610results about "Genetic material administration regime" patented technology

Nucleic acid molecules, compositions, recombinant AAV (rAAV) particles, kits, and methods are described herein for delivering a base editor (or “nucleobase editor”) to cells, e.g., via AAV vectors. In particular, the disclosure provides compositions, methods, and uses for delivery of adenine base editors and cytosine base editors in a single AAV vector (or genome). Further described herein are improved AAV vectors containing size-minimized regulatory components that enable, e.g., the packaging of base editors. Provided herein are methods and compositions for delivering base editor proteins to a cell or tissue in a single recombinant AAV (rAAV) vector. Contemplated herein are improved methods and compositions for delivering these base editors in vivo, in a single rAAV particle. Further provided herein are base editors and compositions and cells comprising these base editors.

The present disclosure is directed to DNA-PK inhibitors having the Formula (I), (I) or a pharmaceutically acceptable salt, stereoisomer, solvate, prodrug, or tautomer thereof, methods of preparing the foregoing, and compositions thereof, as well as methods of use for the compounds of Formula (I), in combination with a DNA cutting agent.

Disclosed herein are molecules and pharmaceutical compositions that induce an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion. Also described herein include methods for treating a disease or disorder that comprises a molecule or a pharmaceutical composition that induces an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion.

Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. The methods use CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting.

Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. Some methods, such as when a subject has preexisting against an immunogen to be administered, use plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting. Other methods, such as when a subject has no preexisting immunity against an immunogen to be administered, use B cell depleting agents (e.g., anti-CD20xCD3 antibody or functional fragment thereof) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting.

Described herein are nucleic acid molecules, compositions, recombinant AAV (rAAV) particles, kits, and methods for delivering a base editor (or a "nucleobase editor") to a cell, for example via an AAV vector. In particular, the present disclosure provides compositions, methods and uses for delivery of adenine base editors and cytosine base editors in a single AAV vector (or genome). Further described herein are improved AAV vectors comprising a minimised size regulatory component, for example, a regulatory component that can enable packaging of a base editor. Provided herein are methods and compositions for delivering a base editor protein to a cell or tissue in a single recombinant AAV (rAAV) vector. Improved methods and compositions for in vivo delivery of these base editors in a single rAAV particle are contemplated herein. Further provided herein are base editors, and compositions and cells comprising these base editors.

Nucleic acid constructs and compositions that allow insertion of a multidomain therapeutic protein (e.g., GAA fusion protein) coding sequence into a target genomic locus such as an endogenous ALB locus and / or expression of the multidomain therapeutic protein (e.g., GAA fusion protein) coding sequence are also provided. The nucleic acid constructs and compositions can be used in methods of integration of a multidomain therapeutic protein (e.g., GAA fusion protein) nucleic acid into a target genomic locus, methods of expression of a multidomain therapeutic protein (e.g., GAA fusion protein) in a cell, methods of reducing glycogen accumulation, methods of treating Pompe disease or GAA deficiency in a subject, and method of preventing or reducing the onset of a sign or symptom of Pompe disease in a subject, including neonatal cells and subjects.

Compositions and methods for treating LaFora disease are provided. The compositions and methods herein comprise AAV9 vector comprising a low expressing promoter driving the expression of a gene for e.g., EPM2A or EPM2B. Further provided are method of administering the composition for reducing one or more symptoms associated with or preventing LaFora disease.

Provided herein is a method of delivering a pharmaceutical composition comprising an AAV vector to an ocular or periocular tissue, comprising injecting the AAV vector periocularly (e.g., periocular injection, optionally peribulbar injection). In some aspects, the method is for use in treating an ophthalmic pathology (e.g., thyroid eye disease (TED) in a subject in need thereof.

The present disclosure provides recombinant adeno-associated virus (rAAV) vectors, methods of treating ocular conditions, pharmaceutical compositions, and other compositions and methods, wherein the rAAV vectors comprise a polynucleotide encoding an oxidoreductase. Methods of treatment may include administration to the lacrimal gland.

