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CN-115052590-B - RNA interference-inducing nucleic acids comprising 8-oxoguanine, modified nucleic acids that bind to microRNAs comprising 8-oxoguanine, and uses thereof

CN115052590BCN 115052590 BCN115052590 BCN 115052590BCN-115052590-B

Abstract

In the present invention, it has been confirmed that various pathophysiological phenomena are induced when an RNA interference-inducing nucleic acid comprising at least one 8-oxo-guanine (o 8 G) from the 1 st to 9 th nucleotides of at least one single strand in a double strand of nucleic acid and a modified nucleic acid specifically binding to microRNA and wherein at least one guanine (G) from the 1 st to 9 th nucleotides from the 5' end is modified with 8-oxo-guanine (o 8 G) is generated and administered to a cell or a mouse. Furthermore, the location where the G > T modification occurs has been identified in cDNA produced by reverse transcription of microRNAs in which guanine (G) is oxidatively modified by 8-oxo-guanine (o 8 G) by oxidative stress in the seed region of microRNAs to confirm the location where oxidative modification to 8-oxo-guanine occurs.

Inventors

  • CHI CHENGXU
  • SHI XIYING

Assignees

  • 高丽大学校产学协力团

Dates

Publication Date
20260505
Application Date
20201130
Priority Date
20191128

Claims (10)

  1. RNA interference inducing nucleic acid comprising the 5' terminal sequence of a polynucleotide selected from the group consisting of 5'p-Uo 8 GGAAUG-3', 5'p-UGo 8 GAAUG-3', and 5'p-UGGAAUo 8 G-3', The RNA interference-inducing nucleic acid induces myocardial hypertrophy when injected into cells or animals.
  2. 2. The RNA interference-inducing nucleic acid of claim 1, comprising a polynucleotide selected from group 2: [ group 2] The polynucleotide of SEQ ID NO. 1 (5'p-Uo 8 GGAAUGUAAAGAAGUAUGUAU-3'); The polynucleotide of SEQ ID NO. 2 (5'p-UGo 8 GAAUGUAAAGAAGUAUGUAU-3'), and The polynucleotide of SEQ ID NO. 3 (5'p-UGGAAUo 8 GUAAAGAAGUAUGUAU-3').
  3. 3. A composition comprising the RNA interference-inducing nucleic acid of claim 1 or claim 2 and an antioxidant.
  4. 4. Modified nucleic acids that specifically bind to the modified microRNAs, Wherein the modified microRNA comprises a polynucleotide selected from group 2: [ group 2] The polynucleotide of SEQ ID NO. 1 (5'p-Uo 8 GGAAUGUAAAGAAGUAUGUAU-3'); The polynucleotide of SEQ ID NO. 2 (5'p-UGo 8 GAAUGUAAAGAAGUAUGUAU-3'), and The polynucleotide of SEQ ID NO. 3 (5'p-UGGAAUo 8 GUAAAGAAGUAUGUAU-3'), and Wherein the modified nucleic acid comprises adenine (a) that binds to at least one 8-oxo-guanine (o 8 G) of nucleotide 2,3, or 7 from the 5' end of the modified microRNA.
  5. 5. The modified nucleic acid of claim 4, wherein The modified nucleic acid comprises any one of the base sequences 5'-ACAUUCA-3', 5'-ACAUUAC-3', or 5 '-AAAUUCC-3'.
  6. 6. A recombinant vector comprising a gene encoding the modified nucleic acid of claim 4 or claim 5.
  7. 7. A pharmaceutical composition for treating cardiac hypertrophy comprising The modified nucleic acid of claim 4 or 5, or a recombinant vector comprising a gene encoding said modified nucleic acid.
  8. 8. The pharmaceutical composition for treating cardiac hypertrophy of claim 7 further comprising an antioxidant.
  9. 9. The pharmaceutical composition of claim 7, wherein the antioxidant is N-acetylcysteine or butylated hydroxyanisole.
  10. 10. Use of an anti-8-oxoguanine (o 8 G) antibody in the manufacture of a kit for diagnosing cardiac hypertrophy, wherein said kit is used in a method comprising: Determining whether one or more guanines (G) in the nucleotides of microRNAs isolated from cardiomyocytes of the animal are modified to 8-oxo-guanine (o 8 G), and When one or more guanines (G) of nucleotides 2, 3 or 7 from the 5' -end of the microRNA are modified to 8-oxo-guanine (o 8 G), cardiac hypertrophy is classified, Wherein the microRNA is miR-1 shown in Seq ID No. 7.

