Compositions and Methods for Treating Neurogenic Disorders of the Pelvic Floor

Abstract

Provided herein are methods for the treatment of bladder dysfunction, including detrusor hyperreflexia and detrusor external sphincter dyssynergia, fecal incontinence, and/or sexual dysfunction in an individual via the use of stably expressed light-responsive opsin proteins capable of selective hyperpolarization or depolarization of the neural cells that innervate the muscles responsible for physiologic functioning of urinary bladder, external urinary sphincter, external anal sphincter, and the male and female genitalia.

1 . A method for treating a bladder dysfunction in an individual in need thereof, the method comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the detrusor muscle and/or a population of neurons responsible for the innervation of the external urinary sphincter muscle in the individual,   wherein the opsin protein induces hyperpolarization of the neurons expressing the opsin protein in response to light, whereby the hyperpolarization of the neurons responsible for the innervation of the detrusor muscle permits storage of urine in the bladder and the hyperpolarization of the neurons responsible for the innervation of the external urinary sphincter muscle permits voiding urine from the bladder,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising: i) a sequence at least 95% identical to the sequence shown in SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:23; ii) an ER export signal; and iii) a membrane trafficking signal.   
     
     
         2 . The method of  claim 1 , further comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the detrusor muscle and/or a population of neurons responsible for the innervation of the external urinary sphincter muscle in the individual,   wherein the opsin protein induces depolarization of the neurons, whereby the depolarization of the neurons responsible for the innervation of the detrusor muscle permits voiding urine from the bladder and the depolarization of the neurons responsible for the innervation of the external urinary sphincter muscle permits storage of urine in the bladder,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising a sequence at least 95% identical to the sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11.   
     
     
         3 . (canceled) 
     
     
         4 . A method for treating a bladder dysfunction in an individual in need thereof, the method comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the detrusor muscle and/or a population of neurons responsible for the innervation of the external urinary sphincter muscle in the individual,   wherein the opsin protein is capable of inducing depolarization of the neurons, whereby the depolarization of the neurons responsible for the innervation of the detrusor muscle permits voiding urine from the bladder and the depolarization of the neurons responsible for the innervation of the external urinary sphincter muscle permits storage of urine in the bladder,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising a sequence at least 95% identical to the sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11.   
     
     
         5 . The method of  claim 1 , wherein the bladder dysfunction is detrusor hyperreflexia and/or detrusor-external sphincter dyssynergia, or wherein the bladder dysfunction is associated with spinal cord injury or multiple sclerosis. 
     
     
         6 . (canceled) 
     
     
         7 . The method of  claim 1 , wherein the population of neurons responsible for the innervation of the detrusor muscle are neurons arising from one or more sacral spinal nerves in the individual. 
     
     
         8 . The method of  claim 1 , wherein the population of neurons responsible for the innervation of the external urinary sphincter muscle comprises neurons of the pudendal nerve in the individual. 
     
     
         9 . The method of  claim 1 , wherein a light source capable of generating light in response to an external signal is placed around the detrusor innervations arising from the sacral spinal nerves and/or around the external urinary sphincter innervations of the pudendal nerve, wherein the light is capable of activating the light-responsive opsin protein expressed on the plasma membrane of the neurons. 
     
     
         10 . (canceled) 
     
     
         11 . The method of  claim 9 , wherein the individual externally controls the polarization state of the neurons responsible for the innervation of the detrusor muscle and/or neurons responsible for the innervation of the external urinary sphincter by activating the light source surrounding the sacral spinal nerve and/or the pudendal nerve. 
     
     
         12 . The method of  claim 1 , wherein the polynucleotide is administered by injection of the polynucleotide into the somatic motor neuron cell body of the sacral spinal nerve, into the pudendal nerve, or into the Onuf's nucleus. 
     
     
         13 . The method of  claim 1 , wherein the polynucleotide is administered by injection of the polynucleotide into the pudendal nerve, or by injection of the polynucleotide into Onuf's nucleus. 
     
     
         14 . (canceled) 
     
     
         15 . A method for treating fecal incontinence in an individual in need thereof, the method comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the external anal sphincter muscle in the individual,   wherein the opsin protein induces hyperpolarization of the neurons expressing the opsin protein in response to light, whereby the hyperpolarization of the neurons responsible for the innervation of the external anal sphincter muscle permits defecation,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising: i) a sequence at least 95% identical to the sequence shown in SEQ ID NO:1, SEQ ID NO:4, or SEQ ID NO:23; ii) an ER export signal; iii) and a membrane trafficking signal.   
     
     
         16 . The method of  claim 15 , further comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the external anal sphincter muscle in the individual,   wherein the opsin protein induces depolarization of the neurons, whereby the depolarization of the neurons responsible for the innervation of the external anal sphincter muscle permits storage of stool in the rectum,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising a sequence at least 95% identical to the sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11.   
     
     
         17 . (canceled) 
     
     
         18 . A method for treating fecal incontinence in an individual in need thereof, the method comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the external anal sphincter muscle in the individual,   wherein the opsin protein is capable of inducing depolarization of the neurons, whereby the depolarization of the neurons responsible for the innervation of the external anal sphincter muscle permits storage of stool in the rectum,   wherein the polynucleotide comprises a nucleotide sequence encoding a light-responsive opsin protein comprising a sequence at least 95% identical to the sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11.   
     
