responsive to an increased paracrine polypeptide level in vivo comprising: providing a paracrine polypeptide-encoding nucleic acid or gene operatively linked to a transcriptional regulatory sequence; or an expression vehicle, a vector, a recombinant virus, or equivalent, having contained therein a paracrine-encoding nucleic acid or gene, and the expression vehicle, vector, recombinant virus, or equivalent can express the paracrine-encoding nucleic acid or gene in a cell or in vivo; and administering or delivering the paracrine polypeptide-encoding nucleic acid or gene operatively linked to a transcriptional regulatory sequence, or the expression vehicle, vector, recombinant virus, or equivalent, to an individual or a patient in need thereof, thereby treating, ameliorating or protecting (preventing) the individual or patient against the disease, infection or condition responsive to an increased paracrine polypeptide level.

1 . A method for treating, ameliorating, protecting or preventing an individual or a patient against a disease, an infection or a condition responsive to an increased or sustained peptide or paracrine polypeptide level in vivo comprising:
 (i) providing a paracrine polypeptide-encoding nucleic acid or gene operatively linked to a transcriptional regulatory sequence; or an expression vehicle, a vector, a recombinant virus, or equivalent, having contained therein a paracrine-encoding nucleic acid or gene, or a paracrine polypeptide-expressing nucleic acid, transcript or message, and the expression vehicle, vector, recombinant virus, or equivalent can express the paracrine-encoding nucleic acid, gene, transcript or message in a cell or in vivo; and   (ii) administering or delivering the paracrine polypeptide-encoding nucleic acid, gene, transcript or message operatively linked to a transcriptional regulatory sequence, or the expression vehicle, vector, recombinant virus, or equivalent, to the cell, or an individual or a patient in need thereof,   thereby treating, ameliorating, protecting or preventing the individual or patient against the disease, infection or condition responsive to an increased or a sustained paracrine polypeptide level.   
     
     
         2 . The method of  claim 1 , wherein:
 (a) the paracrine-encoding nucleic acid or gene operatively linked to the transcriptional regulatory sequence; or the expression vehicle, vector, recombinant virus, or equivalent, is administered or delivered to the individual or a patient in need thereof, by oral, intramuscular (IM) injection, by intravenous (IV) injection, by subcutaneous (SC) or intradermal injection, by intrathecal injection, by intra-arterial (IA) injection, by intracoronary injection, by inhalation, by aerosol, or by a biolistic particle delivery system, or by using a “gene gun”, air pistol or a HELIOS™ gene gun (Bio-Rad Laboratories, Hercules, Calif.); or   (b) the paracrine-encoding nucleic acid or gene operatively linked to the transcriptional regulatory sequence; or the expression vehicle, vector, recombinant virus, or equivalent, is administered or delivered to the individual or a patient in need thereof, by introduction into any tissue or fluid space within the body that is adjacent to or is drained by the bloodstream, such that the encoded protein may be secreted from cells in the tissue and released into the bloodstream.   
     
     
         3 . The method of  claim 1 , wherein the paracrine polypeptide or peptide is or comprises: a mammalian cardiotonic peptide, a growth factor, a Serelaxin, a Relaxin-2, a Urocortin-2 (UCn-2), a Urocortin-1 (UCn-1), a Urocortin-3 (UCn-3), a Brain Natriuretic Peptide, a Prostacyclin Synthase, a Growth Hormone, an Insulin-like Growth Factor-1, or any combination thereof; or, a human cardiotonic peptide, a human growth factor, a Serelaxin, a Relaxin-2, a Urocortin-2, a Urocortin-1, a Urocortin-3, a Brain Natriuretic Peptide, a Prostacyclin Synthase, a Growth Hormone, an Insulin-like Growth Factor-11, or any combination thereof. 
     
     
         4 . The method of  claim 3 , wherein the paracrine polypeptide is a Urocortin, a Urocortin-2, a Urocortin-1, a Urocortin-3, a Relaxin-2 or a Brain Natriuretic Peptide and the disease or condition is a congestive heart failure (CHF); or the paracrine polypeptide is Prostacyclin Synthase and the disease or condition a pulmonary hypertension. 
     
