RNAi inhibition of serum amyloid a for treatment of glaucoma
RNA interference is provided for inhibition of serum amyloid A mRNA expression in glaucomas involving SAA expression.
1 . A method of attenuating expression of serum amyloid A mRNA in an eye of a subject, comprising:
administering to the 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 sequence, an antisense nucleotide sequence, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides between the sense and antisense sequences;
wherein the antisense sequence hybridizes under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3, 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, SEQ ID NO:2, or SEQ ID NO:3, respectively,
wherein the expression of serum amyloid A mRNA is thereby attenuated.
2 . The method of claim 1 wherein the subject has glaucoma.
3 . The method of claim 1 wherein the subject is at risk of developing glaucoma.
4 . The method of claim 1 wherein the antisense sequence has a region of at least near-perfect contiguous complementarity of at least 21 to 23 nucleotides with the hybridizing portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 and comprises an additional TT sequence at the 3′ end of each of the sense and the antisense sequence.
5 . The method of claim 1 wherein the sense nucleotide sequence and the antisense nucleotide sequence are connected by a loop nucleotide sequence.
6 . The method of claim 1 wherein the composition is administered via a topical, intravitreal, or transcleral route.
7 . The method of claim 1 wherein the antisense sequence is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:1 beginning at nucleotide 230, 357, 362, 380, 447, 470, 527, 531, 548, or 557.
8 . The method of claim 1 wherein the antisense sequence is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 43, 170, 175, 193, 260, 283, 339, or 370.
9 . The method of claim 1 wherein the antisense sequence is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 252, 271, 276, 325, 343.
10 . The method of claim 1 wherein the antisense sequence is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:3 beginning at nucleotide 153, 166, 222, 227, 251, 268, 297, 335, 356, 384, 390, 396, 406, or 423.
11 . The method of claim 1 wherein the antisense sequence comprises
CUUUGCCACUCCUGCCCCA,
(SEQ ID NO:37)
UCGGAAGUGAUUGGGGUCU,
(SEQ ID NO:38)
UUUGUCUGAGCCGAUGUAA,
(SEQ ID NO:39)
AACCAGGCCCGUGAGAAGC,
(SEQ ID NO:40)
CUGAGCCGAUGUAAUUGGC,
(SEQ ID NO:41)
GCCACUCCUGCCCCAUUUA,
(SEQ ID NO:69)
CCCCCGAGCAUGGAAGUAU,
(SEQ ID NO:42)
CUCUGGCAUUGCUGAUCAC,
(SEQ ID NO:43)
GCCUGUGAGUCUCUGGAUA,
(SEQ ID NO:44)
GCCACUCCUGCCCCAUUUA,
(SEQ ID NO:45)
GCCAGCAGGUCGGAAGUGA,
(SEQ ID NO:46)
AGUCUCUGGAUAUUCUCUC,
(SEQ ID NO:47)
UUUAUUGGCAGCCUGAUCG,
(SEQ ID NO:48)
UUGCUGAUCACUUCUGCGG,
(SEQ ID NO:49)
CUGGAUAUUCUCUCUGGCA,
(SEQ ID NO:50)
UCUGCCACUCCUGCCCCAU,
(SEQ ID NO:51)
AACCCCUUGGAGAGCCUCC,
(SEQ ID NO:52)
UGCCCAUGUCCCCAACCCC,
(SEQ ID NO:53)
AUAGAGAUAUCUGUUUGAA,
(SEQ ID NO:54)
CGAGCAUAGAGAUAUCUGU,
(SEQ ID NO:55)
CUUUGGGCAGCAUCAUAGU,
(SEQ ID NO:56)
AGACACCCCCAGGUCCUCU,
(SEQ ID NO:57)
CCUGGAACGGCUGAUGAGU,
(SEQ ID NO:58)
CCAAAUAAAUAGUAGUCUA,
(SEQ ID NO:59)
UCCAAUACAGUGCUGCUGU,
(SEQ ID NO:60)
CUCAGCUUUCUCGUUGGAC,
(SEQ ID NO:61)
CCAUUCCUCAGCUUUCUCG,
(SEQ ID NO:62)
CCGGCCGCAUUCCUCAGCU,
(SEQ ID NO:63)
CUUUGCCACUCCGGCCCCA,
(SEQ ID NO:64)
or
UCUGAAGCGGUCGGGGUCU.
(SEQ ID NO:65)
12 . The method of claim 1 wherein the interfering RNA comprises a modification on a base portion, on a sugar portion or on a phosphate portion.
13 . The method of claim 1 further comprising administering to the eye of the subject a second interfering RNA having a length of 19 to 49 nucleotides, and comprising
a sense nucleotide sequence, an antisense nucleotide sequence, and a region of at least near-perfect complementarity of at least 19 nucleotides between the sense and antisense sequences;
wherein the antisense sequence of the second interfering RNA hybridizes under physiological conditions to a second portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 and the antisense sequence 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, SEQ ID NO:2, or SEQ ID NO:3, respectively.