To provide methods of producing and purifying AAV viral vectors having low empty capsids, low host cell proteins, and / or low contaminating DNA, while retaining high potency; and to provide a pharmaceutical composition containing the vector.SOLUTION: A method of manufacturing an AAV viral vector comprises: a. culturing adherent cells; b. transfecting the adherent cells with one or more plasmids to enable production of the AAV viral vector; c. lysing the adherent cells to isolate the AAV viral vector; d. acidifying and clarifying the cell lysate of c; e. purifying the product of d using cation exchange chromatography; f. filtering the product of e using tangential flow filtration; g. ultracentrifuging the product of f in a cesium chloride buffer; h. collecting the AAV viral vector from the product of g; and i. formulating and filling the AAV viral vector collected in h. to produce a final pharmaceutical product.SELECTED DRAWING: None

This application provides improved methods of genome editing. Cas9 molecules can be used to create a break in a genomic region of interest. To increase the likelihood that the break is repaired by HDR (homology-directed repair), the cell can be contacted with molecules that bring a template nucleic acid in close proximity to the break, under conditions that allow the cell to repair the break using the template nucleic acid.

Nucleic acid constructs and compositions that allow insertion of a multidomain therapeutic protein (e.g., GAA fusion protein) coding sequence into a target genomic locus such as an endogenous ALB locus and / or expression of the multidomain therapeutic protein (e.g., GAA fusion protein) coding sequence are also provided. The nucleic acid constructs and compositions can be used in methods of integration of a multidomain therapeutic protein (e.g., GAA fusion protein) nucleic acid into a target genomic locus, methods of expression of a multidomain therapeutic protein (e.g., GAA fusion protein) in a cell, methods of reducing glycogen accumulation, methods of treating Pompe disease or GAA deficiency in a subject, and method of preventing or reducing the onset of a sign or symptom of Pompe disease in a subject, including neonatal cells and subjects.

Provided herein are methods of ex vivo administration of a lipid particle or a payload gene, to a subject. In some embodiments, the methods are in-line methods of administration of a lipid particle or payload gene that are performed in a closed fluid circuit. Also provided are related compositions, containers, and systems in connection with the provided methods.

Disclosed herein are nucleic acid compositions and methods of use. The nucleic acid compositions may have a therapeutic nucleic acid sequence operably linked to a nuclear targeting sequence that increases expression of the therapeutic nucleic acid in a cell by at least 1.25 fold; at least 80% sequence identity to SEQ ID NO: 6; or a first regulatory element comprising a promoter sequence operably linked to a hemoglobin subunit gamma intron (hBGi) sequence, and a second regulatory element comprising a woodchuck hepatitis posttranscriptional regulatory element (WPRE) sequence. The nucleic acid compositions with these features may enhance therapeutic nucleic acid transfection and expression. Also disclosed herein are transgenes optimized for gene therapy applications, including novel FVIII transgene sequences, in which the B domain may be non-naturally occurring the A1 and / or A3 domain may include at least one amino acid substitution.

The invention belongs to the field of preparation of biomedical nano-materials, and particularly relates to a tumor microenvironment responsive bacterium-nano hybrid system as well as a preparation method and application thereof. According to the invention, an engineering bacterium containing an acid response promoter and coding PD-L1 blocking peptide is prepared, and then lipidosome nanoparticles containing calcium carbonate, curcumin and an XIAP / cIAP1 antagonist are attached to the bacterium, so that the bacterium-nano hybrid system with tumor microenvironment response is obtained. The system can realize accurate drug delivery and controllable release in an acidic tumor microenvironment through a unique pH response mechanism, shows remarkable tumor targeting and treatment effects, and can be used as an anti-tumor drug.

A dual vector system for treating ornithine transcarbamylase deficiency is provided. The system includes ((a) a gene editing AAV comprising a first AAV rh79 capsid and a first vector genome comprising a 5′ ITR, a sequence encoding a meganuclease that targets PCSK9 under control of regulatory sequences that direct expression of the meganuclease in a target cell comprising a PCSK9 gene, and a 3′ ITR; and(b) a donor AAV vector comprising a second AAV capsid and a second AAV rh79 vector genome comprising: a 5′ITR, a 5′ homology directed recombination (HDR) arm, a transgene encoding ornithine transcarbamylase (OTC) and regulatory sequences that direct expression of the transgene in the target cell, a 3′ HDR arm, and a 3′ ITR.

Provided herein is a recombinant AAV (rAAV) comprising an AAV capsid and a vector genome packaged therein, wherein the vector genome comprises an AAV 5′ inverted terminal repeat (ITR), an engineered nucleic acid sequence encoding a functional hSGSH, optionally comprising stabilizing amino acid changes, a regulatory sequence which direct expression of hSGSH in a target cell, and an AAV 3′ ITR. Also provided is a pharmaceutical composition comprising a rAAV as described herein in a formulation buffer, and a method of treating a human subject diagnosed with MPS IIIA.