     
         19 . The method of  claim 15 , further comprising administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive cation channel protein to the individual, wherein the light-responsive cation channel protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the muscles responsible for the contraction of the rectum in the individual, wherein the light-responsive cation channel protein is capable of inducing depolarization of the neurons expressing the light-responsive protein in response to light, whereby the depolarization of the neurons responsible for the innervation of the muscles responsible for the contraction of the rectum facilitates defecation. 
     
     
         20 . The method of  claim 15 , wherein the fecal incontinence is associated with childbirth, long-term constipation, stroke, diabetes, multiple sclerosis, or spinal cord injury. 
     
     
         21 . The method of  claim 15 , wherein the population of neurons responsible for the innervation of the external anal sphincter muscle are neurons arising from one or more sacral spinal nerves in the individual, or wherein the population of neurons responsible for the innervation of the external anal sphincter muscle comprises neurons of the inferior hemorrhoidal branch of the pudendal nerve in the individual. 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 15 , wherein a light source capable of generating light in response to an external signal is placed around the external anal sphincter innervations of the sacral spinal nerves and/or around the inferior hemorrhoidal branch of the pudendal nerve, wherein the light is capable of activating the light-responsive opsin protein expressed on the plasma membrane of the neurons. 
     
     
         24 . (canceled) 
     
     
         25 . The method of  claim 23 , wherein the individual externally controls the polarization state of the neurons responsible for the innervation of the external anal sphincter muscle by activating the light source surrounding the external anal sphincter innervations of the sacral spinal nerves and/or around the inferior hemorrhoidal branch of the pudendal nerve. 
     
     
         26 . The method of  claim 15 , wherein the polynucleotide is administered by injection of the polynucleotide into the somatic motor neuron cell body of a sacral spinal nerve, by injection of the polynucleotide into the pudendal nerve, or by injection of the polynucleotide into Onuf s nucleus. 
     
     
         27 - 28 . (canceled) 
     
     
         29 . A method for treating sexual dysfunction in an individual in need thereof, the method comprising:
 administering an effective amount of a polynucleotide comprising a nucleotide sequence encoding a light-responsive opsin protein to the individual, wherein the opsin protein is expressed on the plasma membrane of a population of neurons responsible for the innervation of the genitalia of the individual,   wherein the opsin protein induces depolarization of the neurons expressing the opsin protein in response to light, whereby the depolarization of the neurons responsible for the innervation of the genitalia restores sexual function.   
     
     
         30 . The method of  claim 29 , wherein the restored sexual function is one or more functions selected from the group consisting of erection, orgasm, ejaculation, vaginal lubrication, and tactile sensations to the genitals needed to maintain sexual arousal. 
     
     
         31 . The method of  claim 29 , wherein the sexual dysfunction is associated with diabetic neuropathy, multiple sclerosis, tumor, tertiary syphilis, endocrine disorders, hormonal deficiencies, or spinal cord injury. 
     
     
         32 . The method of  claim 29 , wherein the population of neurons responsible for the innervation of the genitalia comprises: neurons arising from one or more sacral spinal nerves in the individual; neurons of the dorsal nerve of the penis branch of the pudendal nerve in a male individual; or neurons of the dorsal nerve of the clitoris branch of the pudendal nerve in a female individual. 
     
     
         33 - 34 . (canceled) 
     
     
         35 . The method of  claim 29 , wherein a light source capable of generating light in response to an external signal is placed around the sacral spinal nerves and/or around the innervations of the pudendal nerve, wherein the light is capable of activating the light-responsive opsin protein expressed on the plasma membrane of the neurons. 
     
     
         36 . (canceled) 
     
     
         37 . The method of  claim 29 , wherein the individual externally controls the polarization state of the neurons responsible for the innervation of the genitalia by activating the light source surrounding the sacral spinal nerve(s) and/or the pudendal nerve. 
     
     
         38 . The method of  claim 29 , wherein the polynucleotide is administered by injection of the polynucleotide into the somatic motor neuron cell body of one or more sacral spinal nerves, by injection of the polynucleotide into the pudendal nerve, or by injection of the polynucleotide into Onuf s nucleus. 
     
     
         39 .- 40 . (canceled) 
     
     
         41 . The method of  claim 1 , wherein the polynucleotide is a vector. 
     
     
         42 . The method of  claim 41 , wherein the vector is a viral vector selected from the group consisting of an AAV vector, a retroviral vector, an adenoviral vector, an HSV vector, and a lentiviral vector. 
     
     
         43 . (canceled) 
     
     
         44 . The method according to  claim 1 , wherein the light-responsive opsin protein comprises a signal peptide. 
     
     
         45 . The method according to  claim 1 , wherein the ER export signal comprises the amino acid sequence FCYENEV (SEQ ID NO:21). 
     
     
         46 . The method according to  claim 1 , wherein the membrane trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:12). 
     
     
         47 . The method of  claim 1 , wherein the expression of the light-responsive opsin protein is controlled by a promoter selected from the group consisting of a EF1α promoter, a CMV promoter, a CAG promoter, a synapsin promoter, a Thy1 promoter, and a CAMKII promoter. 
     
     
         48 .- 66 . (canceled)