HYPOXIA REGULATED CONDITIONALLY SILENCED AAV EXPRESSING ANGIOGENIC INDUCERS

Methods and compositions for the treatment of hypoxia associated disorders by directional angiogenesis/arteriogenesis. Conditionally silenced vectors expressing a therapeutic molecule under hypoxic conditions avoid chaotic vascularization and allow for the orderly growth of new vessels into damaged tissue. 134-. (canceled)35. A method of treating a patient having a disease or disorder associated with hypoxia or ischemia , comprising administering to the patient a composition comprising a conditionally silenced vector comprising: at least one FROG element , at least one TOAD element , at least one hypoxia response element (HRE) , a gene encoding at least one growth factor , and a promoter upstream of the gene that is in operable linkage with the gene , the at least one FROG element , the at least one TOAD element , and the at least one HRE , such that the conditionally silenced vector induces or promotes directional angiogenesis and the growth factor is not expressed in aerobic tissue but is expressed in ischemic or hypoxic tissue ,wherein expression of the growth factor induces vascularization of a tissue or organ in a desired direction.36. The method of claim 35 , wherein the composition is administered to the patient in a therapeutically effective controlled amount for preventing necrosis in the tissue or organ and increasing blood flow in the tissue or organ.37. The method of claim 35 , wherein the growth factor is not expressed in normoxic brain cells and normoxic stem cells.38. The method of claim 35 , wherein the conditionally silenced vector comprises a combined cassette of HREs and TOAD and FROG elements.39. The method of claim 35 , wherein the at least one FROG element consists of a nucleotide sequence selected from the group consisting of: SEQ ID NOS: 3 and 4 claim 35 , and the at least one TOAD element consists of a nucleotide sequence selected from the group consisting of: SEQ ID NOS: 1 and 2.40. The method of claim 35 , wherein the at least one FROG ...

Tissue specific hypoxia regulated constructs

Methods and compositions relating to chimeric genes containing (i) a tissue-specific promoter and (ii) a hypoxia response enhancer element, both of which are operably linked to a selected gene, such as a reporter gene, therapeutic gene (e.g., bcl-2, NOS, catalase and SOD), or deleterious gene are disclosed. Expression of the selected gene is enhanced in the target tissue under hypoxia conditions, such as conditions encountered during episodes of ischemia and reperfusion. The methods and compositions may be used as therapeutics and/or diagnostics.

COMPOSITIONS, CELLS, KITS AND METHODS FOR AUTOLOGOUS STEM CELL THERAPY

Described herein are compositions, kits and methods for stimulating angiogenic functions of stem cells and/or progenitor cells having pro-angiogenic potential (e.g., endothelial progenitor cells (EPCs), mesenchymal stem cells (MSCs)) before transplantation (e.g., ex vivo cell therapy) based on the discovery that functional recovery of CD34+ cells from coronary artery disease (CAD) patients is improved by transfection of antagomirs against one or more miRs of a plurality of miRs identified to be over-expressed in cells from CAD patients. Described herein are methods to recover the functions of EPCs isolated from patients with cardiovascular disease (e.g., CAD or peripheral artery disease (PAD)) by bioengineering the cells with antagomirs and/or premirs to specific micro-RNAs. The bioengineered cells can then be used to treat patients with ischemic or ischemic-related disease (e.g., CAD or PAD) by autologous stem cell therapy. 1. A composition comprising a plurality of at least one of: stem or somatic cells with pro-angiogenic potential and progenitor cells with pro-angiogenic potential , transduced with at least one nucleic acid encoding at least one of: antagomir to miR-493 , antagomir to miR-515-5p , antagomir to miR-196b , antagomir to miR-1913 , antagomir to miR-520a , antagomir to miR-1281 , antagomir to miR-373 , antagomir to miR-1978 , antagomir to miR-155 , antagomir to miR-92a , antagomir to miR-335 , antagomir to miR-1973 , antagomir to miR-21 , antagomir to miR-26a , antagomir to miR-16 , premir of miR-128 , premir of miR-720 , premir of miR-939 , premir of miR-885-3p , premir of miR-154 , and premir of miR-373;the plurality of cells isolated from a subject and having increased angiogenic function compared to the same cells isolated from the subject that were not transduced with the at least one nucleic acid.2. The composition of claim 1 , wherein the plurality of cells comprise at least one selected from the group consisting of: CD34+ endothelial ...

FROG/TOAD conditionally silenced vectors for hypoxia gene therapy

Methods and compositions for the treatment of hypoxia associated disorders by directional angiogenesis/arteriogenesis. Conditionally silenced vectors expressing a therapeutic molecule under hypoxic conditions avoid chaotic vascularization and allow for the orderly growth of new vessels into damaged tissue.

METHODS, COMPOSITIONS, CELLS, AND KITS FOR TREATING ISCHEMIC INJURY

The methods, compositions, cells and kits described herein are based on the discovery that stem cells, when injected into ischemic tissue of mammals, can be protected by preconditioning of the ischemic tissue with hypoxia-regulated human VEGF and human IGF-1. Methods, compositions, cells and kits for treating tissue injured by ischemia or at risk of ischemic injury in a subject are thus described herein.

