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AssigneesProQR Therapeutics II B.V., Eli Lilly and Company
InventorsAron KOS, Lenka VAN SINT FIET, Lisanne Alieda VAN WISSEN, Marieke HOGERVORST, Ryan Matthew SMITH
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The disclosure relates to the field of diseases caused by a lowered synaptic inhibition, preferably those that are caused by a diminished activity of the potassium (K)/chloride (Cl) cotransporter (KCC2). The disclosure involves oligonucleotides and the use thereof in RNA editing methods in targeting a target adenosine in a codon encoding a phosphorylation site in the KCC2-encoding SLC12A5 pre-mRNA or mRNA, preferably the adenosine in the codon encoding threonine at position 1007 of the KCC2b isoform. Through the editing the threonine is replaced by an alanine, thereby removing the phosphorylation site, and thereby increasing the activity of the KCC2 protein in the process of restoring GABAergic inhibitory tone. The disclosure further relates to oligonucleotides for use in the treatment of chronic pain and epilepsy.
An RNA editing oligonucleotide (EON) capable of forming a double-stranded complex with a region of an endogenous human SLC12A5 transcript molecule in a cell, wherein the region of the SLC12A5 transcript molecule comprises a target adenosine, wherein the nucleotide in the EON that is directly opposite the target adenosine is the orphan nucleotide, wherein the counting of the nucleotides in the EON is such that the orphan nucleotide is number 0 and the nucleotides 5′ from the orphan nucleotide are positively (+) incremented towards the 5′ end and negatively (−) incremented towards the 3′ end, and wherein the double-stranded complex can recruit an endogenous ADAR enzyme to deaminate the target adenosine into an inosine, thereby editing the SLC12A5 transcript molecule.
A vector, preferably a viral vector, more preferably an adeno-associated virus (AAV) vector, comprising a nucleic acid molecule encoding an EON comprising a sequence according to any one of SEQ ID NO:
A method of editing a SLC12A5 polynucleotide, the method comprising contacting the SLC12A5 polynucleotide with an EON capable of effecting an ADAR-mediated adenosine to inosine editing of a target adenosine in a codon encoding an amino acid that is associated with phosphorylation of the SLC12A5-encoded protein KCC2, thereby editing the SLC12A5 polynucleotide.
A method of treating a disorder caused by a diminished GABAergic inhibition, preferably caused by a diminished KCC2 activity, in an individual in need thereof, the method comprising contacting a SLC12A5 polynucleotide in a cell of the subject with an EON capable of effecting an ADAR-mediated adenosine to inosine editing of a target adenosine in a codon encoding an amino acid that is associated with phosphorylation of the SLC12A5-encoded protein KCC2, thereby treating the individual.
A nucleic acid molecule for editing a target adenosine in a human SLC12A5 pre-mRNA or mRNA molecule, wherein the target region is SEQ ID NO:
Dependencies are inferred from the claim text.