RAAV VECTOR COMPOSITIONS, METHODS FOR TARGETING VASCULAR ENDOTHELIAL CELLS AND USE IN TREATMENT OF TYPE I DIABETES

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations that include them. Also disclosed are methods of preparing and using novel capsid-protein-mutated rAAV vector constructs in a variety of diagnostic and therapeutic applications including, inter alia, as delivery agents for diagnosis, treatment, or amelioration of one or more diseases, disorders, or dysfunctions of the mammalian vascular system, and complications from Type I diabetes. Also disclosed are methods for systemic and tissue-localized delivery of therapeutic rAAV-based gene expression cassettes to vascular endothelial cells, tissues, and organs, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and/or in vivo applications including the treatment of vasculitis, and complications arising from Type I diabetes, such as macular edema, nephropathy, diabetic retinopathy, and the like. 1. A recombinant adeno-associated viral (rAAV) expression system comprising:a) a polynucleotide that encodes a modified capsid protein, wherein the modified capsid protein comprises five or more non-native amino acid substitutions at positions corresponding to five or more distinct surface-exposed amino acid residues in the wild-type AAV2 capsid protein, and further wherein the transduction efficiency of a virion comprising the modified capsid protein is higher than that of a virion comprising a corresponding, unmodified wild-type capsid protein; andb) an expression cassette packaged within the virion, that comprises an isolated polynucleotide comprising a nucleic acid segment that encodes or that expresses a diagnostic or a therapeutic molecule in a mammal transformed with the expression system, wherein the nucleic acid segment is operably linked to a promoter or a control region that expresses the nucleic acid segment in one or more vascular endothelial cells of the mammal to produce the diagnostic ...

RIBOSWITCH MODULATED GENE THERAPY FOR RETINAL DISEASES

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated. 1. An exogenous nucleic acid construct for regulating expression of a transgene by modulating the mRNA of the transgene , the nucleic acid encoding:(a) a target gene of interest,(b) at least one smiRNA switch located within the untranslated region of the transgene, wherein the at least one smiRNA switch comprises an aptamer domain capable of binding to a ligand and a pri-miRNA.(c) at least one miRNA target sequence complementary to at least a portion of the pri-miRNA,wherein the at least one smiRNA switch regulates expression of the transgene by both (1) regulation of the cleavage of the mRNA of the transgene, and (b) regulating cleavage of the pri-miRNA from the smiRNA, wherein at least a portion of the cleaved pri-miRNA binds to at least one miRNA target sequences silencing the transgene expression.2. The construct of claim 1 , wherein in the absence of ligand claim 1 , the pri-miRNA is cleaved from the transcript and binds to the at least one miRNA target sequence claim 1 ,wherein binding of the portion of the pri-mRNA to the at least one miRNA target sequence silences the transgene.3. The construct of or claim 1 , wherein binding of the ligand to the aptamer domain alters the conformation of the smiRNA claim 1 , wherein the conformation change inhibits the ability of the pri-miRNA to be cleaved from the transcript claim 1 , and wherein the transgene is able to be translated.4. The construct of any of the proceeding claims claim 1 , wherein the smiRNA switch is encoded by nucleic acid sequence of SEQ ID NO:1.6. The construct of any one of the preceding claims claim 1 , wherein the construct encodes three miRNA target sequences claim 1 , wherein the at least three miRNA target sequences are encoded by SEQ ID NO: 2.7. The construct of any one of the ...

Riboswitch modulated gene therapy for retinal diseases

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

Raav vector compositions, methods for targeting vascular endothelial cells and use in treatment of type i diabetes

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations that include them. Also disclosed are methods of preparing and using novel capsid-protein-mutated rAAV vector constructs in a variety of diagnostic and therapeutic applications including, inter alia , as delivery agents for diagnosis, treatment, or amelioration of one or more diseases, disorders, or dysfunctions of the mammalian vascular system, and complications from Type I diabetes. Also disclosed are methods for systemic and tissue-localized delivery of therapeutic rAAV-based gene expression cassettes to vascular endothelial cells, tissues, and organs, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and/or in vivo applications including the treatment of vasculitis, and complications arising from Type I diabetes, such as macular edema, nephropathy, diabetic retinopathy, and the like.

Treatment of retinitis pigmentosa

A method of treating or preventing retinitis pigmentosa, wherein the method comprises administering an adeno-associated virus (AAV) vector comprising a nucleotide sequence encoding ciliary neurotrophic factor (CNTF) to a subject in need thereof.

