Lipid nanoparticles for gene editing systems

Abstract

The present disclosure provides compositions and methods for treating and preventing local nociceptive feelings, inflammation, or morphological changes associated with joint diseases or disorders, back or spinal conditions or disorders, and musculoskeletal diseases or dysfunctions with an LNP-encapsulated CRISPR / Cas9 gene editing system.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application 63 / 334,476 filed on April 25, 2022, U.S. Provisional Patent Application 63 / 362,858 filed on April 12, 2022, U.S. Provisional Patent Application 63 / 342,471 filed on May 16, 2022, and U.S. Provisional Patent Application 63 / 495,461 filed on April 11, 2023, the contents of which are hereby incorporated by reference in their entirety for all purposes. Background Art

[0003] joint diseases

[0004] Treatment of osteoarthritis, degenerative joint disease, and other joint disorders is complex, with few long-term options for symptom relief or restoration of joint function. Osteoarthritis (OA) is a leading cause of disability due to pain. Neogi, Osteoarthritis Cartilage 2013;21:1145-53. All mammalian species are affected: working animals, livestock, and their owners all experience OA-related discomfort, pain, and disability, depending on the extent of the disease.

[0005] OA is a complex disease characterized by a progressive course of disability. Systemic inflammation is associated with OA and OA disease progression. Inflammation is driven by increased levels of proinflammatory cytokines. New methods and compositions for treating this disease are urgently needed. Disclosed herein are compositions and methods that can be used to treat OA and other inflammatory joint conditions.

[0006] Back and spinal conditions

[0007] Back or spinal conditions or diseases, including low back pain and pain or inflammation associated with discogenic conditions such as degenerative disc disease (DDD) or internal disc rupture (IDD), are major causes of morbidity and disability worldwide, with few long-term options for improvement. Andersson GB. Epidemiological features of chronic low-back pain. Lancet. 1999; 354: 581-585. Currently available treatments include surgery or less invasive treatment options, but these options generally do not provide long-term relief. Ju et al. Global Spiae Journal (2020): 2192568220963058. All vertebrate species, including draft animals, domestic pets, and their owners, are affected by back or spinal conditions or diseases. All species suffer from associated discomfort, pain, and disability, depending on the extent of disease progression.

[0008] Back or spinal conditions or disorders, such as low back pain, are complex diseases characterized by a large input of factors that lead to progressively worsening disability. These factors include morphological abnormalities (e.g., intervertebral disc rupture), inflammation, and changes in the local cellular environment (e.g., vascularization and / or innervation). Peng, Bao-Gan. World Journal of Orthopedics 4.2 (2013): 42. Each factor is driven by the differential expression of various gene products, including at least proinflammatory cytokines, growth factors, and other effector biomolecules. New methods and compositions for treating this disease are urgently needed. Summary of the Invention

[0009] Provided herein are compositions and methods for treating synovial joint dysfunction as described herein. In addition, disclosed herein are compositions and methods for treating or preventing local nociception, inflammation or morphological changes associated with back or spinal symptoms or illness. In addition, described herein are compositions and methods for treating or preventing musculoskeletal diseases and dysfunctions (including, for example, fibrosis and / or scarring of postoperative subjects). In addition, disclosed herein are methods for gene editing cells, including but not limited to synovial cells and / or synovial cells, chondrocytes, synovial macrophages and synovial fibroblasts, and gene-edited synovial cells and / or synovial cells, chondrocytes, synovial macrophages and synovial fibroblasts for the treatment of diseases such as osteoarthritis. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] The embodiments of the present disclosure will be further explained with reference to the accompanying drawings. The drawings shown are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of the embodiments of the present disclosure.

[0011] Figure 1 SEQ ID NOs: 1-48 are described, crRNA sequences generated by the bioinformatics methods described herein that target human ADAM17 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0012] Figure 2 SEQ ID NOs: 49-96 are described, crRNA sequences generated by the bioinformatics methods described herein that target human ADAMTS1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0013] Figure 3 SEQ ID NOs: 97-144 are described, crRNA sequences generated by the bioinformatics methods described herein that target human ADAMTS5 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0014] Figure 4 SEQ ID NOs: 145-192 are described, crRNA sequences generated by the bioinformatics methods described herein that target human ADM to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0015] Figure 5 SEQ ID NOs: 193-240 are described, crRNA sequences generated by the bioinformatics methods described herein that target human ATP1A1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0016] Figure 6 Illustrated are SEQ ID NOs: 241-281, crRNA sequences generated by the bioinformatics methods described herein that target human BDNF to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0017] Figure 7SEQ ID NOs: 282-301 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CALCA to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0018] Figure 8 SEQ ID NOs: 302-318 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CALCB to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0019] Figure 9 SEQ ID NOs: 319-340 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CALCRL to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0020] Figure 10 SEQ ID NOs: 341-357 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCL2 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0021] Figure 11 SEQ ID NOs: 358-374 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCL3 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0022] Figure 12 SEQ ID NOs: 375-391 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCL5 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0023] Figure 13SEQ ID NOs: 392-408 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCL7 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0024] Figure 14 SEQ ID NOs: 409-425 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCL20 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0025] Figure 15 SEQ ID NOs: 426-473 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CCN2 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0026] Figure 16 SEQ ID NOs: 474-517 are described, crRNA sequences generated by the bioinformatics methods described herein that target human CCR7 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0027] Figure 17 SEQ ID NOs: 518-534 are described, crRNA sequences generated by the bioinformatics methods described herein that target human CRCP to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0028] Figure 18 SEQ ID NOs: 535-551 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0029] Figure 19SEQ ID NOs: 552-568 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL2 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0030] Figure 20 SEQ ID NOs: 569-585 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL3 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0031] Figure 21 SEQ ID NOs: 586-602 are described, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL5 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0032] Figure 22 SEQ ID NOs: 603-619 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL6 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0033] Figure 23 SEQ ID NOs: 620-636 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human CXCL8 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0034] Figure 24 SEQ ID NOs: 637-655 are described, crRNA sequences generated by the bioinformatics methods described herein that target human CXCR1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0035] Figure 25SEQ ID NOs: 656-672 are described, crRNA sequences generated by the bioinformatics methods described herein that target human CXCR2 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0036] Figure 26 SEQ ID NOs: 673-720 are described, crRNA sequences generated by the bioinformatics methods described herein that target human FGF2 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0037] Figure 27 SEQ ID NOs: 721-768 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human FGFR1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0038] Figure 28 Illustrated are SEQ ID NOs: 769-786, crRNA sequences generated by the bioinformatics methods described herein, which target human IL1A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0039] Figure 29 Illustrated are SEQ ID NOs: 787-805, crRNA sequences generated by the bioinformatics methods described herein, which target human IL1B to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0040] Figure 30 Illustrated are SEQ ID NOs: 806-839, crRNA sequences generated by the bioinformatics methods described herein that target human IL1R1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0041] Figure 31Illustrated are SEQ ID NOs: 840-887, crRNA sequences generated by the bioinformatics methods described herein, which target human IL1RAP to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0042] Figure 32 SEQ ID NOs: 888-911 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human IL4 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0043] Figure 33 SEQ ID NOs: 912-928 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human IL6 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0044] Figure 34 Illustrated are SEQ ID NOs: 929-963, crRNA sequences generated by the bioinformatics methods described herein that target human IL6R to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0045] Figure 35 SEQ ID NOs: 964-990 are described, crRNA sequences generated by the bioinformatics methods described herein that target human IL6ST to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0046] Figure 36 SEQ ID NOs: 991-1007 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human IL10 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0047] Figure 37Illustrated are SEQ ID NOs: 1008-1055, crRNA sequences generated by the bioinformatics methods described herein that target human IL10RA to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0048] Figure 38 Illustrated are SEQ ID NOs: 1056-1082, crRNA sequences generated by the bioinformatics methods described herein that target human IL10RB to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0049] Figure 39 SEQ ID NOs: 1083-1104 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human IL13 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0050] Figure 40 Illustrated are SEQ ID NOs: 1105-1130, crRNA sequences generated by the bioinformatics methods described herein, that target human IL13RA1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0051] Figure 41 Illustrated are SEQ ID NOs: 1131-1147, crRNA sequences generated by the bioinformatics methods described herein that target human IL13RA2 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0052] Figure 42 Illustrated are SEQ ID NOs: 1148-1173, crRNA sequences generated by the bioinformatics methods described herein, which target human IL17A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0053] Figure 43Illustrated are SEQ ID NOs: 1174-1221, crRNA sequences generated by the bioinformatics methods described herein that target human IL17RA to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0054] Figure 44 SEQ ID NOs: 1222-1238 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human IL18 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0055] Figure 45 SEQ ID NOs: 1239-1262 are described, crRNA sequences generated by the bioinformatics methods described herein that target human IL18R1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0056] Figure 46 Illustrated are SEQ ID NOs: 1263-1310, crRNA sequences generated by the bioinformatics methods described herein, which target human IL18RAP to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0057] Figure 47 Illustrated are SEQ ID NOs: 1311-1343, crRNA sequences generated by the bioinformatics methods described herein, which target human MMP1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0058] Figure 48 Illustrated are SEQ ID NOs: 1344-1391, crRNA sequences generated by the bioinformatics methods described herein that target human MMP2 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0059] Figure 49Illustrated are SEQ ID NOs: 1392-1417, crRNA sequences generated by the bioinformatics methods described herein, which target human MMP3 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0060] Figure 50 Illustrated are SEQ ID NOs: 1418-1436, crRNA sequences generated by the bioinformatics methods described herein that target human MMP7 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0061] Figure 51 Illustrated are SEQ ID NOs: 1437-1474, crRNA sequences generated by the bioinformatics methods described herein that target human MMP8 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0062] Figure 52 Illustrated are SEQ ID NOs: 1475-1497, crRNA sequences generated by the bioinformatics methods described herein that target human MMP10 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0063] Figure 53 Illustrated are SEQ ID NOs: 1498-1541, crRNA sequences generated by the bioinformatics methods described herein that target human MMP12 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0064] Figure 54 Illustrated are SEQ ID NOs: 1542-1568, crRNA sequences generated by the bioinformatics methods described herein, which target human MMP13 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0065] Figure 55SEQ ID NOs: 1569-1585 are described, crRNA sequences generated by the bioinformatics methods described herein that target human MRGPRX2 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0066] Figure 56 SEQ ID NO: 1586-1628, crRNA sequences generated by the bioinformatics methods described herein, which target human NGF to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0067] Figure 57 SEQ ID NOs: 1629-1676 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human NGFR to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0068] Figure 58 SEQ ID NOs: 1677-1724 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human NTF3 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0069] Figure 59 SEQ ID NOs: 1725-1746 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human NTF4 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0070] Figure 60 Illustrated are SEQ ID NOs: 1747-1794, crRNA sequences generated by the bioinformatics methods described herein that target human NTRK1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0071] Figure 61SEQ ID NOs: 1795-1842 are described, crRNA sequences generated by the bioinformatics methods described herein that target human NTRK2 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0072] Figure 62 Illustrated are SEQ ID NOs: 1843-1859, crRNA sequences generated by the bioinformatics methods described herein that target human RAMP1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0073] Figure 63 SEQ ID NOs: 1860-1907 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN1A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0074] Figure 64 SEQ ID NOs: 1908-1955 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human SCN2A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0075] Figure 65 SEQ ID NO: 1956-2003, crRNA sequences generated by the bioinformatics methods described herein, which target human SCN3A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0076] Figure 66 SEQ ID NOs: 2004-2051 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN4A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0077] Figure 67SEQ ID NOs: 2052-2099 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN5A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0078] Figure 68 SEQ ID NOs: 2100-2147 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN8A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0079] Figure 69 SEQ ID NO: 2148-2195, crRNA sequences generated by the bioinformatics methods described herein, which target human SCN9A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0080] Figure 70 SEQ ID NOs: 2196-2243 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN10A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0081] Figure 71 SEQ ID NOs: 2244-2291 are described, crRNA sequences generated by the bioinformatics methods described herein that target human SCN11A to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0082] Figure 72 Illustrated are SEQ ID NOs: 2292-2308, crRNA sequences generated by the bioinformatics methods described herein, which target human TAC1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0083] Figure 73SEQ ID NOs: 2309-2325 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human TAC3 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0084] Figure 74 SEQ ID NOs: 2326-2373 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human TACR1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0085] Figure 75 SEQ ID NOs: 2374-2421 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human TACR2 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0086] Figure 76 SEQ ID NOs: 2422-2469 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human TACR3 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0087] Figure 77 Illustrated are SEQ ID NOs: 2470-2509, crRNA sequences generated by the bioinformatics methods described herein that target human TIMP1 to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0088] Figure 78 Illustrated are SEQ ID NOs: 2510-2557, crRNA sequences generated by the bioinformatics methods described herein that target human TIMP3 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0089] Figure 79SEQ ID NOs: 2558-2574 are illustrated, crRNA sequences generated by the bioinformatics methods described herein that target human TNF to modify and / or eliminate the expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance indicators.

[0090] Figure 80 SEQ ID NOs: 2575-2622 are described, crRNA sequences generated by the bioinformatics methods described herein that target human TNFRSF1A to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0091] Figure 81 Illustrated are SEQ ID NOs: 2623-2670, crRNA sequences generated by the bioinformatics methods described herein that target human TNFRSF1B to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0092] Figure 82 Illustrated are SEQ ID NOs: 2671-2718, crRNA sequences generated by the bioinformatics methods described herein that target human YAP1 to modify and / or eliminate expression of its encoded product. Additional information includes the chromosomal genomic coordinates of the editing site (assembly hg38) and a score summarizing several predicted performance metrics.

[0093] Figure 83A 、 Figure 83B 、 Figure 83C and Figure 83DResults from cell-based and in silico gene editing analyses of crRNA sequences targeting the (A) hIL1A, (B) hIL1B, (C) cIL1A, and (D) cIL1B genes are shown together. '°' indicates the CRISPR cut position within the amino acid (AA) translation frame. '*' indicates the optimization score from Doench, Fusi, et al. (2016). This score is optimized for a 20-bp guide with an NGG PAM. Scores range from 0 to 100. Higher is better. **Specificity score from Hsu et al. (2013). Scores range from 0 to 100. Higher is better. ***This score is based on experiments in U2OS. A high precision score (>0.4) means that DNA repair outcomes are consistent and only a few unique genotypes are enriched. ****This score is based on experiments in U2OS. A high frameshift frequency (>80%) would favor knocking protein-coding genes out of frame. The typical genomic frameshift frequency is higher than 66%, as 1-bp insertions and 1-2 bp deletions are particularly common repair outcomes. ^Combined score = (off-target score + precision score * 100 + frameshift) / 3. The pipe symbol '|' indicates a CRISPR cut site. Curly brackets '{}' indicate insertions. A hyphen '-' indicates a deletion. $ Potential off-target sites. Scoring is based on Hsu et al. (2013). On-target sites have a score of 100.

[0094] Figure 84A 、 Figure 84B 、 Figure 84C and Figure 84D Together, the results of functional assays measuring (A, B) cIL1A and (C, D) cIL1B release in edited or control canine chondrocytes 6 and 24 hours after exposure to PBS or LPS are illustrated.

[0095] Figure 85A 、 Figure 85B 、 Figure 85C and Figure 85D Together, the results of functional assays measuring (A, B) hIL1A and (C, D) cIL1B release in edited and control chondrocytes 6 and 24 hours after exposure to PBS or LPS are illustrated.

[0096] Figure 86 The results of tissue-specific splicing and expression analysis of the hIL1A gene are presented.

[0097] Figure 87 The results of tissue-specific splicing and expression analysis of the hIL1B gene are presented.

[0098] Figure 88The results of computer analysis of crRNA targeting hIL1A or hIL1B are illustrated. On-target scores were optimized for 20-bp gRNAs with an NGG protospacer adjacent motif (PAM) (see Doench et al.). Scores range from 0 to 1. Accuracy scores are based on experiments in U20S cells. A high accuracy score (>0.4) means that the DNA repair results are consistent and only a few unique genotypes are enriched. Frameshift percentages are based on experiments in U2OS cells. A higher (>80%) frameshift frequency will tend to knock protein-coding genes out of frame. Typical genomic frameshift frequencies are above 66% because 1-bp insertions and 1-2bp deletions are particularly common repair outcomes. Off-target scores from CRISPR assess the number of matches in the genome with a given number of mismatches. Mismatches in the seed sequence have a more deleterious effect.

[0099] Figure 89 illustrates the results of splicing and functional analysis of the cIL1A and cIL1B genes. The reference canine genome assembly (CanFam3.1) was used for these analyses.

[0100] Figure 90 Illustrated are the results of in silico analysis of crRNAs targeting the cIL1A or cIL1B gene.

[0101] Figure 91A 、 Figure 9 lB and Figure 91C The knockout efficacy of selected sgRNAs in (A) human chondrocytes, (B) canine chondrocytes, and (C) canine synoviocytes is collectively illustrated.

[0102] Figure 92 Illustrated are the results of in silico analysis of off-target effects of multiple sgRNAs in canine cells.

[0103] Figure 93A and Figure 93B Together illustrate the effect of (A) enhanced-specificity Cas9 (espCas9) in eliminating off-target editing of the indicated sgRNAs in canine cells compared to (B) the effect of standard spCas9.

[0104] Figure 94A and Figure 94B A summary of the editing activity of crRNAs targeting IL1A and IL1B in (A) human and (B) canine chondrocytes is collectively illustrated.

[0105] Figure 95 The results of simultaneous or sequential co-administration of multiple sgRNAs in canine cells are illustrated.

[0106] Figure 96A and Figure 96BSequence alignment of (A) IL1A and (B) IL1B genes from different mammalian species (human, horse, mouse, and dog) are illustrated together. DETAILED DESCRIPTION

[0107] introduction

[0108] Provided herein are compositions and methods for silencing the translation of one or more proteins in an animal in need thereof to treat a disease, disorder, or condition associated with pain.

[0109] In some embodiments, receptor signaling is silenced by CRISPR editing of the gene encoding the receptor. In some embodiments, CRISPR editing results in the ablation of the transmembrane domain (i.e., producing a soluble decoy receptor). In some embodiments, CRISPR editing results in the ablation of the cytoplasmic domain (i.e., producing a membrane-bound decoy receptor). In specific embodiments, compositions and methods for gene editing FGF2, CCN2, ADAMTS5, MMP1 and / or NGF are provided.

[0110] definition

[0111] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0112] In some embodiments, pain is relieved by silencing a nociceptive signaling protein (or its cognate receptor) by CRISPR editing of a gene encoding a protein (or receptor). In some embodiments, CRISPR editing results in ablation of the transmembrane domain of a pain receptor (i.e., generating a soluble decoy receptor). In some embodiments, CRISPR editing results in ablation of the cytoplasmic domain of a pain receptor (i.e., generating a membrane-bound decoy receptor). In certain embodiments, methods are provided for gene editing of (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, or NTRK2), (ii) one or more metalloproteinases or their regulators (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, or TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, , TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, or IL18RAP), (iv) one or more neural signaling regulators (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, or ATP1A1), (v) one or more other cell signaling regulators (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), or (vi) a combination of any of the genes of (i)-(v) to relieve pain.