METHODS, COMPOSITIONS, CELLS, AND KITS FOR TREATING ISCHEMIC INJURY

The methods, compositions, cells and kits described herein are based on the discovery that stem cells, when injected into ischemic tissue of mammals, can be protected by preconditioning of the ischemic tissue with hypoxia-regulated human VEGF and human IGF-1. Methods, compositions, cells and kits for treating tissue injured by ischemia or at risk of ischemic injury in a subject are thus described herein. 135-. (canceled)36. A method of treating tissue injured by ischemia or at risk of ischemic injury in a subject , the method comprising the steps of:a) administering to the subject a therapeutically effective amount of a composition comprising at least one nucleic acid encoding at least one cell survival or growth factor for protecting one or more cell types selected from the group consisting of: somatic cells, stem cells, and progenitor cells, from ischemia in the subject, the at least one nucleic acid operably linked to an inflammation-responsive promoter,wherein the inflammation-responsive promoter comprises at least one inflammatory responsive element (IRE) and optionally, one or more of a silencer element and a Hypoxia Responsive Element (HRE); andb) administering to the subject a therapeutically effective amount of a plurality of at least one of: somatic cells, stem cells, and progenitor cells,wherein administering the at least one nucleic acid followed by administration of the plurality of at least one of: somatic cells, stem cells, and progenitor cells induces at least one of: tissue protection, tissue regeneration, growth of blood vessels, and arteriogenesis at one or more sites of ischemia, ischemic injury, and potential ischemic injury in the subject.37. The method of claim 36 , wherein the at least one cell survival factor is human VEGF (hVEGF).38. The method of claim 37 , wherein the at least one nucleic acid further encodes a second cell survival factor.39. The method of claim 38 , wherein the second cell survival factor is human IGF-1 (hIGF-1).40. The ...

Molecular switch for regulating mammalian gene expression

Expression vectors are disclosed that are comprised of (a) one or more silencer elements and conditionally inducible elements to form silencer-inducible regions and (b) promoters in operative linkage upstream of at least one expressed region. The expression vector thereby regulates expression of at least one downstream region by conditional silencing in which an expressed DNA region of a gene is transcribed to produce RNA transcripts, which may or may not be translated to produce polypeptides. Genetically engineered mammalian cells and non-human mammals can be made using such expression vectors through transfection and transgenesis techniques. Moreover, processes of making and using the aforementioned products are disclosed (e.g., the expression vector may be used diagnostically, therapeutically, or prophylactically).

Hypoxia regulated conditionally silenced aav expressing angiogenic inducers

Methods and compositions for the treatment of hypoxia associated disorders by directional angiogenesis/arteriogenesis. Conditionally silenced vectors expressing a therapeutic molecule under hypoxic conditions avoid chaotic vascularization and allow for the orderly growth of new vessels into damaged tissue.

Methods, compositions, cells, and kits for treating ischemic injury

The methods, compositions, cells and kits described herein are based on the discovery that stem cells, when injected into ischemic tissue of mammals, can be protected by preconditioning of the ischemic tissue with hypoxia-regulated human VEGF and human IGF-1. Methods, compositions, cells and kits for treating tissue injured by ischemia or at risk of ischemic injury in a subject are thus described herein.

A molecular switch for regulating mammalian gene expression

Expression vectors are disclosed that are comprised of (a) one or more silencer elements and conditionally inducible elements to form silencer-inducible regions and (b) promoters in operative linkage upstream of at least one expressed region. The expression vector thereby regulates expression of at least one downstream region by conditional silencing in which an expressed DNA region of a gene is transcribed to produce RNA transcripts, which may or may not be translated to produce polypeptides. Genetically engineered mammalian cells and non-human mammals can be made using such expression vectors through transfection and transgenesis techniques. Moreover, processes of making and using the aforementioned products are disclosed (e.g., the expression vector may be used diagnostically, therapeutically, or prophylactically).

Compositions, cells, kits and methods for autologous stem cell therapy

Hypoxia regulated conditionally silenced aav expressing angiogenic inducers

Methods and compositions for the treatment of hypoxia associated disorders by directional angiogenesis/arteriogenesis. Conditionally silenced vectors expressing a therapeutic molecule under hypoxic conditions avoid chaotic vascularization and allow for the orderly growth of new vessels into damaged tissue.

A molecular switch for regulating mammalian gene expression

Expression vectors are disclosed that are comprised of (a) one or more silencer elements and conditionally inducible elements to form silencer-inducible regions and (b) promoters in operative linkage upstream of at least one expressed region. The expression vector thereby regulates expression of at least one downstream region by conditional silencing in which an expressed DNA region of a gene is transcribed to produce RNA transcripts, which may or may not be translated to produce polypeptides. Genetically engineered mammalian cells and non-human mammals can be made using such expression vectors through transfection and transgenesis techniques. Moreover, processes of making and using the aforementioned products are disclosed (e.g., the expression vector may be used diagnostically, therapeutically, or prophylactically).

Preventing ischemia-induced cell damage

Hypoxia-acidosis-associated cell death is mediated by BNIP3, a member of the Bcl-2 family of apoptosis-regulating proteins. Chronic hypoxia induced the expression and accumulation of BNIP3 mRNA and protein in cardiac myocytes but acidosis was required to activate the death pathway. Acidosis stabilized BNIP3 protein and increased the association with mitochondria. Cell death by hypoxia-acidosis was blocked by pretreatment with antisense BNIP3 oligonucleotides. The pathway included extensive DNA fragmentation and opening of the mitochondrial permeability transition pore but no apparent caspase activation. Overexpression of wild type BNIP3, but not a translocation-defective mutant activated cardiac myocyte death when the myocytes were acidotic or hypoxic.

Preventing ischemia-induced cell damage