PIKFYVE antisense oligonucleotide

Abstract

The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing the same, and methods for treating, inhibiting, suppressing, and preventing neurological disorders using the same. In some aspects, the disclosure provides single-stranded ASOs that suppress expression of PIKFYVE, wherein the ASOs have a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321.

Description

[Technical Field]

[0001] REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 483,517, filed February 6, 2023. The entire disclosures of the above-referenced applications are incorporated herein by reference in their entirety.

[0002] Reference to an electronically submitted sequence listing The contents of the electronically submitted Sequence Listing (Name: 3817_225PC01_SequenceListing_ST26.xml; Size: 5,488,433 bytes; and Created: February 6, 2024) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing same, and methods for treating, inhibiting, suppressing, and preventing neurological or neurodegenerative diseases. [Background technology]

[0004] Many neurodegenerative disorders in patients are difficult to treat effectively, especially when the pathology of the neurodegenerative disorder in a particular patient is not fully understood.

[0005] WO 2016 / 210372 discloses a method for treating neurodegenerative diseases by administering a PIKFYVE inhibitor. There remains a need for effective treatments for many neurodegenerative disorders, such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2016 / 210372 Summary of the Invention [Means for solving the problem]

[0007] The present disclosure relates to PIKFYVE antisense oligonucleotides (ASOs), pharmaceutical compositions containing same, and their use in the treatment of neurodegenerative disorders.

[0008] In some aspects, the disclosure provides a single-stranded ASO that suppresses expression of PIKFYVE, wherein the ASO has a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and an unmodified version of SEQ ID NOs: 1646-2321. In some embodiments, the disclosure provides single-chain ASOs that suppress expression of PIKFYVE, the ASOs being selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1651, 1652, 1653, 1654, 1655, 1656, 1657, 1658, 1659, 1660, 1661, 1662, 1663, 1664, 1665, 1666, 1667, 1668, 1669, 1670, 1671, 1672, 1673, 1674, 1675, 1676, 1677, 1678, 1679, 1680, 1685, 1686, 1687, 1688, 1689, 1690, 1701, 1702, 1703, 1704, 1705, 1706, 1707, 1708, 1710, 1711, 1712, 1713, 171 652~1654, 1660, 1666~1667, 1671~1672, 1674, 1677, 1679, 1684~1687, 1689, 1692, 1694, 1696~1698, 1700, 1702~1703, 1708~1711, 1713, 1716~1717, 1719, 1722, 1725, 1727~1728, 1731, 1740, 1743~1745, 1747~1751, 1754, 1756, 1760, 1762~1764 , 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869 69, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017,2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2150 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0009] The nucleobase sequence of the ASO may include up to 30, 25, 24, 23, 22, 21, or 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. In some embodiments, the nucleobase sequences of the ASOs are selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, as well as SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, and 1673-1674. 72, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788 788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058,2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183, 2185 , 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0010] The ASO may also be any of the unmodified versions of SEQ ID NOs: 1-782, for example, 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and SEQ ID NOs: 1646-2321. In some embodiments, the ASOs also include those of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, as well as those of SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679 , 1684~1687, 1689, 1692, 1694, 1696~1698, 1700, 1702~1703, 1708~1711, 1713, 1716~1717, 1719, 1722, 1725, 1727~1728, 1731, 1740, 1743~1745, 1747~1751, 1754, 1756, 1760, 1762~1764, 1770~1775, 1777~1779, 1782~1784, 1787~1788, 1791~1793, 1795, 1797~1802, 1 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1920 7, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104,It may be any of the unmodified versions of 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0011] In some aspects, the disclosure provides oligonucleotides consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. In some embodiments, the present disclosure provides a method for the preparation of a nucleic acid sequence comprising 12 to 30 linked nucleosides and selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938, 939, 940, 941, , 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1828-1829, 1830-1831, 1832-1833, 1834-1835, 1836-1837, 1838-1839, 1840-1841, 1842-1843, 1844-1845, 1846-1847, 1848-1849, 1850-1851, 1852-1853, 1854-1855, 1856-1857, 1858-1859, 1860-1861, 1862-1863, 1863-1864, 1865-1866, 1867-1868, 1869-1870, 1871-1872, 1873-1874, 1875-1876, 1877-1878, 1879-1880, 1881-1882, 1883-18 30~1832, 1836~1837, 1840~1843, 1845~1853, 1855~1858, 1861~1862, 1864, 1866, 1868~1869, 1871~1881, 1883, 1885~1892, 1895~1897, 1899, 1901~1907, 1909, 1911~1913, 1915, 1917, 1919~1927, 1930, 1932~1936,1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2056-2060 58, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, Oligonucleotides are provided having nucleobase sequences comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the unmodified versions of the nucleobase sequences of 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0012] The oligonucleotide may contain up to 25, 24, 23, 22, 21, or 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. In some embodiments, the oligonucleotides are selected from the group consisting of SEQ ID NOS: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, as well as SEQ ID NOS: 1650, 1652-1654, 1660, 1666-1667, and 1671-1672. 2, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788 88, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1 901~1907, 1909, 1911~1913, 1915, 1917, 1919~1927, 1930, 1932~1936, 1938~1940, 1943, 1945~1953, 1955~1969, 1971~1973, 1976~1980, 1985~1994, 1996~1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058,2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183, 218 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0013] In some embodiments, at least one internucleoside linkage is a modified internucleoside linkage, which may be a phosphorothioate internucleoside linkage or a phosphodiester internucleoside linkage. At least one of the nucleosides may be a modified nucleobase.

[0014] In some embodiments, at least one nucleoside of the ASO can have a modified sugar moiety, which can be a bicyclic sugar moiety or which can include a 2'-O-methoxyethyl group. In certain embodiments, the bicyclic sugar moiety includes a 4'-CH(R)-O-2' bridge, and the R groups are independently selected from H, C, 1-12 alkyl, or a protecting group.

[0015] In some embodiments, the ASO is a gapmer (e.g., an MOE gapmer), where the gap segment can consist of 8-12 linked deoxynucleosides, a 5' wing segment consisting of 3-5 linked nucleosides, and a 3' wing segment consisting of 3-5 linked nucleosides. In certain embodiments, the gap segment can be disposed between the 5' wing segment and the 3' wing segment, and the nucleosides of each wing segment contain modified sugar moieties (e.g., sugar moieties having 2'-O-methoxyethyl groups).

[0016] In some embodiments, the oligonucleotide consists of 12 to 30 linked nucleosides and has a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. In some embodiments, the oligonucleotide consists of 12 to 30 linked nucleosides and is selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 811, 812, 813, 814, 815, 816, 817, 818, 819, 820, 821, 822, 823, 824, 825, 826, 827, 828, 829, 830, 831, 832, 833, 834, 835, 836, 837, 838, 839, 840, 841, 842, 843, 72, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743~1745, 1747~1751, 1754, 1756, 1760, 1762~1764, 1770~1775, 1777~1779, 1782~1784, 1787~1788, 1791~1793, 1795, 1797~1802, 1804~1805, 1809~1811, 1813~1815, 1818~1819, 1822~1824, 1826~1827, 1830~1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940,1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2060-2062 061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~216 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0017] In some embodiments, the disclosure provides pharmaceutical compositions comprising a PIKFYVE ASO of the disclosure and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. In some embodiments, the pharmaceutical compositions are suitable for parenteral administration, e.g., intraventricular injection or intrathecal administration.

[0018] In some aspects, the disclosure provides methods for inhibiting, suppressing, or preventing expression of PIKFYVE in a patient (e.g., a patient having a neurological or neurodegenerative disease) by administering to the patient (e.g., by intracerebroventricular injection or intrathecal administration) a PIKFYVE ASO or a pharmaceutical composition described herein (e.g., an effective amount thereof).

[0019] In some embodiments, the present disclosure provides a method for treating a subject with a neurological or neurodegenerative disease by administering a therapeutically effective amount of PIKFYVE ASO or a pharmaceutical composition described herein. In some embodiments, the disease is amyotrophic lateral sclerosis (ALS) (e.g., C9orf72-associated ALS). In some embodiments, the disease is frontotemporal dementia (FTD), such as FTD with TDP-43 pathology or FTD with tau pathology. In some embodiments, the disease is C9orf72-associated FTD (C9-FTD). In some embodiments, the disease is microtubule-associated protein tau (MAPT)-associated FTD (MAPT-FTD), for example, FTD with a V337M MAPT mutation.

[0020] In some aspects, the disclosure provides methods for treating a subject having a PIKFYVE disease or disorder by administering a therapeutically effective amount of a PIKFYVE ASO or pharmaceutical composition described herein. [Brief explanation of the drawings]

[0021] [Figure 1A] Skipping exon 3 of PIKFYVE is predicted to generate a stop codon, causing premature termination of translation of the PIKFYVE protein. An "*" indicates a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380). [Figure 1B] Skipping exon 3 of PIKFYVE is predicted to generate a stop codon, causing premature termination of translation of the PIKFYVE protein. An "*" indicates a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380).

[0022] [Figure 2A]Skipping exon 5 of PIKFYVE is predicted to result in a major structural change in the PIKFYVE protein. "*" represents a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380). [Figure 2B] Skipping exon 5 of PIKFYVE is predicted to result in a major structural change in the PIKFYVE protein. "*" represents a stop codon. The sequence shown is taken from the Ensembl database (ENST00000264380). DETAILED DESCRIPTION OF THE INVENTION

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present specification, including definitions, will control. Preferred methods and materials are described below; however, methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0024] definition The terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," "may," and variations thereof, as used herein, are intended to be open-ended transitional phrases, terms, or phrases that do not exclude the possibility of additional acts or structures.

[0025] Unless the context clearly indicates otherwise, the singular forms "a," "an," and "the" include plural referents.

[0026] The present disclosure also contemplates other aspects that "comprising," "consisting of," and "consisting essentially of" aspects or elements present herein, whether explicitly stated or not.

[0027] As used herein, an "unmodified version" of an ASO / sequence means an ASO / sequence that contains only DNA or RNA nucleosides linked by phosphodiester bonds.

[0028] As used herein, "2'-deoxynucleoside" refers to a nucleoside containing a 2'-H(H) furanosyl sugar moiety as found in naturally occurring deoxyribonucleic acid (DNA) and nucleobases. In some embodiments, a 2'-deoxynucleoside may contain a modified nucleobase and a furanosyl sugar moiety, or may contain an RNA nucleobase (uracil) and a furanosyl sugar moiety.

[0029] As used herein, "2'-substituted nucleoside" refers to a nucleoside that includes a 2'-substituted sugar moiety. As used herein, "2'-substituted" with respect to the sugar moiety means that the sugar moiety includes at least one 2'-substituent other than H or OH.

[0030] As used herein, "antisense molecule" means an oligomeric nucleic acid or oligomeric duplex capable of achieving at least one antisense activity.

[0031] The modifier "about" used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context (e.g., it includes at least the degree of error associated with measurement of the particular quantity). The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the expression "about 2 to about 4" also discloses a range of "2 to 4." The term "about" can refer to plus or minus 10% of the indicated number. For example, "about 10%" can indicate a range of 9% to 11%, and "about 1" can mean 0.9 to 1.1. Other meanings of "about" are apparent from the context, such as rounding; for example, "about 1" can mean 0.5 to 1.4.

[0032] With respect to the recitation of numerical ranges herein, each intervening number to the same degree of precision is expressly contemplated. For example, in the range of 6 to 9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and in the range of 6.0 to 7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0033] As used herein, "bicyclic sugar" or "bicyclic sugar moiety" refers to a modified sugar moiety comprising two rings, where the second ring is formed via a bridge connecting two of the atoms in the first ring, thereby forming a bicyclic structure. In some embodiments, the first ring of the bicyclic sugar moiety is a furanosyl moiety. In some embodiments, the bicyclic sugar moiety does not comprise a furanosyl moiety. As used herein, "bicyclic nucleoside" or "BNA" refers to a nucleoside comprising a bicyclic sugar moiety.

[0034] As used herein, "chiral enriched population" refers to a plurality of molecules of the same molecular formula, wherein the number or percentage of molecules in the population that contain a specific stereochemical configuration at a specific chiral center is greater than the number or percentage of molecules that are expected to contain the same specific stereochemical configuration at the same specific chiral center in the population, when the specific chiral center is stereorandom. A chiral enriched population of molecules that has multiple chiral centers within each molecule can contain one or more stereorandom chiral centers. In some embodiments, the molecule is a modified oligonucleotide. In some embodiments, the molecule is a compound that includes a modified oligonucleotide.