Description

RNA interference-inducing nucleic acids comprising 8-oxoguanine, modified nucleic acids that bind to microRNAs comprising 8-oxoguanine, and uses thereof Technical Field The present invention relates to an RNA interference-inducing nucleic acid comprising 8-oxoguanine, a modified nucleic acid specifically binding to microRNA comprising 8-oxoguanine, and a pharmaceutical composition, a recombinant animal model, a drug screening method, a disease diagnosis method and a method for controlling pathophysiological phenomenon using the same. Background Oxidative stress typically occurs when cells undergo pathophysiological changes, producing Reactive Oxygen Species (ROS). The generated active oxygen modifies various biological structures due to its high reactivity, with RNA being the easiest to modify. In particular, it is most likely that guanine (G) bases are oxidatively modified and converted to 8-oxo-guanine (o 8G) in oxidatively modified RNA bases. On the other hand, heart disease is one of three causes of death among korean adults including cancer, which starts from various pathological stresses during the course of attacks, resulting in a change in the size of heart tissue and cardiac hypertrophy. Cardiac hypertrophy is characterized by an increase in cardiomyocyte size and an increase in the rate of protein synthesis. Cardiac hypertrophy can gradually induce myocardial fibrosis and heart failure (heart failure), ultimately leading to heart failure (or heart failure). In particular, in the case of heart failure, research is being conducted to finally prevent cardiac hypertrophy or develop a therapeutic method for cardiac hypertrophy due to an extremely high mortality rate of around 50%, and thus it is necessary to build a disease model therefor. Korean patent publication No. 1481007 BRIEF SUMMARY OF THE PRESENT DISCLOSURE Technical problem The present inventors have confirmed that when oxidation modification of guanine (G) base into 8-oxo guanine (o 8 G) occurs due to oxidative stress in the seed region of micrornas, it binds to a target sequence through the o 8 G: a arrangement, and also confirmed that oxidation modification into 8-oxo guanine occurs in the produced cDNA by identifying the position where G > T modification occurs in the cDNA produced by reverse transcription of micrornas. Therefore, it was confirmed that various pathophysiological phenomena are induced when RNA interference-inducing nucleic acid in which guanine (G) in microRNA is replaced with 8-oxo-guanine (o 8 G) is produced and administered to cells or mice, and the present invention was completed based on this. Thus, in the present invention, the 1 st to 9 th nucleotides of the 5' end of at least one single strand of a nucleic acid duplex that induces RNA interference provide an RNA interference-inducing nucleic acid comprising at least one 8-oxo-guanine (o 8 G). Furthermore, the present invention provides a method for identifying the position of 8-oxo-guanine (o 8 G). Furthermore, the present invention provides a modified nucleic acid specifically binding to micrornas in which at least one guanine (G) of nucleotides 1 to 9 of the 5' end is modified to 8-oxo-guanine (o 8 G), and a recombinant vector comprising a gene encoding the modified nucleic acid. Furthermore, the present invention provides a pharmaceutical composition for treating cardiac hypertrophy comprising the modified nucleic acid or recombinant vector and a pharmaceutical composition for treating liver cancer or glioblastoma comprising the RNA interference-inducing nucleic acid. Another embodiment provides a pharmaceutical composition for preventing or treating cardiac hypertrophy comprising an antioxidant as an active ingredient and an information providing method for diagnosing cardiac hypertrophy. Another embodiment provides a method for generating an anti-cardiac hypertrophy animal model and an anti-cardiac hypertrophy animal model. Another embodiment provides a method for screening candidate substances for treating cardiac hypertrophy. Technical objects to be achieved by the present invention are not limited to the above objects, and other objects not mentioned will be clearly understood by those skilled in the art to which the present invention pertains from the following description. Technical proposal The present invention provides an RNA interference inducing nucleic acid comprising at least one 8-oxo-guanine (o 8 G) in the 1 st to 9 th nucleotides of at least one single strand of a nucleic acid duplex. One embodiment of the present invention provides an RNA interference-inducing nucleic acid, wherein the 5' 1 st to 9 th nucleotides comprise a microRNA sequence. In another embodiment of the present invention, the micrornas can be at least one of the following group 1 micrornas: [ group 1] MiR-1, miR-184, let-7f-5p, miR-1-3p, miR-122, let-7 and miR-124. In another embodiment of the invention, the RNA interference inducing nucleic acid may recognize a target site in