RNAi-mediated inhibition of HIF1A for treatment of ocular angiogenesis

Abstract

RNA interference, is provided for inhibition of HIF1A mRNA expression for treating patients with ocular angiogenesis, particularly for treating retinal edema, diabetic retinopathy, sequela associated with retinal ischemia, posterior segment neovascularization (PSNV), and neovascular glaucoma, and for treating patients at risk of developing such conditions.

1 . A method of attenuating expression of HIF1A mRNA of a subject, comprising: 
 administering to the subject a composition comprising an effective amount of interfering RNA having a length of 19 to 49 nucleotides and a pharmaceutically acceptable carrier, the interfering RNA comprising: 
 a sense nucleotide strand, an antisense nucleotide strand, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides;  
 wherein the antisense strand hybridizes under physiological conditions to a portion of MRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2 and has a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with the hybridizing portion of MRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2, respectively,  
   wherein the expression of HIF1A mRNA is attenuated thereby.    
     
     
         2 . The method of  claim 1  wherein the subject is a human and the human has ocular angiogenesis.  
     
     
         3 . The method of  claim 1  wherein the subject is a human and the human is at risk of developing ocular angiogenesis.  
     
     
         4 . The method of  claim 1  wherein the composition is administered via a topical, intravitreal, transcleral, periocular, conjunctival, subtenon, intracameral, subretinal, subconjunctival, retrobulbar, or intracanalicular route.  
     
     
         5 . The method of  claim 1  wherein the antisense strand is designed to target an mRNA corresponding to SEQ ID NO:1 comprising nucleotide 411, 580, 583, 868, 869, 1099, 1100, 1242, 1302, 1371, 1396, 1559, 1560, 1809, 2085, 2087, 2105, 2138, 2256, 2358, 2422, 2636, 2666, 2743, 2858, 2861, 3135, 3544, 3554, 1943, 1791, 2351, or 1408.  
     
     
         6 . The method of  claim 1  wherein the antisense strand is designed to target an mRNA corresponding to SEQ ID NO:2 comprising nucleotide 2360, 2411, 2420, 2536, 2539, 2545, 2616, 2731, 2734, 3008, or 3427.  
     
     
         7 . The method of  claim 1  further comprising administering to the subject a second interfering RNA having a length of 19 to 49 nucleotides, and comprising 
 a sense nucleotide strand, an antisense nucleotide strand, and a region of at least near-perfect complementarity of at least 19 nucleotides;    wherein the antisense strand of the second interfering RNA hybridizes under physiological conditions to a second portion of mRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2 and the antisense strand has a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with the second hybridizing portion of mRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2, respectively.    
     
     
         8 . A method of treating ocular angiogenesis in a subject in need thereof, comprising: 
 administering to an eye of the subject a composition comprising an effective amount of interfering RNA having a length of 19 to 49 nucleotides and a pharmaceutically acceptable carrier, the interfering RNA comprising: 
 a sense nucleotide strand, an antisense nucleotide strand, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides;  
 wherein the antisense strand hybridizes under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2, and has a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with the hybridizing portion of mRNA corresponding to SEQ ID NO:1 or SEQ ID NO:2, respectively,  
   wherein the ocular angiogenesis is treated thereby.    
     
     
         9 . A method of attenuating expression of HIF1A mRNA of a subject, comprising: 
 administering to the subject a composition comprising an effective amount of single-stranded interfering RNA having a length of 19 to 49 nucleotides and a pharmaceutically acceptable carrier, 
 wherein the single-stranded interfering RNA hybridizes under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:1 comprising nucleotide 411, 580, 583, 868, 869, 1099, 1100, 1242, 1302, 1371, 1396, 1559, 1560, 1809, 2085, 2087, 2105, 2138, 2256, 2358, 2422, 2636, 2666, 2743, 2858, 2861, 3135, 3544, 3554, 1943, 1791, 2351, or 1408, and the interfering RNA has a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with the hybridizing portion of mRNA corresponding to SEQ ID NO:1, or  
 wherein the single-stranded interfering RNA hybridizes under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:2 comprising nucleotide 2360, 2411, 2420, 2536, 2539, 2545, 2616, 2731, 2734, 3008, or 3427 and the interfering RNA has a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with the hybridizing portion of mRNA corresponding to SEQ ID NO:2,  
   wherein the expression of HIF1A mRNA is thereby attenuated.    
     
     
         10 . A method of attenuating expression of HIF1A mRNA in a subject, the method comprising: 
 administering to the subject a composition comprising an effective amount of interfering RNA having a length of 19 to 49 nucleotides and a pharmaceutically acceptable carrier, the interfering RNA comprising: 
 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 an mRNA corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:51,  
   wherein the expression of HIFI A mRNA is thereby attenuated.    
     
     
         11 . The method of  claim 10  wherein the interfering RNA comprises: 
 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 an mRNA corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:36.    
     
     
         12 . The method of  claim 10  wherein the interfering RNA comprises: 
 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 an mRNA corresponding to any one of SEQ ID NO:37-SEQ ID NO:51.    
     
     
         13 . The method of  claim 10  wherein the interfering RNA comprises 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 the sequence identified by the sequence identifier.  
     
     
         14 . The method of  claim 10  wherein the interfering RNA comprises 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 the sequence identified by the sequence identifier.  
     