[0035] As used herein, "complementary" with respect to an oligonucleotide means that at least 70% of the nucleobases, or one or more regions thereof, of an oligonucleotide and at least 70% of the nucleobases, or one or more regions thereof, of another nucleic acid can hydrogen bond with each other when the nucleobase sequences of the oligonucleotide and the other nucleic acid are aligned in opposite directions. Complementary nucleobases refer to nucleobases that can form hydrogen bonds with each other. Complementary nucleobase pairs include adenine (A) and thymine (T), adenine (A) and uracil (U), cytosine (C) and guanine (G), and 5-methylcytosine (mC) and guanine (G). Complementary oligonucleotides and / or nucleic acids need not have complementary nucleobases at every nucleoside. Rather, some mismatches are permitted. As used herein, "fully complementary" or "100% complementary" with respect to an oligonucleotide means that the oligonucleotide is complementary to another oligonucleotide or nucleic acid at every nucleoside of the oligonucleotide.

[0036] As used herein, "gapmer" refers to a modified oligonucleotide containing an internal region having multiple nucleosides that support RNase H cleavage, separated by external regions having one or more nucleosides, where the nucleosides comprising the internal region are chemically distinct from the nucleoside or nucleosides comprising the external regions. The internal region may be referred to as the "gap," and the external regions may be referred to as the "wings." Unless otherwise specified, "gapmer" refers to the sugar motif. Unless otherwise specified, the sugar moieties of the nucleosides in the gap are unmodified 2'-deoxyfuranosyl. Thus, the term "MOE gapmer" refers to a gapmer having both wing 2'-MOE nucleoside sugar motifs and a 2'-deoxynucleoside gap. Unless otherwise specified, MOE gapmers may contain one or more modified internucleoside linkages and / or modified nucleobases, and such modifications do not necessarily follow the gapmer pattern of sugar modifications. Exemplary MOE gapmers are shown in Tables 2 and 6 below.

[0037] In some embodiments, oligonucleotides comprise one or more modified and / or unmodified sugar moieties arranged along the oligonucleotide or region thereof in a defined pattern or sugar motif, which in certain instances includes, but is not limited to, any of the sugar modifications discussed herein.

[0038] In some embodiments, modified oligonucleotides comprise or consist of a region having a gap motif defined by two outer regions, or "wings," and a central or internal region, or "gap." The three regions of a gapmer motif include the "5' wing," the "gap," and the "3' wing," which form a continuous sequence of nucleosides, with at least a portion of the sugar moieties of the nucleosides in each wing being different from at least a portion of the sugar moieties of the nucleosides in the gap. Specifically, at least the sugar moieties of the nucleosides in each wing closest to the gap (the 3'-most nucleoside of the 5'-wing and the 5'-most nucleoside of the 3'-wing) are different from the sugar moieties of the adjacent gap nucleosides, thus defining the boundary between the wing and the gap (i.e., the wing / gap junction). In some embodiments, the sugar moieties within the gap are identical to each other. In some embodiments, the gap comprises one or more nucleosides having a sugar moiety that is different from the sugar moieties of one or more other nucleosides in the gap. In some embodiments, the sugar motifs of the two wings are identical to each other (symmetric gapmer). In some embodiments, the sugar motif of the 5'-wing is different from the sugar motif of the 3'-wing (asymmetric gapmer).

[0039] In some embodiments, a gapmer wing comprises 1 to 5 nucleosides. In some embodiments, each nucleoside in each wing of a gapmer is a modified nucleoside.

[0040] In some embodiments, the gapmer gap comprises 7 to 12 nucleosides (e.g., 10 nucleosides). In some embodiments, each nucleoside in the gapmer gap is an unmodified 2'-deoxynucleoside.

[0041] In some embodiments, the gapmer is a deoxygapmer. In some embodiments, the nucleosides on the gap side of each wing / gap junction are unmodified 2'-deoxynucleosides and the nucleosides on the wing side of each wing / gap junction are modified nucleosides. In some embodiments, each nucleoside of the gap is an unmodified 2'-deoxynucleoside. In some embodiments, each nucleoside of each wing of the gapmer is a modified nucleoside.

[0042] In some embodiments, a modified oligonucleotide comprises or consists of a region having a fully modified sugar motif. In some embodiments, each nucleoside of a fully modified region of a modified oligonucleotide comprises a modified sugar moiety. In some embodiments, each nucleoside throughout a modified oligonucleotide comprises a modified sugar moiety. In some embodiments, a modified oligonucleotide comprises or consists of a region having a fully modified sugar motif, and each nucleoside within a fully modified region comprises the same modified sugar moiety, referred to herein as a uniformly modified sugar motif. In some embodiments, a fully modified oligonucleotide is a uniformly modified oligonucleotide. In some embodiments, each nucleoside of a uniformly modified oligonucleotide comprises the same 2' modification.

[0043] As used herein, "inhibit" refers to the ability to substantially antagonize, block, prevent, suppress, arrest, slow, hinder, alter, eliminate, halt, or reverse the progression or severity of the activity of a particular pathogen (e.g., an infectious pathogen) or disease.

[0044] As used herein, "internucleoside bond" refers to the covalent bond between adjacent nucleosides in an oligonucleotide.As used herein, "modified internucleoside bond" refers to any internucleoside bond other than a phosphodiester internucleoside bond.A "phosphorothioate bond" is a modified internucleoside bond in which one of the non-bridging oxygen atoms of a phosphodiester internucleoside bond is replaced with a sulfur atom.

[0045] In some embodiments, the nucleosides of modified oligonucleotides can be linked together using any internucleoside linkage. Two major classes of internucleoside linkage groups are defined by the presence or absence of a phosphorus atom. Representative phosphorus-containing internucleoside linkages include, but are not limited to, phosphodiester linkages (also referred to as unmodified or native linkages), phosphotriesters, methyl phosphonates or other alkyl phosphonates, phosphoramidates, and phosphorothioates, including phosphorodithioates. Representative non-phosphate-containing internucleoside linkage groups include, but are not limited to, methylenemethylimino (-CH2-N(CH3)-O-CH2-), thiodiesters, thionocarbamate (-OC(=O)(NH)-S-), siloxane (-O-SiH2-O-), and N,N'-dimethylhydrazine (-CH2-N(CH3)-N(CH3)-). Modified internucleoside linkages, compared to natural phosphate linkages, can be used to alter (usually increase) the nuclease resistance of oligonucleotides. Methods for preparing phosphorus-containing and non-phosphorus-containing internucleoside linkages are well known to those skilled in the art.

[0046] Representative internucleoside linkages having a chiral center include, but are not limited to, alkylphosphonates and phosphorothioates. Modified oligonucleotides containing internucleoside linkages having a chiral center can be prepared as a population of modified oligonucleotides containing stereorandom internucleoside linkages or as a population of modified oligonucleotides containing phosphorothioate linkages of a specific stereochemical configuration. In some embodiments, the population of modified oligonucleotides contains phosphorothioate internucleoside linkages, where all of the phosphorothioate internucleoside linkages are stereorandom. Such modified oligonucleotides can be produced using synthetic methods that result in random selection of the stereochemistry of each phosphorothioate linkage. Nevertheless, as is well understood by those skilled in the art, each individual phosphorothioate in each individual oligonucleotide molecule has a defined stereochemistry. In some embodiments, the population of modified oligonucleotides is enriched for modified oligonucleotides containing one or more specific phosphorothioate internucleoside linkages of a specific, independently selected stereochemical configuration. In some embodiments, a particular configuration of phosphorothioate linkages is present in at least 65% of the molecules in the population. In some embodiments, a particular configuration of phosphorothioate linkages is present in at least 70% of the molecules in the population. In some embodiments, a particular configuration of phosphorothioate linkages is present in at least 80% of the molecules in the population. In some embodiments, a particular configuration of phosphorothioate linkages is present in at least 90% of the molecules in the population. In some embodiments, a particular configuration of phosphorothioate linkages is present in at least 99% of the molecules in the population.Such chirally enriched populations of modified oligonucleotides can be produced using synthetic methods known in the art, such as those described in Oka et al., JACS 125, 8307 (2003); Wan et al., Nuc. Acid. Res. 42, 13456 (2014); Chapter 10 of Locked Nucleic Acid Aptamers in Nucleic Acid and Peptide Aptamers: Methods and Protocols v 535, 2009 by Barciszewski et al., editor Gunter Mayerand; and WO 2017 / 015555. In some embodiments, the population of modified oligonucleotides is enriched for modified oligonucleotides having at least one designated phosphorothioate in the (Sp) configuration. In some embodiments, the population of modified oligonucleotides is enriched for modified oligonucleotides having at least one designated phosphorothioate in the (Rp) configuration.

[0047] As used herein, "MOE" means methoxyethyl. "2'-MOE" means a -OCH2CH2OCH3 group at the 2' position of the furanosyl ring.

[0048] A "neurological disease" is a disease that causes electrical, biochemical, or structural abnormalities in the brain, spine, or neurons. For example, the neurological disease can be a neurodegenerative disease. The neurodegenerative disease can cause, for example, the degeneration of motor neurons. The neurological disease can be, for example, amyotrophic lateral sclerosis (ALS), Huntington's disease, Alzheimer's disease, or frontotemporal dementia. Further examples of neurological diseases include, but are not limited to, Parkinson's disease, multiple sclerosis, peripheral myopathy, Rasmussen's encephalitis, attention deficit hyperactivity disorder, autism, central pain syndrome, anxiety, and / or depression.

[0049] Neurological disorders may involve abnormal endosomal transport. For example, the endosomal pathway and endosomes are necessary components for the recycling or destruction of membrane-bound proteins, the transport of Golgi-associated proteins, and the extracellular release of proteins in exosomes. These processes support neurotransmission, for example, by promoting the balance between the recycling and degradation of synaptic vesicles or neurotransmitter receptors.

[0050] Neurological diseases may be accompanied by abnormal lysosomal degradation. Altered lysosomal degradation may be present in neurological diseases, such as neurodegenerative diseases. Cathepsin imbalance due to aging and age-related diseases may cause harmful effects on central nervous system (CNS) neurons, and lysosomes may become the site of unfolding and partial degradation of membrane proteins or their precursors, which may then be expelled from the cell or released from dying cells and accumulate as pathological entities.

[0051] A healthcare professional may diagnose a subject as having a disease associated with motor neuron degeneration by assessing one or more symptoms of motor neuron degeneration. To diagnose a neurological disorder, a physical exam may be followed by a thorough neurological examination. A neurological examination may assess motor and sensory skills, nerve function, hearing and speech, vision, coordination and balance, mental status, and mood or behavioral changes. Non-limiting symptoms of disorders associated with neurological disorders include weakness in the arms, legs, feet, or ankles, slurred speech, difficulty lifting the front of the foot and toes, weakness or clumsiness in the hands, muscle paralysis, muscle rigidity, involuntary spasms or writhing movements (chorea), involuntary, persistent muscle contractures (dystonia), bradykinesia, loss of motility, impaired posture and balance, lack of flexibility, tingling in parts of the body, an electric shock sensation when moving the head, and pain in the arms, shoulders, and tongue. Symptoms may include convulsions, difficulty swallowing, difficulty breathing, difficulty chewing, partial or complete loss of vision, double vision, slow or abnormal eye movements, tremors, unsteady gait, fatigue, memory loss, dizziness, difficulty thinking or concentrating, difficulty reading and writing, misunderstanding spatial relationships, disorientation, depression, anxiety, difficulty making decisions and judgments, loss of impulse control, difficulty planning and carrying out routine tasks, aggression, irritability, social withdrawal, mood swings, dementia, changes in sleep habits, wandering, and changes in appetite.

[0052] Tests may be performed to determine diseases and disorders that may mimic neurological disorders, measure muscle involvement, and assess neuronal degeneration. Non-limiting examples of tests include electromyography (EMG), nerve conduction velocity tests, blood, urine, or other substance laboratory tests, magnetic resonance imaging (MRI), magnetic resonance spectroscopy, muscle or nerve biopsy, transcranial magnetic stimulation, genetic screening, X-rays, fluoroscopy, angiography, computed tomography (CT), positron emission tomography, cerebrospinal fluid analysis, intrathecal contrast CT scan, electroencephalography, electronystagmography, evoked responses, polysomnography, thermography, and ultrasound. A medical professional may also assess a patient's family history of diseases associated with motor neuron degeneration and make a diagnosis based in part on the family history of neurological disorders. A health care professional may diagnose a subject's neurological disorder after one or more symptoms appear.

[0053] Neurodegenerative diseases result in the progressive destruction of neurons and affect neuronal signal transduction. For example, neurodegeneration can be amyotrophic lateral sclerosis, Alzheimer's disease, Huntington's disease, Friedreich's ataxia, Lewy body disease, Parkinson's disease, spinal muscular atrophy, primary lateral sclerosis, progressive muscular atrophy, progressive bulbar palsy, and pseudobulbar palsy.

