RNAi-RELATED INHIBITION OF TNFa SIGNALING PATHWAY FOR TREATMENT OF OCULAR ANGIOGENESIS

Abstract

RNA interference is provided for inhibition of tumor necrosis factor α (TNFα) by silencing TNFα cell surface receptor TNF receptor-1 (TNFR1) mRNA expression, or by silencing TNFα converting enzyme (TACE/ADAM17) mRNA expression. Silencing such TNFα targets, in particular, is useful for treating patients having a TNFα-related condition or at risk of developing a TNFα-related condition, such as ocular angiogenesis, retinal ischemia, and diabetic retinopathy.

1 . A method of treating a TNFα-related ocular disorder in a patient in need thereof, comprising administering to the patient an interfering RNA molecule that attenuates expression of the TNFR1 or TACE mRNA via RNA interference, wherein the TNFα-related ocular disorder is ocular angiogenesis, ocular neovascularization, proliferative diabetic retinopathy, sequela associated with retinal ischemia, posterior segment neovascularization (PSNV), or neovascular glaucoma. 
     
     
         2 . The method of  claim 1 , wherein the interfering RNA molecule is double stranded and each strand is independently about 19 to about 27 nucleotides in length. 
     
     
         3 . The method of  claim 2 , wherein each strand is independently about 19 nucleotides to about 25 nucleotides in length. 
     
     
         4 . The method of  claim 2 , wherein each strand is independently about 19 nucleotides to about 21 nucleotides in length. 
     
     
         5 . The method of  claim 2 , wherein the sense and antisense strands are connected by a linker to form a shRNA that can attenuate expression of TACE or TNFR1 mRNA in a patient. 
     
     
         6 . The method of  claim 2 , wherein the interfering RNA molecule has blunt ends. 
     
     
         7 . The method of  claim 2 , wherein at least one strand of the interfering RNA molecule comprises a 3′ overhang. 
     
     
         8 . The method of  claim 7 , wherein the 3′ overhang comprises about 1 to about 6 nucleotides. 
     
     
         9 . The method of  claim 8 , wherein the 3′ overhang comprises 2 nucleotides. 
     
     
         10 . The method of  claim 1 , wherein the interfering RNA molecule is administered via in vivo expression from an expression vector capable of expressing the interfering RNA molecule. 
     
     
         11 . The method of  claim 1 , wherein the patient has or is at risk of developing a TNFα-related ocular disorder. 
     
     
         12 . The method of  claim 1 , wherein the interfering RNA molecule recognizes a portion of TACE mRNA that corresponds to any of SEQ ID NO: 3 and SEQ ID NO: 14-SEQ ID NO: 58. 
     
     
         13 . The method of  claim 1 , wherein the interfering RNA molecule recognizes a portion of TNFR1 mRNA that corresponds to any of SEQ ID NO: 155-SEQ ID NO: 201. 
     
     
         14 . The method of  claim 1 , wherein the interfering RNA molecule recognizes a portion of TACE mRNA, wherein the portion comprises nucleotide 297, 333, 334, 335, 434, 470, 493, 547, 570, 573, 618, 649, 689, 755, 842, 844, 846, 860, 878, 894, 900, 909, 910, 913, 942, 970, 984, 1002, 1010, 1053, 1064, 1137, 1162, 1215, 1330, 1334, 1340, 1386, 1393, 1428, 1505, 1508, 1541, 1553, 1557, 1591, 1592, 1593, 1597, 1604, 1605, 1626, 1632, 1658, 1661, 1691, 1794, 1856, 1945, 1946, 1947, 1958, 2022, 2094, 2100, 2121, 2263, 2277, 2347, 2349, 2549, 2578, 2595, 2606, 2608, 2629, 2639, 2764, 2766, 2767, 2769, 3027, 3028, 3261, 3264, 3284, 3313, 3317, 3332, or 3337 of SEQ ID NO: 1. 
     
     
         15 . The method of  claim 1 , wherein the interfering RNA molecule recognizes a portion of TNFR1 mRNA, wherein the portion comprises nucleotide 124, 328, 387, 391, 393, 395, 406, 421, 423, 444, 447, 455, 459, 460, 467, 469, 470, 471, 475, 479, 513, 517, 531, 543, 556, 576, 587, 588, 589, 595, 601, 602, 611, 612, 651, 664, 667, 668, 669, 677, 678, 785, 786, 788, 791, 792, 804, 813, 824, 838, 843, 877, 884, 929, 959, 960, 961, 963, 964, 965, 970, 973, 974, 1000, 1002, 1013, 1026, 1053, 1056, 1057, 1058, 1161, 1315, 1318, 1324, 1357, 1360, 1383, 1393, 1420, 1471, 1573, 1671, 2044, 2045, 2046, 2047, 2048, 2089, 2090, 2091, or, 2092 of SEQ ID NO: 2. 
     
     
         16 . The method of  claim 1 , wherein the interfering RNA molecule comprises at least one modification. 
     
     
         17 . The method of  claim 1 , wherein the interfering RNA molecule is a shRNA, a siRNA, or a miRNA. 
     
     
         18 . The method of  claim 1 , wherein the interfering RNA molecule is administered via a topical, intravitreal, transcleral, periocular, conjunctival, subtenon, intracameral, subretinal, subconjunctival, retrobulbar, or intracanalicular route. 
     
     
         19 . An interfering RNA molecule having a length of about 19 to about 49 nucleotides, the interfering RNA molecule comprising:
 (a) a region of at least 13 contiguous nucleotides having at least 90% sequence complementarity to, or at least 90% sequence identity with, the penultimate 13 nucleotides of the 3′ end of a mRNA corresponding to any one of SEQ ID NO:3 and SEQ ID NO:14-SEQ ID NO:58;   (b) a region of at least 14 contiguous nucleotides having at least 85% sequence complementarity to, or at least 85% sequence identity with, the penultimate 14 nucleotides of the 3′ end of an mRNA corresponding to any one of SEQ ID NO:3 and SEQ ID NO:14-SEQ ID NO:58; or   (c) a region of at least 15, 16, 17, or 18 contiguous nucleotides having at least 80% sequence complementarity to, or at least 80% sequence identity with, the penultimate 15, 16, 17, or 18 nucleotides, respectively, of the 3′ end of an mRNA corresponding to any one of SEQ ID NO:3 and SEQ ID NO:14-SEQ ID NO:58.   
     
     
         20 . An interfering RNA molecule having a length of about 19 to about 49 nucleotides, the interfering RNA molecule comprising:
 (a) a region of at least 13 contiguous nucleotides having at least 90% sequence complementarity to, or at least 90% sequence identity with, the penultimate 13 nucleotides of the 3′ end of a mRNA corresponding to any one of SEQ ID NO:155-SEQ ID NO:201;   (b) a region of at least 14 contiguous nucleotides having at least 85% sequence complementarity to, or at least 85% sequence identity with, the penultimate 14 nucleotides of the 3′ end of an mRNA corresponding to any one of SEQ ID NO:155-SEQ ID NO:201; or   (d) a region of at least 15, 16, 17, or 18 contiguous nucleotides having at least 80% sequence complementarity to, or at least 80% sequence identity with, the penultimate 15, 16, 17, or 18 nucleotides, respectively, of the 3′ end of an mRNA corresponding to any one of SEQ ID NO:155-SEQ ID NO:201.