Methods and compositions related to induced sensory neurons

Abstract

This invention provides methods of generating induced sensory neurons (iSNs) from non-neuronal cells such as fibroblasts. The invention also provides methods of using iSNs in various therapeutic or non-therapeutic applications, e.g., methods to identify agents or cellular modulations that enhance iSN formation from non-neuronal cells.

We claim: 
     
         1 . A method for generating induced sensory neurons (iSNs), comprising co-expressing Brn3A and Ngn2 genes via one or more expression vectors harboring Brn3A and Ngn1 genomic or cDNA sequences in a non-neuronal cell, thereby generating induced sensory neurons, wherein the non-neuronal cell is a fibroblast or a stem cell from a mammal. 
     
     
         2 . The method of  claim 1 , wherein expression of the Brn3A/Ngn2 genes is temporal. 
     
     
         3 . The method of  claim 2 , wherein temporal expression of the Brn3A/Ngn2 genes is via inducible expression. 
     
     
         4 . The method of  claim 1 , wherein the non-neuronal cell is a fibroblast, an embryonic stem cell (ESC), or an induced pluripotent stem cell (iPSC). 
     
     
         5 . The method of  claim 1 , wherein the non-neuronal cell is an embryonic fibroblast or an adult fibroblast. 
     
     
         6 . The method of  claim 1 , wherein the mammal is human, mouse or rat. 
     
     
         7 . The method of  claim 1 , wherein an expression vector harboring the Brn3A gene and the Ngn2 gene is introduced into the non-neuronal cell. 
     
     
         8 . The method of  claim 7 , wherein the expression vector is a lentiviral vector. 
     
     
         9 . The method of  claim 1 , further comprising detecting in the induced sensory neurons expression of one or more neuronal markers. 
     
     
         10 . An induced sensory neuron generated in accordance with  claim 1 . 
     
     
         11 . An isolated cell comprising one or more expression vectors, wherein the expression vectors express a combination of Brn3A/Ngn1 genes or Brn3A/Ngn2 genes. 
     
     
         12 . A method for treating in a subject a neurological condition or disorder that is associated with or mediated by a loss or degeneration of sensory neurons, comprising (1) obtaining a non-neuronal cell from the subject in need of treatment; (2) generating a population of induced sensory neurons (iSNs) from said non-neuronal cell by co-expressing a combination of Brn3A/Ngn1 genes or Brn3A/Ngn2 genes via one or more expression vectors in the non-neuronal cell; and (3) administering a therapeutically effective amount of the iSN population to the subject, thereby treating the neurological condition or disorder in the subject. 
     
     
         13 . The method of  claim 12 , wherein expression of the Brn3A/Ngn1 genes or Brn3A/Ngn2 genes is temporal. 
     
     
         14 . The method of  claim 13 , wherein temporal expression of the Brn3A/Ngn1 genes or Brn3A/Ngn2 genes is via inducible expression. 
     
     
         15 . The method of  claim 12 , wherein the non-neuronal cell is a fibroblast, an embryonic stem cell (ESC), or an induced pluripotent stem cell (iPSC). 
     
     
         16 . The method of  claim 12 , wherein the non-neuronal cell is an embryonic fibroblast or an adult fibroblast. 
     
     
         17 . The method of  claim 12 , wherein an expression vector harboring the Brn3A/Ngn1 genes or Brn3A/Ngn2 genes is introduced into the non-neuronal cell. 
     
     
         18 . The method of  claim 17 , wherein the expression vector is a lentiviral vector. 
     
     
         19 . The method of  claim 12 , further comprising examining the induced sensory neurons for the presence of a neuronal marker. 
     
     
         20 . The method of  claim 12 , wherein the subject is a human.