[0054] Diseases associated with the degeneration of motor neurons can be pathological conditions that cause the progressive destruction of motor neurons, hindering neuronal signal transmission to muscles, leading to muscle weakness and wasting.In healthy individuals, upper motor neurons transmit signals from the brain to lower motor neurons in the brainstem and spinal cord, and these signals are then transmitted to muscles, resulting in voluntary muscle activity.The destruction of upper and lower motor neurons can affect activities such as breathing, speaking, swallowing, and walking, and over time these functions can be lost.Examples of motor neuron diseases include, but are not limited to, amyotrophic lateral sclerosis, primary lateral sclerosis, progressive muscular atrophy, progressive bulbar palsy, and pseudobulbar palsy.

[0055] Neuronal hyperexcitability can occur when receptors for the excitatory neurotransmitter glutamate (glutamate receptors), such as NMDA and AMPA receptors, are overactivated by excess glutamate or by other compounds or neurotransmitters that act on glutamate receptors. Excitotoxicity can result from neuronal hyperexcitability. Excitotoxicity is a pathological process in which nerve cells are damaged or destroyed by excessive stimulation. Excessive stimulation can cause high levels of calcium ions (Ca 2+ ) can enter the cell. 2+ The influx of ATP activates many enzymes, including phospholipases, endonucleases, and proteases, such as calpain, which can damage cellular structures such as components of the cytoskeleton, membranes, and DNA.

[0056] Neuronal hyperexcitability may be involved in spinal cord injury, stroke, traumatic brain injury, hearing loss (due to noise overexposure or ototoxicity), epilepsy, painful neuropathies, attention deficit hyperactivity disorder, autism, central pain syndromes, neurodegenerative diseases, multiple sclerosis, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, frontotemporal dementia, schizophrenia, Rasmussen's encephalitis, Huntington's disease, alcoholism or alcohol withdrawal, particularly rapid benzodiazepine withdrawal, and Huntington's disease. Another common condition that causes excessive glutamate concentrations around neurons is hypoglycemia. Blood glucose is the primary method of removing glutamate from the intersynaptic space of NMDA and AMPA receptor sites.

[0057] As used herein, "non-bicyclic modified sugar moiety" means a modified sugar moiety that includes modifications, such as substituents, that do not form a bridge between two atoms of the sugar to form a second ring.

[0058] As used herein, "nucleobase" refers to an unmodified nucleobase or a modified nucleobase. As used herein, an "unmodified nucleobase" is adenine (A), thymine (T), cytosine (C), uracil (U), or guanine (G). As used herein, a "modified nucleobase" is an atomic group other than unmodified A, T, C, U, or G that can pair with at least one unmodified nucleobase or modified nucleobase. "5-methylcytosine" or "mC" is a modified nucleobase. A universal base is a modified nucleobase that can pair with any one of the five unmodified nucleobases. As used herein, a "nucleobase sequence" refers to the sequential sequence of nucleobases in a nucleic acid or oligonucleotide, independent of any sugar or nucleoside linkage modifications.

[0059] In some embodiments, modified oligonucleotide comprises one or more nucleosides that comprise unmodified nucleobases.In some embodiments, modified oligonucleotide comprises one or more nucleosides that comprise modified nucleobases.In some embodiments, modified oligonucleotide comprises one or more nucleobases that do not comprise nucleobases, which are referred to as abasic nucleosides.

[0060] In some embodiments, the modified nucleobase is selected from 5-substituted pyrimidines, 6-azapyrimidines, alkyl- or alkynyl-substituted pyrimidines, alkyl-substituted purines, and N-2, N-6, and O-6 substituted purines. In some embodiments, modified nucleobases include 2-aminopropyladenine, 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-propynyl (—C≡C—CH3) uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothymine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, especially 5-bromo, 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazaadenine, 3-deazaguanine, 3-deazaadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, promiscuous bases, size-expanded bases, and fluorinated bases. Further modified nucleobases include tricyclic pyrimidines, such as 1,3-diazaphenoxazin-2-one, 1,3-diazaphenothiazin-2-one, and 9-(2-aminoethoxy)-1,3-diazaphenoxazin-2-one (G-clamp). Modified nucleobases can also include bases in which the purine or pyrimidine base is replaced with other heterocycles, such as 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine, and 2-pyridone.Further nucleobases include those disclosed in U.S. Pat. No. 3,687,808, The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, JI, Ed., John Wiley & Sons, 1990, 858-859, Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613, Sanghvi, YS, Chapter 15, Antisense Research and Applications, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993, 273-288, and Chapters 6 and 15 of Antisense Drug Technology, Crooke ST, Ed., CRC Press, 2008, 163-166, and 442-443.

[0061] As used herein, "nucleoside" refers to a compound containing a nucleobase and a sugar moiety. The nucleobase and sugar moiety are each independently unmodified or modified. As used herein, "modified nucleoside" refers to a nucleoside containing a modified nucleobase and / or a modified sugar moiety. Modified nucleosides include abasic nucleosides, which lack a nucleobase. "Linked nucleosides" are nucleosides linked in a contiguous sequence (i.e., there are no additional nucleosides between the linked nucleosides).

[0062] As used herein, "oligomeric compound" refers to an oligonucleotide and, optionally, one or more additional features, such as a conjugate group or a terminal group. An oligomeric compound may or may not be paired with a second oligomeric compound that is complementary to the first oligomeric compound. A "single-stranded oligomeric compound" is an unpaired oligomeric compound. The term "oligomeric duplex" refers to a duplex formed by two oligomeric compounds having complementary nucleobase sequences. Each oligomeric compound in an oligomeric duplex may be referred to as a "duplexed oligomeric compound."

[0063] As used herein, "oligonucleotide" refers to a single strand of linked nucleosides linked via internucleoside linkages, where each nucleoside and internucleoside linkage may be modified or unmodified. The internucleoside linkage may be any linkage described herein. Unless otherwise specified, an oligonucleotide consists of 8 to 50 linked nucleosides. As used herein, "modified oligonucleotide" refers to an oligonucleotide in which at least one nucleoside or internucleoside linkage is modified. As used herein, "unmodified oligonucleotide" refers to an oligonucleotide that does not contain any nucleoside or internucleoside modifications.

[0064] PIKFYVE, also known in the art as "phosphatidylinositol-3-phosphate 5-kinase type III" or "PIPKIII," is a FYVE finger-containing phosphoinositide kinase encoded by the PIKFYVE gene. PIKFYVE is a highly evolutionarily conserved lipid kinase that also possesses protein kinase activity, regulating endomembrane homeostasis and playing a role in the biogenesis of endosomal carrier vesicles from early endosomes. PIKFYVE-mediated conversion of PI3P to PI(3,5)P2 blocks the recruitment of the protein EEA1. This recruitment is blocked because PIP3 is required to form a platform with RAB5 that allows EEA1 to anchor to early endosomes. EEA1 then promotes fusion with endocytic vesicles and other endosomal vesicles.

[0065] As used herein, "PIKFYVE diseases or disorders" include lysosomal degradation diseases and disorders mediated by PIKFYVE. For example, PIKFYVE diseases or disorders include, but are not limited to, amyloid diseases (such as Alzheimer's disease, Parkinson's disease, Huntington's disease, type 2 diabetes, diabetic amyloidosis, and chronic hemodialysis-associated amyloid), multiple sclerosis, and MPS disorders (such as MPS I, MPS II, MPS IIIA, MPS IIIB, MPS IIIC, MPS HID, MPS IVA, MPS IVB, MPS VI, MPS VII, or MPS IX). In some embodiments, the disease is an autoimmune disease (such as multiple sclerosis, rheumatoid arthritis, juvenile chronic arthritis, ankylosing spondylitis, psoriasis, psoriatic arthritis, adult Still's disease, Behçet's syndrome, familial Mediterranean fever, Crohn's disease, leprosy, osteomyelitis, tuberculosis, chronic bronchiectasis, or Castleman's disease), or a CNS disease (such as spongiform encephalopathies (Creutzfeldt-Jakob disease, kuru, or mad cow disease)). The compositions and methods of the disclosure can be used for treating an individual with a lysosomal storage disease, comprising administering to a subject in need of treatment a therapeutically effective amount of a PIKfyve ASO or pharmaceutical composition described herein. In some embodiments, the ASOs and compositions of the disclosure reduce or inhibit the activity of PIKfyve and alter the biogenesis, function, or dynamics of the endosomal or lysosomal system in a manner that reduces or inhibits the activity of PIKfyve and decreases the amount of material abnormally stored in lysosomes in lysosomal storage diseases. In some embodiments, the ASOs and compositions target, reduce, or inhibit the activity of PIKfyve, thus altering the biogenesis, function, or dynamics of the endoplasmic reticulum or Golgi apparatus in a manner that reduces the amount of material abnormally stored in lysosomes in lysosomal storage diseases. In some embodiments, the disease is a neurological disorder.

[0066] As used herein, "sugar moiety" refers to an unmodified sugar moiety or a modified sugar moiety. A superscript prime (') is used to describe the numbering of the sugar of a nucleoside or nucleotide (nucleobase positions are numbered without a prime). When describing the sugar alone, no prime is used. As used herein, "unmodified sugar moiety" refers to a 2-OH(H) furanosyl moiety found in RNA (an "unmodified RNA sugar moiety") or a 2-H(H) moiety found in DNA (an "unmodified DNA sugar moiety"). An unmodified sugar moiety has one hydrogen at each of positions 1, 3, and 4, one oxygen at position 3, and two hydrogens at position 5. As used herein, "modified sugar moiety" or "modified sugar" refers to a modified furanosyl sugar moiety or sugar surrogate. As used herein, a modified furanosyl sugar moiety refers to a furanosyl sugar containing a non-hydrogen substituent in place of at least one hydrogen of an unmodified sugar moiety. In some embodiments, the modified furanosyl sugar moiety is a 2-substituted sugar moiety. Such modified furanosyl sugar moieties include bicyclic and non-bicyclic sugars.

[0067] In some embodiments, the modified sugar moiety is a non-bicyclic modified sugar moiety comprising a furanosyl ring bearing one or more substituents, none of which bridges two atoms of the furanosyl ring to form a bicyclic structure. Such non-bridging substituents may be located at any position on the furanosyl, including, but not limited to, substituents at the 2-, 4-, and / or 5-positions. In some embodiments, one or more of the non-bridging substituents on the non-bicyclic modified sugar moiety is branched. Examples of suitable 2-substituents on non-bicyclic modified sugar moieties include, but are not limited to, 2-F, 2-OCH ("OMe" or "O-methyl"), and 2-O(CH)OCH ("MOE"). In some embodiments, the 2'-substituent is halo, allyl, amino, azido, SH, CN, OCN, CF, OCF, OC, or OC. 1-10 Alkoxy, OC 1-10 Substituted alkoxy, OC 1-10 Alkyl, OC 1-10 Substituted alkyl, S-alkyl, N(R m)-alkyl, O-alkenyl, S-alkenyl, N(R m )-alkenyl, O-alkynyl, S-alkynyl, N(Rm)-alkynyl, O-alkyleneyl-O-alkyl, alkynyl, alkaryl, aralkyl, O-alkaryl, O-aralkyl, O(CH2)2SCH3, O(CH2)2ON(R m )(R n ) or OCH2C(=O)-N(R m )(R n ), wherein each R m and R n are independently H, an amino protecting group, or a substituted or unsubstituted C 1-10 and alkyl, wherein the 2-substituent may be further substituted independently with one or more substituents selected from hydroxyl, amino, alkoxy, carboxy, benzyl, phenyl, nitro(NO), thiol, thioalkoxy, thioalkyl, halogen, alkyl, aryl, alkenyl, and alkynyl. Examples of suitable 4'-substituents for non-bicyclic modified sugar moieties include, but are not limited to, alkoxy (e.g., methoxy), and alkyl. Examples of suitable 5-substituents for non-bicyclic modified sugar moieties include, but are not limited to, 5-methyl (R or S), 5-vinyl, and 5-methoxy. In some embodiments, non-bicyclic modified sugar moieties include multiple non-bridging sugar substituents, such as 2-F-5-methyl sugar moieties.

[0068] In some embodiments, the 2'-substituted non-bicyclic modified nucleoside is selected from the group consisting of F, NH, N, OCF, OCH, O(CH)NH, CHCH=CH, OCHCH=CH, OCHCHOCH, O(CH)SCH, O(CH)ON(R m )(R n ), O(CH2)2O(CH2)2N(CH3)2, and N-substituted acetamides (OCH2C(=O)-N(R m )(R n )) wherein each R m and R n are independently H, an amino protecting group, or a substituted or unsubstituted C1-10 It is alkyl.