[0113] definition

[0114] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications mentioned herein are incorporated by reference in their entirety.

[0115] The term "FGF2 gene" refers to a mammalian gene encoding a fibroblast growth factor 2 polypeptide. Non-limiting examples of FGF2 genes include: NCBI Gene ID: 2247 [Human], NCBI Gene ID: 403857 [Canine], NCBI Gene ID: 100033955 [Equine], NCBI Gene ID: 100135772 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by FGF2 genes include: UniProt: P09038; NP_001348594.1 [Human], XP_038421156.1 [Canine], NP_001182150.1 [Equine], XP_044911834.1 [Feline]), and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by the genes listed above, in addition to strong binding to heparin and integrins, also act as ligands for the FGF receptors FGFR1, FGFR2, FGFR3, and FGFR4. Furthermore, FGF2 signaling is thought to influence local nociception, at least through its pro-angiogenic activity, and is implicated in pain perception associated with at least IVD degeneration and joint damage. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of a particular species (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0116] The term "FGFR1 gene" refers to a mammalian gene encoding a fibroblast growth factor receptor 1 polypeptide. Non-limiting examples of FGFR1 genes include: NCBI Gene ID: 2260 [Human], NCBI Gene ID: 100856477 [Canine], NCBI Gene ID: 100057614 [Equine], NCBI Gene ID: 101086055 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by FGFR1 genes include: UniProt: P11362; NP_001167534.1 [Human], XP_038545782.1 [Canine], XP_023486323.1 [Equine], XP_011279822.1 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by the genes listed above is a tyrosine kinase, which serves as a cell surface receptor for fibroblast growth factor. In this effect, they play an important role in the regulation of embryonic development, cell proliferation, differentiation and migration. In some cases, and only for the purpose of eliminating ambiguity, prefixes (h, c, e and f refer to the forms of humans, dogs, horses and cats, respectively) are added when referring to proteins or genes of specific species.

[0117] The term "CCN2 gene" refers to a mammalian gene encoding a cell communication network factor 2 polypeptide. Non-limiting examples of CCN2 genes include: NCBI gene ID: 1490 [human], NCBI gene ID: 476202 [canine], NCBI gene ID: 100073098 [equine], NCBI gene ID: 101094598 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCN2 genes include: UniProt: P29279; NP_001892.2 [human], XP_038321343.1 [canine], XP_023506869.1 [equine], XP_023110145.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by the genes listed above are mitogens secreted by vascular endothelial cells and are involved in chondrocyte proliferation and differentiation, cell adhesion in many cell types. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0118] The term "ADAMTS5 gene" refers to a mammalian gene encoding an ADAM metallopeptidase having a thrombospondin type 1 motif 5 polypeptide. Non-limiting examples of ADAMTS5 genes include: NCBI gene ID: 11096 [human], NCBI gene ID: 487713 [canine], NCBI gene ID: 100066005 [equine], NCBI gene ID: 101085063 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the ADAMTS5 gene include: UniProt: Q9UNA0; NP_008969.2 [human], XP_038299214.1 [canine], XP_023485737.1 [equine], XP_023094603.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. The member of this family typically shares several different protein modules, comprises propeptide region, metalloproteinase domain, disintegrin-like domain and type 1 thrombospondin (TS) motif, and each member of this family is different in the quantity of C-terminal TS motif.ADAMTS5 has two unique C-terminal domains. Once proteolytic processing produces ripe enzyme, ADAMTS5 just is used as aggrecanase, and it cuts aggrecan (the main proteoglycan of cartilage), and may mediate the cartilage destruction in osteoarthritis. In some cases, and only for elimination of ambiguity, when referring to the protein of particular species or gene, add prefix (h, c, e and f refer to people, dog, horse and cat respectively).

[0119] The term "ADAMTS1 gene" refers to a mammalian gene encoding an ADAM metallopeptidase having a thrombospondin type 1 motif 1 polypeptide. Non-limiting examples of ADAMTS1 genes include: NCBI gene ID: 9510 [human], NCBI gene ID: 100686153 [canine], NCBI gene ID: 791251 [equine], NCBI gene ID: 101085309 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by ADAMTS1 genes include: UniProt: Q9UHI8; NP_008919.3 [human], XP_038374156.1 [canine], XP_023485736.1 [equine], XP_019695041.3 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, members of this family share several different protein modules, including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a type 1 thrombospondin (TS) motif, with individual members of the family differing in the number of C-terminal TS motifs. ADAMTS1 contains two disintegrin loops and three C-terminal TS motifs. This protein has anti-angiogenic activity and acts as an aggrecanase, which cuts aggrecan (the major proteoglycan of cartilage) and may be involved in its conversion and is associated with various inflammatory processes. In some cases, and simply to eliminate ambiguity, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0120] The term "MMP1 gene" refers to a mammalian gene encoding a matrix metallopeptidase 1 polypeptide. Non-limiting examples of MMP1 genes include: NCBI Gene ID: 4312 [human], NCBI Gene ID: 489428 [canine], NCBI Gene ID: 100033896 [equine], NCBI Gene ID: 101084217 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP1 genes include: UniProt: P03956; NP_001139410.1 [human], XP_038521018.1 [canine], NP_001075316.1 [equine], XP_003992365.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, MMP1 is proteolytically processed from a preproprotein to produce a mature protease. This secreted protease breaks down interstitial collagens, including types I, II, and III. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f refer to human, canine, equine, and feline, respectively).

[0121] The term "MMP2 gene" refers to a mammalian gene encoding a matrix metallopeptidase 2 polypeptide. Non-limiting examples of MMP2 genes include: NCBI Gene ID: 4313 [human], NCBI Gene ID: 403733 [canine], NCBI Gene ID: 100033948 [equine], NCBI Gene ID: 101098838 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP2 genes include: UniProt: P08253; NP_001121363.1 [human], XP_038515255.1 [canine], XP_023492775.1 [equine], XP_003998091.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP2 is a gelatinase A, type IV collagenase that contains three type II fibronectin repeats in its catalytic site, allowing denatured type IV and type V collagen and elastin to bind. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0122] The term "MMP3 gene" refers to a mammalian gene encoding a matrix metallopeptidase 3 polypeptide. Non-limiting examples of MMP3 genes include: NCBI Gene ID: 4314 [Human], NCBI Gene ID: 403733 [Canine], NCBI Gene ID: 100034195 [Equine], NCBI Gene ID: 493666 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP3 genes include: UniProt: P08254; NP_002413.1 [Human], NP_001002967.1 [Canine], NP_001075964.1 [Equine], XP_003992356.2 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave extracellular matrix components. MMP3 is an enzyme that degrades fibronectin, laminin, collagens III, IV, IX, and X, and cartilage proteoglycans, and is thought to be involved in wound repair, progression of atherosclerosis, and tumor initiation. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0123] The term "MMP7 gene" refers to a mammalian gene encoding a matrix metallopeptidase 7 polypeptide. Non-limiting examples of MMP7 genes include: NCBI Gene ID: 4316 [Human], NCBI Gene ID: 489432 [Canine], NCBI Gene ID: 100068985 [Equine], NCBI Gene ID: 727698 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP7 genes include: UniProt: P09237; NP_002414.1 [Human], NP_001229655.1 [Canine], XP_001498859.1 [Equine], XP_003992352.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP7 is proteolytically processed to produce a mature protease that, in addition to activating procollagenase, also breaks down proteoglycans, fibronectin, elastin, and casein. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0124] The term "MMP8 gene" refers to a mammalian gene encoding a matrix metallopeptidase 8 polypeptide. Non-limiting examples of MMP8 genes include: NCBI Gene ID: 4317 [human], NCBI Gene ID: 489429 [canine], NCBI Gene ID: 100069005 [equine], NCBI Gene ID: 101080995 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP8 genes include: UniProt: P22894; NP_001291370.1 [human], XP_038521019.1 [canine], XP_005611595.1 [equine], XP_003992354.3 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP8 is an enzyme that degrades interstitial collagen. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0125] The term "MMP10 gene" refers to a mammalian gene encoding a matrix metallopeptidase 10 polypeptide. Non-limiting examples of MMP10 genes include: NCBI Gene ID: 4319 [human], NCBI Gene ID: 100146442 [horse], NCBI Gene ID: 101081247 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by MMP10 genes include: UniProt: P09238; NP_002416.1 [human], XP_005614947.1 [horse], XP_003992355.2 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave extracellular matrix components. MMP10 is an enzyme that degrades fibronectin and types I, III, IV, and V gelatin. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a particular species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0126] The term "MMP12 gene" refers to a mammalian gene encoding a matrix metallopeptidase 12 polypeptide. Non-limiting examples of MMP12 genes include: NCBI Gene ID: 4321 [human], NCBI Gene ID: 611789 [canine], NCBI Gene ID: 100069047 [equine], NCBI Gene ID: 101084472 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP12 genes include: UniProt: P39900; NP_002417.2 [human], NP_001274067.1 [canine], XP_001498924.2 [equine], XP_003992366.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP12 is an enzyme with significant elastolytic activity and may be involved in tissue injury and remodeling. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0127] The term "MMP13 gene" refers to a mammalian gene encoding a matrix metallopeptidase 13 polypeptide. Non-limiting examples of MMP13 genes include: NCBI Gene ID: 4322 [Human], NCBI Gene ID: 403763 [Canine], NCBI Gene ID: 100009711 [Equine], NCBI Gene ID: 493679 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by MMP13 genes include: UniProt: P45452; NP_002418.1 [Human], XP_038521017.1 [Canine], NP_001075273.1 [Equine], XP_023094811.2 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene belongs to a broader family of zinc-dependent enzymes that cleave components of the extracellular matrix. MMP13 is an enzyme that degrades various types of collagen and is involved in wound healing, tissue remodeling, cartilage degradation, bone development, bone mineralization, and ossification. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0128] The term "TIMP1 gene" refers to a mammalian gene encoding a TIMP metallopeptidase inhibitor 1 polypeptide. Non-limiting examples of TIMP1 genes include: NCBI Gene ID: 7076 [human], NCBI Gene ID: 403816 [canine], NCBI Gene ID: 100034220 [equine], NCBI Gene ID: 101095886 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TIMP1 genes include: UniProt: P01033; NP_003245.1 [human], NP_001003182.1 [canine], XP_023488949.1 [equine], XP_023105059.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene exerts its effect by forming a one-to-one complex with a target metalloproteinase (such as collagenase), which is irreversibly inactivated by binding to its catalytic zinc cofactor. TIMP1 acts on MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, and MMP16, but not on MMP14, and has been shown to act as a growth factor that regulates cell differentiation, migration, and cell death. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0129] The term "TIMP3 gene" refers to a mammalian gene encoding a TIMP metallopeptidase inhibitor 3 polypeptide. Non-limiting examples of TIMP3 genes include: NCBI Gene ID: 7078 [human], NCBI Gene ID: 481289 [canine], NCBI Gene ID: 100033947 [equine], NCBI Gene ID: 101091215 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TIMP3 genes include: UniProt: P35625; NP_000353.1 [human], NP_001271368.1 [canine], NP_001075339.1 [equine], XP_003989265.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene complexes with metalloproteinases (such as collagenases), irreversibly inactivating them by binding to their catalytic zinc cofactor. TIMP3 is known to act on MMP1, MMP2, MMP3, MMP7, MMP9, MMP13, MMP14, and MMP15. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0130] The term "CXCL1 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 1 polypeptide. Non-limiting examples of CXCL1 genes include: NCBI Gene ID: 2919 [human], NCBI Gene ID: 100034121 [horse], NCBI Gene ID: 102901432 [cat], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL1 gene include: UniProt: P09341; NP_001502.1 [human], NP_001296409.1 [horse], XP_023108817.2 [cat], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene has chemotactic activity for neutrophils and may play a role in inflammation. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a particular species (h, e, e, and f for human, canine, equine, and feline, respectively).

[0131] The term "CXCL2 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 2 polypeptide. Non-limiting examples of CXCL2 genes include: NCBI gene ID: 2920 [human], NCBI gene ID: 100233237 [horse], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL2 gene include: UniProt: P19875, Q9UPB8; NP_002080.1 [human], NP_001137427.1 [horse], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene has antibacterial function through its regulation of inflammatory and immunoregulatory processes. CXCL2 is expressed at sites of inflammation and has been shown to inhibit the proliferation of hematopoietic progenitor cells. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0132] The term "CXCL3 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 3 polypeptide. Non-limiting examples of CXCL3 genes include: NCBI gene ID: 2921 [human] NCBI gene ID: 100056258 [horse], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL3 gene include: UniProt: P19876, Q4W5H9; NP_002081.2 [human], NP_001137265.1 [horse], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene is a secreted growth factor that signals through the G protein-coupled receptor CXCR2 and plays a role in inflammation and acts as a chemoattractant for neutrophils. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0133] The term "CXCL5 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 5 polypeptide. Non-limiting examples of CXCL5 genes include: NCBI gene ID: 6374 [human] and its synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by the CXCL5 gene include: UniProt: P19876, Q4W5H9; NP_002081.2 [human], UniProt: P97885 [rat], UniProt: P50228 [mouse] and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene is thought to interact with the G protein-coupled receptor CXCR2 to promote angiogenesis, remodel connective tissue, and recruit neutrophils. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0134] The term "CXCL6 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 6 polypeptide. Non-limiting examples of CXCL6 genes include: NCBI Gene ID: 6372 [human], NCBI Gene ID: 106557449 [canine], NCBI Gene ID: 100033988 [equine], NCBI Gene ID: 101094593 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CXCL6 genes include: UniProt: P80162; NP_002984.1 [human], XP_038541813.1 [canine], NP_001075355.2 [equine], XP_003985379.3 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by the gene is a chemotactic factor for neutrophils and exhibits antimicrobial activity. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0135] The term "CXCL8 gene" refers to a mammalian gene encoding a CXC motif chemokine ligand 8 polypeptide. Non-limiting examples of CXCL8 genes include: NCBI Gene ID: 3576 [Human], NCBI Gene ID: 403850 [Canine], NCBI Gene ID: 100037400 [Equine], NCBI Gene ID: 493836 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CXCL8 genes include: UniProt: P10145; NP_000575.1 [Human], NP_001003200.1 [Canine], NP_001077420.2 [Equine], NP_001009281.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene is secreted by mononuclear macrophages, neutrophils, eosinophils, T lymphocytes, epithelial cells and fibroblasts and is used as a chemotactic factor to guide neutrophils to the site of infection. CXCL8 also participates in proinflammatory signaling cascades with other cytokines. In some cases, and simply to eliminate ambiguity, prefixes are added when referring to proteins or genes of specific species (h, e and f refer to humans, dogs, horses and cats, respectively).

[0136] The term "CCL2 gene" refers to a mammalian gene encoding a C-C motif chemokine ligand 2 polypeptide. Non-limiting examples of CCL2 genes include: NCBI gene ID: 6347 [human], NCBI gene ID: 403981 [canine], NCBI gene ID: 100034136 [equine], NCBI gene ID: 100127112 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCL2 genes include: UniProt: P13500; NP_002973.1 [human] ], NP_001003297.1 [Canine], NP_001075400.1 [Equine], XP_003996605.1 [Feline], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene acts as a ligand for CCR2, which induces chemotactic activity of monocytes and basophils (but not neutrophils or eosinophils). In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, dog, horse, and cat, respectively).

[0137] The term "CCL3 gene" refers to a mammalian gene encoding a C-C motif chemokine ligand 3 polypeptide. Non-limiting examples of CCL3 genes include: NCBI Gene ID: 6348 [Human], NCBI Gene ID: 448787 [Canine], NCBI Gene ID: 100057909 [Equine], NCBI Gene ID: 100302540 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCL3 genes include: UniProt: P10147; NP_002974.1 [Human], NP_001005251.2 [Canine], NP_001108413.1 [Equine], NP_001157129.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene plays a role in inflammatory responses by binding to the receptors CCR1, CCR4, and CCR5. In some cases, and solely to disambiguate, a prefix is ​​added when referring to a protein or gene of a particular species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0138] The term "CCL5 gene" refers to a mammalian gene encoding a C-C motif chemokine ligand 5 polypeptide. Non-limiting examples of CCL5 genes include: NCBI Gene ID: 6352 [Human], NCBI Gene ID: 403522 [Canine], NCBI Gene ID: 100033925 [Equine], NCBI Gene ID: 493689 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCL5 genes include: UniProt: P13501; NP_001265665.1 [Human], NP_001003010.1 [Canine], NP_001075332.1 [Equine], NP_001009827.1 [Feline]), as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene acts as a chemoattractant for blood monocytes, memory T helper cells, and eosinophils, induces histamine release from basophils, and activates eosinophils. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0139] The term "CCL7 gene" refers to a mammalian gene encoding a C-C motif chemokine ligand 7 polypeptide. Non-limiting examples of CCL7 genes include: NCBI Gene ID: 6354 [human], NCBI Gene ID: 491148 [canine], NCBI Gene ID: 100071714 [equine], NCBI Gene ID: 101096931 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCL7 genes include: UniProt: P80098; NP_006264.2 [human], NP_001010960.1 [canine], XP_005597638.1 [equine], XP_044900774.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene is a secreted chemokine that attracts macrophages during inflammation and metastasis and is an in vivo substrate of MMP2. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0140] The term "CCL20 gene" refers to a mammalian gene encoding a C-C motif chemokine ligand 20 polypeptide. Non-limiting examples of CCL20 genes include: NCBI Gene ID: 6364 [human], NCBI Gene ID: 448790 [canine], NCBI Gene ID: 100629808 [equine], NCBI Gene ID: 101089032 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the CCL20 gene include: UniProt: P78556; NP_001123518.1 [human], NP_001005254.1 [canine], XP_003365179.2 [equine], XP_003991274.2 [feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene is involved in inflammatory processes and exhibits chemotactic activity for lymphocytes. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, e, and f for human, canine, equine, and feline, respectively).