Disclosed herein is a method for the treatment of Pompe Disease comprising administering a recombinant AAV (rAAV) vector comprising a rAAV genome comprising a heterologous nucleic acid encoding a heterologous BM40 signal peptide and an acid alpha-glucosidase (GAA) polypeptide, or N-terminal truncation thereof, where the heterologous nucleic acid is operatively linked to a liver-specific promoter, where the nucleic acid encoding GAA polypeptide can be wild type nucleic acid sequence, or modified nucleic acid sequence, or a codon optimized nucleic acid sequence, and can optionally be modified to reduce or completely eliminate CG and CpG dinucleotides and, optionally eliminated alternative reading frames (ARF) content.

To provide a pharmaceutical composition comprising the recombinant nucleic acid, and to provide a method of treating Leber's hereditary optic neuropathy (LHON) using the pharmaceutical composition.SOLUTION: Provided is a pharmaceutical composition for use in combination with a second pharmaceutical composition to treat Leber's hereditary optic neuropathy (LHON) in a patient in need thereof, the pharmaceutical composition comprising a nucleic acid sequence encoding a mitochondrial-tropic peptide and comprising a therapeutically effective amount of an adeno-associated virus (AAV) comprising a recombinant nucleic acid to which a promoter is operably linked, the second pharmaceutical composition comprising a steroid, the steroid being methylprednisolone or prednisone, the steroid being administered daily for two days prior to the administration of the pharmaceutical composition.SELECTED DRAWING: Figure 8

Provided herein is a method for improving delivery of a pharmaceutical composition to the central nervous system in a subject in need thereof, the method comprising administering to the subject, in combination with the pharmaceutical composition, an agent that enhances glymphatic influx.

To provide compositions and methods for regulating protein production in target cells. [Solution] The compositions and methods disclosed herein can improve diseases associated with protein or enzyme deficiency. The present invention provides, for example, a composition comprising (a) at least one mRNA molecule that is at least partly encoding a functionally secreted polypeptide, and (b) a transport vehicle comprising lipid nanoparticles. The present invention also provides a method for treating a subject deficient in a functional polypeptide, comprising administering a composition comprising (a) at least one mRNA that is at least partly encoding the functionally secreted polypeptide, and (b) a transport vehicle comprising lipid nanoparticles, wherein after administration of the composition, the mRNA is expressed in a target cell to produce the functionally secreted polypeptide.

To provide a gene therapy for treating mucopolysaccharidosis type I.SOLUTION: Provided is a treatment regimen useful for the treatment of α-L-iduronidase deficiency in human patients, the regimen comprising administering to the patient: (a) a recombinant AAV having a nucleic acid, comprising an AAV9 capsid and a sequence encoding human α-L-iduronidase under the control of regulatory sequences that direct its expression in the patient; (b) a first immunosuppressant selected from glucocorticoids, steroids, antimetabolites, T-cell inhibitors, macrolides, or cytostatics; and (c) a second immunosuppressant selected from glucocorticoids, steroids, antimetabolites, T-cell inhibitors, macrolides, or cytostatics.SELECTED DRAWING: Figure 22

The present invention relates to systems for expressing antibodies or antigen-binding fragments in a subject, and methods of treating diseases therewith.

Disclosed herein in one aspect is a method of reducing immunogenicity comprising administering an effective amount of a non-depleting B-cell inhibitor to a patient who is receiving or has received BTHS gene therapy. Related compositions are also provided.

Provided are recombinant adeno-associated virus (rAAV) vectors, methods of treating a condition (e.g., an ocular condition), pharmaceutical compositions, and other compositions and methods, in which the rAAV vector comprises a polynucleotide encoding diamine oxidase (DAO1). Methods of treatment may include administration to the lacrimal gland.

The disclosure relates to methods of delivering a gene therapy agent to a subject, treating an individual in need of a gene therapy agent, increasing expression of a gene therapy agent, reducing an immune response to a gene therapy agent, and preventing immune-related adverse events in a subject by administering a gene therapy agent and an IL-2 conjugate to the subject. The IL-2 conjugate has at least one amino acid residue replaced by an unnatural amino acid linked to a conjugating moiety. The disclosure further relates to methods of selecting a subject for treatment with a gene therapy agent and an IL-2 conjugate.