[0069] In some embodiments, the 2'-substituted nucleoside non-bicyclic modified nucleoside comprises a sugar moiety that includes a non-linear 2'-substituent selected from F, OCF3, OCH3, OCH2CH2OCH3, O(CH2)2SCH3, O(CH2)2ON(CH3)2, O(CH2)2O(CH2)2N(CH3)2, and OCH2C(=O)-N(H)CH3 ("NMA").

[0070] In some embodiments, the 2'-substituted non-bicyclic modified nucleoside comprises a sugar moiety comprising a non-bridging 2'-substituent selected from F, OCH3, and OCH2CH2OCH3.

[0071] Certain modified sugar moieties include a substituent that bridges two atoms of a furanosyl ring to form a second ring, resulting in a bicyclic sugar moiety. In some embodiments, the bicyclic sugar moiety includes a bridge between four and two furanose ring atoms. Examples of such four to two bridged sugar substituents include 4-CH2-2, 4-(CH2)2-2, 4-(CH2)3-2, 4-CH2-O-2 ("LNA"), 4-CH2-S-2, 4-(CH2)2-O-2 ("ENA"), 4-CH(CH3)-O-2 (referred to as "constrained ethyl" or "cEt"), 4-CH2-O-CH2-2, 4-CH2-N( R)-2, 4-CH(CH2OCH3)-O-2 ("constrained MOE" or "cMOE") and its analogs, 4-C(CH3)(CH3)-O-2 and its analogs, 4-CH2-N(OCH3)-2 and its analogs, 4-CH2-ON(CH3)-2, 4-CH2-C(H)(CH3)-2, 4-CH2-C(=CH2)-2 and its analogs, 4-C(R a R b )-N(R)-O-2,4-C(R a R b )-ON(R)-2, 4-CH2-ON(R)-2, and 4-CH2-N(R)-O-2, wherein each R, R a , and R b are independently H, a protecting group, or C 1-12It is alkyl.

[0072] In some embodiments, such 4 to 2 bridges are independently —[C(R a )(R b )] n -, -[C(R a )(R b )] n -O-, -C(R a )=C(R b )-, -C(R a )=N-, -C(=NR a )-, -C(=O)-, -C(=S)-, -O-, -Si(R a )2-, -S(=O) x- , and -N(R a )-, x is 0, 1, or 2, n is 1, 2, 3, or 4, and each R a and R b are independently H, a protecting group, a hydroxyl, or C 1-12 Alkyl, substituted C 1-12 Alkyl, C 1-12 Alkenyl, substituted C 2-12 Alkenyl, C 2-12 Alkynyl, substituted C 2-12 Alkynyl, C 5-20 Aryl, substituted C 5-20 Aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C 5-7 Alicyclic radicals, substituted C5-7 alicyclic radicals, halogen, OJ1, NJ1J2, SJ1, N3, COOJ1, acyl (C(=O)-H), substituted acyl, CN, sulfonyl (S(=O)2-J1), or sulfoxyl (S(=O)-J1), each J1 and J2 independently being H, C 1-12 Alkyl, substituted C 1-12 Alkyl, C 2-12 Alkenyl, substituted C 2-12 Alkenyl, C 2-12 Alkynyl, substituted C 2-12 Alkynyl, C 5-20 Aryl, substituted C 5-20 Aryl, acyl (C(=O)-H), substituted acyl, heterocyclic radical, substituted heterocyclic radical, C1-12 Aminoalkyl, substituted C 1-12 aminoalkyl, or a protecting group.

[0073] The addition of the bicyclic sugar moiety is a well-known technical field, and the technical field is well-known. al., Chem. Commun., 1998, 4, 455-456; Koshkin et al., Tetrahedron, 1998, 54, 3607-3630; Kumar et al., Bioorg. Med. Chem. Lett., 1998, 8, 2219-2222; Singh et al. al., J.Org.Chem., 1998, 63, 10035-10039; Srivastava et al., J.Am.Chem.Soc., 20017, 129, 8362-8379; Wengel et al., U.S. Patent No. 7,053,207; Imanishi et al., U.S. Patent No. 6,268,490; Imanishi et al., U.S. Patent No. 6,770,748; Imanishi et al., USRE44,779; Wengel et al., U.S. Patent No. 6,794,499; Wengel et al., U.S. Patent No. 6,670,461; Wengel et al., U.S. Patent No. 7,034,133; Wengel et et al., U.S. Patent No. 8,080,644; Wengel et al., U.S. Patent No. 8,034,909; Wengel et al., U.S. Patent No. 8,153,365; Wengel et al., U.S. Patent No. 7,572,582; and Ramasamy et al., U.S. Patent No. 6,525,191; Torsten et al., WO2004 / 106356; Wengel et al., WO1999 / 014226; Seth et al., WO2007 / 134181; Seth et al., U.S. Patent No. 7,547,684; Seth et al., U.S. Patent No. 7,666,854; Seth et al., U.S. Patent No. 8,088,746; Seth et Seth et al., U.S. Patent No. 7,750,131; Seth et al., U.S. Patent No. 8,030,467; Seth et al., U.S. Patent No. 8,268,980; Seth et al.See, e.g., U.S. Patent No. 8,546,556; Seth et al., U.S. Patent No. 8,530,640; Migawa et al., U.S. Patent No. 9,012,421; Seth et al., U.S. Patent No. 8,501,805; and Allerson et al., U.S. Patent Publication No. US2008 / 0039618 and Migawa et al., U.S. Patent No. US2015 / 0191727.

[0074] As used herein, "subject" and "patient" refer to any vertebrate, including, but not limited to, mammals (e.g., cows, pigs, camels, llamas, horses, goats, rabbits, sheep, hamsters, guinea pigs, cats, dogs, rats, and mice, non-human primates (e.g., monkeys such as cynomolgus or rhesus monkeys, chimpanzees, etc.), and humans). In some embodiments, the subject may be human or non-human. In some embodiments, the subject or patient is human. The subject or patient may also undergo other forms of treatment. In some embodiments, the patient has a neurological disorder due to a mutation in the C9ORF72 gene (e.g., the patient may be haploinsufficient for the C9ORF72 gene (e.g., reducing C9ORF72 protein activity by 50% or more), or the C9ORF72 gene contains a GGGGCC repeat expansion (e.g., (GGGGCC) within C9ORF72). n (SEQ ID NO: 1565) hexanucleotide extension). The variable "n" may be at least 30.

[0075] As used interchangeably herein, "therapeutically effective amount," "effective dose," or "effective amount," unless otherwise defined, refers to a dose of a drug effective over a period of time necessary to achieve a desired therapeutic result. An effective dose may be determined by one of ordinary skill in the art and may vary depending on factors such as the individual's disease state, age, sex, and weight, as well as the ability of the drug to elicit a desired response in the individual. As used herein, the term may also refer to an amount effective to produce a desired in vivo effect in an animal, mammal, or human, such as reducing and / or inhibiting receptor function. A therapeutically effective amount may be administered in one or more administrations (e.g., the agent may be administered as a preventative treatment or therapeutically at any stage of disease progression, before or after symptoms appear, etc.), application, or dose, and is not intended to be limited to a particular formulation, combination, or route of administration. It is within the scope of the present disclosure that a drug may be administered at various times during the course of treatment of a subject. The administration time and dose used will depend on several factors, such as the goal of treatment (e.g., treatment or prevention), the condition of the subject, and can be readily determined by one of ordinary skill in the art.

[0076] As used herein, the terms "treating" or "treating" a subject refers to administering a composition or agent described herein to a subject so that at least one symptom of a disease or disorder is cured, alleviated, mitigated, altered, treated, reduced, ameliorated, or improved. Treatment includes administering an amount effective to alleviate, alleviate, alter, treat, reduce, ameliorate, and / or improve one or more symptoms associated with a disease or disorder. The treatment may prevent the worsening or exacerbation of symptoms associated with a disease or disorder.

[0077] The therapeutic methods described herein may include administering to a subject in need thereof a composition comprising an effective amount of one or more antisense oligonucleotides that treats a neurological disease by inhibiting or suppressing PIKFYVE expression. The one or more antisense oligonucleotides may reduce or inhibit neurodegeneration. The one or more antisense oligonucleotides may reduce neuronal hyperexcitability. Reducing PIKFYVE mRNA and PIKFYVE protein levels inhibits neurodegeneration by promoting toxic TDP-43 aggregates, DPR aggregates (e.g., in C9ORF72-ALS patients), and TDP-43 nuclear retention. Delivery of an ASO targeting PIKFYVE mRNA described herein reduces PIKFYVE protein levels.

[0078] The composition may inhibit kinase activity by inhibiting kinase expression. The composition of the present invention may inhibit the activity or expression of PIKFYVE kinase. One or more antisense oligonucleotides may be combined with a small molecule therapeutic agent (e.g., apilimod and / or YM201636).

[0079] The present disclosure provides oligonucleotides (modified or unmodified) that can be used to regulate PIKFYVE expression. Table 1 shows the general base sequences (5' to 3') for PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. Table 6 shows the modified base sequences (5' to 3') for PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. The unmodified sequences in Table 6 are further contemplated as the general base sequences for PIKFYVE antisense oligonucleotides or inhibitory nucleic acids of the present disclosure. The ASO sequences of SEQ ID NOS: 1-49 are predicted to efficiently skip exon 3 of PIKFYVE, generating a stop codon that causes premature termination of PIKFYVE translation, as shown in Figures 1A and 1B. The ASO sequences of SEQ ID NOS: 50-80 are predicted to efficiently skip exon 5 of PIKFYVE, causing a major structural change in the protein, rendering it non-functional, as shown in Figures 2A and 2B. ASO sequences SEQ ID NOS: 81-328, 330-373, 376-420, and 423-479 have low acute neurotoxicity scores as measured by the method described in Hagedorn et al., Nucleic Acid Therapeutics, 2022, 32(3):151-162 (DOI: 10.1089 / nat / 2021 / 0071). Unmodified versions of ASO sequences SEQ ID NOS: 307, 308, 330, 424, 480-540, 542-619, 621-685, 688-782, and SEQ ID NOS: 1646-2321 have low predicted off-target binding sites as measured by the GGGenome search tool using two mismatches / gaps. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] Table 1-5 Table 1-6 Table 1-7 Table 1-8 Table 1-9 Table 1-10 Table 1-11 Table 1-12 Table 1-13 Table 1-14 Table 1-15 Table 1-16 Table 1-17 Table 1-18 Table 1-19 Table 1-20 Table 1-21

[0080] In some aspects, the disclosure provides modified oligonucleotides consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleotide bases of any of the nucleobase sequences of SEQ ID NOs: 1-782 in Table 1, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. In some embodiments, the present disclosure provides a nucleic acid sequence comprising 12 to 30 linked nucleosides and SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790, 900, 901, 902, 903, 904, 905, 906, 907, 908, 909, 910, 911, 912, 913, 914, 915, 916, 917, 918, 919, 920, 921, 922, 923, 924, 925, 926, 927, 928, 929, 930, 931, 932, 933, 934, 935, 936, 937, 938, 939, 940, 941, 942, 943, 94 , 772, 781, or as well as SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826- 1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930,1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2048 055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-216 and 2319-2321.

[0081] In some embodiments, the modified oligonucleotide is at least 80% to 100% identical (i.e., 80%, 82%, 84%, 86%, 88%, 90%, 92%, 94%, 96%, 98%, or 100%, or any numerical range or value between any of the foregoing values) to any of the sequences comprising or consisting of unmodified versions of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and SEQ ID NOs: 1646-2321. In some embodiments, the modified oligonucleotides are selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1761 1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1863-1864, 1865-1866, 1866-1867, 1868-1869, 1870-1871, 1872-1873, 1874-1875, 1876-1877, 1878-1879, 1880-1881, 1882-1883, 1884-1885, 1886-1887, 1888-1889, 1890-1891, 1892-1893, 1894-1895, 1896-1900, 1902-1903, 1904-1905, 1906-1907, 1908-1910, 1911-1912, 1913-1914, 1915-1916, 1917-1918, 1919-1920, 192 862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994,1996~1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095 , 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183 , 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

[0082] Unmodified versions of the sequences shown in Table 1 and the sequences in Table 6 can be used to design antisense molecules for the inhibition of PIKFYVE expression. For example, gapmer oligonucleotides can be designed using unmodified versions of the sequences in Table 1 and the sequences in Table 6 and can include a 5' wing of about 3-5 nucleotides, a 3' wing of about 3-5 nucleotides, and a gap region comprising 8-12 consecutive deoxyribonucleosides of any one of the sequences in Table 1 and the unmodified versions of the sequences in Table 6.

