Uses of modified mullerian inhibiting substance (mis) proteins for the treatment of neurodegenerative diseases

Abstract

The present invention relates to methods to treat a neurodegenerative disease or disorder, e.g., a motor neuron disease in a subject, whereby the subject is administered a recombinant human Mullerian Inhibiting Substance (MIS) protein as disclosed herein, wherein the recombinant human MIS protein comprises a modified Kex cleavage site for increased cleavage. The recombinant human MIS protein can be produced from a pre-proprotein comprising a non-MIS leader sequence or a functional fragment thereof in place of the MIS leader sequence.

1 . A method for treating a subject with a neurodegenerative disease or disorder, comprising administering a composition comprising a recombinant Mullerian Inhibiting Substance (MIS) protein, wherein the recombinant MIS protein comprises at least one of: (i) a modification of amino acid 450 of SEQ ID NO: 1 from Q to R to increase cleavage as compared to in the absence of such a modification, and (ii) a modification of amino acid 452 of SEQ ID NO: 1 from S to R to increase cleavage as compared to in the absence of such a modification, and wherein the vector does not include a nucleic acid encoding a FLAG tag. 
     
     
         2 .- 4 . (canceled) 
     
     
         5 . The method of  claim 1 , wherein one or more neurons from the subject express the MIS type II receptor or a homologue or functional fragment thereof. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the neurodegenerative disease is selected from the group of: amyotrophic lateral sclerosis (ALS), progressive bulbar palsy, pseudobulbar palsy; primary lateral sclerosis (PLS); progressive muscular atrophy; spinal muscular atrophy (SMA, including SMA type I, SMA type II, and SMA type III); Fazio-Londe disease; progressive spinobulbar muscular atrophy; congenital SMA with arthrogryposis or post-polio syndrome (PPS). 
     
     
         8 . The method of  claim 1 , wherein the neurodegenerative disease is selected from the group of: Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), Friedreich's ataxia, cerebellar ataxia, other brain disorders such as bipolar disorder, epilepsy, schizophrenia, depression, mania, autism, ADHD, brain trauma injuries or stroke. 
     
     
         9 .- 24 . (canceled) 
     
     
         25 . The method of  claim 1 , wherein the recombinant MIS protein is encoded by a viral vector or expression vector. 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 25 , wherein the viral vector is selected from the group consisting of an adenoviral vector, adeno-associated vector (AAV), a poxvirus vector or a lentiviral vector. 
     
     
         29 . (canceled) 
     
     
         30 . The method of  claim 28 , wherein the adeno-associated vector (AAV) is AAV9. 
     
     
         31 . The method of  claim 25 , wherein the vector comprises a nucleic acid sequence that has at least 95% sequence identity to the nucleic acid sequence of SEQ ID NO: 4 or SEQ ID NO: 5, wherein the nucleic acid sequence is operatively linked to a tissue- or cell-type specific promoter. 
     
     
         32 .- 44 . (canceled) 
     
     
         45 . The method of  claim 31 , wherein the nucleic acid encodes a non-MIS leader sequence in place of the MIS leader sequence of amino acids 1-25 of SEQ ID NO: 1. 
     
     
         46 . The method of  claim 45 , wherein the non-MIS leader sequence comprises at least 10 amino acids of SEQ ID NO: 6 or a variant that is at least 80% homologous thereto. 
     
     
         47 . The method vector of  claim 25 , wherein the vector comprises a nucleic acid sequence of nucleotides 79-1680 of SEQ ID NO: 4. 
     
     
         48 . The method of  claim 31 , wherein the nucleic acid sequence comprises SEQ ID NO: 4. 
     
     
         49 . The method of  claim 1 , wherein the recombinant MIS protein comprises the amino acid residues 25-559 of SEQ ID NO: 2. 
     
     
         50 . The method of  claim 45 , wherein the non-MIS leader sequence is selected from any in the group consisting of: a human serum albumin (HSA) leader sequence, an immunoglobulin signal peptide fused to a tissue-type plasminogen activator propeptide (IgSP-tPA), murine immunoglobulin signal peptide (IgSP), a MPIF-1 signal sequence (MKVSVAALSCLMLVTALGSQA (SEQ ID NO: 15)); a stanniocalcin signal sequence (MLQNSAVLLLLVISASA (SEQ ID NO:16)); an invertase signal sequence (MLLQAFLFLLAGFAAKISA (SEQ ID NO:17)); a yeast mating factor alpha signal sequence ( K. lactis  killer toxin leader sequence); a hybrid signal sequence (MKWVSFISLLFLFSSAYSRSLEKR, (SEQ ID NO:18)); a HSA/MFα-1 hybrid signal sequence (MKWVSFISLLFLFSSAYSRSLDKR (SEQ ID NO:19)); a  K. lactis  killer/MFα-1 fusion leader sequence (MNIFYIFLFLLSFVQGSLDKR (SEQ ID NO:20)); an immunoglobulin Ig signal sequence (MGWSCIILFLVATATGVHS (SEQ ID NO:21)); a Fibulin B precursor signal sequence (MERAAPSRRVPLPLLLLGGLALLAAGVDA (SEQ ID NO:22)); a clusterin precursor signal sequence (MMKTLLLFVGLLLTWESGQVLG (SEQ ID NO: 23)); and the insulin-like growth factor-binding protein 4 signal sequence (MLPLCLVAALLLAAGPGPSLG (SEQ ID NO:24)). 
     
     
         51 . The method of  claim 50 , wherein a HSA leader sequence is selected from the group consisting of: MKWVTFISLLFLFSSAYS (SEQ ID NO: 13); MKWVTFISLLFLFSSAYSRGVFRR (SEQ ID NO: 6); MKWVSFISLLFLFSSAYS (SEQ ID NO: 14).