     
         15 . The method of  claim 10  wherein the composition further comprises a second interfering RNA having a length of 19 to 49 nucleotides and comprising a region of at least 13 contiguous nucleotides having at least 90% complementarity to, or at least 90% sequence identity with, the penultimate 13 nucleotides of the 3′ end of a second mRNA corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:51.  
     
     
         16 . A method of treating ocular angiogenesis in a subject in need thereof, the method comprising: 
 administering to an eye of the subject a composition comprising an effective amount of interfering RNA having a length of 19 to 49 nucleotides and a pharmaceutically acceptable carrier, the interfering RNA comprising:    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 an mRNA corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:51, wherein the ocular angiogenesis is treated thereby.    
     
     
         17 . The method of  claim 16  wherein the interfering RNA comprises: 
 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 an mRNA corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:36.    
     
     
         18 . The method of  claim 16  wherein the interfering RNA comprises: 
 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 an mRNA corresponding to any one of SEQ ID NO:37-SEQ ID NO:51.    
     
     
         19 . The method of  claim 16  wherein the subject has retinal edema, retinal ischemia or diabetic retinopathy.  
     
     
         20 . The method of  claim 1  wherein the sense nucleotide strand and the antisense nucleotide strand are connected by a hairpin loop.  
     
     
         21 . The method of  claim 8  wherein the sense nucleotide strand and the antisense nucleotide strand are connected by a hairpin loop.  
     
     
         22 . The method of  claim 10  wherein the interfering RNA is an shRNA.  
     
     
         23 . The method of  claim 16  wherein the interfering RNA is an shRNA.  
     
     
         24 . The method of  claim 10  wherein the interfering RNA is an siRNA.  
     
     
         25 . The method of  claim 16  wherein the interfering RNA is an siRNA.  
     
     
         26 . The method of  claim 10  wherein the interfering RNA is an miRNA.  
     
     
         27 . The method of  claim 16  wherein the interfering RNA is an miRNA.  
     
     
         28 . The method of  claim 8  wherein the composition is administered via a topical, intravitreal, transcleral, periocular, conjunctival, subtenon, intracameral, subretinal, subconjunctival, retrobulbar, or intracanalicular route.  
     
     
         29 . The method of  claim 16  wherein the composition is administered via a topical, intravitreal, transcleral, periocular, conjunctival, subtenon, intracameral, subretinal, subconjunctival, retrobulbar, or intracanalicular route.  
     
     
         30 . The method of  claim 8  wherein the composition is administered via in vivo expression from an interfering RNA expression vector.  
     
     
         31 . The method of  claim 16  wherein the composition is administered via in vivo expression from an interfering RNA expression vector.  
     
     
         32 . The method of  claim 8  wherein the subject has retinal edema, retinal ischemia or diabetic retinopathy.  
     
     
         33 . A method of treating ocular angiogenesis in a subject in need thereof, comprising: 
 administering to the subject a composition comprising a double stranded siRNA molecule that down regulates expression of a HIF1A gene via RNA interference,    wherein:    each strand of the siRNA molecule is independently about 19 to about 27 nucleotides in length; and    one strand of the siRNA molecule comprises a nucleotide sequence having substantial complementarity to an mRNA corresponding to the HIF1A gene so that the siRNA molecule directs cleavage of the mRNA via RNA interference.    
     
     
         34 . The method of  claim 33  wherein the composition is administered via an aerosol, buccal, dermal, intradernal, inhaling, intramuscular, intranasal, intraocular, intrapulmonary, intravenous, intraperitoneal, nasal, ocular, oral, otic, parenteral, patch, subcutaneous, sublingual, topical, or transdermal route.  
     
     
         35 . The method of  claim 33  wherein the interfering RNA is administered via in vivo expression from an expression vector capable of expressing the interfering RNA.  
     
     
         36 . The method of  claim 33  wherein the interfering RNA is an miRNA.  
     
     
         37 . The method of  claim 33  wherein each strand of the siRNA molecule is independently about 19 nucleotides to about 25 nucleotides in length.  
     
     
         38 . The method of  claim 33  wherein each strand of the siRNA molecule is independently about 19 nucleotides to about 21 nucleotides in length.  
     
     
         39 . A composition comprising an interfering RNA having a length of 19 to 49 nucleotides and having a nucleotide sequence corresponding to any one of SEQ ID NO:3, and SEQ ID NO:9-SEQ ID NO:51, or a complement thereof, and a pharmaceutically acceptable carrier.  
     
     
         40 . The composition of  claim 39  wherein the interfering RNA is an shRNA.  
     
     
         41 . The composition of  claim 39  wherein the interfering RNA is an siRNA.  
     
     
         42 . The composition of  claim 39  wherein the interfering RNA is an miRNA.  
     
     
         43 . A composition comprising a double stranded siRNA molecule that down regulates expression of a HIF1A gene via RNA interference, wherein: 
 each strand of the siRNA molecule is independently about 19 to about 27 nucleotides in length; and    one strand of the siRNA molecule comprises a nucleotide sequence having substantial complementarity to an mRNA corresponding to the HIF1A gene so that the siRNA molecule directs cleavage of the mRNA via RNA interference.    
     
     
         44 . The composition of  claim 43  wherein each strand of the siRNA molecule is independently about 19 nucleotides to about 25 nucleotides in length.  
     
     
         45 . The composition of  claim 43  wherein each strand of the siRNA molecule is independently about 19 nucleotides to about 21 nucleotides in length.