[0083] In some embodiments, oligonucleotides of the present disclosure comprise gapmers having a gap segment (adjacent to 5' and 3' wing segments) of at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 contiguous nucleotide bases of the nucleobase sequence of any of SEQ ID NOS: 1-782 in Table 1, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and an unmodified version of SEQ ID NOS: 1646-2321, where the gap segment is positioned between the 5' and 3' wing segments and each wing segment comprises a modified sugar. In some embodiments, the gap segment is 8 to 10 nucleosides in length and each wing segment is 3 to 5 modified nucleosides in length. In some embodiments, oligonucleotides of the disclosure comprise modified sugars and a 5' wing segment having the first 3 to 5 nucleobases of the nucleobase sequence of any of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321, followed by a 5' wing segment having the first 3 to 5 nucleobases of any of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and unmodified versions of SEQ ID NOs: 1646-2321. and a 3' wing segment having the nucleobase sequence of the last 3 to 5 nucleobases of the same sequence corresponding to an unmodified version of SEQ ID NOs: 1-782, e.g., 1-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and SEQ ID NOs: 1646-2321. Tables 2 and 6 show MOE gapmers of the present disclosure.

[0084] In some embodiments, the oligonucleotides of the disclosure are selected from the group consisting of SEQ ID NOS: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781 in Table 1, as well as SEQ ID NOS: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1690, 1691, 1692, 1693, 1694, 1695, 1696, 1697, 1698, 1699, 2000, 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2100, 2110, 2111, 2112, 2113, 2114, 2115, 2116, 2117, 2118, 2119, 2200, 2219, 2221, 2222, 2230, 2231, 2232, 692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1 760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1823 824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895- 1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985~1994, 1996~1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177,and 2319-2321. These nucleobase sequences include gapmers having at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 gap segments (adjacent to 5' and 3' wing segments) of at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 of the contiguous nucleotide bases of any of the unmodified versions of the nucleobase sequences of 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the gap segment is positioned between the 5' and 3' wing segments and each wing segment comprises a modified sugar. In some embodiments, the gap segment is 8-10 nucleosides in length and each wing segment is 3-5 modified nucleosides in length. In some embodiments, the oligonucleotides of the present disclosure comprise modified sugars and are selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-743, 746-753, 754-763, 765-770, 766-780, 771-782, 772-784, 773-790, 775-800, 776-810, 777-811, 778-822, 779-823, 780-824, 781-825, 782-826, 783-827, 784-828, 785-830, 786-831, 787-832, 788-833, 789-834, 790-901, 801-812, 802-813, 803-814, 804-815, 805-816, 806-817, 807-818, 808-819, 819-824, 819-825, 826-827, 828-831, 829-8 734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1778 79, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843,1845~1853, 1855~1858, 1861~1862, 1864, 1866, 1868~1869, 1871~1881, 1883, 1885~1892, 1895~1897, 1899, 1901~1907, 1909, 1911~1913, 1915, 1917, 1919~1927, 1930, 1932~1936, 1938~1940, 1943, 1945~1953, 1955~1969, 1971~1973, 1976~1980, 1985~1994, 1996~1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183, 2185, 2187~2205, 2207~2216, 2218, 2220~ 2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and an unmodified version of 2319-2321, followed by a 5' wing segment having the nucleobase sequence of the first 3 to 5 nucleobases of any of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 753, 754, 755, 756, 757, 758, 759, 760, 761, 762, 763, 764, 765, 766, 767, 768, 769, 770, 771, 772, 773, 774, 775, 776, 777, 778, 779, 780, 781, 782, 783, 784, 785, 786, 787, 788, 789, 790, 800, 801, 802, 803, 804, 805, 806, 807, 808, 809, 810, 9, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775,1777~1779, 1782~1784, 1787~1788, 1791~1793, 1795, 1797~1802, 1804~1805, 1809~1811, 1813~1815, 1818~1819, 1822~1824, 1826~1827, 1830~1832, 1836~1837, 1840~1843, 1845~1853, 1855~1858, 1861~1862, 1864, 1866, 1868~1869, 1871~1881, 1883, 1885~1892, 1895~1897, 1899, 1901~1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999- 2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, The subsequent 8 to 12 unmodified amino acids of the same sequence correspond to the unmodified versions of 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321. a gap of modified nucleotides followed by a modified sugar, and including SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689;1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1818 15, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945 ~1953, 1955~1969, 1971~1973, 1976~1980, 1985~1994, 1996~1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2125~2126, 2127~2128, 2129~2130, 2131~2132, 2133~2134, 2135~2136, 2137~2138, 2139~2140, 2141~2142, 2143~2144, 2145~2146, 2147~2148, 2149~2150, 2151~2152, 2153~2154, 2155~2156, 2157~2158, 2159~2160, 2161~2162, 2163~2164, 2165~2166, 2167~2168, 2170~2171, 2 and 3' wing segments having nucleobase sequences of the last 3 to 5 nucleobases of the same sequence corresponding to unmodified versions of 126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321. Tables 2 and 6 show MOE gapmers of the present disclosure.

[0085] The 5' and / or 3' wings may comprise the following compounds: 2'-OMe, 2'-MOE, LNA, or DNA, used alone or in combination with each other. The backbone linkages of the 5' and / or 3' wings may be phosphorothioate or a mixture of phosphodiester and phosphorothioate. The linkages in the gap region may be phosphorothioate.

[0086] In some embodiments, the oligonucleotide is single-stranded, hi some embodiments, the oligonucleotide comprises or is complexed with a moiety that neutralizes the charge on the oligonucleotide to facilitate uptake and transport across a cell membrane.

[0087] In some embodiments, the ASO sequences of SEQ ID NOs: 1-80 in Table 1 contain the following 5-8-5 motif: 2MOE * 2MOE-2MOE-2MOE-2MOE-N * N * N * N * N * N * N * N * 2MOE-2MOE-2MOE * 2MOE * 2MOE, (i) 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, and (iii) an asterisk ( * ) indicates a phosphorothioate bond, and (iv) a dash (-) indicates a phosphodiester bond. Table 2 below illustrates this motif on SEQ ID NOs: 1-80 (here, SEQ ID NOs: 783-862). In some embodiments, the ASO sequences of SEQ ID NOs: 783-831 efficiently skip exon 3 of PIKFYVE. In some embodiments, the ASO sequences of SEQ ID NOs: 832-862 efficiently skip exon 5 of PIKFYVE.

[0088] In some embodiments, the ASO sequences of unmodified versions of SEQ ID NOs: 1-782 in Table 1, e.g., 81-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782, and SEQ ID NOs: 1646-2321, contain the following 5-10-5 motif: 2MOE * 2MOE-2MOE-2MOE-2MOE-N * N * N * N * N * N * N * N * N * N * 2MOE-2MOE-2MOE * 2MOE * 2MOE, (i) 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, and (iii) an asterisk ( * (iv) a dash (-) indicates a phosphorothioate bond, and (iv) a dash (-) indicates a phosphodiester bond. Table 2 below shows this motif on SEQ ID NOs: 1-782, e.g., 81-328, 330-373, 376-420, 423-540, 542-619, 621-685, 688-782 (here, SEQ ID NOs: 783-1564, e.g., 863-1110, 1112-1155, 1158-1202, 1205-1322, 1324-1401, 1403-1467, 1470-1564).

[0089] In some embodiments, the amino acids selected from SEQ ID NOS: 81-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781 in Table 1, as well as SEQ ID NOS: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1699, 2000-2001, 2002-2003, 2004-2005, 2006-2007, 2008-2009, 2010-2011, 2012-2013, 2014-2015, 2016-2017, 2018-2019, 2020-2021, 2022-2023, 2024-2025, 2026-2027, 2028-2029, 2030-2031, 2030-2032, 2030-2033, 2030-203 1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770~1775, 1777~1779, 1782~1784, 1787~1788, 1791~1793, 1795, 1797~1802, 1804~1805, 1809~1811, 1813~1815, 1818~1819, 1822~1824, 1826~1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901- 1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996- 1997, 1999~2010, 2012~2015, 2017, 2019~2021, 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183, 2185,The unmodified versions of the ASO sequences 2187–2205, 2207–2216, 2218, 2220–2232, 2234, 2236–2245, 2247–2268, 2270–2289, 2291–2300, 2302, 2304–2316, and 2319–2321 contain the following 5-10-5 motifs: 2MOE, * 2MOE-2MOE-2MOE-2MOE-N * N * N * N * N * N * N * N * N * N * 2MOE-2MOE-2MOE * 2MOE * 2MOE, (i) 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), (ii) N is a nucleobase, and (iii) an asterisk ( * ) indicates a phosphorothioate bond, and (iv) a dash (-) indicates a phosphodiester bond.

[0090] Table 2: Base sequence in PIKFYVE antisense oligonucleotide (ASO) (gapmer design: 5'-5 2'-methoxyethyl ribose nucleotides-10 DNA nucleotides-5 2'-methoxyethyl ribose nucleotides-3', uppercase letters indicate 2'-methoxyethyl ribose nucleosides, lowercase letters indicate DNA nucleosides, asterisks ( * ) are phosphorothioate linkages, and linkages without an asterisk are phosphodiester linkages) (note that the table below shows 2'MOE wings, but alternative wings including 2'-OMe or LNA (locked nucleic acid) are also envisioned). [Table 2-1] [Table 2-2] [Table 2-3] Table 2-4 Table 2-5 Table 2-6 Table 2-7 Table 2-8 Table 2-9 Table 2-10 Table 2-11 Table 2-12 Table 2-13 Table 2-14 Table 2-15 Table 2-16 Table 2-17 Table 2-18 Table 2-19 Table 2-20 [Table 2-21]

[0091] In some embodiments, the ASO sequences of SEQ ID NOs: 1-49 in Table 1 contain the following motif: 2MOE * 2MOE-2MOE-2MOE-2MOE-2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE-2MOE-2MOE * 2MOE * 2MOE, (i) the 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), and (ii) an asterisk ( * ) indicates a phosphorothioate bond, and (iii) a dash (-) indicates a phosphodiester bond. Table 3 below illustrates this motif on SEQ ID NOS: 1-49 (here, SEQ ID NOS: 1566-1614). In some embodiments, the ASO sequences in Table 3 efficiently skip exon 3 of PIKFYVE. In some embodiments, the ASO sequences set forth in SEQ ID NOS: 1-49 efficiently skip exon 3 of PIKFYVE.

[0092] Table 3: Base sequence in PIKFYVE antisense oligonucleotide (ASO) (Design: 5'-18 2'-methoxyethyl ribose nucleotides-3', capital letters indicate 2'-methoxyethyl ribose nucleosides, asterisks ( * ) are phosphorothioate linkages, and linkages without an asterisk are phosphodiester linkages) (note that the table below shows 2'MOE wings, but alternative wings including 2'-OMe or LNA (locked nucleic acid) are also envisioned). [Table 3-1] [Table 3-2]

[0093] In some embodiments, the ASO sequences of SEQ ID NOs: 50-80 in Table 1 contain the following motif: 2MOE * 2MOE-2MOE-2MOE-2MOE-2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE * 2MOE-2MOE-2MOE * 2MOE * 2MOE, (i) the 2MOE is a nucleobase having a 2'-OCH2CH2-OCH3 group (i.e., 2'-MOE), and (ii) an asterisk ( * ) indicates a phosphorothioate bond, and (iii) a dash (-) indicates a phosphodiester bond. Table 4 below shows this motif on SEQ ID NOs: 50-80 (here, SEQ ID NOs: 1615-1645). In some embodiments, the ASO sequences in Table 4 efficiently skip exon 5 of PIKFYVE. In some embodiments, the ASO sequences set forth in SEQ ID NOs: 50-80 efficiently skip exon 5 of PIKFYVE.

[0094] Table 4: Base sequence in PIKFYVE antisense oligonucleotide (ASO) (Design: 5'-18 2'-methoxyethyl ribose nucleotides-3', capital letters indicate 2'-methoxyethyl ribose nucleosides, asterisks ( * ) are phosphorothioate linkages, and linkages without an asterisk are phosphodiester linkages) (note that the table below shows 2'MOE wings, but alternative wings including 2'-OMe or LNA (locked nucleic acid) are also envisioned). [Table 4]

[0095] The PIKFYVE kinase antisense or inhibitory nucleic acids of the present disclosure can inhibit the expression and thus activity associated with PIKFYVE. The PIKFYVE kinase antisense or inhibitory nucleic acids can include any combination of oligonucleotides set forth in Tables 1, 2, 3, 4, 5, and 6, and sequences that are 98% to 99% identical thereto.