[0141] The term "CXCR1 gene" refers to a mammalian gene encoding a CXC motif chemokine receptor 1 polypeptide. Non-limiting examples of CXCR1 genes include: NCBI Gene ID: 3577 [human], NCBI Gene ID: 478906 [canine], NCBI Gene ID: 100058291 [equine], NCBI Gene ID: 101085650 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CXCR1 genes include: UniProt: P25024; NP_000625.1 [human], XP_038303849.1 [canine], XP_001491062.1 [equine], XP_011283865.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene is a receptor for IL-8 and transduces signaling to mediate migration of neutrophils to sites of inflammation, among other activities. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0142] The term "CXCR2 gene" refers to a mammalian gene encoding a CXC motif chemokine receptor 2 polypeptide. Non-limiting examples of CXCR2 genes include: NCBI Gene ID: 3579 [human], NCBI Gene ID: 478905 [canine], NCBI Gene ID: 100055552 [equine], NCBI Gene ID: 101085396 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CXCR2 genes include: UniProt: P25025; NP_001161770.1 [human], NP_001003151.2 [canine], XP_005610662.1 [equine], XP_044890398.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by this gene is a receptor for IL-8 and transduces signaling to mediate migration of neutrophils to sites of inflammation, among other activities. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0143] The term "CCR7 gene" refers to a mammalian gene encoding a C-C motif chemokine receptor 7 polypeptide. Non-limiting examples of CCR7 genes include: NCBI Gene ID: 1236 [human], NCBI Gene ID: 491011 [canine], NCBI Gene ID: 100067673 [equine], NCBI Gene ID: 101084327 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CCR7 genes include: UniProt: P32248; NP_001288643.1 [human], XP_038403305.1 [canine], XP_001500231.1 [equine], XP_003996882.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene controls the migration of memory T cells to inflamed tissues and stimulates the maturation of dendritic cells. Signals mediated by this receptor may also play a role in the pathogenesis of chronic inflammation. In some cases, and simply to eliminate ambiguity, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0144] The term "ADAM17 gene" refers to a mammalian gene encoding an ADAM metallopeptidase domain 17 polypeptide. Non-limiting examples of ADAM17 genes include: NCBI gene ID: 6868 [human], NCBI gene ID: 475662 [canine], NCBI gene ID: 100072496 [equine], NCBI gene ID: 101089004 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the ADAM17 gene include: UniProt: P78536; NP_001369706.1 [human], NP_001273795.1 [canine], NP_001295481.1 [equine], XP_003984558.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are proteolytically processed to produce a mature protease that acts by shedding the extracellular domain of tumor necrosis factor-alpha, thereby releasing soluble tumor necrosis factor-alpha from its membrane-bound precursor. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0145] The term "TNF gene" refers to a mammalian gene encoding a tumor necrosis factor polypeptide. Non-limiting examples of TNF genes include: NCBI Gene ID: 7124 [human], NCBI Gene ID: 403922 [canine], NCBI Gene ID: 100033834 [equine], NCBI Gene ID: 493755 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TNF genes include: UniProt: P01375; NP_000585.2 [human], NP_001003244.4 [canine], NP_001075288.2 [equine], NP_001009835.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by these genes is a multifunctional proinflammatory cytokine that is primarily secreted by macrophages and can bind to (and thus act through) their receptors TNFRSF1A and TNFRSF1B. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0146] The term "TNFRSF1A gene" refers to a mammalian gene encoding a tumor necrosis factor receptor 1 polypeptide. Non-limiting examples of TNFRSF1A genes include: NCBI Gene ID: 7132 [human], NCBI Gene ID: 403634 [canine], NCBI Gene ID: 100059548 [equine], NCBI Gene ID: 493957 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TNFRSF1A genes include: UniProt: P19438; NP_001056.1 [human], XP_038295153.1 [canine], XP_023498787.1 [equine], NP_001009361.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by the genes listed above are transmembrane receptor proteins that are capable of binding their primary ligands, tumor necrosis factor alpha (TNFA) or lymphotoxin alpha (LTA). Upon binding to TNFA, the receptors trimerize and become activated, transmitting intracellular signaling cascades and playing a role in various processes, including apoptosis and inflammation. See generally Ward-Kavanagh, LK. et al. (2016). Immunity, 44(5), 1005-1019. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, dog, horse, and cat, respectively).

[0147] The term "TNFRSF1B gene" refers to a mammalian gene encoding a tumor necrosis factor receptor 2 polypeptide. Non-limiting examples of TNFRSF1B genes include: NCBI Gene ID: 7133 [human], NCBI Gene ID: 487437 [canine], NCBI Gene ID: 100055840 [equine], NCBI Gene ID: 101080392 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TNFRSF1B genes include: UniProt: P20333; XP_011540362.1 [human], XP_038387905.1 [canine], XP_023491528.1 [equine], XP_023113905.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are transmembrane receptor proteins that can bind to TNFA or LTA and participate in pro-survival pathways through downstream activation of the NFkB pathway. See generally Ward-Kavanagh, LK et al. (2016). Immunity, 44(5), 1005-1019. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, dog, horse, and cat, respectively).

[0148] The term "IL4 gene" refers to a mammalian gene encoding an interleukin-4 polypeptide. Non-limiting examples of IL4 genes include: NCBI Gene ID: 3565 [Human], NCBI Gene ID: 403785 [Canine], NCBI Gene ID: 100034225 [Equine], NCBI Gene ID: 751514 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL4 genes include: UniProt: P05112; NP_000580.1 [Human], NP_001003159.1 [Canine], NP_001075988.1 [Equine], NP_001036804.1 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are pleiotropic cytokines produced by activated T cells and are considered important cytokines for tissue repair, counteracting the effects of proinflammatory type 1 cytokines, although they also promote allergic airway inflammation and mediate acute inflammation, among other activities. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0149] The term "IL4R gene" refers to a mammalian gene encoding an interleukin-4 receptor polypeptide. Non-limiting examples of IL4R genes include: NCBI Gene ID: 3566 [human], NCBI Gene ID: 489957 [canine], NCBI Gene ID: 791252 [equine], NCBI Gene ID: 101096277 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL4R genes include: UniProt: P24394; NP_000409.1 [human], NP_001003159.1 [canine], XP_005598791.2 [equine], XP_023102076.2 [feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are type I transmembrane proteins that can bind interleukin-4 and interleukin-13 to regulate IgE production and promote the differentiation of Th2 cells, among other activities. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0150] The term "IL6 gene" refers to a mammalian gene encoding an interleukin-6 polypeptide. Non-limiting examples of IL6 genes include: NCBI Gene ID: 3569 [Human], NCBI Gene ID: 403985 [Canine], NCBI Gene ID: 100034196 [Equine], NCBI Gene ID: 493687 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL6 genes include: UniProt: P05231; NP_000591.1 [Human], NP_001003301.1 [Canine], NP_001075965.2 [Equine], NP_001009211.2 [Feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are cytokines that play a role in inflammation and B cell maturation, are produced primarily at sites of acute and chronic inflammation, are secreted into the serum at these sites, and induce transcriptional inflammatory responses via the interleukin 6 receptor. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of a particular species (h, e, and f refer to humans, dogs, horses, and cats, respectively).

[0151] The term "IL6R gene" refers to a mammalian gene encoding an interleukin-6 receptor polypeptide. Non-limiting examples of IL6R genes include: NCBI Gene ID: 3560 [human], NCBI Gene ID: 612271 [canine], NCBI Gene ID: 102148787 [equine], NCBI Gene ID: 101085689 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL6R genes include: UniProt: P08887; CAA41231.1 [human], XP_038527979.1 [canine], XP_023496854.1 [equine], XP_023103841.2 [feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are transmembrane proteins that bind to their natural ligand, interleukin-6. This binding event triggers intracellular signaling events, leading to a pro-inflammatory response. See generally Wolf, J. et al. (2014). Cytokine, 70(1), 11-20. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, dog, horse, and cat, respectively).

[0152] The term "IL6ST gene" refers to a mammalian gene encoding a signaling polypeptide of the interleukin-6 cytokine family. Non-limiting examples of IL6ST genes include: NCBI Gene ID: 3572 [Human], NCBI Gene ID: 403545 [Canine], NCBI Gene ID: 100051700 [Equine], NCBI Gene ID: 101089832 [Feline], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by IL6ST genes include: UniProt: P40189; NP_001177910.1 [Human], NP_001273950.1 [Canine], XP_023481030.1 [Equine], XP_011281205.1 [Feline]), and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by the genes listed above are signal transducers shared by many cytokines, including interleukin 6 (IL6), ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), and oncostatin M (OSM), and function as part of a cytokine receptor complex. The activation of this protein depends on the binding of cytokines to their receptors (e.g., IL6 to IL6R). Knockout studies in mice have shown that this gene plays a key role in regulating myocyte apoptosis. See generally Martínez-Pérez, C. et al. (2021). Journal of Personalized Medicine, 11(7), 618. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0153] The term "IL10 gene" refers to a mammalian gene encoding an interleukin-10 polypeptide. Non-limiting examples of IL10 genes include: NCBI Gene ID: 3586 [Human], NCBI Gene ID: 403628 [Canine], NCBI Gene ID: 100034187 [Equine], NCBI Gene ID: 493683 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL10 genes include: UniProt: P22301; NP_000563.1 [Human], NP_001003077.1 [Canine], NP_001075959.1 [Equine], NP_001009209.1 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by these genes is a pleiotropic cytokine that regulates inflammation and acts on many immune cell types by binding to a heterodimeric receptor composed of IL10RA and IL10RB, thereby activating downstream signaling cascades such as the JAK-STAT pathway. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0154] The term "IL10RA gene" refers to a mammalian gene encoding an interleukin-10 receptor alpha polypeptide. Non-limiting examples of IL10RA genes include: NCBI Gene ID: 3587 [human], NCBI Gene ID: 610823 [canine], NCBI Gene ID: 100071172 [equine], NCBI Gene ID: 101087601 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL10RA genes include: UniProt: Q13651; NP_001549.2 [human], XP_038520677.1 [canine], XP_014596783.1 [equine], XP_003992449.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes act as regulators of proinflammatory signaling by binding to its ligand, IL-10, when forming heterodimers with IL10RB. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0155] The term "IL10RB gene" refers to a mammalian gene encoding an interleukin-10 receptor beta polypeptide. Non-limiting examples of IL10RB genes include: NCBI Gene ID: 3588 [human], NCBI Gene ID: 478404 [canine], NCBI Gene ID: 100052549 [equine], NCBI Gene ID: 101090038 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL10RB genes include: UniProt: Q08334; NP_000619.3 [human], XP_038299308.1 [canine], XP_023485821.1 [equine], XP_003991512.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes act as regulators of proinflammatory signaling by binding to its ligand, IL-10, when forming heterodimers with IL10RA. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0156] The term "IL13 gene" refers to a mammalian gene encoding an interleukin-13 polypeptide. Non-limiting examples of IL13 genes include: NCBI Gene ID: 3596 [Human], NCBI Gene ID: 442990 [Canine], NCBI Gene ID: 100034113 [Equine], NCBI Gene ID: 101084678 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL13 genes include: UniProt: P35225; NP_001341920.1 [Human], NP_001003384.1 [Canine], NP_001137263.1 [Equine], NP_001009209.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes regulate the production of proinflammatory cytokines and chemokines. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0157] The term "IL13RA1 gene" refers to a mammalian gene encoding an interleukin-13 receptor alpha 1 polypeptide. Non-limiting examples of IL13RA1 genes include: NCBI Gene ID: 3597 [human], NCBI Gene ID: 403623 [canine], NCBI Gene ID: 100055312 [equine], NCBI Gene ID: 101091351 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL13RA1 genes include: UniProt: P78552; NP_001551.1 [human], XP_038306633.1 [canine], XP_023490026.1 [equine], XP_023104651.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are low-affinity binding partners for IL13 and contain a functional receptor that once associates with IL13RA2. Once bound to IL13, the receptor complex stimulates the production of proinflammatory cytokines and chemokines. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f refer to human, canine, equine, and feline, respectively).

[0158] The term "IL13RA2 gene" refers to a mammalian gene encoding an interleukin-13 receptor α2 polypeptide. Non-limiting examples of IL13RA2 genes include: NCBI Gene ID: 3598 [human], NCBI Gene ID: 403622 [canine], NCBI Gene ID: 100057673 [equine], NCBI Gene ID: 101100114 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL13RA2 genes include: UniProt: Q14627; NP_000631.1 [human], NP_001003075.1 [canine], XP_023489189.1 [equine], XP_044906881.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are high-affinity binding partners of IL-13 but lack the cytoplasmic domain. This protein, together with IL-13RA1, forms a functional receptor that stimulates the production of proinflammatory cytokines and chemokines. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0159] The term "IL17A gene" refers to a mammalian gene encoding an interleukin-17A polypeptide. Non-limiting examples of IL17A genes include: NCBI Gene ID: 3605 [human], NCBI Gene ID: 481837 [canine], NCBI Gene ID: 100034142 [equine], NCBI Gene ID: 101095339 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL17A genes include: UniProt: Q16552; NP_002181.1 [human], NP_001159350.1 [canine], NP_001137264.1 [equine], XP_006931878.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are inflammatory cytokines that activate the NFκB signaling pathway by interacting with the heterodimeric receptor complex of IL17RA and IL17RC, thereby activating the transcription of various chemokines, cytokines, and other factors. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, canine, equine, and feline, respectively).

[0160] The term "IL17RA gene" refers to a mammalian gene encoding an interleukin-17 receptor A polypeptide. Non-limiting examples of IL17RA genes include: NCBI Gene ID: 23765 [human], NCBI Gene ID: 486759 [canine], NCBI Gene ID: 100055511 [equine], NCBI Gene ID: 101095588 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL17RA genes include: UniProt: Q96F46; NP_001276834.1 [human], XP_038295433.1 [canine], XP_005610881.1 [equine], XP_023112364.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are transmembrane proteins that bind to IL17RA with low affinity as part of a multimeric receptor complex. IL17RA and its ligands are associated with many inflammatory conditions. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0161] The term "IL18 gene" refers to a mammalian gene encoding an interleukin-18 polypeptide. Non-limiting examples of IL18 genes include: NCBI Gene ID: 3606 [Human], NCBI Gene ID: 403796 [Canine], NCBI Gene ID: 100034216 [Equine], NCBI Gene ID: 493688 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL18 genes include: UniProt: Q14116; NP_001230140.1 [Human], XP_038520002.1 [Canine], XP_005611483.1 [Equine], NP_001009213.2 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by these genes is a proinflammatory cytokine that, when bound to its receptor and co-receptor IL18R1 and IL18RAP, regulates inflammatory signaling through the NFκB pathway. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0162] The term "IL18R1 gene" refers to a mammalian gene encoding an interleukin-18 receptor 1 polypeptide. Non-limiting examples of IL18R1 genes include: NCBI Gene ID: 8809 [human], NCBI Gene ID: 611438 [canine], NCBI Gene ID: 100058269 [equine], NCBI Gene ID: 493938 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL18R1 genes include: UniProt: Q13478; NP_001269328.1 [human], XP_038536128.1 [canine], XP_023474273.1 [equine], NP_001009863.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are essential components for transducing IL18-mediated pro-inflammatory signaling. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0163] The term "IL18RAP gene" refers to a mammalian gene encoding an interleukin-18 receptor accessory protein polypeptide. Non-limiting examples of IL18RAP genes include: NCBI Gene ID: 8807 [human], NCBI Gene ID: 481327 [canine], NCBI Gene ID: 100050212 [equine], NCBI Gene ID: 101084868 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL18RAP genes include: UniProt: Q53TU5; NP_001380415.1 [human], XP_038536125.1 [canine], XP_014586460.1 [equine], XP_019682529.2 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by these genes is an accessory protein that enhances signal transduction of IL18-mediated proinflammatory signaling. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0164] The term "NGF gene" refers to a mammalian gene encoding a nerve growth factor polypeptide. Non-limiting examples of NGF genes include: NCBI gene ID: 4803 [human], NCBI gene ID: 403402 [canine], NCBI gene ID: 100065669 [equine], NCBI gene ID: 100144611 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by NGF genes include: UniProt: P01138; NP_002497.2 [human], XP_038546347.1 [canine], XP_001496237.2 [equine], XP_044889256.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, this secreted protein forms functional homodimers that are incorporated into larger complexes and have nerve growth stimulating activity. This complex is also involved in the regulation of the growth and differentiation of sympathetic and certain sensory neurons. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0165] The term "NGFR gene" refers to a mammalian gene encoding a nerve growth factor receptor polypeptide. Non-limiting examples of NGFR genes include: NCBI Gene ID: 4804 [human], NCBI Gene ID: 491071 [canine], NCBI Gene ID: 100069694 [equine], NCBI Gene ID: 101101519 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by NGFR genes include: UniProt: P08138; NP_002498.1 [human], XP_038531049.1 [canine], XP_023508464.1 [equine], XP_023099534.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes contain four 40-amino acid repeats in their extracellular domain, with six cysteine ​​residues at conserved positions, followed by a serine / threonine-rich region. This cysteine-rich region contains a nerve growth factor binding domain that, once bound, allows signal transduction. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0166] The term "NTF3 gene" refers to a mammalian gene encoding a neurotrophin-3 polypeptide. Non-limiting examples of NTF3 genes include: NCBI Gene ID: 4908 [Human], NCBI Gene ID: 493963 [Canine], NCBI Gene ID: 100051839 [Equine], NCBI Gene ID: 486731 [Feline], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by NTF3 genes include: UniProt: P20783; NP_001096124.1 [Human], XP_038293846.1 [Canine], XP_023498780.1 [Equine], NP_001009367.1 [Feline], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes control the survival and differentiation of neurons. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a particular species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0167] The term "NTF4 gene" refers to a mammalian gene encoding a neurotrophin-4 polypeptide. Non-limiting examples of NTF4 genes include: NCBI Gene ID: 4909 [human], NCBI Gene ID: 611987 [canine], NCBI Gene ID: 100054859 [equine], NCBI Gene ID: 101100428 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by NTF4 genes include: UniProt: P34130; NP_001382418.1 [human], NP_001177358.2 [canine], XP_023505846.1 [equine], XP_023101354.2 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are proteolytically processed into mature forms that can promote neuronal survival by binding to their cognate receptors. Disregulation of this protein has been observed in various neurological disorders. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0168] The term "NTRK1 gene" refers to a mammalian gene encoding a neurotrophic receptor tyrosine kinase 1 polypeptide. Non-limiting examples of NTRK1 genes include: NCBI Gene ID: 4914 [human], NCBI Gene ID: 490404 [canine], NCBI Gene ID: 100064594 [equine], NCBI Gene ID: 101081603 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by NTRK1 genes include: UniProt: P04629; NP_001007793.1 [human], XP_038527745.1 [canine], XP_023496742.1 [equine], XP_023103311.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by these genes is a membrane-bound receptor that binds to neutrophils and signals through the MAPK pathway to regulate cell differentiation, among other functions. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0169] The term "NTRK2 gene" refers to a mammalian gene encoding a neurotrophic receptor tyrosine kinase 2 polypeptide. Non-limiting examples of NTRK2 genes include: NCBI Gene ID: 4915 [human], NCBI Gene ID: 484147 [canine], NCBI Gene ID: 100061700 [equine], NCBI Gene ID: 101101347 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by NTRK2 genes include: UniProt: Q16620; NP_001007098.1 [human], XP_038510982.1 [canine], XP_023482906.1 [equine], XP_023097987.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by these genes is a membrane-bound receptor that binds to neutrophils and signals through the MAPK pathway to regulate cell differentiation, among other functions. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0170] The term "BDNF gene" refers to a mammalian gene encoding a brain-derived neurotrophic factor polypeptide. Non-limiting examples of BDNF genes include: NCBI Gene ID: 627 [Human], NCBI Gene ID: 403461 [Canine], NCBI Gene ID: 100009689 [Equine], NCBI Gene ID: 493690 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by BDNF genes include: UniProt: P23560; NP_001137277.1 [Human], NP_001002975.1 [Canine], NP_001075256.1 [Equine], NP_001009828.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are proteolytically processed into mature forms that can promote neuronal survival by binding to their cognate receptors. Disregulation of this protein has been observed in various neurological disorders. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0171] The term "SCN1A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α1 polypeptide. Non-limiting examples of SCN1A genes include: NCBI Gene ID: 6323 [human], NCBI Gene ID: 478775 [canine], NCBI Gene ID: 100052059 [equine], NCBI Gene ID: 101081823 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN1A gene include: UniProt: P35498; NP_001159435.1 [human], XP_038302870.1 [canine], XP_023478839.1 [equine], XP_019693764.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, these genes encode proteins that mediate voltage-dependent sodium ion permeability of excitable membranes and are involved in the sensory perception of mechanical pain (i.e., activation of somatosensory neurons has been shown to induce pain without neurogenic inflammation). In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0172] The term "SCN2A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α2 polypeptide. Non-limiting examples of SCN2A genes include: NCBI Gene ID: 6326 [human], NCBI Gene ID: 478773 [canine], NCBI Gene ID: 100051816 [equine], NCBI Gene ID: 101080472 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN2A gene include: UniProt: Q99250; NP_001035232.1 [human], XP_038302857.1 [canine], XP_023478830.1 [equine], XP_023115179.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes mediate voltage-dependent sodium ion permeability of excitable membranes. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0173] The term "SCN3A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α3 polypeptide. Non-limiting examples of SCN3A genes include: NCBI Gene ID: 6328 [human], NCBI Gene ID: 478772 [canine], NCBI Gene ID: 100061941 [equine], NCBI Gene ID: 101082587 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN3A gene include: UniProt: Q9NY46; NP_001075145.1 [human], XP_038302852.1 [canine], XP_023478823.1 [equine], XP_019693750.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are subunits of voltage-gated sodium channels and are responsible for the propagation of action potentials in neurons and muscle tissue. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0174] The term "SCN4A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α4 polypeptide. Non-limiting examples of SCN4A genes include: NCBI Gene ID: 6328 [human], NCBI Gene ID: 119873250 [canine], NCBI Gene ID: 100049793 [equine], NCBI Gene ID: 101098669 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN4A gene include: UniProt: Q9NY46; NP_001075145.1 [human], XP_038531923.1 [canine], NP_001075230.2 [equine], XP_006940553.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are subunits of voltage-gated sodium channels and are responsible for the propagation of action potentials in neurons and muscle tissue. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0175] The term "SCN5A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α5 polypeptide. Non-limiting examples of SCN5A genes include: NCBI Gene ID: 6331 [human], NCBI Gene ID: 403497 [canine], NCBI Gene ID: 100034027 [equine], NCBI Gene ID: 101100994 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN5A gene include: UniProt: Q14524; NP_000326.2 [human], NP_001002994.1 [canine], NP_001157367.1 [equine], XP_044893792.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are subunits of voltage-gated sodium channels and are found primarily in cardiac muscle and are responsible for the initial upstroke of the action potential in the electrocardiogram. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0176] The term "SCN8A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α8 polypeptide. Non-limiting examples of SCN8A genes include: NCBI gene ID: 6335 [human], NCBI gene ID: 477604 [canine], NCBI gene ID: 100052777 [equine], NCBI gene ID: 101096578 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN8A gene include: UniProt: Q9UQD0; NP_001171455.1 [human], XP_038294063.1 [canine], XP_023499351.1 [equine], XP_023112849.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are ion pore subunits of voltage-gated sodium channels and are essential for rapid membrane depolarization during neuronal action potentials. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0177] The term "SCN9A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α9 polypeptide. Non-limiting examples of SCN9A genes include: NCBI gene ID: 6335 [human], NCBI gene ID: 100855710 [canine], NCBI gene ID: 100052120 [equine], NCBI gene ID: 101082841 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN9A gene include: UniProt: Q15858; NP_001352465.1 [human], XP_038302872.1 [canine], XP_023478844.1 [equine], XP_044889827.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are voltage-dependent sodium ion channels, which are involved in various pain conditions, especially in the development of inflammatory pain. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0178] The term "SCN10A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α10 polypeptide. Non-limiting examples of SCN10A genes include: NCBI Gene ID: 6336 [human], NCBI Gene ID: 477026 [canine], NCBI Gene ID: 100055493 [equine], NCBI Gene ID: 101085569 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN10A gene include: UniProt: Q9Y5Y9; NP_001280235.2 [human], NP_001003203.1 [canine], XP_014587037.1 [equine], XP_044893784.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are transmembrane subunits of voltage-dependent sodium channels, which may be involved in the onset of pain associated with neuropathy. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0179] The term "SCN11A gene" refers to a mammalian gene encoding a sodium voltage-gated channel α11 polypeptide. Non-limiting examples of SCN11A genes include: NCBI gene ID: 11280 [human], NCBI gene ID: 485593 [canine], NCBI gene ID: 100068480 [equine], NCBI gene ID: 101085312 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the SCN11A gene include: UniProt: Q9UI33; NP_001336182.1 [human], XP_038426400.1 [canine], XP_001916634.3 [equine], XP_044893782.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are transmembrane subunits of voltage-dependent sodium channels and are highly expressed in the nociceptive neurons of the dorsal root ganglion and trigeminal ganglion. Mutations in the SCN11A gene are associated with various pain conditions. In some cases, and only for the purpose of eliminating ambiguity, prefixes (h, c, e, and f refer to humans, dogs, horses, and cats, respectively) are added when referring to proteins or genes of specific species.

