Optimized Aga Genes and Expression Cassettes and their Use

Content Provider	The Lens
Description	La présente invention concerne des polynucléotides comprenant des cadres de lecture ouverts de l'aspartylglucosaminidase (AGA) optimisés et des vecteurs et des cellules les comprenant. L'invention concerne en outre des méthodes d'utilisation de celle-ci pour l'administration du cadre de lecture ouvert à une cellule ou à un sujet et des méthodes de traitement de l'aspartylglucosaminurie (AGU) chez un sujet.
Abstract	This invention relates to polynucleotides comprising optimized aspartylglucosaminidase (AGA) open reading frames and vectors and cells comprising the same. The invention further relates to methods of using the same for delivery of the open reading frame to a cell or a subject and methods for treating aspartylglucosaminuria (AGU) in a subject.
Related Links	https://www.lens.org/lens/patent/009-570-396-848-309/frontpage
Language	English
Publisher Date	2019-05-16
Access Restriction	Open
Alternative Title	Gènes D'aga Optimisés Et Cassettes D'expression Optimisées Et Leur Utilisation
Content Type	Text
Resource Type	Patent
Date Applied	2018-03-22
Agent	Schwartzman, Robert A.
Applicant	Univ North Carolina Chapel Hill
Application No.	2018023727
Claim	That which is claimed is: A polynucleotide comprising a human AGA open reading frame, wherein the nucleotide sequence has been codon-optimized for expression in human cells. The polynucleotide of claim 1 , wherein said polynucleotide comprises the nucleotide sequence of SEQ ID NO: 1 or a sequence at least about 90% identical \ thereto. An expression cassette comprising a polynucleotide comprising a human AGA open reading frame. The expression cassette of claim 3, wherein the polynucleotide is the polynucleotide of claim 1 or 2. The expression cassette of claim 3 or 4, wherein the polynucleotide is operably linked to a promoter. The expression cassette of claim 5, wherein the promoter is a chicken beta actin promoter. The expression cassette of any one of claims 3-6, wherein the polynucleotide is operably linked to an enhancer. The expression cassette of claim 7, wherein the enhancer is a cytomegalovirus (CMV) enhancer. The expression cassette of any one of claims 3-8, wherein the polynucleotide is operably linked to an intron. The expression cassette of claim 9, wherein the intron is a hybrid/modified MVM intron. The expression cassette of any one of claims 3-10, wherein the polynucleotide is operably linked to a polyadenylation signa , The expression cassette of claim 11, wherein the polyadenylation signal is a bovine growth hormone polyadenylation signa The expression cassette of any one of claims 3-12, further comprising at least one adeno-associated virus (AAV) inverted terminal repeat (ITR). The expression cassette of claim 13, wherein the expression cassette comprises two AAV ITRs. The expression cassette of claim 14, wherein the two AAV ITRs have the same nucleotide sequence. The expression cassette of claim 14, wherein the two AAV ITRs have different nucleotide sequences. The expression cassette of any one of claims 13-16, wherein the AAV ITRs are AAV2 ITRs. The expression cassette of any one of claims 3-17, wherein the expression cassette is a self-complementary AAV genome. The expression cassette of any one of claims 3-18, wherein the expression cassette comprises an enhancer, a promoter, an intron, a human AGA open reading frame, and a polyadenylation site. The expression cassette of claim 19, wherein the expression cassette comprises an AAV ITR, an enhancer, a promoter, an intron, a human AGA open reading frame, a polyadenylation site, and an AAV ITR. The expression cassette of any one of claims 3-18, wherein the expression cassette comprises a CMV enhancer, a chicken beta actin promoter, a hybrid/modified MVM intron, a human AGA open reading frame, and a bovine growth hormone polyadenylation site. The expression cassette of claim 21, wherein the expression cassette comprises a mutant AAV ITR, a CMV enhancer, a chicken beta actin promoter, a hybrid/modified MVM intron, a human AGA open reading frame, a bovine growth hormone polyadenylation site, and a wild-type AAV ITR. The expression cassette of claim 22, comprising the nucleotide sequence of SEQ ID NO: 7 or a