[0096] The PIKFYVE ASOs described herein, such as those set forth in SEQ ID NOs: 783-1564, e.g., 783-1110, 1112-1155, 1158-1202, 1205-1322, 1324-1401, 1403-1467, 1470-1564, and 1566-2321, suppress expression of PIKFYVE mRNA while minimizing off-target binding.

[0097] In some embodiments, SEQ ID NOs: 783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, and 1566-2321 The PIKFYVE ASOs described herein, such as those set forth in SEQ ID NOs: 863-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, and 1563, suppress PIKFYVE mRNA expression with minimal off-target binding.

[0098] Therapeutic methods can include any number of ways of administering the disclosed compositions. Administration methods can include aqueous, lipid, oily, or other solutions, emulsions such as simulated cerebrospinal fluid solutions, oil-in-water emulsions, liposomes, aqueous, or oily suspensions. Typically, the ASOs of the present disclosure are administered directly to the CNS of the subject. Therefore, the formulations or compositions are sterile, and more preferably suitable for injection. The following formulations and methods are merely examples and are in no way limiting.

[0099] Preparations suitable for parenteral administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostatic agents, and solutes that make the preparation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions, which may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives.Preparations may be provided in unit-dose or multi-dose sealed containers such as ampoules and vials, and may be stored as liquids that only require the addition of sterile liquid excipients, such as water, immediately before use for injection, or in freeze-dried (lyophilized) form.Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets.Preparations may also be provided in pre-filled syringes.

[0100] The additional therapeutic agent(s) may be administered simultaneously or sequentially with one or more of the disclosed antisense or inhibitory nucleic acids and compositions. Sequential administration includes administration before or after one or more of the disclosed antisense or inhibitory nucleic acids or compositions. In some embodiments, the additional therapeutic agent(s) may be administered in the same composition as one or more of the disclosed antisense or inhibitory nucleic acids. In some embodiments, there may be a time interval between the administration of the additional therapeutic agent and the administration of one or more of the disclosed antisense or inhibitory nucleic acids. In some embodiments, administering an additional therapeutic agent together with one or more of the disclosed antisense or inhibitory nucleic acids may allow for lower doses and / or less frequent administration of the other therapeutic agent(s). When used in combination with one or more other active ingredients, one or more of the disclosed antisense or inhibitory nucleic acids and the other active ingredients may each be used in lower doses than when used alone. Thus, the pharmaceutical compositions of the present disclosure include those containing one or more of the disclosed antisense or inhibitory nucleic acids as well as one or more other active ingredients. The above combination includes the combination of one or more antisense or inhibitory nucleic acids of the present disclosure with not only one other active compound, but also two or more other active compounds.For example, the compound of the present disclosure can be combined with various drugs for treating neurological diseases.Antisense oligonucleotides can also be covalently linked with other oligonucleotides, for example, targets other than PIKFYVE.Antisense oligonucleotides can also be covalently linked with antibodies.

[0101] One or more of the disclosed antisense or inhibitory nucleic acids can be combined with drugs including, but not limited to, anticholinergics, anticonvulsants, antidepressants, benzodiazepines, decongestants, muscle relaxants, analgesics, and / or stimulants. Additional types of therapies and treatments include, but are not limited to, digital communication devices, feeding tubes, mechanical ventilation, nutritional support, deep brain stimulation, occupational therapy, physical therapy, and / or speech therapy.

[0102] The disclosed composition(s) may be incorporated into a pharmaceutical composition suitable for administration to a subject (e.g., a patient, which may be human or non-human). The pharmaceutical composition may include a carrier (e.g., a pharmaceutically acceptable carrier). Any suitable carrier may be used in connection with the present disclosure, and such carriers are well known in the art. The choice of carrier will be determined, in part, by the particular use of the composition (e.g., administration to an animal) and the particular method used to administer the composition. Accordingly, there are a wide variety of suitable formulations of the compositions of the present disclosure.

[0103] The pharmaceutical composition may contain a therapeutically effective amount or a prophylactically effective amount of antisense oligonucleotide. The therapeutically effective amount of a composition may be determined by a person skilled in the art and may vary depending on factors such as the individual's disease state, age, sex, and weight, as well as the ability of the composition to induce a desired response in the individual. A therapeutically effective amount is also an amount in which the toxic or harmful effects of one or more antisense or inhibitory nucleic acids of the present disclosure outweigh the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at a dosage and for a period of time necessary, to achieve the desired preventative result. Typically, a prophylactic dose is used in subjects before or at an early stage of disease, so the prophylactically effective amount will be less than the therapeutically effective amount.

[0104] Pharmaceutical compositions may contain one or more pharmaceutically acceptable carriers.As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid, or liquid filler, diluent, encapsulating material, or any kind of formulation aid.Some examples of substances that can function as pharmaceutically acceptable carriers include sugars, such as but not limited to lactose, glucose, and sucrose; starches, such as but not limited to corn starch and potato starch; cellulose and its derivatives, such as but not limited to sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; excipients, such as but not limited to powdered tragacanth, malt, gelatin, talc, cocoa butter, and suppository wax; peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. Oils, including but not limited to, glycols such as propylene glycol, esters, such as but not limited to ethyl oleate and ethyl laurate, buffers, including but not limited to agar, magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and phosphate buffer, as well as other non-toxic compatible lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as release agents, coating agents, preservatives, and antioxidants may also be present in the composition, at the discretion of the formulator.

[0105] The route of administration of one or more of the disclosed antisense or inhibitory nucleic acids and the form of the composition will determine the type of carrier used.

[0106] The pharmaceutical compositions of the present disclosure can be administered in a number of ways, depending on whether local or systemic treatment is desired and the area to be treated. Administration can be parenteral, including intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial, e.g., intrathecal, intraventricular, or intraventricular administration. In some embodiments, the antisense or inhibitory nucleic acid is administered as an intravenous injection, intraperitoneal injection, or bolus injection, or administered directly to the target organ. In some embodiments, the antisense or inhibitory nucleic acid is administered intrathecally or intraventricularly as a bolus injection.

[0107] Carriers for systemic administration generally include at least one of a solvent, diluent, lubricant, binder, disintegrant, colorant, flavoring, sweetener, antioxidant, preservative, glidant, vehicle, suspending agent, wetting agent, surfactant, combinations thereof, etc. All carriers are optional in the composition.

[0108] Suitable diluents include sugars such as glucose, lactose, dextrose, and sucrose; diols such as propylene glycol; calcium carbonate, sodium carbonate, sugar alcohols such as glycerin, mannitol, and sorbitol.

[0109] Suitable lubricants include liquid lubricants such as silica, talc, stearic acid and its magnesium and calcium salts, calcium sulfate, and polyethylene glycol, as well as vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and theobroma oil. The amount of lubricant(s) in a systemic or topical composition is typically about 5 to about 10%.

[0110] Suitable binders include polyvinylpyrrolidone, magnesium aluminum silicate, starches such as corn starch and potato starch, gelatin, tragacanth, and cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose, and sodium carboxymethylcellulose. The amount of binder(s) in a systemic composition is typically about 5 to about 50%.

[0111] Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixture, croscarmellose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays, and ion exchange resins. The amount of one or more disintegrant(s) in the systemic composition generally ranges from about 0.1% to about 10%.

[0112] Suitable coloring agents include colorants such as the FD&C dyes. If used, the amount of coloring agent in a systemic or topical composition is typically about 0.005 to about 0.1%.

[0113] Suitable flavorings include menthol, peppermint, and fruit flavors. When used in systemic or topical compositions, the amount of flavoring(s) is typically about 0.1 to about 1.0%.

[0114] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of antioxidant(s) in a systemic or topical composition is typically from about 0.1 to about 5%.

[0115] Suitable preservatives include benzalkonium chloride, methylparaben, and sodium benzoate. The amount of preservative(s) in a systemic or topical composition is typically about 0.01 to about 5%.

[0116] Suitable lubricants include silicon dioxide. The amount of lubricant(s) in a systemic or topical composition is typically about 1 to about 5%.

[0117] Suitable solvents include water, isotonic saline, ethyl oleate, glycerin, hydroxylated castor oil, alcohols such as ethanol, and phosphate buffer solutions. The amount of solvent in a systemic or topical composition is typically from about 0 to about 100%.

[0118] Suitable suspending agents include AVICEL RC-591 (FMC Corporation, Philadelphia, PA) and sodium alginate. The amount of suspending agent(s) in a systemic or topical composition is typically about 1 to about 8%.

[0119] Suitable surfactants include lecithin, polysorbate 80, and sodium lauryl sulfate, including TWEEN® manufactured by Atlas Powder Company of Wilmington, Delaware. Suitable surfactants include those disclosed in CTFA Cosmetic Ingredient Handbook, 1992, pp. 587-592; Remington's Pharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon's Volume 1, Emulsifiers & Detergents, 1994, North American Edition, pp. 236-239. The amount of surfactant(s) in a systemic or topical composition is typically about 0.1% to about 5%.

[0120] Compositions and formulations for parenteral, intrathecal, intraventricular, or intracerebroventricular administration may also include sterile aqueous solutions, which may further contain other suitable additives, including, but not limited to, buffers, diluents, and penetration enhancers, carrier compounds, and other pharmaceutically acceptable carriers or excipients. For example, an intrathecal cerebrospinal fluid (CSF) catheter can be used to deliver the antisense formulations of the present disclosure. The catheter can be inserted into the L3 or L4 vertebra. The distal end of the catheter is advanced intrathecally to approximately the LI vertebra. The antisense oligonucleotide is dissolved in saline, sterilized by filtration, and administered at 0.33 ml / min in a volume of 1.0 ml, followed by a 0.5 ml rinse with sterile water. The total infusion time is 4.5 minutes.

[0121] Compositions for parenteral administration usually contain 0.1% to 10% of active substance and 90% to 99.9% of carriers, including diluents and solvents.

[0122] The amount of carrier used with the disclosed compounds is sufficient to provide a practical amount of composition for administration per unit dose of drug.Techniques and compositions for preparing dosage forms useful in the disclosed methods are described in the following references:Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd Ed., (1976).

[0123] In vivo testing of candidate antisense or inhibitory nucleic acids may be performed by means known to those skilled in the art. For example, one or more candidate antisense or inhibitory nucleic acids may be administered to a mammal, such as a mouse or rabbit. A dose of the candidate antisense or inhibitory nucleic acid may be administered to the mammal by any route deemed appropriate. The animal may then be monitored for signs of reduced or improved motor neuron activity and / or PIKFYVE gene or protein expression or activity, respectively, using conventional methods and criteria. If desired, results obtained in the presence of the candidate antisense or inhibitory nucleic acid may be compared with results in control animals not treated with the candidate antisense or inhibitory nucleic acid. Administration tests may be performed using or in conjunction with the methods described herein to identify one or more antisense or inhibitory nucleic acids capable of treating a neurological disorder and / or to perform any subsequent testing of the candidate antisense or inhibitory nucleic acid in vivo. Those skilled in the art will be able to determine the appropriate dose of one or more antisense or inhibitory nucleic acids. The dose may be determined by monitoring the subject for signs of disease suppression or improvement. The dose may be increased or decreased to achieve the desired treatment frequency. The toxicity and efficacy of one or more antisense or inhibitory nucleic acids can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, by determining the lethal dose (LD50) for 50% of a population and the therapeutically effective dose (ED50) for 50% of a population. The dose ratio LD50 / ED50 is the therapeutic index, which indicates the ratio between toxicity and therapeutic effect. Delivery systems may be designed to help prevent adverse side effects by delivering one or more antisense or inhibitory nucleic acids to specific targets, for example, neurons in the motor nervous system or central nervous system. The optimal dose of one or more antisense or inhibitory nucleic acids may be determined based on the results of clinical electrophysiology or electromyography, for example, to analyze peripheral nerve excitability.

[0124] Dosages for use in humans may be determined by evaluating data obtained from animal studies and cell culture assays. A dose that exhibits little or no toxicity and includes the ED50 is preferred. This dose may vary depending on the dosage form and route of administration. For any antisense or inhibitory nucleic acid used in the methods described herein, the dose may be initially estimated in cell culture. Doses may be formulated in animal models containing the concentration of the test compound that achieves half-maximal suppression of symptoms (LD50), as determined in cell culture. Such information obtained from cell culture and animal models may be used to more accurately determine effective doses in humans.

[0125] The present disclosure has multiple aspects, illustrated by the following non-limiting examples. [Example]

[0126] Small molecule inhibitors of the PIKFYVE kinase and antisense oligonucleotides (ASOs) that suppress PIKFYVE expression can prevent degeneration of neurons in humans and mice carrying mutations in the C9ORF72 gene that cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD).