[0180] The term "TAC1 gene" refers to a mammalian gene encoding a tachykinin precursor 1 polypeptide. Non-limiting examples of TAC1 genes include: NCBI Gene ID: 6863 [Human], NCBI Gene ID: 475239 [Canine], NCBI Gene ID: 100052324 [Equine], NCBI Gene ID: 101095481 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TAC1 genes include: UniProt: P20366; NP_003173.1 [Human], XP_038541905.1 [Canine], XP_014594521.1 [Equine], XP_003982840.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are precursors to four products of the tachykinin peptide hormone family: substance P, neurokinin A, neuropeptide K, and neuropeptide gamma. These hormones are thought to function as neurotransmitters that interact with nerve receptors and smooth muscle cells. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0181] The term "TAC3 gene" refers to a mammalian gene encoding a tachykinin precursor 3 polypeptide. Non-limiting examples of TAC3 genes include: NCBI Gene ID: 6866 [Human], NCBI Gene ID: 607315 [Canine], NCBI Gene ID: 100052722 [Equine], NCBI Gene ID: 101089368 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TAC3 genes include: UniProt: Q9UHF0; NP_001171525.1 [Human], UniProt: A0A8I3N7Z8; NP_001362511.2 [Canine], XP_023499603.1 [Equine], XP_019690663.1 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are proteolytically processed to generate mature peptides, which are mainly expressed in the central and peripheral nervous systems and play a role as neurotransmitters. This peptide is a ligand for the neurokinin-3 receptor. These hormones are thought to play the role of neurotransmitters that interact with nerve receptors and smooth muscle cells. In some cases, and only for the purpose of eliminating ambiguity, prefixes are added when referring to proteins or genes of specific species (h, c, e and f refer to humans, dogs, horses and cats, respectively).

[0182] The term "TACR1 gene" refers to a mammalian gene encoding a tachykinin receptor 1 polypeptide. Non-limiting examples of TACR1 genes include: NCBI Gene ID: 6869 [human], NCBI Gene ID: 403815 [canine], NCBI Gene ID: 100053491 [equine], NCBI Gene ID: 101090094 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TACR1 genes include: UniProt: P25103; NP_001049.1 [human], NP_001012637.1 [canine], XP_001499730.1 [equine], XP_003984209.1 [feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are receptors for the tachykinin substance P, also known as neurokinin 1. TACR1 activates the phosphatidylinositol-calcium second messenger system and can also bind with lower affinity substance K and neuromedin-K. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0183] The term "TACR2 gene" refers to a mammalian gene encoding a tachykinin receptor 2 polypeptide. Non-limiting examples of TACR2 genes include: NCBI Gene ID: 6865 [human], NCBI Gene ID: 489020 [canine], NCBI Gene ID: 100034168 [equine], NCBI Gene ID: 101094541 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the TACR2 gene include: UniProt: P21452; NP_001048.2 [human], NP_001012635.1 [canine], XP_001502752.2 [equine], XP_044896003.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are receptors for tachykinin substance K, also known as neurokinin A. TACR2 activates the phosphatidylinositol-calcium second messenger system and can also bind with lower affinity to neuromedin-K and substance P. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0184] The term "TACR3 gene" refers to a mammalian gene encoding a tachykinin receptor 3 polypeptide. Non-limiting examples of TACR3 genes include: NCBI Gene ID: 6870 [Human], NCBI Gene ID: 403814 [Canine], NCBI Gene ID: 100073088 [Equine], NCBI Gene ID: 101093603 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by TACR3 genes include: UniProt: P29371; NP_001050.1 [Human], NP_001091010.1 [Canine], XP_023492571.1 [Equine], XP_003985169.3 [Feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the proteins encoded by these genes are receptors for the tachykinin neurokinin 3, also known as neurokinin B or neuromedin-K. TACR3 activates the phosphatidylinositol-calcium second messenger system and can also bind substance K and substance P with lower affinity. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0185] The term "MRGPRX2 gene" refers to a mammalian gene encoding a MAS-associated GPR family member X2 polypeptide. Non-limiting examples of MRGPRX2 genes include: NCBI gene ID: 117194 [human], NCBI gene ID: 485410 [canine], NCBI gene ID: 100071950 [equine], NCBI gene ID: 101097092 [feline]) or encoded gene products (e.g., UniProt: Q96LB1; NP_001290544.1 [human], XP_038285538.1 [canine], XP_02350 1936.1 [Equine], XP_003993155.1 [Feline], as well as sequence variants, isoforms encoded by alternative splicing, and their various glycoforms. Typically, the proteins encoded by these genes are capable of G protein-coupled receptor activity and neuropeptide binding activity, and are involved in the positive regulation of mast cell degranulation and cytokinesis. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to human, canine, equine, and feline forms, respectively).

[0186] The term "ATP1A1 gene" refers to a mammalian gene encoding the ATPase Na+ / K+ transport subunit α1 polypeptide. Non-limiting examples of ATP1A1 genes include: NCBI Gene ID: 476 [Human], NCBI Gene ID: 403992 [Canine], NCBI Gene ID: 100034139 [Equine], NCBI Gene ID: 101083695 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by the ATP1A1 gene include: UniProt: P05023; NP_000692.2 [Human], NP_001376153.1 [Canine], NP_001108004.2 [Equine], XP_011283388.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are integral membrane protein subunits of complexes responsible for establishing and maintaining the electrochemical gradients of Na and K ions across the plasma membrane, which are essential for osmoregulation and electrical excitability of nerves and muscles. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0187] The term "CALCA gene" refers to a mammalian gene encoding a calcitonin-related polypeptide alpha polypeptide. Non-limiting examples of CALCA genes include: NCBI Gene ID: 796 [Human], NCBI Gene ID: 403946 [Canine], NCBI Gene ID: 100033906 [Equine], NCBI Gene ID: 101095582 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CALCA genes include: UniProt: P01258; NP_001029124.1 [Human], NP_001300719.1 [Canine], NP_001075323.1 [Equine], XP_019667660.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the gene encodes a variety of gene products, such as calcitonin, calcitonin gene-related peptide, and procalcitonin, by tissue-specific alternative RNA splicing of the gene transcript and cleavage of an inactive precursor protein. These proteins are involved in calcium regulation, regulating phosphorus metabolism, and acting as vasodilators, among other functions. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0188] The term "CALCB gene" refers to a mammalian gene encoding a calcitonin-related polypeptide beta polypeptide. Non-limiting examples of CALCB genes include: NCBI Gene ID: 797 [Human], NCBI Gene ID: 403415 [Canine], NCBI Gene ID: 100034126 [Equine], NCBI Gene ID: 101094539 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CALCB genes include: UniProt: P10092; NP_000719.1 [Human], NP_001002948.1 [Canine], NP_001075397.1 [Equine], XP_044894937.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes act as vasodilators and neurotransmitters, among other functions. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0189] The term "CALCRL gene" refers to a mammalian gene encoding a calcitonin receptor-like receptor polypeptide. Non-limiting examples of CALCRL genes include: NCBI Gene ID: 10203 [human], NCBI Gene ID: 488438 [canine], NCBI Gene ID: 100054281 [equine], NCBI Gene ID: 101086333 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CALCRL genes include: UniProt: Q16602; NP_001258680.1 [human], XP_038303202.1 [canine], XP_023477941.1 [equine], XP_011283721.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes include the receptor for CGRP (RAMP1) and the receptor for ADM (with RAMP2 / 3), and activate adenylate cyclase. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0190] The term "RAMP1 gene" refers to a mammalian gene encoding a receptor activity-modifying protein 1 polypeptide. Non-limiting examples of RAMP1 genes include: NCBI Gene ID: 10267 [human], NCBI Gene ID: 607163 [canine], NCBI Gene ID: 100066550 [equine], NCBI Gene ID: 101092133 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by RAMP1 genes include: UniProt: O60894; NP_005846.1 [human], XP_038291846.1 [canine], XP_023498460.1 [equine], XP_044890618.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by these genes are required for transport of calcitonin receptor-like receptors (CRLRs) to the plasma membrane and function together with the CRLRs as CGRP receptors. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0191] The term "ADM gene" refers to a mammalian gene encoding an adrenomedullin polypeptide. Non-limiting examples of ADM genes include: NCBI Gene ID: 133 [Human], NCBI Gene ID: 403817 [Canine], NCBI Gene ID: 100033857 [Equine], NCBI Gene ID: 101087095 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by ADM genes include: UniProt: P35318; NP_001115.1 [Human], NP_001003183.1 [Canine], NP_001157351.1 [Equine], XP_044894880.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the protein encoded by the gene is a 52-aa peptide with multiple functions, including vasodilation, regulation of hormone secretion, and promotion of angiogenesis. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene from a specific species (h, e, and f for human, dog, horse, and cat, respectively).

[0192] The term "CRCP gene" refers to a mammalian gene encoding a CGRP receptor component polypeptide. Non-limiting examples of CRCP genes include: NCBI Gene ID: 27297 [human], NCBI Gene ID: 479705 [canine], NCBI Gene ID: 100061681 [equine], NCBI Gene ID: 101084503 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by CRCP genes include: UniProt: O75575; NP_001035737.1 [human], XP_038523718.1 [canine], XP_001493592.3 [equine], XP_044903465.1 [feline], and sequence variants thereof, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by this gene is an accessory protein for the CGRP receptor, which regulates CGRP responsiveness in a variety of tissues. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0193] The term "YAP1 gene" refers to a mammalian gene encoding a Yes1-related protein polypeptide. Non-limiting examples of YAP1 genes include: NCBI Gene ID: 10413 [Human], NCBI Gene ID: 479465 [Canine], NCBI Gene ID: 100068834 [Equine], NCBI Gene ID: 101101408 [Feline], and their synonymous and non-synonymous sequence variants. Non-limiting examples of gene products encoded by YAP1 genes include: UniProt: P46937; NP_001123617.1 [Human], XP_038521022.1 [Canine], XP_023500466.1 [Equine], XP_044894121.1 [Feline], and their sequence variants, isoforms encoded by alternative splicing, and various glycoforms. Typically, the protein encoded by the gene is involved in development, growth, repair, and homeostasis. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a particular species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0194] The term "IL1RAP gene" refers to a mammalian gene encoding an interleukin-1 receptor accessory protein polypeptide. Non-limiting examples of ILRAP1 genes include: NCBI Gene ID: 3556 [human], NCBI Gene ID: 488126 [canine], NCBI Gene ID: 100068726 [equine], NCBI Gene ID: 101094125 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL1RAP genes include: UniProt: Q9NPH3; NP_002173.1 [human], XP_038318680.1 [canine], XP_001498597.2 [equine], XP_044893081.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are capable of associating with IL1R1 bound to IL1 to form a high-affinity interleukin-1 receptor complex that mediates interleukin-1-dependent activation of NF-κB and other signaling pathways via TIR-TIR interactions with the cytoplasmic domains of the receptor / co-receptor subunits by recruiting adaptor molecules (such as TOLLIP, MYD88, and IRAK1 or IRAK2). In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e, and f refer to humans, dogs, horses, and cats, respectively).

[0195] The term "IL1R1 gene" refers to a mammalian gene encoding an interleukin-1 receptor type 1 polypeptide. Non-limiting examples of IL1R1 genes include: NCBI Gene ID: 3554 [human], NCBI Gene ID: 481328 [canine], NCBI Gene ID: 100009699 [equine], NCBI Gene ID: 101080705 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by ILR1 genes include: UniProt: P14778; NP_001307909.1 [human], XP_038536135.1 [canine], NP_001075263.2 [equine], XP_023107327.2 [feline], as well as sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are capable of binding to all forms of the proinflammatory cytokine interleukin 1 (IL1 or IL1) to mediate interleukin 1-dependent activation of NF-κB, MAPK and other signaling pathways. This intracellular signaling involves the recruitment of adaptor molecules (such as TOLLIP, MYD88 and IRAK1 or IRAK2) via TIR-TIR interactions with the cytoplasmic domains of receptor / co-receptor subunits. IL1R1 can also bind to interleukin 1 receptor antagonists (IL1Ra or IL1Ra or IL1RN), thereby preventing the formation of a complex with signaling capability and association with IL1RAP. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of specific species (h, c, e and f refer to humans, dogs, horses and cats, respectively).

[0196] The term "IL1A gene" refers to a mammalian gene encoding an interleukin-1α polypeptide. Non-limiting examples of IL1A genes include: NCBI Gene ID: 3552 [Human], NCBI Gene ID: 403782 [Canine], NCBI Gene ID: 100064969 [Equine], NCBI Gene ID: 493944 [Feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL1A genes include: UniProt: P01583; NP_000566.3 [Human], NP_001003157.2 [Canine], NP_001075969.2 [Equine], NP_001009351.1 [Feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are proinflammatory cytokines that signal through interaction with IL1R1 and IL1RAP to activate various pathways, including MAPK, JNK, and NF-κB. In some cases, and solely for the purpose of disambiguation, prefixes are added when referring to proteins or genes of a specific species (h, c, e, and f refer to the human, canine, equine, and feline forms, respectively).

[0197] The term "IL1B gene" refers to a mammalian gene encoding an interleukin-1 beta polypeptide. Non-limiting examples of IL1B genes include: NCBI Gene ID: 3553 [human], NCBI Gene ID: 403974 [canine], NCBI Gene ID: 100034237 [equine], NCBI Gene ID: 768274 [feline], and synonymous and non-synonymous sequence variants thereof. Non-limiting examples of gene products encoded by IL1B genes include: UniProt: P01584; NP_000567.1 [human], NP_001033060.1 [canine], NP_001075995.1 [equine], NP_001070882.1 [feline], and sequence variants, isoforms encoded by alternative splicing, and various glycoforms thereof. Typically, the proteins encoded by the genes listed above are major mediators of inflammatory responses and pyrogens, signaling through interactions with IL1R1 and IL1RAP. In the central nervous system (CNS), IL1B has been shown to contribute to inflammatory pain hypersensitivity and other pathologies. In some cases, and solely for the purpose of disambiguation, a prefix is ​​added when referring to a protein or gene of a specific species (h, c, e, and f for human, canine, equine, and feline, respectively).