14 . The method of claim 1 wherein the composition comprises an effective amount of a mixture of at least four interfering RNAs, each interfering RNA having a length of 19 to 49 nucleotides, and a pharmaceutically acceptable carrier, each interfering RNA comprising:
a sense nucleotide sequence, an antisense nucleotide sequence, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides between the sense and antisense sequences of each of the four interfering RNAs;
wherein the antisense sequences of the mixture hybridize under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 175, 252, 276, and 325, respectively, and have 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 beginning at nucleotide 175, 252, 276, and 325, respectively.
15 . A method of attenuating expression of serum amyloid A mRNA in an eye of a subject, comprising:
administering to the 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 nucleotide sequence having a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with a hybridizing portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3,
wherein the expression of serum amyloid A mRNA is thereby attenuated.
16 . The method of claim 15 wherein the composition is administered via a topical, intravitreal, or transcleral route.
17 . The method of claim 15 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:1 beginning at nucleotide 230, 357, 362, 380, 447, 470, 527, 531, 548, or 557.
18 . The method of claim 15 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 43, 170, 175, 193, 260, 283, 339, or 370.
19 . The method of claim 15 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 252, 271, 276, 325, 343.
20 . The method of claim 15 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:3 beginning at nucleotide 153, 166, 222, 227, 251, 268, 297, 335, 356, 384, 390, 396, 406, or 423.
21 . The method of claim 15 further comprising administering to the eye of the subject a second interfering RNA having a length of 19 to 49 nucleotides, and comprising
a second nucleotide sequence having a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with a second hybridizing portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.
22 . The method of claim 15 wherein the composition comprises an effective amount of a mixture of at least four interfering RNAs, each interfering RNA having a length of 19 to 49 nucleotides, and the mixture comprising:
a first, second, third and fourth nucleotide sequence having 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 beginning at nucleotide 175, 252, 276, and 325, respectively.
23 . A method of treating a serum amyloid A-associated glaucoma in a subject in need thereof, comprising:
administering to the 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 sequence, an antisense nucleotide sequence, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides between the sense and antisense sequences;
wherein the antisense sequence hybridizes under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 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, SEQ ID NO:2, or SEQ ID NO:3, respectively,
wherein the serum amyloid A-associated glaucoma is treated thereby.
24 . The method of claim 23 wherein the composition is administered via a topical, intravitreal, or transcleral route.
25 . The method of claim 23 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:1 beginning at nucleotide 230, 357, 362, 380, 447, 470, 527, 531, 548, or 557.
26 . The method of claim 23 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 43, 170, 175, 193, 260, 283, 339, or 370.
27 . The method of claim 23 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 252, 271, 276, 325, 343.
28 . The method of claim 23 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:3 beginning at nucleotide 153, 166, 222, 227, 251, 268, 297, 335, 356, 384, 390, 396, 406, or 423.
29 . The method of claim 23 further comprising administering to the eye of the subject a second interfering RNA having a length of 19 to 49 nucleotides, and comprising
a sense nucleotide sequence, an antisense nucleotide sequence, and a region of at least near-perfect complementarity of at least 19 nucleotides between the sense and antisense sequences;
wherein the antisense sequence of the second interfering RNA hybridizes under physiological conditions to a second portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3 and the antisense sequence 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, SEQ ID NO:2, or SEQ ID NO:3, respectively.
30 . The method of claim 23 wherein the composition comprises an effective amount of a mixture of at least four interfering RNAs, each interfering RNA having a length of 19 to 49 nucleotides, and a pharmaceutically acceptable carrier, each interfering RNA comprising:
a sense nucleotide sequence, an antisense nucleotide sequence, and a region of at least near-perfect contiguous complementarity of at least 19 nucleotides between the sense and antisense sequences of each of the four interfering RNAs;
wherein the antisense sequences of the mixture hybridize under physiological conditions to a portion of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 175, 252, 276, and 325, respectively, and have 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 beginning at nucleotide 175, 252, 276, and 325, respectively.
31 . A method of treating a serum amyloid A-associated glaucoma in a subject in need thereof, comprising:
administering to the 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 nucleotide sequence having a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with a hybridizing portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3,
wherein the expression of serum amyloid A mRNA is thereby attenuated.
32 . The method of claim 31 wherein the composition is administered via a topical, intravitreal, or transcleral route.
33 . The method of claim 31 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:1 beginning at nucleotide 230, 357, 362, 380, 447, 470, 527, 531, 548, or 557.
34 . The method of claim 31 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 43, 170, 175, 193, 260, 283, 339, or 370.
35 . The method of claim 31 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:2 beginning at nucleotide 252, 271, 276, 325, 343.
36 . The method of claim 31 wherein the interfering RNA is designed to target a nucleotide sequence of mRNA corresponding to SEQ ID NO:3 beginning at nucleotide 153, 166, 222, 227, 251, 268, 297, 335, 356, 384, 390, 396, 406, or 423.
37 . The method of claim 31 further comprising administering to the eye of the subject a second interfering RNA having a length of 19 to 49 nucleotides, and comprising
a second nucleotide sequence having a region of at least near-perfect contiguous complementarity of at least 19 nucleotides with a second hybridizing portion of mRNA corresponding to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.
38 . The method of claim 31 wherein the composition comprises an effective amount of a mixture of at least four interfering RNAs, each interfering RNA having a length of 19 to 49 nucleotides, and the mixture comprising:
a first, second, third and fourth nucleotide sequence having 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 beginning at nucleotide 175, 252, 276, and 325, respectively.