Silencing RNA molecules and their use in bone formation
The present invention is directed to siRNA molecules that down-regulate the expression of proteins that inhibit bone formation. In another aspect, the instant invention is directed to compositions and/or implants comprising in combination such siRNA molecules in a pharmaceutically acceptable carrier or implant.
1 . An siRNA molecule comprising a double stranded RNA portion having about 5 to 40 bp said molecule down-regulating the expression in a mammal of a protein inhibitor of osteogenesis or chondrogenesis.
2 . The siRNA molecule of claim 1 , wherein said mammal is human.
3 . The siRNA molecule of claim 2 , wherein the double stranded RNA portion consists of 13 to 30 bp.
4 . The siRNA molecule of claim 3 , wherein the double stranded RNA portion consists of 19 to 23 bp.
5 . The siRNA molecule of claim 3 , wherein said protein inhibitor is a protein inhibitor is selected from the group consisting of HDAC-3, STAT-1, SMURF-1, SMURF-3, SMURF-8, Tob, Calponin, and noggin.
6 . The siRNA molecule of claim 5 , wherein said protein inhibitor is HDAC-3 or SMURF-1.
7 . The siRNA molecule of claim 6 , wherein said protein inhibitor is HDAC-3.
8 . The siRNA molecule of claim 7 , wherein the double stranded RNA portion is selected from the group consisting of the following combinations of sense and anti-sense strands: SEQ ID NO: 9/SEQ ID NO: 10; SEQ ID NO: 11/SEQ ID NO: 12; SEQ ID NO: 13/SEQ ID NO: 14; and SEQ ID NO: 15/SEQ ID NO: 16.
9 . The siRNA molecule of claim 7 , wherein the double stranded RNA portion comprises:
a first RNA strand selected from the group consisting SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16;
and a second RNA strand that is a complement of the first RNA strand.
10 . The siRNA molecule of claim 6 , wherein said protein inhibitor is SMURF-1.
11 . The siRNA molecule of claim 10 , wherein the double stranded RNA portion is selected from the group consisting of the following combinations of sense and anti-sense strands: SEQ ID NO: 1/SEQ ID NO: 2; SEQ ID NO: 3/SEQ ID NO: 4; SEQ ID NO: 5/SEQ ID NO: 6; and SEQ ID NO: 7/SEQ ID NO: 8.
12 . The siRNA molecule of claim 10 , wherein the double stranded RNA portion comprises:
a first RNA strand selected from the group consisting SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8;
and a second RNA strand that is a complement of the first RNA strand.
13 . An siRNA molecule comprising a double stranded RNA portion having about 5 to 40 bp said molecule down-regulating the expression in a mammal of a protein inhibitor of osteogenesis or chondrogenesis.
14 . An siRNA molecule comprising a double stranded RNA portion and two single stranded DNA portions, the double stranded RNA portion having about 15-40 bp, the two single stranded DNA portions may be the same or different, each DNA portion comprising two deoxynucleotides which may be the same or different, each DNA portion being attached to the 3′ end of each RNA strand, the siRNA molecule down-regulating the expression in a mammal of a protein inhibitor of osteogenesis.
15 . The siRNA molecule of claim 14 wherein the patient is human.
16 . An implantable bone graft comprising in combination a sterilized segment of allograft or xenograft bone in combination with an siRNA molecule of claim 1 .
17 . An implantable bone graft comprising in combination a sterilized segment of allograft or xenograft bone in combination with an siRNA molecule of claim 9 .
18 . An implantable bone graft comprising in combination a sterilized segment of allograft or xenograft bone in combination with an siRNA molecule of claim 1 .
19 . A method of enhancing osteogenesis in a mammalian patient at the site of an osteo implant comprising
a. providing an osteo implant to a site in a mammalian patient in need of an osteo implant;
b. providing to the patient at the site of the implant an siRNA molecule of claim 14 whereby an inhibitor to osteoblastic activity is down-regulated.
20 . An implantable composition for use in a mammal comprising in combination a demineralized bone matrix and an siRNA molecule of claim 1.