MIRI26-5P for treating motor neuron diseases
The present invention relates to methods for treating motor neuron diseases and neuromuscular junction abnormalities. Particularly, the methods comprise increasing mi RNA 126-5p expression in skeletal muscle cells and/or motor neurons, and/or inhibiting mi RNA 126-5p expression in glial cells, thereby spatially up-regulating and/or down-regulating mi RNA 126-5p levels, and thus treating amyotrophic lateral sclerosis.
1 . A method for treating a motor neuron disease (MND), the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a pharmaceutical combination comprising:
(a) a first pharmaceutical composition comprising a therapeutically effective amount of an agent selected from the group consisting of: (i) microRNA (miR) 126-5p, a precursor, or a homolog thereof; (ii) a polynucleotide encoding miR126-5p, a precursor or a homolog thereof; and (iii) an expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the polynucleotide and the expression vector comprising the polynucleotide further comprises a muscle specific promoter, and wherein the first pharmaceutical composition is effective to increase miR126-5p expression in a skeletal muscle cell; and
(b) a second pharmaceutical composition comprising a therapeutically effective amount of an agent selected from the group consisting of: (i) miR126-5p, a precursor or a homolog thereof; (ii) a polynucleotide encoding miR126-5p, a precursor or a homolog thereof; and (iii) an expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the polynucleotide and the expression vector comprising the polynucleotide further comprises a neuron specific promoter, and wherein the second pharmaceutical composition is effective to increase miR126-5p expression in a motor neuron;
wherein the MND is selected from the group consisting of Amyotrophic Lateral Sclerosis (ALS), primary lateral sclerosis, progressive muscular atrophy, pseudobulbar palsy and progressive bulbar palsy;
wherein the miR126-5p consists of the nucleotide sequence set forth in SEQ ID NO: 1, and wherein a homolog thereof comprises a seed region identical to positions 2-8 of SEQ ID NO: 1, and is at least 90% identical to SEQ ID NO: 1.
2 . The method according to claim 1 ,
wherein the pharmaceutical compositions are administered simultaneously or sequentially.
3 . The method according to claim 1 , wherein the pharmaceutical combination comprises the following pharmaceutical compositions: (a) a first pharmaceutical composition comprising a therapeutically effective amount of the expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the first pharmaceutical composition is effective to increase miR126-5p expression in a skeletal muscle cell; and (b) a second pharmaceutical composition comprising a therapeutically effective amount of the expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the second pharmaceutical composition is effective to increase miR126-5p expression in a motor neuron.
4 . The method according to claim 1 , wherein at least one of the expression vectors is a viral vector.
5 . The method according to claim 4 , wherein the viral vector is selected from the group consisting of lentiviral vectors and adeno-associated viral vectors.
6 . The method according to claim 5 , wherein the lentiviral vector is selected from the group consisting of an HIV-based lentiviral vector, an EIAV-based lentiviral vector, and self-inactivating (SIN) lentiviral vector.
7 . The method according to claim 1 , wherein the muscle specific promoter is a skeletal muscle specific promoter.
8 . The method according to claim 1 , wherein the muscle specific promoter is selected from the group consisting of skeletal muscle α-actin promoter, myogenin promoter, and muscle creatine kinase promoter.
9 . The method according to claim 1 , wherein the neuron specific promoter is selected from the group consisting of neurofilament promoter, HB9 promoter, Thy-1 promoter, and synapsin promoter.
10 . The method according to claim 1 , wherein the muscle specific promoter is a skeletal muscle α-actin promoter, and wherein the neuron specific promoter is a neurofilament promoter.
11 . The method according to claim 1 , wherein the first pharmaceutical composition is formulated for intramuscular, intravenous, or intra-arterial injection.
12 . The method according to claim 1 , wherein the second pharmaceutical composition is formulated for injection or infusion into the spinal cord or CNS.
13 . The method according to claim 1 , wherein the pharmaceutical combination comprises:
(a) the first pharmaceutical composition comprising an expression vector comprising SEQ ID NO: 1 operably linked to a skeletal muscle α-actin promoter, and wherein the first pharmaceutical composition is formulated for injection into a skeletal muscle tissue; and
(b) the second pharmaceutical composition comprising an expression vector comprising SEQ ID NO: 1 operably linked to a neurofilament promoter, and wherein the second pharmaceutical composition is formulated for injection or infusion into the spinal cord or CNS.
14 . A method of treating a motor neuron disease (MND), the method comprising administering to a subject having the MND at least one pharmaceutical composition selected from:
a first pharmaceutical composition comprising an agent selected from the group consisting of: (i) miR126-5p, a precursor or a homolog thereof; (ii) a polynucleotide encoding miR126-5p, a precursor or a homolog thereof; and (iii) an expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the polynucleotide and the expression vector comprising the polynucleotide further comprises a muscle specific promoter, and wherein the first pharmaceutical composition is effective to increase miR126-5p expression in a skeletal muscle cell; and
a second pharmaceutical composition comprising an agent selected from the group consisting of: (i) miR126-5p, a precursor or a homolog thereof; (ii) a polynucleotide encoding miR126-5p, a precursor or a homolog thereof; and (iii) an expression vector comprising a polynucleotide encoding miR126-5p, a precursor or a homolog thereof, wherein the polynucleotide and the expression vector comprising the polynucleotide further comprises a neuron specific promoter, and wherein the second pharmaceutical composition is effective to increase miR126-5p expression in a motor neuron; and
wherein the MND is selected from the group consisting of Amyotrophic Lateral Sclerosis (ALS), primary lateral sclerosis, progressive muscular atrophy, pseudobulbar palsy and progressive bulbar palsy; wherein the miR126-5p consists of the nucleotide sequence set forth in SEQ ID NO: 1, and wherein a homolog thereof comprises a seed region identical to positions 2-8 of SEQ ID NO: 1, and is at least 90% identical to SEQ ID NO: 1.