[0198] The term "treat" refers to obtaining a desired pharmacological and / or physiological effect. The effect may be preventive, in terms of preventing a disease or its symptoms in whole or in part, and / or therapeutic, in terms of partially or completely curing a disease and / or attributing to adverse effects of the disease. For example, the compositions, methods or systems of the present disclosure may be administered as a preventive treatment to a subject who is prone to a given condition (e.g., arthritis). As used herein, "treat" encompasses any treatment of a disease in a mammal (particularly in humans, canines, felines or equines), and includes: (a) preventing the occurrence of the disease in a subject who may be susceptible to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, i.e., preventing the development or progression of the disease; and (c) alleviating the disease, i.e., causing regression of the disease and / or alleviating one or more symptoms of the disease.

[0199] "Treatment" is also meant to encompass the delivery of an agent to provide a pharmacological effect even in the absence of a disease or condition. For example, "treatment" encompasses the delivery of a composition that can elicit an immune response or confer immunity in the absence of a disease condition, such as in the case of a vaccine. It should be understood that the compositions and methods of the present disclosure are suitable for treating all mammals, including but not limited to human, canine, feline, equine, and bovine subjects.

[0200] The term "therapeutically effective" refers to an amount of a composition or combination of compositions as described herein that is sufficient to achieve the intended application, including but not limited to the treatment of a disease. A therapeutically effective amount can vary depending on the intended application (in vitro or in vivo), or the subject and disease condition being treated (e.g., the subject's weight, age, and sex), the severity of the disease condition, or the mode of administration. The term also applies to a dose that will induce a specific response in the target cell (e.g., reducing platelet adhesion and / or cell migration). The specific dosage will vary depending on the specific composition selected, the dosing regimen followed, whether the composition is administered in combination with other compositions or compounds, the timing of administration, the tissue to be administered, and the physical delivery system in which the composition is carried.

[0201] "Spinal conditions or disorders" include, but are not limited to, low back pain, neck pain, discogenic disease, adolescent idiopathic scoliosis, adult degenerative scoliosis, cervical degenerative disc disease, cervical disc herniation, cervical spondylotic myelopathy, cervical stenosis, compression fractures, degenerative spondylolisthesis, isthmic spondylolisthesis, lumbar sprains and strains, lumbar degenerative disc disease, lumbar disc herniation, lumbar stenosis, neck sprains (whiplash) and strains, neck strain, osteoporosis, and whiplash. Generally, such disorders or conditions result in or cause local nociception, inflammation, or morphological changes (e.g., fibrosis, degeneration, osteolysis, osteogenesis) in the cervical, thoracic, lumbar, or sacral spine or surrounding tissues.

[0202] " low back pain " is defined as measurable or discernible pain or discomfort (chronic or sporadic) in a given subject (comprising at least the lumbar region of a mammal).Pain can manifest as pain (e.g., muscle pain) confined to the waist or as a disseminated pain sensation of hitting, burning, tingling and / or spreading over the back and / or limbs of the whole subject.Pain can be idiopathic, or can be associated with one or more (diagnosed or undiagnosed) potential conditions of illness, including but not limited to degenerative disc disease, chronic inflammation, arthritis, osteoporosis, trauma (e.g., postoperatively), infection (e.g., spondylitis intervertebral disc), neuropathy, musculoskeletal abnormalities (e.g., herniated disc or spinal stenosis or spondylolisthesis), herniated nucleus pulposus (HNP), annular ligament tear, facet joint arthritis, root nerve compression and / or other degenerative conditions.

[0203] "Neck pain" is defined as measurable or discernible pain or discomfort associated with the cervical spine or adjacent ligaments, muscles, and / or tendons. The pain can manifest as localized pain in the neck or as throbbing, tingling, burning, and / or diffuse pain sensations throughout the back or extremities (including, but not limited to, the subject's head, shoulders, arms, legs, and / or back). Neck pain can be idiopathic or associated with one or more underlying conditions (diagnosed or undiagnosed), including, but not limited to, degenerative disc disease, rheumatoid arthritis, osteoporosis, fibromyalgia, chronic inflammation, infection (e.g., disc spondylitis), herniated disc, spondylosis, spinal stenosis, cervical compressive myelopathy, whiplash, and / or other conditions.

[0204] The terms "polynucleotide," "nucleotide," and "nucleic acid" are used interchangeably herein to refer to all forms of nucleic acids, oligonucleotides, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). Polynucleotides include genomic DNA, cDNA, and antisense DNA, as well as spliced ​​or unspliced ​​mRNA, rRNA, tRNA, lncRNA, RNA antagomirs, and inhibitory DNA or RNA (RNAi, e.g., small or short hairpin (sh) RNA, microRNA (miRNA), aptamers, small or short interfering (si) RNA, trans-splicing RNA, or antisense RNA). Polynucleotides also include non-coding RNA, including but not limited to RNAi, miRNA, lncRNA, RNA antagomirs, aptamers, and any other non-coding RNA known to those skilled in the art. Polynucleotides include naturally occurring, synthetic, and intentionally altered or modified polynucleotides, as well as analogs and derivatives. The term "polynucleotide" also refers to a polymeric form of nucleotides of any length, including deoxyribonucleotides or ribonucleotides or analogs thereof, and is synonymous with a nucleic acid sequence. Polynucleotide can comprise modified nucleotide, such as methylated nucleotide and nucleotide analog, and can be interrupted by non-nucleotide component.If exist, then can give the modification of nucleotide structure before or after the assembly of polymer.As used herein, term polynucleotide refers to double-stranded molecule and single-stranded molecule interchangeably.Unless otherwise specified or required, otherwise any embodiment of the polynucleotide as described herein covers double-stranded form and known or predicted to constitute each of two complementary single-stranded forms of double-stranded form.Polynucleotide can be single-stranded, double-stranded or triple-stranded, linear or circular, and can be any length.When discussing polynucleotide, this paper can be based on the conventional description of specific polynucleotide that provides sequence in 5 ' to 3 ' direction or structure.

[0205] The term "gene" or "nucleotide sequence encoding a polypeptide" refers to a segment of DNA involved in producing a polypeptide chain. A DNA segment may include regions preceding and following the coding region (leader and trailer) involved in the transcription / translation and regulation of transcription / translation of a gene product, as well as intervening sequences (introns) between individual coding segments (exons). For example, a gene comprises a polynucleotide containing at least one open reading frame that is capable of encoding a specific protein or polypeptide after being transcribed and translated.

[0206] The terms "extracellular domain" and "ectodomain" are used interchangeably when referring to a transmembrane cellular receptor and are defined as the portion of a protein that is exposed to the extracellular environment and is capable of engaging and / or binding a ligand.

[0207] The terms "cytoplasmic domain" and "intracellular domain" are used interchangeably when referring to transmembrane receptors to define the portion of the protein that is exposed to the cytoplasm. In many cases, these portions of the protein contain signaling domains that recruit and associate with various intracellular factors. Upon engagement of a ligand via the extracellular domain, changes in the interaction occur that can result in new associations, dissociations, or recruitment of various cytoplasmic factors that contribute to signal transduction.

[0208] The term "transmembrane domain" (which may be abbreviated as "TM") refers to a transmembrane receptor and is defined as the portion of a protein that is embedded within the plasma membrane (i.e., not exposed to the extracellular environment or cytosol). The transmembrane domain generally has a more hydrophobic nature than the extracellular or cytosolic portions and generally adopts a higher-order helical structure. Although the primary role of ligand-induced conformational changes in a particular receptor is anchoring, it has been demonstrated that this change can affect the transmembrane domain, allowing it to become part of subsequent intracellular signaling.

[0209] The term "receptor" refers to a protein that is able to bind to another homologous protein (i.e., its ligand) with high affinity. This receptor-ligand interaction can be 1:1, or result in multimerization, in which multiple protein aggregates bind to one or more ligands. Receptors are typically present on the cell surface so that they can most efficiently encounter ligands and initiate intracellular signaling.

[0210] The term "intracellular signaling" refers to changes in cells that occur as a result of events occurring at the cell surface. Typically, a soluble ligand binds to a cell surface receptor, inducing changes in the receptor that affect related intracellular factors. These factors can then influence other factors within the cell, and this cascade continues. In many cases, a specific factor can alter gene expression in the cell nucleus in response to a surface stimulus.

[0211] The term "RNA-guided nuclease" refers to an enzyme that can break the backbone of, for example, a DNA molecule. The activity of the RNA-guided nuclease is guided by a nucleic acid molecule (i.e., a guide RNA). Once the enzyme is correctly oriented to form a functional ribonucleoprotein complex, it is positioned to a specific position within the target nucleic acid (e.g., a gene or locus) by sequence complementarity with a portion of the guide RNA. Non-exhaustive examples of RNA-guided nucleases include Cas9, Cas12, and Cas12a (referred to as Cpf1 above).

[0212] The term "Cas9" refers to a double-stranded DNA binding nuclease protein or a nickase protein guided by RNA, or a variant thereof, and can be used to refer to naturally occurring or recombinant Cas9 nuclease variants (e.g., ES-Cas9, HF-Cas9, PE-Cas9, and AR-Cas9). Wild-type Cas9 nuclease has two functional domains, e.g., RuvC and HNH, which simultaneously cut two chains of double-stranded DNA, thereby causing double-strand breaks. The Cas9 enzyme described herein can include an HNH or HNH-like nuclease domain and / or a RuvC or RuvC-like nuclease domain without affecting the ability to induce double-strand breaks in genomic DNA (e.g., at the target locus) when both functional domains are active. The Cas9 enzyme may comprise one or more catalytic domains of a Cas9 protein derived from a bacterium belonging to the group consisting of Corynebacter, Sutterella, Legionella, Treponema, Filifactor, Eubacterium, Streptococcus, Lactobacillus, Mycoplasma, Bacteroides, In some embodiments, the two catalytic domains are derived from different bacterial species.

[0213] As used herein, "PAM" refers to the protospacer adjacent motif, which is necessary for RNA-guided nucleases to bind to target nucleic acids. In many cases, the PAM is directly adjacent to the complementary sequence in the target. For example, naturally occurring Cas9 molecules can recognize specific PAM sequences (see, for example, Table 1). In some embodiments, the Cas9 molecule has the same PAM specificity as the naturally occurring Cas9 molecule. In other embodiments, the Cas9 molecule has a PAM specificity that is unrelated to the naturally occurring Cas9 molecule. In other embodiments, the PAM specificity of the Cas9 molecule is unrelated to the naturally occurring Cas9 molecule to which it has the closest sequence homology. For example, the naturally occurring Cas9 molecule can be changed so that the PAM sequence recognition is changed to reduce off-target sites, improve specificity, or eliminate PAM recognition requirements. In one embodiment, the Cas9 molecule can be changed (for example, to extend the PAM recognition sequence, improve the Cas9 specificity to a high level of identity, reduce off-target sites, and / or increase specificity). In one embodiment, the length of the PAM recognition sequence is at least 4, 5, 6, 7, 8, 9, 10, or 15 amino acids. In some embodiments, the Cas9 molecule can be changed to ablate PAM recognition.

[0214] The terms "guide RNA," "gRNA," or "sgRNA" are used interchangeably to refer to an RNA molecule, preferably a synthetic RNA molecule, consisting of a targeting (crRNA) sequence and a scaffold. Once these molecules are attached to a functional RNA-guided nuclease, they can guide sequence-specific cleavage of the target nucleic acid.

[0215] sgRNA can be administered or formulated as, for example, synthetic RNA, or a nucleic acid comprising a sequence encoding the gRNA, which is then expressed in the target cell. It will be apparent to one of ordinary skill in the art that various tools can be used to design and / or optimize sgRNAs, for example, to increase the specificity and / or accuracy of genome editing at a specific site.

[0216] In general, candidate sgRNA can be designed and identified by the following steps: first, a suitable PAM is positioned within the genomic sequence. Additional calculations can then be used to predict on-target efficiency and off-target efficiency. Available web-based tools that contribute to the initial design and modeling of candidate sgRNA include, but are not limited to, CRISPRseek, CRISPR DesignTool, Cas-OFFinder, E-CRISP, ChopChop, CasOT, CRISPR direct, CRISPOR, BREAKING-CAS, CrispRGold, and CCTop. See, for example, Safari, F. et al. (2017). Current Pharmaceutical Biotechnology, 18 (13): 1038-54, which is incorporated herein by reference for all purposes. Such tools are also, for example, published in PCT WO2014093701A1 and Liu, G. et al. (2020). Computational approaches for effective CRISPR guide RNA design and evaluation are described in Computational and Structural Biotechnology Journal, 18: 35-44, each of which is incorporated herein by reference in its entirety for all purposes. Candidate sgRNAs can be further evaluated by experimental screening or other methods.

[0217] The term "CRISPR RNA" or "crRNA" refers to the portion of a sgRNA molecule that is complementary to a target nucleic acid.

[0218] The phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0219] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and inert ingredients. The use of such pharmaceutically acceptable carriers or pharmaceutically acceptable excipients for active pharmaceutical ingredients is well known in the art. Except where any conventional pharmaceutically acceptable carrier or pharmaceutically acceptable excipient is incompatible with the active pharmaceutical ingredient, its use in the therapeutic compositions of the present disclosure is contemplated. Additional active pharmaceutical ingredients, such as other pharmaceutical agents, may also be incorporated into the described compositions and methods.

[0220] The term "pharmaceutically acceptable excipient" is intended to include media and carriers that can be co-administered with a compound to facilitate its intended function. The use of such media for pharmaceutically active substances is well known in the art. Examples of such media and carriers include solutions, solvents, dispersion media, delay agents, emulsions, and the like. Any other conventional carrier suitable for use with multi-binding compounds also falls within the scope of this disclosure.

[0221] As used herein, the terms "a," "an," or "the" are generally construed to cover both the singular and the plural.

[0222] The terms "about" and "approximately" mean within a statistically significant range of values. This range can be within the magnitude of a given value or range, preferably within 50%, more preferably within 20%, more preferably still within 10%, and even more preferably within 5%. The permissible variations encompassed by the terms "about" or "approximately" depend on the specific system under study and can be readily understood by those of ordinary skill in the art. In addition, as used herein, the terms "about" and "approximately" mean that the composition, amount, formula, parameter, shape, and other quantities and features are not precise and do not need to be precise, but can be approximate and / or larger or smaller as needed, reflecting tolerances, conversion factors, rounding, measurement errors, etc., as well as other factors known to those skilled in the art. In general, dimensions, sizes, formulas, parameters, shapes, or other quantities or features are "about" or "approximately", regardless of whether they are explicitly stated as such. It should be noted that embodiments of very different sizes, shapes, and dimensions can employ the described arrangements.

[0223] As used herein, the term "substantially" may refer to a majority or majority, such as at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more.

[0224] When used in the appended claims, in both their original and amended forms, the transition terms "comprising," "consisting essentially of," and "consisting of" define the scope of the claims with respect to which unrecited appended claim elements or steps, if any, are excluded from the scope of the claims. The term "comprising" is intended to be inclusive or open-ended and does not exclude any additional, unrecited elements, methods, steps, or materials. The term "consisting of" excludes any elements, steps, or materials other than those specified in the claim and, in the latter case, impurities normally associated with the specified materials. The term "consisting essentially of" limits the scope of the claims to the specified elements, steps, or materials, and those that do not materially affect the basic and novel characteristics of the claimed methods and compositions. In alternative embodiments, all compositions, methods, and kits described herein embodying the present disclosure may be more specifically defined by any of the transition terms "comprising," "consisting essentially of," and "consisting of."

[0225] As used herein, the term "delivery" means providing an entity to a destination. For example, delivering a therapeutic and / or prophylactic agent to a subject can involve administering a nanoparticle composition comprising the therapeutic and / or prophylactic agent to the subject (e.g., via intravenous, intramuscular, intradermal, subcutaneous, intraarticular, or intradiscal routes). Administering a nanoparticle composition to a mammal or mammalian cell can involve contacting one or more cells with the nanoparticle composition.

[0226] As used herein, "naturally occurring" means occurring in nature without human assistance.

[0227] As used herein, "PEG lipid" or "PEGylated lipid" refers to a lipid comprising a polyethylene glycol component. These lipids may also be referred to as PEG-modified lipids.

[0228] As used herein, " phospholipid " is a lipid comprising a phosphate moiety and one or more carbon chains (e.g., unsaturated fatty acid chains). Phospholipid can comprise one or more multiple bonds (e.g., double bonds or triple bonds) (e.g., one or more unsaturated bonds). Specific phospholipid can promote fusion with membrane. For example, cationic phospholipid can interact with one or more negatively charged phospholipids of membrane (e.g., cell membrane or intracellular membrane). The fusion of phospholipid with membrane can allow one or more elements of the composition comprising lipid to pass through the membrane, thereby allowing, for example, one or more elements to be delivered to the cell.

[0229] Any of the compositions disclosed herein can be administered to non-human subjects, such as laboratory or farm animals. Non-limiting examples of non-human subjects include laboratory or research animals, pets, wild or domestic animals, farm animals, etc., for example, dogs, goats, guinea pigs, hamsters, mice, pigs, non-human primates (e.g., gorillas, apes, orangutans, lemurs, baboons, etc.), rats, sheep, horses, cattle, etc. As used herein, a "lipid component" is a component of a nanoparticle composition that includes one or more lipids. For example, a lipid component can include one or more cationic / ionizable, pegylated, structural lipids or other lipids, such as phospholipids.

[0230] method

[0231] CRISPR

[0232] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to the methods and materials described herein can be used to practice or test this disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In addition, materials, methods, and examples are illustrative only and are not intended to be limiting.

[0233] In one aspect, the present disclosure encompasses compositions and related methods, components, and compositions involving clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated RNA-guided nucleases (hereinafter referred to as CRISPR / Cas systems). Such systems require at least one isolated or non-naturally occurring RNA-guided nuclease (e.g., Cas9 protein) and at least one isolated or non-naturally occurring guide RNA (e.g., sgRNA) to achieve amplification of nucleic acid sequences (e.g., genomic DNA).

[0234] In some embodiments, the CRISPR / Cas system effects alteration of a target gene or locus in a eukaryotic cell by altering the sequence at the target location (e.g., by generating insertions or deletions (collectively referred to as indels) that result in loss of function (i.e., knockout) of the affected gene or allele; for example, nucleotide substitutions that result in truncation, nonsense mutations, or other types of loss of function of the encoded product of, for example, (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2),

[0235] (ii) one or more metalloproteinases or their regulators (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more neuronal signaling regulators (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN 8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (v) one or more other cell signaling regulators (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) any combination of genes of (i)-(v), i.e., mRNA or protein; resulting in, for example, one or more encoded products of FGF2, CCN2, ADAMTS5, MMP1, or NGF genes. The deletion of one or more nucleotides that cause truncation, nonsense mutation or other types of loss of function, such as loss of function of an mRNA or protein caused by a single nucleotide, dinucleotide or other frameshift deletion, or a deletion that produces a premature stop codon; or the insertion of a truncation, nonsense mutation or other type of loss of function of an encoding gene product (such as, for example, one or the encoding gene product of FGF2, CCN2, ADAMTS5, MMP1 or NGF gene) (i.e., mRNA or protein); for example, a single nucleotide, dinucleotide or other frameshift insertion, or an insertion that produces a premature stop codon. In some embodiments, the CRISPR / Cas system of the present disclosure provides for the alteration of genes and / or the encoded products of genes such that the altered products have the resulting loss of function and become dominant negative products or decoys (e.g., transmembrane receptors or soluble receptors that cannot trigger intracellular signaling). In some embodiments, the CRISPR / Cas system of the present disclosure is encapsulated in an LNP system described herein, such as, but not limited to, any one of the LNP systems LNP001 to LNP240.