[0127] ASOs are an attractive treatment option for neurodegenerative diseases because they are easily delivered to the central nervous system and have relatively low peripheral exposure. These properties maximize target engagement in the central nervous system and minimize unwanted target engagement or off-target effects in the periphery.

[0128] This disclosure provides novel antisense oligonucleotide (ASO) sequences targeting the PIKFYVE gene that can suppress PIKFYVE expression in human cells. PIKFYVE ASOs can also restore the survival of motor neurons from sporadic ALS patients. Furthermore, PIKFYVE ASOs can reduce the levels of neurotoxic dipeptide repeat protein aggregates derived from C9ORF72 repeat sequence expansions in mice in vivo.

[0129] Example 1 To identify ASO sequences that suppress PIKFYVE expression in human cells, ASOs were designed and synthesized as MOE gapmers containing sugar and linkage modifications that increase nuclease resistance and melting temperature while maintaining the ability to serve as RNase H substrates (see Tables 2, 3, 4, and 6). The ability of each ASO to suppress PIKFYVE RNA levels was tested by transfecting them into human embryonic kidney 293T cells with Lipofectamine 2000 at a concentration of 100 nM and measuring PIKFYVE expression 7 days after transfection. A control (NCASO) was used. Relative PIKFYVE expression shown is the average of three technical replicates, and values ​​are calculated by normalizing to the GAPDH control.

[0130] Example 2 In the study, newborn transgenic hPIKFYVE BAC mice are administered 25 μg of negative control ASO or test compound by intracerebroventricular (ICV) injection at P1 (postnatal day 1), and tissue samples are collected 14 days after treatment.

[0131] Example 3 In silico testing of the ASOs of SEQ ID NOs: 863-1261 was performed to determine the ASO acute neurotoxicity score for each ASO. Acute neurotoxicity scores were generated using the method described in Hagedorn et al. 2022 (DOI: 10.1089 / nat / 2021 / 0071). The ASOs of SEQ ID NOs: 863-1261 were determined to have low acute neurotoxicity scores. The results of the testing are shown in Table 5 below. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7] [Table 5-8] [Table 5-9] [Table 5-10] [Table 5-11]

[0132] In some embodiments, ASOs of the sequences set forth in SEQ ID NOs: 81-306, 310-328, 332-373, 376-420, 423, and 425-479 have a low acute neurotoxicity score.

[0133] Example 4 For various ASOs described herein, in silico suppression of off-target genes, such as CNTN5, was predicted. In silico testing of various ASOs was performed to determine the number of off-target binding sites for various ASOs. The GGGenome search tool was used to generate the number of off-target binding sites using two mismatches / gaps. The results of the testing are shown in Table 6 below.

[0134] The ASOs in Table 6 have the following gapmer design: 5' - 5 2'-methoxyethyl ribose nucleotides - 10 DNA nucleotides - 5 2'-methoxyethyl ribose nucleotides - 3' (the 1st, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 18th, and 19th linkages are phosphorothioate linkages, and the 2nd, 3rd, 4th, 5th, 16th, and 17th linkages are phosphodiester linkages (same gapmer design as in Table 2)). Note that the table below provides a 2'MOE wing. However, alternative wings including 2'-OMe or LNA (locked nucleic acid) are contemplated. Additionally, note that although the table below shows gapmer ASOs, ASOs with 20 DNA nucleotides (without a 2'MOE wing) and the full phosphodiester backbone (unmodified version) of the SEQ ID NOs in Table 6 are contemplated. [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6] [Table 6-7] [Table 6-8] [Table 6-9] [Table 6-10] Table 6-11 Table 6-12 Table 6-13 Table 6-14 Table 6-15 Table 6-16 Table 6-17 Table 6-18 Table 6-19 Table 6-20 Table 6-21 Table 6-22 Table 6-23 Table 6-24 Table 6-25

[0135] In some embodiments, the amino acids selected from SEQ ID NOs: 480-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, as well as SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, and 1716, ~1717, 1719, 1722, 1725, 1727~1728, 1731, 1740, 1743~1745, 1747~1751, 1754, 1756, 1760, 1762~1764, 1770~1775, 1777~1779, 1782~1784, 1787~1788 , 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853 , 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927 , 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021 , 2023, 2026~2029, 2031~2042, 2044~2055, 2058, 2061~2070, 2072~2083, 2085~2090, 2092~2095, 2097, 2099~2104, 2106~2109, 2111~2116, 2118, 2120 ~2121, 2123~2124, 2126~2128, 2130, 2132~2143, 2146~2147, 2149~2164, 2166~2177, 2179~2183, 2185, 2187~2205, 2207~2216, 2218, 2220~2232, 2234,ASOs of the sequences set forth in 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and the unmodified versions of 2319-2321 contain fewer off-target binding sites.

[0136] The present disclosure provides ASOs that suppress PIKFYVE expression in human cells, and accompanying data are consistent with these ASOs having the ability to prevent, inhibit, or slow neurodegeneration in ALS and FTD patients.

[0137] The foregoing description and drawings should be considered merely as illustrative of the principles of the present disclosure. The present disclosure is not intended to be limited by its embodiments, which may be implemented in various ways apparent to those skilled in the art. Many applications of the present disclosure will readily occur to those skilled in the art. Therefore, it is not desired to limit the disclosure to the specific embodiments disclosed or to the exact construction and operation shown and described. Rather, all suitable modifications and equivalents may be utilized within the scope of the present disclosure. All references cited herein are incorporated by reference in their entirety.

Claims

1. A single-stranded antisense oligonucleotide that inhibits the expression of PIKFYVE, wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1673-1674, 1675-1676, 1677-1678, 1679-1680, 1681-1682, 1682-1683, 1683-1684, 1685-1686, 1687-1688, 1689-1690, 1691-1692, 1693-1694, 1695-1696, 1697-1698, 1699-2000, 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2100, 2110, 2111, 2120, 2121, 2 74, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1 743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1810 811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143,the antisense oligonucleotide comprising a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any of the nucleobase sequences set forth in 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and an unmodified version of 2319-2321.

2. The antisense oligonucleotides are selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762- 1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826- 1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183,The antisense oligonucleotide of claim 1, comprising the nucleobase sequence set forth in any one of unmodified versions of 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

3. The antisense oligonucleotide of claim 1 or 2, wherein the antisense oligonucleotide has 18 to 20 linked nucleosides.

4. 10. The antisense oligonucleotide of any of the preceding claims, wherein at least one internucleoside linkage is a modified internucleoside linkage.

5. 5. The antisense oligonucleotide of claim 4, wherein at least one modified internucleoside linkage is a phosphorothioate internucleoside linkage.

6. 5. The antisense oligonucleotide of claim 4, wherein each modified internucleoside linkage is a phosphorothioate internucleoside linkage.

7. 10. The antisense oligonucleotide of any preceding claim, wherein at least one internucleoside linkage is a phosphodiester internucleoside linkage.

8. 8. The antisense oligonucleotide of claim 7, wherein at least one internucleoside linkage is a phosphorothioate linkage and at least one internucleoside linkage is a phosphodiester linkage.

9. 10. The antisense oligonucleotide of any preceding claim, wherein at least one nucleoside comprises a modified nucleobase.

10. The antisense oligonucleotide of claim 9, wherein the modified nucleobase is 5-methylcytosine.

11. 10. The antisense oligonucleotide of claim 1, wherein at least one nucleoside of the antisense oligonucleotide comprises a modified sugar moiety.

12. 12. The antisense oligonucleotide of claim 11, wherein the modified sugar moiety comprises a 2'-O-methoxyethyl group.

13. 10. The antisense oligonucleotide of any preceding claim, wherein the antisense oligonucleotide is a gapmer.

14. the antisense oligonucleotide a gap segment consisting of 8 to 12 linked deoxynucleosides; a 5' wing segment consisting of 3 to 5 linked nucleosides, and a 3' wing segment consisting of 3 to 5 linked nucleosides; 14. The antisense oligonucleotide of claim 13, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise modified sugar moieties.

15. 15. The antisense oligonucleotide of claim 14, wherein each nucleoside of each wing segment comprises a modified sugar moiety.

16. 15. The antisense oligonucleotide of claim 14, wherein the nucleosides comprising each wing segment contain at least two different modified sugar moieties.

17. 15. The antisense oligonucleotide of claim 14, wherein the nucleosides comprising each wing segment contain the same modified sugar moiety.

18. 16. The antisense oligonucleotide of claim 15, wherein the modified sugar moiety comprises a 2'-O-methoxyethyl group.

19. 10. The antisense oligonucleotide of any of the preceding claims, wherein the antisense oligonucleotide comprises 15 to 50 nucleosides.

20. The antisense oligonucleotides are selected from the group consisting of SEQ ID NOs: 783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1678, 1680, 1681, 1682, 1683, 1684, 1685, 1686, 1687, 1688, 1689, 1690, 1691, 1692, 1693, 1694, 1695, 1696, 1697, 1698, 1699, 1700, 1701, 1702, 1703, 1704, 1705, 1706, 1707, 1708, 1709, 1710, 1711, 1712, 1713, 1714, 1715, 1716, 1717, 79, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1 747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1818 815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147,2. An antisense oligonucleotide according to any of the preceding claims, comprising a nucleobase sequence comprising at least 12 or 15 consecutive nucleobases of any of the nucleobase sequences set forth in 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

21. The antisense oligonucleotides are selected from the group consisting of SEQ ID NOs: 783-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1678, 1680, 1681, 1682, 1683, 1684, 1685, 1686, 1687, 1688, 1689, 1690, 1691, 1692, 1693, 1694, 1695, 1696, 1697, 1698, 1699, 1700, 1701, 1702, 1703, 1704, 1705, 1706, 1707, 1708, 1709, 1710, 1711, 1712, 1713, 1714, 1715, 1716, 1717, 79, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1 747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1818 815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147,2. An antisense oligonucleotide according to any one of the preceding claims, comprising a sequence as set forth in any one of 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321.

22. A pharmaceutical composition comprising an antisense oligonucleotide according to any one of the preceding claims and a pharmaceutically acceptable carrier, diluent and / or excipient.

23. 23. The pharmaceutical composition of claim 22, wherein the pharmaceutical composition is formulated for parenteral administration.

24. 23. The pharmaceutical composition of claim 22, wherein the pharmaceutical composition is formulated for intracerebroventricular injection.

25. 26. A method for treating a subject having a neurological or neurodegenerative disease in need of such treatment, comprising administering a therapeutically effective amount of the antisense oligonucleotide of any one of claims 1 to 21 or the pharmaceutical composition of any one of claims 22 to 24.

26. 26. The method of claim 25, wherein the neurological disorder is associated with neuronal hyperexcitability.

27. 26. The method of claim 25, wherein the neurological disorder is associated with abnormal endosomal trafficking.

28. 26. The method of claim 25, wherein the neurological disorder is associated with abnormal lysosomal trafficking.

29. 26. The method of claim 25, wherein the neurological disease is selected from the group consisting of familial and sporadic amyotrophic lateral sclerosis (ALS), familial and sporadic frontotemporal dementia (FTD), progressive supranuclear palsy, Alzheimer's disease, chronic traumatic encephalopathy, Parkinson's disease, Charcot-Marie-Tooth disease types 2A and 4B, Huntington's disease, dementia, transmissible spongiform encephalopathies, spinal-bulbar muscular atrophy, dentatorubral-pallidoluysian atrophy, spinocerebellar ataxia, and Creutzfeldt-Jakob disease.

30. 26. The method of claim 25, wherein the neurological disease is familial amyotrophic lateral sclerosis.

31. 26. The method of claim 25, wherein the neurological disease is sporadic amyotrophic lateral sclerosis.

32. 26. The method of claim 25, wherein the neurological disease is familial frontotemporal dementia.

33. 26. The method of claim 25, wherein the neurological disease is sporadic frontotemporal dementia.

34. 26. The method of claim 25, wherein the neurological disease is frontotemporal dementia with TDP-43 pathology.

35. 26. The method of claim 25, wherein the neurological disease is frontotemporal dementia with tau pathology.

36. The method of any one of claims 25 to 35, wherein the subject is haploinsufficient for the C9ORF72 gene.

37. The method of any one of claims 25 to 35, wherein the subject has a GGGGCC repeat sequence expansion in C90RF72.

38. 36. The method of any one of claims 25-35, wherein the subject has a (GGGGCC)n (SEQ ID NO: 1565) hexanucleotide expansion in C90RF72, wherein n is at least 30.

39. 36. The method of any one of claims 25 to 35, wherein the subject has C9orf72-associated frontotemporal dementia.

40. 36. The method of any one of claims 25 to 35, wherein the subject has microtubule-associated protein tau (MAPT)-related frontotemporal dementia.