     
         5 . The method of  claim 1 , wherein:
 (a) the individual, patient or subject is administered a stimulus or signal that induces expression of the paracrine-expressing nucleic acid or gene, or induces or activates a promoter operably linked to the paracrine-expressing nucleic acid or gene that induces expression of the paracrine-expressing nucleic acid or gene;   (b) the individual, patient or subject is administered a stimulus or signal that induces synthesis of an activator of a promoter,   optionally a paracrine-expressing nucleic acid or gene-specific promoter operably linked to the paracrine-expressing nucleic acid or gene;   (c) the individual, patient or subject is administered a stimulus or signal that induces synthesis of a natural or a synthetic activator of the paracrine-expressing nucleic acid or gene or the paracrine-expressing nucleic acid or gene-specific promoter,   wherein optionally the natural activator is an endogenous transcription factor;   (d) the method of (c), wherein the synthetic activator is a zinc-finger DNA binding protein designed to specifically and selectively turn on an endogenous or exogenous target gene, wherein optionally the endogenous target is a gene paracrine-expressing nucleic acid or gene or an activator of a paracrine-expressing nucleic acid or gene, or an activator of a promoter operatively linked to a paracrine-expressing nucleic acid or gene;   (e) the method of any of (a) to (c), wherein the stimulus or signal comprises a biologic, a light, a chemical or a pharmaceutical stimulus or signal;   (f) the individual, patient or subject is administered a stimulus or signal that stimulates or induces expression of a post-transcriptional activator of a paracrine-expressing nucleic acid or gene, or an activator of a promoter operatively linked to a paracrine-expressing nucleic acid or gene, or   (g) the individual, patient or subject is administered a stimulus or signal that inhibits or induces inhibition of a transcriptional repressor or a post-transcriptional repressor of a paracrine-expressing nucleic acid or gene.   
     
     
         6 . The method of  claim 5 , wherein the chemical or pharmaceutical that induces expression of the paracrine-expressing nucleic acid or gene, or induces expression of the regulated or inducible promoter operatively linked to the paracrine-expressing nucleic acid or gene, is an oral antibiotic, a doxycycline or a rapamycin; or a tet-regulation system using doxycycline is used to induce expression of the paracrine-expressing nucleic acid or gene, or an equivalent thereof. 
     
     
         7 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent, is formulated in a liquid, a gel, a hydrogel, a powder or an aqueous or a saline formulation. 
     
     
         8 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent, is formulated in a vesicle, liposome, nanoparticle or nanolipid particle (NLP). 
     
     
         9 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent, is formulated in an isolated or cultured cell, and optionally the cell is a mammalian cell, a cardiac cell, or a human cell, a non-human primate cell, a monkey cell, a mouse cell, a rat cell, a guinea pig cell, a rabbit cell, a hamster cell, a goat cell, a bovine cell, an equine cell, an ovine cell, a canine cell or a feline cell. 
     
     
         10 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent, is formulated as a pharmaceutical or sterile. 
     
     
         11 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent, is formulated or delivered with, on, or in conjunction with a product of manufacture, an artificial organ or an implant. 
     
     
         12 . The method of  claim 1 , wherein the paracrine-expressing nucleic acid or gene or the expression vehicle, vector, recombinant virus, or equivalent expresses a paracrine polypeptide in vitro or ex vivo. 
     
     
         13 . A method for treating, ameliorating, protecting or preventing an individual or a patient against a paracrine-responsive pathology, infection, disease, illness, or condition, comprising practicing the method of  claim 1 . 
     
     
         14 . A method for treating, ameliorating, protecting or preventing a cardiac contractile dysfunction; a congestive heart failure (CHF); a cardiac fibrosis; a cardiac myocyte disease, dysfunction or apoptosis; a pulmonary hypertension; a heart, skin, liver, lung, muscle, nerve, brain or kidney disease, cancer or dysfunction; a cancer or a neoplasia; or, a hemophilia or a Hemophilia B, comprising practicing the method of  claim 1 . 
     
     
         15 . A treating, ameliorating, protecting or preventing a diabetes or a pre-diabetes in a patient or an individual comprising:
 (a) practicing the method of  claim 1 , wherein the paracrine polypeptide or peptide comprises or consists of a urocortin-2 (UCn-2); and   (b) administering a urocortin-2 (UCn-2) peptide or polypeptide, or a nucleic acid, gene, message or transcript encoding a urocortin-2 (UCn-2) to an individual or patient in need thereof,   wherein optionally the urocortin-2 (UCn-2) peptide or polypeptide is an isolated, a recombinant, a synthetic and/or a peptidomimetic peptide or polypeptide or variant thereof,   thereby treating, ameliorating, protecting or preventing a the diabetes or pre-diabetes in the patient or individual.   
     