[0236] In one aspect, the CRISPR / Cas system achieves changes in nucleic acid sequences through nuclease activity. For example, in the case of genomic DNA, RNA-guided nucleases locate the target position within the target gene or locus by sequence complementarity with the target genomic sequence (e.g., CRISPR RNA (crRNA) or the complementary component of a synthetic single guide RNA (sgRNA)) and cut the genomic DNA after recognizing a specific nuclease-specific motif (called a protospacer adjacent motif (PAM)). See generally Collias, D., & Beisel, CL (2021). Nature Communications, 12 (1), 1-12.

[0237] In the case of genomic DNA, nuclease activity (i.e., cutting) induces double-strand breaks (DSBs). Due to the interference of the components from the CRISPR / Cas system, the endogenous cellular mechanisms of DSB repair, i.e., non-homologous end joining (NHEJ), microhomology-mediated end joining (MMEJ) and homologous recombination, result in error repair at a given target position with a certain calculable frequency, thereby introducing substitution or insertion and deletion into genomic DNA. Generally referring to Scully, R. et al. (2019). "Nature Reviews Molecular Cell Biology (Nature Reviews Molecular Cell Biology)", 20 (11), 698-714. At a certain frequency, these insertions and deletions and / or replacements may result in frameshifts, nonsense mutations (i.e., early stop codons) or truncation affecting the availability of gene products such as mRNA and / or proteins. In certain embodiments, the CRISPR / Cas system can be provided to cells as part of the system together with nuclease and gRNA using a template (e.g., HDR template) to induce homology-directed repair (HDR) mechanism, resulting in the insertion of non-random sequences at the target position. See Bloh, K., & Rivera-Torres, N. (2021). International Journal of Molecular Sciences, 22(8), 3834.

[0238] In general, the minimum requirements of a CRISPR / Cas system will depend on the nuclease (i.e., Cas protein) provided with it. In this regard, these bacterially derived nucleases have been functionally divided into types I, III, and V (all of which belong to class 1) and types II, IV, and VI (which are grouped into class 2).

[0239] Type 1 CRISPR / Cas system:

[0240] The exact components, compositions and methods for using Class 1 CRISPR / Cas systems to achieve changes in the targeted nucleic acid sequence will vary, but should minimally include: a nuclease (selected from at least Type I and Type III), at least one guide RNA selected from a combination of 1) sgRNA or 2) crRNA and tracrRNA. These CRISPR / Cas systems are collectively classified as Class 1 CRISPR / Cas systems due to their similarities in requirements and modes of action within eukaryotic cells. For this reason, the compositions, components and methods in Class 1 components can be considered functionally interchangeable, and the following details provided for exemplary purposes only do not represent an exhaustive list of class members. In some embodiments, the CRISPR / Cas system of the present disclosure is encapsulated in an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0241] Cas3 (see Table 1) is a prototype type I DNA nuclease that is used as an effector protein as part of a larger complex (a Cascade complex comprising Cse1 and Cse2) capable of genome editing. See generally He, L. et al. (2020). Genes, 11(2), 208. Unlike other CRISPR / Cas systems, type I systems are located on DNA targets without Cas3 nucleases via the Cascade complex, which then recruits Cas3 to cut DNA when it binds and locates 3' PAM. The Cascade complex is also responsible for processing crRNA so that it can be used to guide it to the target position. Due to this function, the Cascade has the ability to process multiple array crRNAs from a single molecule. See Luo, M. (2015), Nucleic Acids Research, 43(1), 674-681. Therefore, type I systems can be used to edit multiple targeted genes or loci from a single molecule.

[0242] Because the natural Cas3 substrate is ssDNA, its function in genome editing is thought to be a nicking enzyme; however, when targeted in tandem, the resulting edit is a blunt-end cleavage of the opposite strand to approximate the results of a blunt-cutting nuclease such as Cas9. See Pickar-Oliver, A., & Gersbach, CA (2019). Nature Reviews Mol Cell Biology, 20(8), 490-507.

[0243] Like type I nucleases, type III systems rely on protein complexes to achieve nucleic acid cutting. In particular, Cas10 has the nuclease activity of cutting ssDNA in prokaryotes. See Tamulaitis, G. Trends in Microbiology, 25 (1), 49-61 (2017). Interestingly, the CRISPR / Cas system (natural to archaea) shows dual specificity and targets both ssDNA and ssRNA. In addition to this change, the function of the system is very similar to type I, because crRNA targets the effector complex in a sequence-dependent manner (similar to cascade). Similarly, the effector complex processes crRNA before associating. The dual nature of this nuclease makes its application in genome editing potentially more powerful, because genomic DNA and, in some cases, mRNA with the same sequence can be targeted to silence specific target genes.

[0244] Type 2 CRISPR / Cas systems:

[0245] The exact components, compositions and methods for using Class 2 CRISPR / Cas systems to achieve changes in the targeted nucleic acid sequence will vary, but should minimally include: a nuclease (selected from at least Type II and Type V), at least one guide RNA selected from a combination of 1) sgRNA or 2) crRNA and tracrRNA. These CRISPR / Cas systems are collectively classified as Class 2 CRISPR / Cas systems due to their similarities in requirements and modes of action within eukaryotic cells. For this reason, the compositions, components and methods in Class 2 components can be considered functionally interchangeable, and the following details, provided for exemplary purposes only, do not represent an exhaustive list of class members:

[0246] Type II nucleases are the best characterized CRISPR / Cas systems, specifically the prototypical genome editing nuclease, Cas9 (see Table 1). A variety of Cas9 proteins have been isolated from various bacterial species. The primary difference between these nucleases is the PAM, a recognition site within the dsDNA required for targeting. Upon association with the gRNA molecule, the crRNA (or targeting domain of the sgRNA) directs the nuclease to the appropriate location, but recognition of the PAM by the protein is responsible for inducing cleavage events near this site, resulting in blunt DSBs.

[0247] In addition to naturally derived Cas9 proteins, several engineered variants have also been reported. These range from Cas9 with enhanced specificity (i.e., lower off-target activity), such as espCas9. Others have been catalytically modified via point mutations in RuvC (e.g., D10A) and HNH (e.g., H840A) domains, such that they only induce single-strand breaks (i.e., Cas9 nickases). See Frock, R. et al. (2015). Nature Biotechnology, 33(2), 179-186. These variants have also been shown to be less prone to error when editing. When the desired insertion (i.e., knock-in mutation, wherein the desired nucleotide sequence is introduced into the target nucleic acid molecule) is selected rather than deleted, this type of mitigation of off-target effects becomes crucial. In fact, fewer off-target effects can contribute to a preferred DNA repair mechanism (HDR, in most cases for knock-in mutations). See generally Naeem, M. et al. (2020). Cells, 9(7), 1608.

[0248] Other exemplary additional engineered variants (e.g., mutants, chimeras) of typical Cas proteins, and include the following patents (each of which is hereby incorporated by reference in its entirety for all purposes): WO2015035162A2, WO2019126716A1, WO2019126774A1, WO2014093694A1, WO2014150624A1, US20190225955A1, U.S. Patent 11427818, U.S. Patent 11242542, U.S. Patent 11098297, U.S. Patent 10876100, U.S. Patent 10767193, U.S. Patent 10494621 and U.S. Patent 10100291.

[0249] For the avoidance of doubt, SpCas9 is collectively referred to as any one of the group consisting of: espCas9 (also referred to herein as ES-Cas9 or esCas9), HF-Cas9, PE-Cas9, ARCas9 (also referred to as AR-Cas9), SpCas9-D1135E, SpCas9-HF1, HypaCas9, HiFiCas9, xCas9-3.6, xCas9-3.7, Sniper-Cas9, evoCas9, SpartaCas, LZ3Cas9, miCas9, and SuperFi-Cas9. Additional examples of Cas9 variants disclosed below are hereby incorporated by reference in their entirety for all purposes: Huang, X. et al. (2022). Cells, 11(14), 2186.

[0250] Like the typical Cas9 system, type V nucleases only require synthetic sgRNAs with targeting domains complementary to the genomic sequence for genome editing. These nucleases contain the RuvC domain but lack the HNH domain of type II nucleases. In addition, for example, Cas12 leaves staggered cuts in the dsDNA substrate distal to the PAM, compared to the blunt cut of Cas9 next to the PAM. Both Cas12a (also known as Cpf1) and Cas12b (also known as C2c1) (see Table 1) act as part of a larger complex of two gRNA-associated nucleases that act on dsDNA in a quaternary structure, cutting each strand simultaneously. See Zetsche, B. et al. (2015). Cell, 163(3):759-771; see also Liu, L. et al. (2017). Molecular Cell, 65(2):310-322. In addition, Cas12b (C2c1) is a highly accurate nuclease with little tolerance for mismatches. See Yang, H. et al. (2016). Cell, 167(7): 1814-1828.e12.

[0251] Table 1. Exemplary list of Cas nucleases and their requirements

[0252]

[0253] See generally Wang, J., Zhang, C., & Feng, B. (2020). Journal of Cellular and Molecular Medicine, 24(6), 3256-3270, where N = any nucleotide; R = any purine (A or G); Y = any pyrimidine (C or T); W = A or T; V = A, C or G.

[0254] In one aspect, the CRISPR / Cas system of the present disclosure comprises at least one RNA-guided nuclease (e.g., Cas protein) derived from one or more of the following selected bacterial genera: Corynebacterium, Sutterella, Legionella, Treponema, Filamentosa, Eubacterium, Streptococcus, Lactobacillus, Mycoplasma, Bacteroides, Flavobacterium, Spirochaete, Azospirillum, Gluconoacetobacter, Neisseria, Rose, Parvibacter, Nitrobacter, Campylobacter, Pseudomonas, Streptomyces, Staphylococcus, Francisella, Acidobacterium, Lachnospira, Leptotrichia, and Prevotella. In some embodiments, the Cas protein is derived from a δ-Proteobacteria or Planctomycetes bacterial species.

[0255] Some aspects of the present disclosure provide strategies, methods, compositions and treatments for changing the target sequence in the locus (e.g., changing the sequence of the wild type and / or mutant sequence in a cell or mammal) by inserting or deleting one or more nucleotides mediated by an RNA-guided nuclease and one or more guide RNAs (gRNAs) so that the target gene product undergoes a loss of function. In some embodiments, loss of function "knocks out" the gene of interest (i.e., produces a "knockout") by ablating gene expression. In some embodiments, loss of function produces a non-functional gene product (i.e., a gene product that does not have all the functions of a wild-type gene product). In some embodiments, loss of function triggers the expression of a gene product with different characteristics (e.g., different binding affinities or different cellular localizations).

[0256] In certain embodiments, the targeted gene is selected from (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or their regulatory factors (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL 18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more neuronal signaling regulators (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (v) one or more other cell signaling regulators (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) a combination of any of the genes of (i)-(v). In some embodiments, any region of the target gene (e.g., promoter region, 5' untranslated region, 3' untranslated region, exon, intron, or exon / intron boundary) can be targeted by RNA-guided nucleases to alter the gene. In some embodiments, non-coding regions of the target gene (e.g., enhancer region, promoter region, intron, 5'UTR, 3'UTR, polyadenylation signal) are targeted to alter the gene.

[0257] CRISPR guide RNA:

[0258] In one aspect, the CRISPR / Cas system of the present disclosure also provides a gRNA molecule (for example, an isolated or non-naturally occurring RNA molecule) interacting with the nuclease guided by RNA. In certain embodiments, gRNA is an sgRNA comprising a crRNA sequence, and the crRNA sequence comprises a nucleotide sequence complementary to the sequence in the target nucleic acid. In some embodiments, sgRNA also includes an RNA scaffold portion (i.e., tracrRNA) interacting with the nuclease guided by RNA, so that crRNA is positioned to scan the target nucleic acid for complementarity. In some embodiments, the system is further optionally composed of oligonucleotides, which are HDR templates with homology to either side of the target position. See Bloh, K., & Rivera-Torres, N. (2021). International Journal of Molecular Sciences, 22 (8): 3834.

[0259] In one embodiment, the RNA-guided nuclease and sgRNA are configured to direct related nucleases so that a cleavage event (e.g., a double-strand break or a single-strand break) occurs at a position sufficiently close to the complementary sequence in the target nucleic acid, thereby promoting changes in the nucleic acid sequence. In some embodiments, the crRNA is 20 nucleotides in length. In some embodiments, the crRNA is 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

[0260] In some embodiments, the crRNA directs the RNA-guided nuclease so that the cleavage event occurs within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of the complementary sequence in the targeting nucleic acid. Double-strand breaks or single-strand breaks can be located upstream or downstream of the complementary sequence in the targeting nucleic acid. In some embodiments, the cleavage event occurs within the target gene. In some embodiments, the cleavage event occurs upstream of the target gene.

[0261] In certain embodiments, the second gRNA molecule comprising the second crRNA directs the nuclease guided by the second RNA so that the cleavage event occurs at a position close enough to the complementary sequence in the target nucleic acid, thereby promoting the change of the nucleic acid sequence. In some embodiments, the first gRNA and the second gRNA promote the cleavage event in a single target gene. In some embodiments, the first gRNA and the second gRNA promote the cleavage event in different target genes. In some embodiments, the second crRNA has a length of 20 nucleotides. In some embodiments, the second crRNA has a length of 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides.

[0262] In some embodiments, the second crRNA directs the RNA-guided nuclease so that the cleavage event occurs within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175 or 200 nucleotides of the complementary sequence in the targeting nucleic acid. Double-strand breaks or single-strand breaks can be located upstream or downstream of the complementary sequence in the targeting nucleic acid. In some embodiments, the cleavage event occurs in the target gene. In some embodiments, the cleavage event occurs upstream of the target gene.

[0263] In some embodiments, the targeting domains of the first gRNA and the second gRNA are configured such that the cleavage event is independently located within 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 nucleotides of the other cleavage event for each gRNA molecule. In some embodiments, the first gRNA molecule and the second gRNA molecule change the target nucleic acid sequence simultaneously. In some embodiments, the first gRNA molecule and the second gRNA molecule change the target nucleic acid sequence sequentially.

[0264] In some embodiments, single-strand break is accompanied by the second single-strand break, which is positioned by the crRNA of the first gRNA and the second gRNA respectively.For example, crRNA can direct the nuclease guided by the relevant RNA so that cutting events (for example, two single-strand breaks) are positioned in 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175 or 200 nucleotides of each other.In some embodiments, the first crRNA and the second crRNA are configured to direct the nuclease guided by the relevant RNA so that, for example, two single-strand breaks occur in the same position on the relative chain of genomic DNA, or in 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50 nucleotides of each other, thereby substantially approximating to double-strand break.

[0265] In some embodiments, the nucleic acid encodes a second sgRNA molecule. In some embodiments, the nucleic acid encodes a third sgRNA molecule. In some embodiments, the nucleic acid encodes a fourth sgRNA molecule.

[0266] In certain embodiments, the nucleic acid may comprise (a) a sequence encoding a first sgRNA comprising a crRNA complementary to a sequence in a target gene, (b) a sequence encoding a second sgRNA comprising a crRNA complementary to a sequence in a second target gene, and (c) a sequence encoding an RNA-guided nuclease (e.g., Cas9). Optionally, (d) and (e) are sequences encoding a third sgRNA and a fourth sgRNA, respectively. In some embodiments, the second target gene is the same as the first target gene. In other embodiments, the second target gene is different from the first target gene. In some embodiments, (a), (b), and (c) are encoded within the same nucleic acid molecule (e.g., the same vector). In some embodiments, (a) and (b) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), and (d) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), (d), and (e) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), (d), and (e) are encoded within the same nucleic acid molecule. In some embodiments, (a), (b), and (c) are encoded within separate nucleic acid molecules. When more than two sgRNAs are used, any combination of (a), (b), (c), (d), and (e) can be encoded within a single or separate nucleic acid molecule.

[0267] In one aspect, nucleic acid molecules (i.e., those encoding (a), (b), (c), (d), or (e)) are delivered to target cells (i.e., any combination of the RNA-guided nuclease encoded by (c) and at least one gRNA molecule encoded by (a), (b), (d), or (e) contacts the target cell). In some embodiments, the nucleic acid molecules are delivered to the target cell in vivo. In other embodiments, the nucleic acid molecules are delivered to the target cell ex vivo. In some embodiments, the nucleic acid molecules are delivered to the target cell in vitro. In certain embodiments, the nucleic acid molecules are delivered to the target cell as DNA. In other embodiments, the nucleic acid molecules are delivered to the target cell as RNA (e.g., mRNA). In some embodiments, the products of the nucleic acid molecules are delivered as assembled ribonucleoproteins (RNPs).

[0268] In some embodiments, contacting the target cell comprises delivering the RNA-guided nuclease of (c) as a protein having at least one of the nucleic acid molecules selected from (a), (b), (d), and (e). In some embodiments, contacting the target cell comprises delivering the encoded RNA-guided nuclease of (c) as a DNA having at least one of the nucleic acid molecules selected from (a), (b), (d), and (e). In some embodiments, contacting the target cell comprises delivering the encoded RNA-guided nuclease of (c) as an mRNA having at least one of the nucleic acid molecules selected from (a), (b), (d), and (e).

[0269] In certain embodiments, CRISPR components are delivered to target cells via nanoparticles. Exemplary nanoparticles that can be used with all CRISPR / Cas systems disclosed herein include at least lipid nanoparticles or liposomes, hydrogel nanoparticles, metal organic nanoparticles, gold nanoparticles, magnetic nanoparticles, and virus-like particles. See generally Xu, CF et al. (2021). Advanced Drug Delivery Reviews, 168: 3-29. In some embodiments, the CRISPR components of the present disclosure are encapsulated in LNP systems as described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0270] TALEN

[0271] In one aspect, the present disclosure contemplates the use of methods, components, and compositions involving transcription activator-like effector nucleases (TALENs) to achieve enhancement of nucleic acid sequences (eg, target genes).

[0272] TALE stands for "transcription activator-like effector" protein, which includes TALEN ("transcription activator-like effector nuclease"). The method of gene editing using the TALE system may also be referred to as the TALE method in this article. TALE is a naturally occurring protein from the plant pathogenic bacteria genus Xanthomonas and contains a DNA binding domain consisting of a series of 33-35 amino acid repeat domains that each recognizes a single base pair. TALE specificity is determined by two hypervariable amino acids called repeat variable diresidues (RVDs). Modular TALE repeats are linked together to recognize adjacent DNA sequences. Specific RVDs in the DNA binding domain recognize bases in the target locus, providing structural features for assembling predictable DNA binding domains. The DNA binding domain of TALE is fused to the catalytic domain of the type IIS FokI endonuclease to form a targetable TALE nuclease. In order to induce site-specific mutations, two separate TALEN arms separated by 14-20 base pair spacers bring the FokI monomers into close proximity to dimerize and produce targeted double-strand breaks.

