Gene editing to improve joint function
Inventors
Millett, Peter J. • Russell, Iain Alasdair • Allen, Matthew J.
Assignees
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Publication Number
US-11033590-B2
Publication Date
2021-06-15
Expiration Date
Abstract
The present invention provides compositions and methods for treating joint disorders that are characterized by an inflammatory component. In some aspects, the compositions and methods are to prevent the progression of osteoarthritis and other arthritides and to treat osteoarthritis and other arthritides in a mammalian joint.
Core Innovation
The invention relates to a pharmaceutical composition for the treatment or prevention of a joint disease or condition. The composition comprises a therapeutically effective amount of a recombinant adeno-associated virus of serotype 5 (AAV5) or serotype 6 (AAV6) comprising one or more nucleic acids encoding a CRISPR gene-editing system, the system comprising a Cas9 protein and at least one guide RNA targeting an IL-1α gene or an IL-1β gene. The composition is capable of reducing inflammation in a joint following injection of the pharmaceutical composition into the joint.
For IL-1α targeting, the at least one guide RNA comprises a crRNA sequence complementary to a target sequence in exon 4 of the IL-1α gene. The crRNA sequence forms no nucleotide mismatches with the target sequence, and the target sequence is adjacent to a protospacer adjacent motif (PAM) sequence for the Cas9 protein. For IL-1β targeting, the at least one guide RNA comprises a crRNA sequence complementary to a target sequence in exon 4 of the IL-1β gene, with no nucleotide mismatches, and the target sequence is adjacent to a PAM sequence for the Cas9 protein.
The disclosed approach is positioned as reducing inflammatory cytokines in joint cells, including synovial cells (synoviocytes), synovial fibroblasts, chondrocytes, and synovial macrophages. The inflammation targets described include IL-1α and IL-1β, with additional targets described in the disclosure including TNF-α, IL-6, IL-8, IL-18, MMPs, and NLRP3 inflammasome components. Programmable nuclease systems are described, including CRISPR-Cas9, and delivery concepts are described using AAV vectors, including AAV serotypes and variants such as AAV5 and AAV6.
Claims Coverage
The independent claims are directed to a pharmaceutical composition for a joint disease or condition that includes an AAV5 or AAV6 vector carrying a CRISPR gene-editing system. The claim coverage focuses on the combination of 2 inventive features: an AAV5/AAV6 recombinant vector and a Cas9/guide RNA system targeting exon 4 of IL-1α or IL-1β with strict sequence and PAM adjacency constraints.
AAV5 or AAV6 carrying CRISPR gene-editing system targeting IL-1α exon 4
A pharmaceutical composition comprising a therapeutically effective amount of recombinant AAV5 or AAV6 comprising one or more nucleic acids encoding a CRISPR gene-editing system, the system comprising Cas9 protein and at least one guide RNA targeting an IL-1α gene, wherein the guide RNA comprises a crRNA complementary to a target sequence in exon 4 of the IL-1α gene, the crRNA sequence forms no nucleotide mismatches with the target sequence, and the target sequence is adjacent to a protospacer adjacent motif (PAM) sequence for the Cas9 protein, wherein the composition is capable of reducing inflammation in a joint following injection.
AAV5 or AAV6 carrying CRISPR gene-editing system targeting IL-1β exon 4
A pharmaceutical composition comprising a therapeutically effective amount of recombinant AAV5 or AAV6 comprising one or more nucleic acids encoding a CRISPR gene-editing system, the system comprising Cas9 protein and at least one guide RNA targeting an IL-1β gene, wherein the guide RNA comprises a crRNA complementary to a target sequence in exon 4 of the IL-1β gene, the crRNA sequence forms no nucleotide mismatches with the target sequence, and the target sequence is adjacent to a protospacer adjacent motif (PAM) sequence for the Cas9 protein, wherein the composition is capable of reducing inflammation in a joint following injection.
Across the independent claims, the key inventive feature is a joint-injection pharmaceutical composition using recombinant AAV5 or AAV6 to deliver a CRISPR gene-editing system with Cas9 and an exon 4 IL-1α- or IL-1β-targeting guide RNA. The guide RNA is constrained to a crRNA complementary to exon 4 with no nucleotide mismatches and with the target adjacent to a Cas9 PAM, and the claimed composition is capable of reducing inflammation after injection into the joint.
Stated Advantages
Capable of reducing inflammation in a joint following injection of the pharmaceutical composition into the joint.
Documented Applications
Treatment or prevention of a joint disease or condition by injection of the pharmaceutical composition into a joint.
Reducing inflammation in a joint following injection of the pharmaceutical composition into the joint.
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