JP-2022515211-A5 -

Dates

Publication Date

20221223

Application Date

20191219

Description

Another aspect of the present invention is a method for determining whether a subject is at risk of cardiovascular disease or is suffering from cardiovascular disease, (A) Provide a sample from an individual suspected of having cardiovascular disease: (B) If applicable, RNA may be extracted from the sample: (C) A method comprising the step of measuring the expression level of hsa-miR-297 or hsa-miR-574-3p by qRT-PCR, wherein a lower expression level of hsa-miR-297 or hsa-miR-574-3p compared to a control sample indicates the presence of or susceptibility to cardiovascular disease. Further aspects of the present invention are described in the following sections: [Section 1] A synthetic miRNA molecule having at least 80% sequence identity with SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 15, or SEQ ID NO: 16 or its complementary sequence. [Section 2] A synthetic miRNA molecule according to item 1 above, wherein the seed sequence includes GAUGUGU, UACAUAC, or UGUAUGUG. [Section 3] The synthetic miRNA molecule according to item 1 or 2 above, wherein the nucleic acid molecule has a length of 14 to 30 nucleotides. [Section 4] The synthetic miRNA molecule according to any one of items 1 to 3 above, wherein the nucleic acid molecule binds to the promoter region or 3'UTR of the target gene. [Section 5] A synthetic miRNA molecule as described in item 4 above, wherein the target gene is VEGFA, VEGFD, or HIF1A. [Section 6] A synthetic miRNA molecule as described in item 5 above, wherein the target gene is VEGFA. [Section 7] A synthetic miRNA molecule as described in any of items 1 to 6 above, for use in the treatment of cardiovascular disease. [Section 8] A pharmaceutical composition comprising at least one synthetic miRNA molecule as described in any of items 1 to 7 above, and a carrier or excipient. [Section 9] The pharmaceutical composition according to item 8, wherein the carrier or excipient is a pharmaceutically acceptable carrier or excipient suitable for medical treatment. [Section 10] A pharmaceutical composition according to item 8 or 9 above, for use in the treatment of cardiovascular disease. [Section 11] A recombinant expression vector comprising at least one synthetic miRNA molecule as described in any of items 1 to 7 above. [Section 12] Cells containing the recombinant expression vector described in item 11 above. [Section 13] A method for regulating the expression of a target gene in humans, comprising administering a synthetic miRNA molecule as described in any of items 1 to 7 above. [Section 14] The method according to item 13 above, which increases the expression of a target gene. [Section 15] The method according to item 13 or 14, wherein the method comprises administering to a subject an effective amount at least one synthetic miRNA having at least 80% sequence identity with SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 15, or SEQ ID NO: 16. [Section 16] A method for determining whether a subject is at risk of or has cardiovascular disease includes the following steps: (A) Provide a sample from an individual suspected of having cardiovascular disease; (B) If applicable, RNA may be extracted from the sample; (C) The expression level of hsa-miR-297 or hsa-547-3p is measured by qRT-PCR; here, a lower expression level of hsa-miR-297 or hsa-547-3p compared to the control sample indicates the presence of cardiovascular disease or a predisposition to cardiovascular disease.