[0273] Several large-scale systematic studies using various assembly methods have shown that TALE repeats can be combined to recognize almost any user-defined sequence. Custom-designed TALE arrays can also be commercially available from Cellectis Bioresearch (Paris, France), Transposagen Biopharmaceuticals (Lexington, KY, USA), and Life Technologies (Grand Island, NY, USA). TALE and TALEN methods suitable for use in the present disclosure are described in U.S. Patent Application Publication Nos. US 2011 / 0201118 A1; US ​​2013 / 0117869 A1; US ​​2013 / 0315884 A1; US ​​2015 / 0203871 A1, and US 2016 / 0120906 A1, which are incorporated herein by reference.

[0274] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing by the TALE approach include (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or their regulators (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, I , TACR1, TACR2, TACR3, ATP1A1), (v) one or more other cellular signaling regulators (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) a combination of any of the genes listed in (i)-(v). In one aspect, the present disclosure provides compositions for upregulating protein receptors (including wild-type or gene-edited), including compositions bound to anti-inflammatory cytokines via a TALE approach.

[0275] Examples of systems, methods, and compositions for altering the expression of target gene sequences via the TALE approach and that can be used according to embodiments of the present disclosure are described in U.S. Patent No. 8,586,526, which is incorporated herein by reference.

[0276] Zinc finger nucleases (ZFNs)

[0277] In one aspect, the present disclosure contemplates the use of methods, components, and compositions involving zinc finger nucleases (ZFNs) to achieve enhancement of a nucleic acid sequence (eg, a target gene).

[0278] Each zinc finger contains approximately 30 amino acids in a conserved ββα configuration. Several amino acids on the surface of the α-helix typically contact 3 bp in the major groove of DNA, with varying levels of selectivity. Zinc fingers have two protein domains. The first domain is the DNA-binding domain, which contains eukaryotic transcription factors and contains the zinc finger. The second domain is the nuclease domain, which contains the FokI restriction enzyme and is responsible for catalytic DNA cleavage.

[0279] The DNA binding domain of a single ZFN typically contains three to six separate zinc finger repeats, and each zinc finger repeat can recognize 9 to 18 base pairs. If the zinc finger domain is specific for its intended target site, then in theory even a pair of 3-finger ZFNs that recognize a total of 18 base pairs can target a single locus in the mammalian genome. One way to generate new zinc finger arrays is to combine smaller zinc finger "modules" of known specificity. The most common modular assembly process involves combining three separate zinc fingers, each of which can recognize a 3 base pair DNA sequence, to generate a 3-finger array that can recognize a 9 base pair target site. Alternatively, selection-based methods such as oligomer pool engineering (OPEN) can be used to select new zinc finger arrays from a random library that takes into account context-dependent interactions between adjacent fingers. Engineered zinc fingers are commercially available; Sangamo Biosciences (Richmond, CA, USA) has collaborated with Sigma-Aldrich (St. Louis, MO, USA) to develop a proprietary platform for zinc finger construction.

[0280] Non-limiting examples of genes that can be silenced or inhibited by permanent gene editing by zinc finger methods include (i) one or more growth factors or growth factor receptors (e.g., FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2), (ii) one or more metalloproteinases or their regulators (e.g., ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3), (iii) one or more cytokines, chemokines, or cytokine / chemokine receptors (e.g., CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL2 13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP), (iv) one or more neuronal signaling regulators (e.g., SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1), (v) one or more other cell signaling regulators (e.g., CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2), and (vi) a combination of any of the genes of (i)-(v). Non-limiting examples of genes that can be enhanced by permanent gene editing by zinc finger methods (so that their resulting products serve as decoys or dominant negative products) include. In one aspect, the present disclosure provides compositions for upregulating protein receptors (including wild-type or gene-edited), including compositions that bind to anti-inflammatory cytokines by zinc finger methods.

[0281] Examples of systems, methods, and compositions for altering the expression of target gene sequences by zinc finger approaches that can be used in accordance with embodiments of the present disclosure are described in U.S. Patent Nos. 6,534,261;

[0282] Other examples of systems, methods, and compositions for altering expression of target gene sequences through zinc finger approaches that can be used in accordance with embodiments of the present disclosure are described in Beane et al., Mol. Therapy, 2015, 23 1380-1390, the disclosure of which is incorporated herein by reference.

[0283] Joint diseases or conditions

[0284] introduction

[0285] As described herein, embodiments of the present disclosure provide compositions and methods for improving joint function and treating joint diseases. In specific embodiments, there is provided gene editing of synovial fibroblasts, synoviocytes, chondrocytes, tissue (resident) macrophages or other cells to reduce the compositions and methods of pro-inflammatory signaling mediated by inflammatory cytokines (including but not limited to IL1α, IL1β, TNFα, IL6, IL8, IL18, IL33, matrix metalloproteinases (MMP), TGFβ1, TGFβ2 and combinations thereof) in conjunction with their cognate receptors. Some embodiments can be used to treat various forms of arthritis and other inflammatory joint diseases. Some embodiments can also be used to treat dog lameness caused by osteoarthritis. Some embodiments can also be used to treat horse lameness caused by joint disease. Some embodiments can also be used to treat cat lameness caused by joint disease. Some embodiments can also be used to treat post-traumatic arthritis, gout, pseudogout, psoriatic arthritis and other inflammatory-mediated or immune-mediated joint diseases. Some embodiments are further useful as they relate to packaging in an LNP system described herein, such as, but not limited to, any of LNP systems LNP001 through LNP240.

[0286] The treatment of osteoarthritis, degenerative joint disease, and other joint dysfunctions is complex, and there are few therapeutic options for long-term symptom relief or restoration of joint function. Osteoarthritis (OA) is a leading cause of disability due to pain. See Neogi, T. (2013). Osteoarthritis Cartilage, 21(9): 1145-53. OA and similar diseases affect all mammalian species, including draft animals, domestic pets, and their owners. A common mechanism of joint disease is the presence of acute or chronic inflammation, which is driven by elevated levels of proinflammatory cytokine signaling. Joint disease tends to be progressive, with discomfort, pain, and (especially in OA) disability, depending on the extent of disease progression.

[0287] Psoriatic arthritis (PsA) is another chronic inflammatory joint disease in which joint symptoms are accompanied by skin lesions, such as those commonly associated with psoriasis. See Boehncke, W. et al. (2014). British Journal of Dermatology, 170(4):772-786. As with other forms of arthritis (such as OA), PsA is caused by proinflammatory signaling of multiple cytokines, including IL1. In fact, the pathogenesis of PsA has been shown to be associated with single nucleotide polymorphisms (SNPs) that affect the activity of the IL1 gene locus. See Rahman, P. et al. (2006). Arthritis and Rheumatism, 54(7):2321–2325. These studies have also generally been associated with inflammatory cytokine signaling, more specifically IL1, in disease progression.

[0288] Gout is a chronic inflammatory condition that affects the joints. The underlying cause is the deposition of monosodium urate (MSU) crystals and the resulting host response, particularly in joint structures (as well as in subcutaneous tissue and other sites). See Dalbeth, N. and Stamp, L. (2014). Annals of the Rheumatic Diseases, 73(9): 1598-1600. Clinical manifestations include recurrent acute attacks of severe inflammatory arthritis and tendon bursitis. IL1 and other proinflammatory mediators are major contributors to this host response. See Dinarello, CA (2014). Molecular Medicine, 20(1): S43-S58. For this reason, effective blockade of these signaling pathways could provide relief for patients with gout.

[0289] The current standard of care for many patients with joint diseases includes anti-inflammatory drugs (e.g., NSAIDs) or anti-rheumatic drugs (e.g., methotrexate [an inhibitor of AICAR] or adalimumab [an anti-TNFα monoclonal antibody]). See Friedman, B. and Cronstein, B. (2019). Joint Bone Spine, 86(3): 301–307. All of these treatments require repeated dosing to maintain their effects, which may lead to toxicity or tolerance over time. Therefore, new methods and compositions for treating joint diseases and conditions are urgently needed to treat these chronic conditions.

[0290] In one aspect, the compositions and methods described herein are intended to treat a joint disease or condition in a mammal in need thereof. In some embodiments, the joint disease or condition is osteoarthritis. In some embodiments, the joint disease or condition is psoriatic arthritis. In some embodiments, the joint disease or condition is gout.

[0291] The advantages of the present disclosure over currently available treatments for mammals suffering from one or more joint diseases or conditions include a period of symptom remission. After genetic editing of cells within the joint, pro-inflammatory signaling can be silenced by the target gene for the entire lifespan of the cell and any mitotic progeny. In contrast, biological treatments require regular administration, which may exacerbate the effects of a variety of potentially serious side effects. Among various genetic approaches, the present disclosure also has advantages, one of which is that it combats leakage by modifying protein receptors rather than eliminating the expression of ligands, which may lead to compensatory effects (e.g., the accumulation of other factors due to the lack of negative feedback).

[0292] In some embodiments, the present disclosure includes a method for treating or preventing a joint disease or condition in a subject in need thereof, the method comprising administering a pharmaceutical composition to the subject's joint, the pharmaceutical composition comprising a therapeutically effective amount of a clustered regularly interspaced short palindromic repeat (CRISPR) gene editing system comprising (i) a CRISPR-associated (Cas) protein; and (ii) at least one guide RNA targeting the IL1R1 gene, IL1RAP gene, TGFBR1 gene, TGFBR2 gene, IL6R gene, IL6ST gene, TNFRSF1A gene, TNFRSF1B gene, TNFRSF3 gene, TNFRSF4 gene, or TNFRSF11A gene, or a combination thereof. In some embodiments, the joint disease or condition is osteoarthritis. In some embodiments, the joint disease or condition is psoriatic arthritis. In some embodiments, the joint disease or condition is gout.

[0293] In some embodiments, the present disclosure includes methods for treating or preventing arthritis. Non-limiting examples of arthritis that can be treated using the compositions and methods described herein include post-traumatic arthritis, osteoarthritis (a degenerative condition that affects joints, most commonly the hips, knees, and hands), rheumatoid arthritis (an autoimmune disease that causes inflammation of the joints and surrounding tissues), psoriatic arthritis (a type of arthritis that occurs in patients with psoriasis, a skin disease characterized by scaly, red patches), gout (a type of arthritis caused by a buildup of uric acid crystals in the joints), lupus (a chronic autoimmune disease that can cause inflammation and damage to the joints and other organs), ankylosing spondylitis (an arthritis that primarily affects the spine, causing inflammation and stiffness), reactive arthritis (an arthritis that occurs as a response to an infection in the body), septic arthritis (an arthritis caused by a joint infection), juvenile idiopathic arthritis (an arthritis that affects children under 16 years of age), and fibromyalgia (a chronic pain condition that can cause widespread pain and stiffness, including in the joints).

[0294] In some embodiments, the present disclosure includes methods for treating or preventing pseudogout, crystal arthropathy (caused by crystal formation in the joints, such as gout and pseudogout), or CPPD disease (calcium pyrophosphate deposition disease, also known as chondrocalcinosis).

[0295] In some embodiments, the present disclosure includes methods for treating or preventing rheumatoid arthritis, psoriasis, asthma, inflammatory bowel disease, multiple sclerosis, Alzheimer's disease, type 2 diabetes, cardiovascular disease, or cancer. In some embodiments, these conditions are treated by administering a CRISPR composition targeting an IL1 receptor (e.g., IL1R1 or IL1RAP) as described herein.

[0296] osteoarthritis

[0297] In one aspect, the present disclosure encompasses the treatment of osteoarthritis (OA). In some embodiments, OA treatment includes a therapeutically effective amount of a clustered regularly interspaced short palindromic repeat (CRISPR) gene editing system comprising: (i) CRISPR-associated (Cas) protein; and (ii) at least one guide RNA targeting IL1R1. In some embodiments, OA treatment includes a CRISPR gene editing system targeting hIL1R1. In some embodiments, OA treatment includes a CRISPR gene editing system targeting cIL1R1. In some embodiments, OA treatment includes a CRISP gene editing system targeting eIL1R1 R. In some embodiments, OA treatment includes a CRISPR gene editing system targeting fIL1R1. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of IL1R1 for treating OA is delivered by one or more lipid nanoparticles (LNPs), and the LNP includes an LNP system as described herein, such as, but not limited to, any one of the LNP systems LNP001 to LNP240.

[0298] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 8 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 9 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 10 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 11 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 12 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 13 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 14 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of hIL1R1.In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 19 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 20 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 21 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hIL1R1is is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0299] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 8 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 9 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 10 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 11 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 12 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 13 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 14 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of cIL1R1.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of cIL1R1is for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0300] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 8 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 9 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 10 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 11 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 12 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 13 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 14 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eIL1R1 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0301] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 8 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 9 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 10 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 11 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 12 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 13 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 14 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of fIL1R1.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of fIL1R1 for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0302] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of IL1RAP is delivered by one or more lipid nanoparticles (LNPs), the LNP comprising an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0303] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of hIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of hIL1RAP.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of hIL1RAP for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0304] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of cIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of cIL1RAP.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of cIL1RAP for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0305] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of eIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of eIL1RAP. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of eIL1RAP for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0306] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of fIL1RAP. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of fIL1RAP. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of fIL1RAP for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0307] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TGFBR1 is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of LNP systems LNP001 to LNP240.

[0308] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hTGFBR1is is delivered by one or more lipid nanoparticles (LNPs), wherein the LNP comprises an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0309] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTGFBR1 is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0310] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of eTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTGFBR1 is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0311] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of fTGFBR1. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTGFBR1 is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0312] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TGFBR2 is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of LNP systems LNP001 to LNP240.

[0313] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hTGFBR2 is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0314] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTGFBR2 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0315] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTGFBR2 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0316] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTGFBR2. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTGFBR2 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0317] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL6R. In some embodiments, the CRISPR gene editing system comprising one or more sgRNAs targeting exons of IL6R for treating OA is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of LNP systems LNP001 to LNP240.

[0318] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL6R. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of hIL6R for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0319] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of cIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cIL6R is delivered by one or more lipid nanoparticles (LNPs), wherein the LNP comprises an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0320] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eIL6R is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0321] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fIL6R. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fIL6R is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0322] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting IL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fIL6ST. In some embodiments, the CRISPR gene editing system comprising one or more sgRNAs targeting exons of IL6ST for treating OA is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of the LNP systems LNP001 to LNP240.

[0323] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of hIL6ST.In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 19 of hIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hIL6ST is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0324] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of cIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of cIL6ST.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of cIL6ST for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0325] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of eIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of eIL6ST.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting an exon of eIL6ST for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0326] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 13 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 14 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 15 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 16 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 17 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 18 of fIL6ST.In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 19 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 20 of fIL6ST. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fIL6ST is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0327] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TNFRSF1A is delivered by one or more lipid nanoparticles (LNPs), the LNP comprising an LNP system described herein, for example, but not limited to any one of LNP systems LNP001 to LNP240.

[0328] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hTNFRSF1A is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0329] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTNFRSF1A is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0330] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTNFRSF1A is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0331] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF1A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTNFRSF1A is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0332] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TNFRSF1B is delivered by one or more lipid nanoparticles (LNPs), wherein the LNP comprises an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0333] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 8 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 9 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 10 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 11 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 12 of hTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 13 of hTNFRSF1B. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of hTNFRSF1B for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0334] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTNFRSF1B is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0335] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of eTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTNFRSF1B is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0336] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF1B. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTNFRSF1B is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0337] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF3. In some embodiments, the CRISPR gene editing system comprising one or more sgRNAs targeting exons of TNFRSF3 for treating OA is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of the LNP systems LNP001 to LNP240.

[0338] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hTNFRSF3 is delivered by one or more lipid nanoparticles (LNPs), wherein the LNP comprises an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0339] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTNFRSF3 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0340] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTNFRSF3 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0341] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF3. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTNFRSF3 is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0342] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TNFRSF4 is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of LNP systems LNP001 to LNP240.

[0343] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of hTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of hTNFRSF4. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of hTNFRSF4 for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0344] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF4. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of cTNFRSF4 for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0345] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTNFRSF4. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of eTNFRSF4 for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0346] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF4. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF4. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of fTNFRSF4 for treating OA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0347] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting TNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of TNFRSF11A is delivered by one or more lipid nanoparticles (LNPs), and the LNP comprises an LNP system described herein, for example, but not limited to any one of the LNP systems LNP001 to LNP240.

[0348] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 12 of hTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of hTNFRSF11A is delivered by one or more lipid nanoparticles (LNPs), the LNP comprising an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0349] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of cTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of cTNFRSF11A is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0350] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 1 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 2 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 3 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 4 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 5 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 6 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs that target exon 7 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of eTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of eTNFRSF11A is delivered by one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0351] In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 1 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 2 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 3 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 4 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 5 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 6 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 7 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 8 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 9 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 10 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprises one or more sgRNAs targeting exon 11 of fTNFRSF11A. In some embodiments, the CRISPR gene editing system for treating OA comprising one or more sgRNAs targeting exons of fTNFRSF11A is delivered by one or more lipid nanoparticles (LNPs), comprising an LNP system described herein, for example, but not limited to, any one of LNP systems LNP001 to LNP240.

[0352] Psoriatic arthritis

[0353] In one aspect, the present disclosure covers the treatment of psoriatic arthritis (PsA). In some embodiments, psoriatic arthritis treatment includes a therapeutically effective amount of a clustered regularly interspaced short palindromic repeats (CRISPR) gene editing system comprising: (i) CRISPR-associated (Cas) protein; and (ii) at least one guide RNA targeting IL1R1. In some embodiments, psoriatic arthritis treatment includes a CRISPR gene editing system targeting hIL1R1. In some embodiments, psoriatic arthritis treatment includes a CRISPR gene editing system targeting cIL1R1. In some embodiments, psoriatic arthritis treatment includes a CRISP gene editing system targeting eIL1R1. In some embodiments, psoriatic arthritis treatment includes a CRISPR gene editing system targeting fIL1R1. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of IL1R1 for treating PsA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0354] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of hIL1R1.In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs that target exon 17 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs that target exon 18 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs that target exon 19 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs that target exon 20 of hIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs that target exon 21 of hIL1R1. In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of hIL1R1 for treating PsA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0355] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of cIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of cIL1R1.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of cIL1R1 for treating PsA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0356] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of eIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of eIL1R1.In some embodiments, a CRISPR gene editing system comprising one or more sgRNAs targeting exons of eIL1R1 for treating PsA is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0357] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 1 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 2 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 3 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 4 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 5 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 6 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 7 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 8 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 9 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 10 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 11 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 12 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 13 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 14 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 15 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 16 of fIL1R1.In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 17 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting exon 18 of fIL1R1. In some embodiments, the CRISPR gene editing system for treating PsA comprising one or more sgRNAs targeting exons of fIL1R1 is delivered via one or more lipid nanoparticles (LNPs) comprising an LNP system described herein, such as, but not limited to, any one of LNP systems LNP001 to LNP240.

[0358] In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs targeting IL1RAP. In some embodiments, the CRISPR gene editing system for treating psoriatic arthritis comprises one or more sgRNAs...