sequence at least about 90% identical thereto. A vector comprising the polynucleotide of claim 1 or 2 or the expression cassette of any one of claims 3-16. The vector of claim 24, wherein the vector is a viral vector. The vector of claim 25, wherein the vector is an AAV vector. The vector of claim 26, wherein the AAV vector is an AAV9 vector. The vector of claim 27, wherein the AAV vector comprises wild-type capsid proteins. The vector of claim 27, wherein the AAV vector comprises a modified capsid protein with altered tropism compared to a wild-type capsid protein. The vector of claim 29, wherein the modified capsid protein is liver- detargeted. A transformed cell comprising the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, and/or the vector of any one of claims 24-30. The transformed cell of claim 31, wherein the polynucleotide, expression cassette, and/or vector is stably incorporated into the cell genome. A transgenic animal comprising the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, the vector of any one of claims 24-30, and/or the transformed cell of claim 31 or 32. A pharmaceutical formulation comprising the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, the vector of any one of claims 24- 30, and/or the transformed cell of claim 31 or 32 in a pharmaceutically acceptable carrier. The pharmaceutical formulation of claim 34, wherein the pharmaceutically acceptable carrier is phosphate buffered saline with 5% D-sorbito A method of expressing an AGA open reading frame in a cell, comprising contacting the cell with the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, and/or the vector of any one of claims 24-30, thereby expressing the AGA open reading frame in the cel A method of expressing an AGA open reading frame in a subject, comprising delivering to the subject the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, the vector of any one of claims 24-30, and/or the transformed cell of claim 31 or 32, thereby expressing the AGA open reading frame in the subject. A method of treating a disorder associated with aberrant expression of an AGA gene or aberrant activity of an AGA gene product in a subject in need thereof, comprising delivering to the subject a therapeutically effective amount of the polynucleotide of claim 1 or 2, the expression cassette of any one of claims 3-23, the vector of any one of claims 24-30, and/or the transformed cell of claim 31 or 32, thereby treating the disorder associated with aberrant expression of the AGA gene in the subject or aberrant activity of the AGA gene product. The method of claim 38, wherein the disorder associated with aberrant expression of the AGA gene or aberrant activity of the AGA gene product is aspartylglucosaminuria. The method of any one of claims 37-39, wherein the polynucleotide, expression cassette, vector, and/or transformed cell is delivered intravenously. The method of any one of claims 37-39, wherein the polynucleotide, expression cassette, vector, and/or transformed cell is delivered intrathecally.
CPC Classification	Preparations For Medical; Dental Or Toiletry Purposes MICROORGANISMS OR ENZYMES;COMPOSITIONS THEREOF;PROPAGATING; PRESERVING; OR MAINTAINING MICROORGANISMS;MUTATION OR GENETIC ENGINEERING;CULTURE MEDIA Animal Husbandry; Aviculture; Apiculture; Pisciculture; Fishing; Rearing Or Breeding Animals; Not Otherwise Provided For; New Breeds Of Animals Specific Therapeutic Activity Of Chemical Compounds Or Medicinal Preparations
Extended Family	070-345-995-521-173 007-701-960-409-089 081-268-509-208-601 083-359-258-767-500 192-292-947-295-586 172-538-896-258-92X 061-749-315-804-458 135-048-108-061-242 009-570-396-848-309
Patent ID	2019094061
Inventor/Author	Gray Steven James
IPC	C12N15/861 A01K67/027 A61K31/711 A61K48/00 A61P25/28 C12N5/10 C12N15/63 C12N15/79
Status	Pending
Simple Family	070-345-995-521-173 007-701-960-409-089 081-268-509-208-601 083-359-258-767-500 192-292-947-295-586 172-538-896-258-92X 061-749-315-804-458 135-048-108-061-242 009-570-396-848-309
CPC (with Group)	A61K48/005 A61K48/0075 A61K31/711 C12N9/80 C12N15/86 A01K2217/075 A01K2227/105 A01K2267/0306 C12N2750/14143 C12N2800/22 A61P25/28 C12N15/63 C12N15/79
Issuing Authority	United States Patent and Trademark Office (USPTO)
Kind	Patent Application Publication