     
         16 . A method of treating, ameliorating, protecting or preventing an obesity in a patient or an individual comprising:
 (a) practicing the method of  claim 1 , wherein the paracrine polypeptide or peptide comprises or consists of a urocortin-2 (UCn-2); and   (b) administering a urocortin-2 (UCn-2) peptide or polypeptide, or a nucleic acid, gene, message or transcript encoding a urocortin-2 (UCn-2) to an individual or patient in need thereof,   wherein optionally the urocortin-2 (UCn-2) peptide or polypeptide is an isolated, a recombinant, a synthetic and/or a peptidomimetic peptide or polypeptide or variant thereof,   thereby treating, ameliorating, protecting or preventing a the obesity in the patient or individual.   
     
     
         17 . A method of suppressing weight gain, or suppressing the appetite, or stimulating or initiating weight loss, in a patient or an individual comprising:
 (a) practicing the method of  claim 1 , wherein the paracrine polypeptide or peptide comprises or consists of a urocortin-2 (UCn-2); and   (b) administering a urocortin-2 (UCn-2) peptide or polypeptide, or a nucleic acid, gene, message or transcript encoding a urocortin-2 (UCn-2) to an individual or patient in need thereof,   wherein optionally the urocortin-2 (UCn-2) peptide or polypeptide is an isolated, a recombinant, a synthetic and/or a peptidomimetic peptide or polypeptide or variant thereof,   thereby suppressing weight gain, or suppressing the appetite, or stimulating or initiating weight loss, in the patient or individual.   
     
     
         18 . The method of  claim 10 , wherein the urocortin-2 (UCn-2) peptide or polypeptide is formulated in or as a vesicle, liposome, nanoparticle or nanolipid particle (NLP), or is formulated for: oral administration, intramuscular (IM) injection, intravenous (IV) injection, subcutaneous (SC) or intradermal injection, intrathecal injection, intra-arterial (IA) injection, intracoronary injection, inhalation, or administration by aerosol. 
     
     
         19 . The method of  claim 1 , wherein the expression vehicle, vector, recombinant virus, or equivalent is or comprises:
 an adeno-associated virus (AAV), a lentiviral vector or an adenovirus vector,   an AAV serotype AAV5, AAV6, AAV8 or AAV9,   a rhesus-derived AAV, or the rhesus-derived AAV AAVrh.10hCLN2,   an AAV capsid mutant or AAV hybrid serotype,   an organ-tropic AAV, or a cardiotropic AAV, or a cardiotropic AAVM41 mutant,   wherein optionally the AAV is engineered to increase efficiency in targeting a specific cell type that is non-permissive to a wild type (wt) AAV and/or to improve efficacy in infecting only a cell type of interest,   and optionally the hybrid AAV is retargeted or engineered as a hybrid serotype by one or more modifications comprising: 1) a transcapsidation, 2) adsorption of a bi-specific antibody to a capsid surface, 3) engineering a mosaic capsid, and/or 4) engineering a chimeric capsid.   
     
     
         20 . The method of  claim 1 , wherein:
 (a) the paracrine-encoding nucleic acid, gene, transcript or message is operatively linked to a regulated or inducible transcriptional regulatory sequence;   (b) the regulated or inducible transcriptional regulatory sequence is a regulated or inducible promoter,   wherein optionally a positive or an activator and/or a negative or a repressor modulator of transcription and/or translation is operably linked to the paracrine polypeptide-encoding nucleic acid, gene, transcript or message;   (c) administering the paracrine polypeptide-encoding nucleic acid, gene, transcript or message operatively linked to a transcriptional regulatory sequence, or the expression vehicle, vector, recombinant virus, or equivalent, to an individual or a patient in need thereof results in a paracrine protein being released into the bloodstream or general circulation, or an increased or sustained expression of the paracrine protein in the cell,   wherein optionally the release or increased or sustained expression of the paracrine protein is dependent on activation of an inducible promoter, or de-repression of a repressor, operably linked to the paracrine polypeptide-encoding nucleic acid, gene, transcript or message; or   (d) the method of any of (a) to (c), wherein the disease, infection or condition responsive to an increased paracrine polypeptide level in vivo is a cardiac contractile dysfunction; a congestive heart failure (CHF); a cardiac fibrosis; a cardiac myocyte disease, dysfunction or apoptosis; a pulmonary hypertension; a heart, skin, liver, lung, muscle, nerve, brain or kidney disease, cancer or dysfunction; a cancer or a neoplasia; or, a hemophilia or a Hemophilia B.