Claims

1. A pharmaceutical composition for treating a disease, the composition comprising a plurality of lipid nanoparticles (LNPs), wherein the plurality of lipid nanoparticles encapsulate the following substances: (i) an RNA-guided nuclease or a nucleic acid encoding an RNA-guided nuclease; and (ii) at least one guide RNA or a nucleic acid encoding at least one guide RNA, wherein the at least one guide RNA targets FGF2, CCN2, NGF, NTF3, NTF4, BDNF, FGFR1, NGFR, NTRK1, NTRK2, ADAM17, ADAMTS1, ADAMTS5, MMP1, MMP2, MMP3, MMP7, MMP8, MMP10, MMP12, MMP13, TIMP1, TIMP3 CXCL1, CXCL2, CXCL3, CXCL5, CXCL6, CXCL8, CCL2, CCL3, CCL5, CCL7, CCL20, IL1A, IL1B, IL4, IL6, IL10, IL13, IL17A, IL18, TNF, CXCR1, CXCR2, CCR7, TNFRSF1A, TNFRSF1B, IL1R1, IL1RAP, IL4R, IL6R, IL6ST Genes for IL10RA, IL10RB, IL13RA1, IL13RA2, IL17RA, IL18R1, IL18RAP, SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SCN10A, SCN11A, TAC1, TAC3, TACR1, TACR2, TACR3, ATP1A1, CALCA, CALCB, CALCRL, RAMP1, ADM, CRCP, YAP1, MRGPRX2, TGFB, TGFBR1, and TGFBR2.

2. The pharmaceutical composition of claim 1, wherein the at least one guide RNA comprises a crRNA sequence selected from any one of SEQ ID NOs: 1-2731.

3. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a musculoskeletal disorder.

4. The pharmaceutical composition of claim 3, wherein the musculoskeletal disorder is selected from the group consisting of rheumatoid arthritis, gout, osteoarthritis, osteoporosis, intervertebral disc disease (IVDD), psoriatic arthritis (PsA), arthritis, polymyositis, proliferative synovitis, malignant osteophytes, sarcoid myopathy, cortical bone disease, idiopathic scoliosis, tendinopathy, myofibrillar myopathy, enthesis-related arthritis, ankylosing spondylitis, degenerative polyarthritis, arthropathy, osteitis deformans, lumbar disc prolapse, polymyositis ossificans, idiopathic polymyositis, Luft's disease, adult-onset Still's disease, osteoarthritis deformans, and Behcet's disease.

5. The pharmaceutical composition of claim 3, wherein the musculoskeletal disorder is selected from the group consisting of rheumatoid arthritis, idiopathic osteoporosis, postmenopausal osteoporosis, Paget's disease, osteoarthritis, juvenile idiopathic arthritis (JIA), Still's disease, ankylosing spondylitis, polymyalgia rheumatica, arthritis, secondary malignant neoplasms of bone type II, mucolipidosis, Sjögren's syndrome, psoriatic arthritis, rheumatism, Castleman's disease, degenerative polyarthritis, arthropathy, osteophytes, osteoporosis, massive osteolysis, fracture healing, systemic sclerosis, systemic juvenile idiopathic arthritis, and synovitis.

6. The pharmaceutical composition of claim 3, wherein the musculoskeletal disorder is selected from the group consisting of arthritis, infectious intermittent joint effusion, ankylosing spondylitis, arthritis, osteoarthritis, spondyloarthritis, plantar fasciitis, degenerative polyarthritis, hemophilic arthritis, inflammatory myopathy with abundant macrophages, polymyositis, tendinopathy, malignant osteophytes, osteoporosis, psoriatic arthritis, rheumatism, rheumatoid arthritis, adult-onset Still's disease, juvenile arthritis, early rheumatoid arthritis, relapsing rheumatic disease, gout, infectious arthritis, myotonic dystrophy, dermatomyositis, hemophilic arthropathy, osteopenia, Sjögren's syndrome, juvenile idiopathic arthritis, myasthenia gravis, osteolysis, inflammation, degenerative polyarthritis, osteitis deformans, pigmented villonodular synovitis, or hyperphosphatasia with bone disease.

7. The pharmaceutical composition of claim 3, wherein the musculoskeletal disorder is selected from the group consisting of Lloyd-Dietz syndrome, osteoarthrosis, Marfan syndrome, aortic aneurysm (familial thoracic 3), and craniofacial abnormalities.

8. The pharmaceutical composition of claim 1 or 2, wherein the disorder is neoplasia.

9. The pharmaceutical composition of claim 8, wherein the neoplasia is selected from the group consisting of osteosarcoma, colon cancer, metastasis (general), lung cancer, multiple myeloma, breast cancer, solid tumors, lymphoma, pancreatic cancer, gastric cancer, epithelial ovarian cancer, breast neoplasms, oropharyngeal cancer, renal cell carcinoma, chondrosarcoma, esophageal neoplasms, B-cell lymphoma, cutaneous T-cell lymphoma, leukemia, thyroid cancer, skin carcinogenesis, bile duct cancer, glioma, hepatocarcinoma, melanoma, neuroblastoma, polycystic ovary syndrome, glioblastoma, prostate cancer, cervical cancer, ovarian cancer, bladder cancer, squamous cell carcinoma, Kaposi's sarcoma, oral cancer, leiomyosarcoma, malignant peripheral nerve sheath tumor, and Ewing's sarcoma.

10. The pharmaceutical composition of claim 8, wherein the neoplasia is selected from the group consisting of: multiple myeloma, lung cancer, gastric cancer, breast cancer, kidney cancer, neoplasia metastasis, colorectal neoplasm, ovarian cancer, osteosarcoma, bile duct cancer, leukemia, prostate cancer, plasmacytoma, pancreatic cancer, cervical cancer, lymphoma, liver neoplasm, neuroblastoma, melanoma, mastocytosis, endometrial cancer, bladder cancer, squamous cell carcinoma, gallbladder cancer, adenocarcinoma, thyroid Carcinoma, Prostate Neoplasm, Gastric Cancer, Liver Cancer, Pituitary Neoplasm, Hepatoblastoma, Multiple Myeloma, Hepatocellular Adenoma, Breast Cancer, Carcinogenesis, Leukemia, Colon Cancer, Melanoma, Metastasis (General), Lung Cancer, Pancreatic Cancer, Kaposi's Sarcoma, Medulloblastoma, Bladder Cancer, Squamous Cell Carcinoma, Mast Cell Neoplasm, Glioma, Mast Cell Tumor, Brain Neoplasm, Ovarian Neoplasm, Osteoporosis, Rhabdomyosarcoma, Solid Tumors, Metastatic Renal Cancer, and Metastatic Renal Cell Carcinoma.

11. The pharmaceutical composition of claim 8, wherein the neoplasia is selected from the group consisting of neoplasms, glioblastomas, astrocytomas, adenocarcinomas, osteosarcomas, squamous cell carcinomas, metastases, ameloblastomas, bile duct carcinomas, choriocarcinomas, ovarian cancers, prostate cancers, lung cancers, laryngeal cancers, breast cancers, lip and oral cancers, squamous intraepithelial lesions, neuroblastomas, non-Hodgkin's lymphomas, malignant neoplasms, skin neoplasms, neuroblastomas, neoplasm metastases, colorectal cancers, fibrosarcomas, myeloid leukemias, Blood disease, myelofibrosis, Hodgkin's lymphoma, non-small cell lung cancer, cervical cancer, liver cancer, extrapulmonary small cell carcinoma, basal cell carcinoma, kidney cancer, pancreatic cancer, mesothelioma, gastric cancer, polycystic ovary syndrome, chordoma, bile duct cancer, malignant ascites, nasopharyngeal cancer, head and neck cancer, cancer metastasis, prostate cancer, fibrosarcoma, liver cancer, bladder cancer, T-cell lymphoblastic leukemia, bladder neoplasm, melanoma, leukemia, acute myeloid leukemia, hepatocellular carcinoma, colorectal cancer, lymphoma, mycosis fungoides and Sezary syndrome.

12. The pharmaceutical composition of claim 8, wherein the neoplasia is selected from the group consisting of pancreatic cancer, multiple spontaneously healing squamous neoplasia (Ferguson-Smith disease), pancreatic cancer, gastrointestinal stromal tumors (GIST), hereditary nonpolyposis colorectal cancer (Lynch syndrome), metastasis, colorectal cancer, bone neoplasms, anaplastic carcinoma, spindle cell carcinoma, malignant bone neoplasms, lung neoplasms, and malignant brain neoplasms.

13. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a neurological disorder.

14. The pharmaceutical composition of claim 13, wherein the neurological disorder is selected from the group consisting of acute thrombotic stroke, epilepsy, multiple sclerosis and Alzheimer's disease.

15. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a cardiac disorder.

16. The pharmaceutical composition of claim 15, wherein the cardiac disorder is selected from the group consisting of acute myocardial infarction, refractory heart failure, arrhythmia, pericarditis, myocarditis, sepsis-induced cardiomyopathy, acute decompensated heart failure, refractory idiopathic pericarditis, atherosclerosis, coronary artery disease, myocardial infarction, and cardiac remodeling.

17. The pharmaceutical composition of claim 15, wherein the cardiac disorder is atherosclerosis or abdominal aortic aneurysm.

18. The pharmaceutical composition of claim 1 or 2, wherein the disorder is an inflammatory disorder.

19. The pharmaceutical composition of claim 18, wherein the inflammatory condition is selected from the group consisting of autoinflammatory disease (AID), cryptopyrin-associated periodic syndrome (CAPS), familial Mediterranean fever (FMF), TNF receptor-associated periodic syndrome (TRAPS), hyper-IgD syndrome (HIDS), systemic lupus erythematosus (SLE), and fibrosis.

20. The pharmaceutical composition of claim 18, wherein the inflammatory disorder is selected from the group consisting of cytokine storm, acute local inflammation, inflammatory bowel disease, Crohn's disease, sepsis, experimental sepsis, or Castleman's disease.

21. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a digestive system disorder.

22. The pharmaceutical composition of claim 21, wherein the digestive system disorder is selected from the group consisting of inflammatory bowel disease (IBD), gastroesophageal reflux disease (GERD) or non-HP related peptic ulcer disease.

23. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a respiratory disorder.

24. The pharmaceutical composition of claim 23, wherein the respiratory disorder is asthma or pulmonary fibrosis.

25. The pharmaceutical composition of claim 1 or 2, wherein the disorder is a renal disorder, a metabolic disorder, or an ocular disorder.

26. The pharmaceutical composition of claim 25, wherein the renal disorder, metabolic disorder or ocular disorder is selected from the group consisting of chronic kidney disease, type 2 diabetes, ocular disease, uveitis, scleritis, Sjögren's syndrome and dry eyes.

27. The pharmaceutical composition of claim 1 or 2, wherein the condition is an autoimmune disease.

28. The pharmaceutical composition of claim 27, wherein the autoimmune disease is systemic lupus erythematosus.

29. The pharmaceutical composition of claim 1 or 2, wherein the condition is acute synovitis, chronic synovitis, inflammatory arthritis, or immune-mediated arthritis.

30. The pharmaceutical composition of claim 1 or 2, wherein the disorder is an autoimmune disorder or an inflammatory disorder.

31. The pharmaceutical composition of claim 29, wherein the autoimmune or inflammatory disorder is selected from the group consisting of allergy, asthma, multiple sclerosis, psoriasis, inflammatory bowel disease, autoimmune experimental uveitis, colitis, hypersensitivity diseases, diabetes, Crohn's disease, systemic lupus erythematosus, pulmonary sarcoidosis, leishmaniasis, experimental autoimmune encephalomyelitis, HTLV-1 associated myelopathy, tropical spastic paraplegia, scleroderma, idiopathic inflammatory myopathy, polymyositis, and dermatomyositis.

32. The pharmaceutical composition of any one of claims 1 to 31, wherein the RNA-guided nuclease or a nucleic acid encoding an RNA-guided nuclease is the RNA-guided nuclease.

33. The pharmaceutical composition of any one of claims 1 to 31, wherein the RNA-guided nuclease or nucleic acid encoding the RNA-guided nuclease is a DNA encoding the RNA-guided nuclease.

34. The pharmaceutical composition of any one of claims 1 to 31, wherein the RNA-guided nuclease or nucleic acid encoding the RNA-guided nuclease is an mRNA encoding the RNA-guided nuclease.

35. A pharmaceutical composition as described in any one of claims 1 to 34, wherein the RNA-guided nuclease is a Cas protein.

36. A pharmaceutical composition as described in claim 35, wherein the Cas protein is a Cas9 protein.

37. A pharmaceutical composition as described in claim 35, wherein the Cas9 protein is a Streptococcus pyogenes Cas9 polypeptide.

38. The pharmaceutical composition of claim 35, wherein the Cas9 protein is selected from the group consisting of esCas9, hfCas9, peCas9, and ARCas9.

39. The pharmaceutical composition of any one of claims 1 to 38, wherein the at least one guide RNA or a nucleic acid encoding at least one guide RNA is the at least one guide RNA.

40. The pharmaceutical composition of any one of claims 1 to 38, wherein the at least one guide RNA or a nucleic acid encoding at least one guide RNA is a DNA encoding the at least one guide RNA.

41. The pharmaceutical composition of any one of claims 1 to 38, comprising a nucleic acid encoding both the RNA-guided nuclease and the at least one guide RNA.

42. The pharmaceutical composition of any one of claims 1 to 41, wherein the at least one guide RNA is a single guide RNA (sgRNA).

43. The pharmaceutical composition of any one of claims 1 to 42, wherein the at least one guide RNA targets a human gene.

44. The pharmaceutical composition of any one of claims 1 to 42, wherein the at least one guide RNA targets a canine gene.

45. The pharmaceutical composition of any one of claims 1 to 42, wherein the at least one guide RNA targets an equine gene.

46. ​​The pharmaceutical composition of any one of claims 1 to 42, wherein the at least one guide RNA targets a feline gene.

47. The pharmaceutical composition of any one of claims 1 to 42, wherein the at least one guide RNA targets a mammalian gene.

48. A pharmaceutical composition as described in any one of claims 1 to 47, wherein the composition is formulated for parenteral administration.

49. A pharmaceutical composition as described in any one of claims 1 to 47, wherein the composition is formulated for intra-articular injection into a joint of a subject.

50. The pharmaceutical composition of any one of claims 1 to 47, wherein the composition is formulated for intradiscal injection.

51. The pharmaceutical composition of any one of claims 1 to 47, wherein the composition is formulated for intradiscal injection.

52. A pharmaceutical composition as described in any one of claims 1 to 47, wherein the composition is formulated for intraspinal injection.

53. A pharmaceutical composition as described in any one of claims 1 to 47, wherein the composition is formulated for epidural injection.

54. A pharmaceutical composition as described in any one of claims 1 to 47, wherein the composition is formulated for injection into a facet joint of the spine.

55. The pharmaceutical composition of any one of claims 1 to 54, wherein the one or more LNPs comprise one or more ionizable lipids selected from the group consisting of 3-(didodecylamino)-N1, N1, 4-tridodecyl-1-piperazineethylamine (KL10), N1-[2-(didodecylamino)ethyl]-N1, N4, N4-tridodecyl-1, 4-piperazinediethylamine (KL22), 14,25-tricosyl-15,18,21,24-tetraaza-triacontane (KL25 ), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLin-DMA), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA), heptathriacontane-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butyrate (DLin-MC3-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLin-KC2-DMA), 1,2-di Oleyloxy-N,N-dimethylaminopropane (DODMA), 2-({8-[(3.β.)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propan-1-amine (Octyl-CLinDMA), (2R)-2-({8-[(3.β.)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propan-1-amine -9,12-dien-1-yloxy]propan-1-amine (Octyl-CLinDMA(2R)), (2S)-2-({8-[(3.β.)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z-,12Z)-octadec-9,12-dien-1-yloxy]propan-1-amine (Octyl-CLinDMA(2S)), LP01, ALC-0315, SM-102, lipids containing cyclic amine groups, and mixtures thereof.

56. pharmaceutical composition as claimed in claim 55, wherein said ionizable lipid is DLin-KC2-DMA.

57. pharmaceutical composition as claimed in claim 55, wherein said ionizable lipid is DLin-MC3-DMA.

58. The pharmaceutical composition of claim 55, wherein the ionizable lipid is SM-102.

59. The pharmaceutical composition of claim 55, wherein the ionizable lipid is ALC-0315.

60. The pharmaceutical composition of claim 55, wherein the ionizable lipid is LP01.

61. The pharmaceutical composition of any one of claims 1 to 54, wherein the one or more LNPs comprise one or more auxiliary lipids selected from the group consisting of 1,2-dilinoleoyl-sn-glyceryl-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glyceryl-phosphocholine (DMPC), 1,2-dioleoyl-sn-glyceryl-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glyceryl-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glyceryl-3-phosphocholine (DSPC), 1,2-heneicosanoyl-sn-glyceryl-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glyceryl-3-phosphocholine (POPC), 1,2-di-O-octadecene-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine (18:0Diether-1-yl)-sn-glyceryl-3-phosphocholine PC), 1-oleoyl-2-cholesterol hemisuccinyl-sn-glyceryl-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glyceryl-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glyceryl-3-phosphocholine, 1,2-diamidonoyl-sn-glyceryl-3-phosphocholine, 1,2-docosahexaenoyl-sn-glyceryl-3-phosphocholine, 1,2-dioleoyl-sn-glyceryl-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glyceryl-3-phosphoethanolamine (ME16.0PE), 1,2-distearoyl-sn-glyceryl-3-phosphocholine Acid ethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dialinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-docosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), sphingomyelin (SM) and mixtures thereof.

62. pharmaceutical compositions as described in any one in claim 1 to 54, wherein the one or more LNPs comprise one or more PEG lipids selected from: PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol and their mixture.For example, the PEG lipid can be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, PEG-DMA or PEG-DSPE lipid.

63. A pharmaceutical composition as described in any one of claims 1 to 54, wherein the one or more LNPs comprise one or more structural lipids selected from the group consisting of cholesterol, coprosterol, stigmasterol, stigmasterol, sitosterol, β-sitosterol, lupeol, betulin, ursolic acid, oleanolic acid, campesterol, fuccasterol, brassicasterol, ergosterol, 9,11-dehydroergosterol, tomatine, tomatine, alpha-tocopherol, and mixtures thereof.

64. The pharmaceutical composition of claim 63, wherein the structured lipid is cholesterol.

65. The pharmaceutical composition of claim 63, wherein the one or more LNPs comprise a structured lipid and a corticosteroid (e.g., prednisolone, dexamethasone, prednisone, and hydrocortisone) or a combination thereof.

66. The pharmaceutical composition of claim 65, wherein the corticosteroid is dexamethasone.

67. A pharmaceutical composition as described in any one of claims 1 to 66, wherein the LNP comprises a plurality of particles having a diameter exceeding 100 nm.

68. A pharmaceutical composition as described in any one of claims 1 to 66, wherein the LNP comprises a plurality of particles having a diameter of about 60 nm to about 120 nm.

69. The pharmaceutical composition of any one of claims 1 to 68, wherein the plurality of LNPs comprises an LNP system selected from any one of LNP001 to LNP240.

70. A method for treating a disease or condition in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition as described in any one of claims 1 to 69.

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