41. 41. The method of claim 40, wherein the patient has a V337M MAPT mutation.

42. A method for inhibiting or suppressing the expression of PIKFYVE in a patient with a neurological or neurodegenerative disease, the method comprising administering an effective amount of the antisense oligonucleotide of any one of claims 1 to 21 or the pharmaceutical composition of any one of claims 22 to 24.

43. consisting of 12 to 30 linked nucleosides, SEQ ID NOs: 1 to 306, 310 to 328, 332 to 373, 376 to 420, 423, 425 to 498, 508, 554, 586, 600 to 601, 619, 639, 667, 673, 680, 685, 699, 717, 733 to 734, 749 to 750, 752, 759, 762, 764, 770, 772, 781, 783 to 1088, 1092 to 1110, 1114 to 1155, 1158 to 1202, 1205, 1207 to 1280, 1290, 1336, 1368, 1382 to 1383, 1401, 1421 , 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672 , 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740 , 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809 ~1811, 1813~1815, 1818~1819, 1822~1824, 1826~1827, 1830~1832, 1836~1837, 1840~1843, 1845~1853, 1855~1858, 1861~1862, 1864, 1866, 1868~1869 , 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953 , 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058,2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143 , 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-230 0, 2302, 2304-2316, and 2319-2321, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1765-1766, 1767-1768, 1769-1770, 1771-1772, 1772-1774, 1773-1775, 1774-1776, 1775-1777, 1778-1779, 1780-1781, 1782-1783, 1784-1785, 1786-1787, 1788-1789, 1790-1791, 1792-1793, 1794-1795, 1796-1797, 1798-2000, 1799-2001, 2002-2003, 70-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1 830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1910 907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996- 1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097,2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-23 an oligonucleotide comprising a nucleobase sequence comprising at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 consecutive nucleobases of any of the nucleobase sequences set forth in unmodified versions of 2300, 2302, 2304-2316, and 2319-2321;

44. The antisense oligonucleotide of claim 1, comprising a sequence set forth in SEQ ID NOs: 1 to 80.

45. The antisense oligonucleotide of claim 1, which inhibits the expression of PIKFYVE by at least 80%.

46. A single-stranded antisense oligonucleotide that inhibits the expression of PIKFYVE, wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 1-306, 310-328, 332-373, 376-420, 423, 425-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, and SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1673-1674, 1675-1676, 1677-1678, 1679-1680, 1681-1682, 1682-1683, 1683-1684, 1685-1686, 1687-1688, 1689-1690, 1691-1692, 1693-1694, 1695-1696, 1697-1698, 1699-2000, 2001, 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2100, 2110, 2111, 2120, 2121, 2 74, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1 743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1810 811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143,an antisense oligonucleotide having a nucleobase sequence comprising a sequence set forth in any of the following unmodified versions: 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the first 3-5 nucleosides of the 5' end (the "5' wing segment") comprise modified sugars, the last 3-5 nucleosides of the 3' end (the "3' wing segment") comprise modified sugars, and the remaining nucleosides comprise a gap segment.

47. 47. The antisense oligonucleotide of claim 46, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

48. 48. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises 2'-OMe.

49. 48. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises 2'-MOE.

50. 48. The antisense oligonucleotide of claim 47, wherein at least one of the modified sugars comprises an LNA.

51. 47. The antisense oligonucleotide of claim 46, wherein the backbone linkages of the 5' wing segment, the 3' wing segment, and the gap segment comprise a mixture of phosphorothioate and phosphodiester linkages.

52. A single-stranded antisense oligonucleotide that inhibits the expression of PIKFYVE, wherein the antisense oligonucleotide is selected from the group consisting of SEQ ID NOs: 863-1088, 1092-1110, 1114-1155, 1158-1202, 1205, 1207-1280, 1290, 1336, 1368, 1382-1383, 1401, 1421, 1449, 1455, 1462, 1467, 1481, 1499, 1515-1516, 1531-1532, 1534, 1541, 1544, 1546, 1552, 1554, 1563, 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1711, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1784, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1843, 1845-1853, 1855-1858, 1861- 1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111-2116, 2118, 2120-2121, 2123-2124,The antisense oligonucleotide comprises a sequence set forth in any one of 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218, 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the backbone linkages of the antisense oligonucleotide comprise a mixture of phosphorothioate and phosphodiester linkages.

53. 53. The antisense oligonucleotide of claim 52, wherein the 1st, 6th, 7th, 8th, 9th, 10th, 11th, 12th, 13th, 14th, 15th, 18th, and 19th bonds are phosphorothioate bonds and the 2nd, 3rd, 4th, 5th, 16th, and 17th bonds are phosphodiester bonds.

54. 47. The antisense oligonucleotide of claim 46, wherein at least one nucleoside comprises a modified nucleobase.

55. 47. The antisense oligonucleotide of claim 46, wherein the modified nucleobase is 5-methylcytosine.

56. 47. The antisense oligonucleotide of claim 46, wherein the antisense oligonucleotide comprises a moiety that neutralizes the charge of the antisense oligonucleotide.

57. 47. The antisense oligonucleotide of claim 46, which inhibits the expression of PIKFYVE by at least 80%.

58. 47. A pharmaceutical composition comprising the antisense oligonucleotide of claim 46 and a pharmaceutically acceptable carrier, diluent, and / or excipient.

59. 53. A pharmaceutical composition comprising the antisense oligonucleotide of claim 52 and a pharmaceutically acceptable carrier, diluent, and / or excipient.

60. 54. A pharmaceutical composition comprising the antisense oligonucleotide of claim 53 and a pharmaceutically acceptable carrier, diluent, and / or excipient.

61. 61. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition is formulated for parenteral delivery or intracerebroventricular injection.

62. 62. The pharmaceutical composition of claim 61, wherein the pharmaceutical composition is formulated for parenteral delivery or intraventricular injection.

63. the antisense oligonucleotide a gap segment consisting of eight linked deoxynucleosides; a 5' wing segment consisting of five linked nucleosides, and a 3' wing segment consisting of 5 linked nucleosides; 15. The antisense oligonucleotide of claim 14, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise modified sugar moieties.

64. the antisense oligonucleotide a gap segment consisting of 10 linked deoxynucleosides; a 5' wing segment consisting of five linked nucleosides, and a 3' wing segment consisting of 5 linked nucleosides; 15. The antisense oligonucleotide of claim 14, wherein the gap segment is positioned between the 5' wing segment and the 3' wing segment, and wherein the nucleosides of each wing segment comprise modified sugar moieties.

65. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises a nucleic acid base sequence set forth in any one of SEQ ID NOs: 1 to 49, and the antisense oligonucleotide skips exon 3 of the PIKFYVE protein.

66. 66. The antisense oligonucleotide of claim 65, wherein the oligonucleotide comprises one or more modified sugars.

67. 67. The antisense oligonucleotide of claim 66, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

68. 68. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises 2'-OMe.

69. 68. The antisense oligonucleotide of claim 67, wherein each nucleoside of the antisense oligonucleotide comprises a 2'-MOE modified sugar.

70. 68. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises 2'-MOE.

71. 68. The antisense oligonucleotide of claim 67, wherein at least one of the modified sugars comprises an LNA.

72. 66. The antisense oligonucleotide of claim 65, wherein the backbone-linked oligonucleotide comprises a mixture of phosphorothioate and phosphodiester linkages.

73. 73. The antisense oligonucleotide of claim 72, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, sixteenth, and seventeenth bonds are phosphorothioate bonds, and the second, third, fourth, fifth, fourteenth, and fifteenth bonds are phosphodiester bonds.

74. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises a nucleic acid base sequence set forth in any one of SEQ ID NOs: 50 to 80, and the antisense oligonucleotide skips exon 5 of the PIKFYVE protein.

75. 75. The antisense oligonucleotide of claim 74, wherein the oligonucleotide comprises one or more modified sugars.

76. 76. The antisense oligonucleotide of claim 75, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

77. 77. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises 2'-OMe.

78. 77. The antisense oligonucleotide of claim 76, wherein each nucleoside of the antisense oligonucleotide comprises a 2'-MOE modified sugar.

79. 77. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises 2'-MOE.

80. 77. The antisense oligonucleotide of claim 76, wherein at least one of the modified sugars comprises an LNA.

81. 75. The antisense oligonucleotide of claim 74, wherein the backbone-linked oligonucleotide comprises a mixture of phosphorothioate and phosphodiester linkages.

82. 82. The antisense oligonucleotide of claim 81, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, sixteenth, and seventeenth bonds are phosphorothioate bonds, and the second, third, fourth, fifth, fourteenth, and fifteenth bonds are phosphodiester bonds.

83. 2. The antisense oligonucleotide of claim 1, wherein the antisense oligonucleotide comprises a nucleobase sequence set forth in any one of SEQ ID NOs: 81-306, 310-328, 332-373, 376-420, 423, and 425-479, and wherein the antisense oligonucleotide has a low acute neurotoxicity score.

84. 84. The antisense oligonucleotide of claim 83, wherein the oligonucleotide comprises one or more modified sugars.

85. 85. The antisense oligonucleotide of claim 84, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

86. 86. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises 2'-OMe.

87. 86. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises 2'-MOE.

88. 86. The antisense oligonucleotide of claim 85, wherein at least one of the modified sugars comprises an LNA.

89. 84. The antisense oligonucleotide of claim 83, wherein the backbone-linked oligonucleotide comprises a mixture of phosphorothioate and phosphodiester linkages.

90. 90. The antisense oligonucleotide of claim 89, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth bonds are phosphorothioate bonds, and the second, third, fourth, fifth, sixteenth, and seventeenth bonds are phosphodiester bonds.

91. The antisense oligonucleotides are selected from the group consisting of SEQ ID NOs: 480-498, 508, 554, 586, 600-601, 619, 639, 667, 673, 680, 685, 699, 717, 733-734, 749-750, 752, 759, 762, 764, 770, 772, and 781, as well as SEQ ID NOs: 1650, 1652-1654, 1660, 1666-1667, 1671-1672, 1674, 1677, 1679, 1684-1687, 1689, 1692, 1694, 1696-1698, 1700, 1702-1703, 1708-1710, 1, 1713, 1716-1717, 1719, 1722, 1725, 1727-1728, 1731, 1740, 1743-1745, 1747-1751, 1754, 1756, 1760, 1762-1764, 1770-1775, 1777-1779, 1782-1783 84, 1787-1788, 1791-1793, 1795, 1797-1802, 1804-1805, 1809-1811, 1813-1815, 1818-1819, 1822-1824, 1826-1827, 1830-1832, 1836-1837, 1840-1841 43, 1845-1853, 1855-1858, 1861-1862, 1864, 1866, 1868-1869, 1871-1881, 1883, 1885-1892, 1895-1897, 1899, 1901-1907, 1909, 1911-1913, 1915, 1 917, 1919-1927, 1930, 1932-1936, 1938-1940, 1943, 1945-1953, 1955-1969, 1971-1973, 1976-1980, 1985-1994, 1996-1997, 1999-2010, 2012-2015, 2 017, 2019-2021, 2023, 2026-2029, 2031-2042, 2044-2055, 2058, 2061-2070, 2072-2083, 2085-2090, 2092-2095, 2097, 2099-2104, 2106-2109, 2111- 2116, 2118, 2120-2121, 2123-2124, 2126-2128, 2130, 2132-2143, 2146-2147, 2149-2164, 2166-2177, 2179-2183, 2185, 2187-2205, 2207-2216, 2218,2. The antisense oligonucleotide of claim 1, comprising the nucleobase sequence set forth in any one of unmodified versions of 2220-2232, 2234, 2236-2245, 2247-2268, 2270-2289, 2291-2300, 2302, 2304-2316, and 2319-2321, wherein the antisense oligonucleotide comprises a small number of off-target binding sites.

92. 92. The antisense oligonucleotide of claim 91, wherein the oligonucleotide comprises one or more modified sugars.

93. 93. The antisense oligonucleotide of claim 92, wherein the modified sugar comprises 2'-OMe, 2'-MOE, LNA, or any combination thereof.

94. 94. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises 2'-OMe.

95. 94. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises 2'-MOE.

96. 94. The antisense oligonucleotide of claim 93, wherein at least one of the modified sugars comprises an LNA.

97. 92. The antisense oligonucleotide of claim 91, wherein the backbone-linked oligonucleotide comprises a mixture of phosphorothioate and phosphodiester linkages.

98. The antisense oligonucleotide of claim 97, wherein the first, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, eighteenth, and nineteenth bonds are phosphorothioate bonds, and the second, third, fourth, fifth, sixteenth, and seventeenth bonds are phosphodiester bonds.

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