Compositions and methods for neurological diseases

Abstract

Compositions and methods are provided for modulating the activity of cells using engineered receptors, polynucleotide encoded engineered receptors, and gene therapy vectors comprising polynucleotides encoding engineered receptors. These compositions and methods find particular use in modulating the activity of neurons, for example in the treatment of disease or in the study of neuronal circuits.

1 - 29 . (canceled) 
     
     
         30 . An engineered receptor, wherein the engineered receptor is a chimeric ligand gated ion channel (LGIC) receptor and comprises
 (a) a ligand binding domain derived from a human α7 nicotinic acetylcholine receptor (α7-nAChR) and comprising a Cys-loop domain from a second LGIC receptor; and   (b) an ion pore domain derived from the second LGIC receptor,   wherein the second LGIC receptor is a Cys-loop receptor, and wherein the second LGIC receptor is not an α7-nAChR or a serotonin receptor.   
     
     
         31 . The engineered receptor according to  claim 30 , wherein the ligand binding domain shares a sequence identity of 85% or more to a ligand binding domain of a wild type human α7-nAChR according to SEQ ID NO: 4. 
     
     
         32 . The engineered receptor according to  claim 30 , wherein the second LGIC receptor is a Glycine receptor (GlyR) or a λ-aminobutyric acid (GABA) receptor. 
     
     
         33 . The engineered receptor according to  claim 30 , wherein the ligand binding domain comprises one or more amino acid substitutions at a residue corresponding to a residue of the wild type human α7-nAChR selected from the group consisting of W77, Y94, R101, W108, Y115, T128, N129, V130, L131, Q139, L141, Y151, S170, W171, S172, S188, Y190, Y210, C212, C213 and Y217. 
     
     
         34 . The engineered receptor according to  claim 30 , wherein the ligand binding domain comprises one or more amino acid substitutions at a residue corresponding to a residue of the wild type human α7-nAChR selected from the group consisting of R101, Y115, L131, S170, S172, and Y210. 
     
     
         35 . The engineered receptor according to  claim 34 , wherein the amino acid substitution is selected from a substitution corresponding to Y115D, L131S, L131T, L131D, S170T, or S172D of the wild type human α7-nAChR. 
     
     
         36 . The engineered receptor according to  claim 30 , wherein the second LGIC receptor is a Glycine receptor (GlyR), and the engineered receptor shares a sequence identity of 85% or more to SEQ ID NO: 33. 
     
     
         37 . The engineered receptor according to  claim 36 , wherein the ligand binding domain comprises one or more amino acid substitutions at a residue corresponding to a residue of the wild type human α7-nAChR selected from the group consisting of R101, Y115, L131, S170, S172, and Y210. 
     
     
         38 . The engineered receptor according to  claim 37 , wherein the amino acid substitution is selected from a substitution corresponding to Y115D, L1315, L131T, L131D, S170T, or S172D of the wild type human α7-nAChR. 
     
     
         39 . The engineered receptor according to  claim 30 , wherein the ligand binding domain further comprises a β1-2 loop domain from the second LGIC receptor. 
     
     
         40 . The engineered receptor according to  claim 39 , wherein the ligand binding domain comprises one or more amino acid substitutions at a residue corresponding to a residue of the wild type human α7-nAChR selected from the group consisting of R101, Y115, L131, S170, S172, and Y210. 
     
     
         41 . The engineered receptor according to  claim 40 , wherein the amino acid substitution is selected from a substitution corresponding to Y115D, L1315, L131T, L131D, S170T, or S172D of the wild type human α7-nAChR. 
     
     
         42 . A polynucleotide encoding the engineered receptor according to  claim 30 . 
     
     
         43 . A vector comprising the polynucleotide according to  claim 42 . 
     
     
         44 . The vector according to  claim 43 , wherein the vector is a viral vector selected from the group consisting of an adenoviral vector, a retroviral vector, an adeno-associated viral (AAV) vector, and a herpes simplex-1 viral vector (HSV-1). 
     
     
         45 . A pharmaceutical composition comprising the vector according to  claim 43  and a pharmaceutically acceptable vehicle. 
     
     
         46 . A method of ameliorating a neurological disorder in a subject in need thereof comprising: administering to the subject a polynucleotide encoding the engineered ligand-gated ion channel (LGIC) receptor of  claim 30 , and administering to the subject a binding agent, wherein the binding agent acts as an agonist of the engineered LGIC receptor. 
     
     
         47 . The method according to  claim 46 , wherein the polynucleotide is delivered to the subject in a viral vector. 
     
     
         48 . The method according to  claim 47 , wherein the viral vector is an adenoviral vector, a retroviral vector, an adeno-associated viral (AAV) vector, or a herpes simplex-1 viral vector (HSV-1). 
     
     
         49 . The method according to  claim 48 , wherein the AAV vector is AAV5 or a variant thereof, AAV6 or a variant thereof, or AAV9 or a variant thereof. 
     
     
         50 . The method according to  claim 46 , wherein the polynucleotide is administered to the subject by a non-viral method. 
     
     
         51 . The method according to  claim 50 , wherein the non-viral method is lipofection, nanoparticle delivery, particle bombardment, electroporation, sonication, or microinjection. 
     
     
         52 . The method according to  claim 46 , wherein the polynucleotide encoding the engineered receptor is operably linked to a promoter that is active in an excitable cell. 
     
     
         53 . The method according to  claim 52 , wherein the excitable cell is a neuron or a myocyte. 
     
     
         54 . The method according to  claim 53 , wherein the neuron is a dorsal root ganglion, a motor neuron, an excitatory neuron, an inhibitory neuron, or a sensory neuron. 
     
     
         55 . The method according to  claim 54 , wherein the polynucleotide is administered subcutaneously, orally, intrathecally, topically, intravenously, intraganglionically, intraneurally, intracranially, intraspinally, or to the cisterna magna. 
     
     
         56 . The method according to  claim 46 , wherein the binding agent is selected from the group consisting of AZD0328, ABT-126, TC6987, and Facinicline/RG3487. 
     
     
         57 . The method according to  claim 56 , wherein the binding agent is administered orally, subcutaneously, topically, or intravenously. 
     
     
         58 . The method according to  claim 46 , wherein the subject suffers from pain, a seizure disorder, a movement disorder, an eating disorder, a spinal cord injury, neurogenic bladder, a spasticity disorder, or pruritus. 
     
     
         59 . The method according to  claim 58 , wherein the subject is a human.