Modified ligand-gated ion channels and methods of use
AssigneeHoward Hughes Medical Institute
InventorsScott Sternson, Peter Lee, Christopher Magnus
Life of this patent
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Abstract
This document relates to materials and methods for controlling ligand gated ion channel (LGIC) activity. For example, modified LGICs including at least one LGIC subunit having a modified ligand binding domain (LBD) and/or a modified ion pore domain (IPD) are provided. Also provided are exogenous LGIC ligands that can bind to and activate the modified LGIC, as well as methods of modulating ion transport across the membrane of a cell of a mammal, methods of modulating the excitability of a cell in a mammal, and methods of treating a mammal having a channelopathy.
Claims43
A method of treating neuropathic pain in a mammal, the method comprising:
- administering to the mammal a nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising:
- (a) a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) having:
- (i) at least 93 percent sequence identity to the sequence set forth in residues 23-224 of SEQ ID NO:1, residues 23-229 of SEQ ID NO:2, or residues 23-233 of SEQ ID NO:11;
- and
- (ii) an L131G amino acid substitution as numbered in SEQ ID NOs:
- 1, 2, or 11;
- and
- (b) a human glycine receptor (GlyR) ion pore domain (IPD),
- wherein an exogenous LGIC ligand selectively binds to and activates the modified LGIC;
- and administering the exogenous ligand to the mammal.
A method of modulating the activity of a cell in a mammal, said method comprising:
- administering to the cell a nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising:
- (a) a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) comprising:
- (i) at least 93 percent sequence identity to the sequence set forth in residues 23-224 of SEQ ID NO:1, residues 23-229 of SEQ ID NO:2, or residues 23-233 of SEQ ID NO:11;
- and
- (ii) an L131G amino acid substitution as numbered in SEQ ID NOs:
- 1, 2, or 11;
- and
- (b) a human glycine receptor (GlyR) ion pore domain (IPD),
- wherein an exogenous LGIC ligand selectively binds to and activates the modified LGIC;
- and administering the exogenous ligand to the mammal.
A method of treating neuropathic pain in a mammal, the method comprising:
- administering to the mammal a nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising:
- (a) a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) comprising:
- the amino acid sequence of SEQ ID NO:2 having (i) an L131G amino acid substitution, (ii) an L131G amino acid substitution and a Q139L amino acid substitution, (iii) an L131G amino acid substitution and a Y217F amino acid substitution, or (iv) an L131G amino acid substitution, a Q139L amino acid substitution, and a Y217F amino acid substitution;
- and
- (b) a human glycine receptor (GlyR) ion pore domain (IPD),
- wherein an exogenous LGIC ligand selectively binds to and activates the modified LGIC;
- and
- administering the exogenous ligand to the mammal.
A method of treating neuropathic pain in a mammal, the method comprising:
- administering to the mammal a nucleic acid encoding a modified ligand gated ion channel (LGIC) comprising at least one modified LGIC subunit, said modified LGIC subunit comprising a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) and a glycine receptor (GlyR) ion pore domain (α7-GlyR LGIC subunit), wherein said α7-GlyR LGIC subunit comprises the sequence set forth in SEQ ID NO:
- 7 having an L131G amino acid substitution, a Q139L amino acid substitution, and a Y217F amino acid substitution as numbered in SEQ ID NO:
- 7.
A method of treating neuropathic pain in a mammal, the method comprising:
- administering to the mammal a nucleic acid encoding a modified ligand gated ion channel (LGIC) subunit, said modified LGIC subunit comprising a modified alpha7 nicotinic acetylcholine receptor (α7-nAChR) ligand binding domain (LBD) and a glycine receptor (GlyR) ion pore domain (α7-GlyR LGIC subunit), wherein said α7-GlyR LGIC subunit comprises the sequence set forth in SEQ ID NO:
- 7 having an L131G amino acid substitution, a Q139L amino acid substitution, and a Y217F amino acid substitution as numbered in SEQ ID NO:
- 7.
Dependencies are inferred from the claim text.