網膜疾患のためのリボスイッチ調節遺伝子治療

【課題】対象内のトランスジーンの発現を制御するための改変リボスイッチを含むコンストラクトを提供する。【解決手段】トランスジーンのｍＲＮＡを調節することによりトランスジーンの発現を制御するための外来核酸コンストラクトであって、前記核酸は、（ａ）目的の標的遺伝子、（ｂ）前記トランスジーンの非翻訳領域内に位置する少なくとも１つのｓｍｉＲＮＡスイッチであって、リガンドに結合できるアプタマードメイン及びｐｒｉ－ｍｉＲＮＡを含む、少なくとも１つのｓｍｉＲＮＡスイッチ、（ｃ）前記ｐｒｉ－ｍｉＲＮＡの少なくとも一部に相補的な少なくとも１つのｍｉＲＮＡ標的配列、をコードし、前記少なくとも１つのｓｍｉＲＮＡスイッチは、（１）前記トランスジーンのｍＲＮＡの切断の制御、及び（２）前記ｓｍｉＲＮＡからの前記ｐｒｉ－ｍｉＲＮＡの切断の制御、の両方によって前記トランスジーンの発現を制御する、コンストラクトである。【選択図】図１

Laser-Speckle Contrast Imaging System and Method

Laser-Speckle Contrast imaging apparatus configured to assess and quantify motion associated with an object and, in a specific case of an eye—retinal vascular anatomy and hemodynamics and generate substantially contrast-free maps of retinal blood flow over a wide field-of-view at up to 590 fps and under short exposure durations (>50 μs), is applicable for diagnosis, study, and management of neurodegenerative conditions (i.e. mild cognitive impairment and Alzheimer's disease) and systemic cardiovascular diseases (i.e. athero- and arteriosclerosis, coronary artery occlusion, and hypertension). The apparatus employs a) a set of apertures substantially blocking light, delivered from a source of light to an illumination arm of the apparatus, from impinging onto an axial point of the front surface of the lens of the illumination arm, and b) polarization gating between the illumination and light-collecting arms of the apparatus. In one implementation, the apparatus is configured to allow for irradiation of the object with an optical field a degree of coherence and/or spectral content of which are varied delivered through the same optical train including the set of apertures.

Laser-speckle contrast imaging system and method

Laser-Speckle Contrast imaging apparatus configured to assess and quantify motion associated with an object and, in a specific case of an eye—retinal vascular anatomy and hemodynamics and generate substantially contrast-free maps of retinal blood flow over a wide field-of-view at up to 590 fps and under short exposure durations (>50 μs), is applicable for diagnosis, study, and management of neurodegenerative conditions (i.e. mild cognitive impairment and Alzheimer's disease) and systemic cardiovascular diseases (i.e. athero- and arteriosclerosis, coronary artery occlusion, and hypertension). The apparatus employs a) a set of apertures substantially blocking light, delivered from a source of light to an illumination arm of the apparatus, from impinging onto an axial point of the front surface of the lens of the illumination arm, and b) polarization gating between the illumination and light-collecting arms of the apparatus. In one implementation, the apparatus is configured to allow for irradiation of the object with an optical field a degree of coherence and/or spectral content of which are varied delivered through the same optical train including the set of apertures.

Riboswitch modulated gene therapy for retinal diseases

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

Laser-speckle contrast imaging system and method

Laser-Speckle Contrast imaging apparatus configured to assess and quantify motion associated with an object and, in a specific case of an eye - retinal vascular anatomy and hemodynamics and generate substantially contrast-free maps of retinal blood flow over a wide field-of-view at up to 590 fps and under short exposure durations (> 50µs), is applicable for diagnosis, study, and management of neurodegenerative conditions (i.e. mild cognitive impairment and Alzheimer's disease) and systemic cardiovascular diseases (i.e. athero- and arteriosclerosis, coronary artery occlusion, and hypertension). The apparatus employs a) a set of apertures substantially blocking light, delivered from a source of light to an illumination arm of the apparatus, from impinging onto an axial point of the front surface of the lens of the illumination arm, and b) polarization gating between the illumination and light-collecting arms of the apparatus. In one implementation, the apparatus is configured to allow for irradiation of the object with an optical field a degree of coherence and/or spectral content of which are varied delivered through the same optical train including the set of apertures.

Riboswitch modulated gene therapy for retinal diseases

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

Riboswitch modulated gene therapy for retinal diseases

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

Riboswitch modulated gene therapy for retinal diseases

The present invention provides constructs comprising modified riboswitches to regulate expression of a transgene within a subject. Methods of treating a disease, specifically an eye disease, are also contemplated.

Riboswitch modulated gene therapy for retinal diseases

Laser-Speckle Contrast Imaging System and Method

Laser-Speckle Contrast imaging apparatus configured to assess and quantify motion associated with an object and, in a specific case of an eye-retinal vascular anatomy and hemodynamics and generate substantially contrast-free maps of retinal blood flow over a wide field-of-view at up to 590 fps and under short exposure durations (>50 μs), is applicable for diagnosis, study, and management of neurodegenerative conditions (i.e. mild cognitive impairment and Alzheimer's disease) and systemic cardiovascular diseases (i.e. athero- and arteriosclerosis, coronary artery occlusion, and hypertension). The apparatus employs a) a set of apertures substantially blocking light, delivered from a source of light to an illumination arm of the apparatus, from impinging onto an axial point of the front surface of the lens of the illumination arm, and b) polarization gating between the illumination and light-collecting arms of the apparatus. In one implementation, the apparatus is configured to allow for irradiation of the object with an optical field a degree of coherence and/or spectral content of which are varied delivered through the same optical train including the set of apertures.