Methods and compositions for delivery of exogenous factors to nervous system sites

Abstract

The present invention relates to treatment methods and methods for sustained delivery of one or more exogenous factors to desired nervous system sites. In certain embodiments, the invention relates to the use of biodegradable microspheres to deliver exogenous factors, such as the morphogenic factor, sonic hedgehog (Shh), to the site of spinal cord injury. In certain embodiments, the Shh-releasing microspheres are administered together with stem cells, which may be spinal cord neural stem cells. In certain embodiments, the invention relates to regrowth of neural cells in both the central and peripheral nervous systems.

1 . A method for treating a nervous system injury in a mammal comprising administering to a mammal in need of such treatment an effective amount for treating the nervous system injury of a pharmaceutical formulation comprising at least one exogenous factor, said pharmaceutical formulation providing sustained delivery of the at least one exogenous factor for at least about 7 days. 
     
     
         2 . The method of  claim 1 , wherein the mammal is a human. 
     
     
         3 . The method of  claim 1 , wherein the nervous system injury is selected from the group consisting of spinal cord injury, amyotrophic lateral sclerosis (ALS), peripheral nerve injury, and spinal nerve injury. 
     
     
         4 . The method of  claim 1 , wherein the sustained delivery composition comprises a plurality of biodegradable microspheres. 
     
     
         5 . The method of  claim 1 , wherein the pharmaceutical formulation further comprises an effective amount of stem cells, wherein the amount of stem cells in combination with said at least one exogenous factor is effective for treating a nervous system injury. 
     
     
         6 . The method of  claim 1 , wherein the at least one exogenous factor is a growth factor selected from the group consisting of Nerve Growth Factor (NGF), Glial Cell Line-Derived Growth Factor (GDNF), Neurotrophin (NT) 3, NT 4/5, NT 6, Ciliary Neurotrophic Factor (CNTF), Leukemia Inhibitory Factor (LIF), Interleukin 6 (IL6), Interleukin 11(IL11), Cardiotrophin 1, a growth factor hormone, hyaluronidase, chondroitinase ABC(CABC), basic fibroblast growth factor (bFGF), insulin-related growth factor (IGF-I), brain-derived neurotrophic factor (BDNF), epidermal growth factor (EGF), and sonic hedgehog (Shh). 
     
     
         7 . The method of  claim 5 , wherein the stem cells are neural stem cells. 
     
     
         8 . The method of  claim 1 , wherein the pharmaceutical formulation is administered by at least one injection, wherein a first injection is administered at the site of nervous system injury. 
     
     
         9 . The method of  claim 8 , wherein a second injection is administered at a site rostral to the site of nervous system injury. 
     
     
         10 . The method of  claim 1 , wherein the stem cells are endothelial-expanded stem cells. 
     
     
         11 . The method of  claim 10 , wherein the endothelial-expanded stem cells are pre-treated with sonic hedgehog. 
     
     
         12 . The method of  claim 10 , wherein the endothelial-expanded stem cells are pre-treated with sonic hedgehog and retinoic acid. 
     
     
         13 . A method for increasing neural cell growth or regenerating neural cells in a mammal which comprises administering to a mammal in need of such treatment an effective amount for increasing neural cell growth or regenerating neuronal cells of a sustained release pharmaceutical formulation comprising at least one exogenous factor, said pharmaceutical formulation continuously delivering the at least one exogenous factor for at least about 7 days. 
     
     
         14 . The method of  claim 13 , wherein the mammal is a human. 
     
     
         15 . The method of  claim 13 , wherein the neural cell is selected from the group consisting of neurons, glial cells, and progenitor cells. 
     
     
         16 . The method of  claim 13 , wherein the sustained delivery composition comprises a plurality of biodegradable microspheres. 
     
     
         17 . The method of  claim 13 , wherein the pharmaceutical formulation further comprises an effective amount of stem cells, wherein said amount in combination with said at least one exogenous factor is effective for increasing neural cell growth or regenerating neural cells. 
     
     
         18 . The method of  claim 13 , wherein the at least one exogenous factor is a growth factor selected from the group consisting of Nerve Growth Factor (NGF), Glial Cell Line-Derived Growth Factor (GDNF), Neurotrophin (NT) 3, NT 4/5, NT 6, Ciliary Neurotrophic Factor (CNTF), Leukemia Inhibitory Factor (LIF), Interleukin 6 (IL6), Interleukin 11(IL 11), Cardiotrophin 1, a growth factor hormone, hyaluronidase, chondroitinase ABC(CABC), basic fibroblast growth factor (bFGF), insulin-related growth factor (IGF-1), brain-derived neurotrophic factor (BDNF), epidermal growth factor (EGF), and sonic hedgehog (Shh). 
     
     
         19 . The method of  claim 17 , wherein the stem cells are neural stem cells. 
     
     
         20 . The method of  claim 13 , which comprises administering at least one injection of the pharmaceutical formulation at a site at which it is desired to increase neural cell growth or regenerate neural cells. 
     
     
         21 . The method of  claim 20 , which comprises administering a second injection containing said pharmaceutical formulation at a site rostral to the first site of injection. 
     
     
         22 . The method of  claim 13 , wherein the stem cells are endothelial-expanded stem cells. 
     
     
         23 . The method of  claim 22 , wherein the endothelial-expanded stem cells are pre-treated with sonic hedgehog. 
     
     
         24 . The method of  claim 22 , which comprises pre-treating endothelial-expanded stem cells with sonic hedgehog and retinoic acid. 
     
     
         25 . A pharmaceutical formulation which comprises an effective amount for increasing neuronal cell growth or regenerating neuronal cells of at least one sustained delivery composition comprising at least one exogenous factor selected from the group consisting of Nerve Growth Factor (NGF), Glial Cell Line-Derived Growth Factor (GDNF), Neurotrophin (NT) 3, NT 4/5, NT 6, Ciliary Neurotrophic Factor (CNTF), Leukemia Inhibitory Factor (LIF), Interleukin 6 (IL6), Interleukin 11(IL11), Cardiotrophin 1, a growth factor hormone, hyaluronidase, chondroitinase ABC(CABC), basic fibroblast growth factor (bFGF), insulin-related growth factor (IGF-1), brain-derived neurotrophic factor (BDNF), epidermal growth factor (EGF), and sonic hedgehog (Shh), wherein the composition provides sustained delivery of at least one exogenous factor for at least 7 days, and a pharmaceutically acceptable excipient. 
     
     
         26 . The pharmaceutical formulation of  claim 25 , further comprising an effective amount of stem cells, wherein said amount in combination with said at least one exogenous factor is effective for increasing neural cell growth or regenerating neural cells. 
     
     
         27 . The pharmaceutical formulation of  claim 25 , wherein the at least one sustained delivery composition comprises a plurality of biodegradable microspheres. 
     
     
         28 . The pharmaceutical formulation of  claim 26 , wherein the stem cell is a neural stem cell. 
     
     
         29 . The pharmaceutical formulation of  claim 26 , wherein the stem cell is an endothelial-expanded stem cell. 
     
     
         30 . The pharmaceutical formulation of  claim 25 , wherein the endothelial-expanded stem cell is pre-treated with sonic hedgehog. 
     
     
         31 . The pharmaceutical formulation of  claim 25 , wherein the endothelial-expanded stem cell is pre-treated with sonic hedgehog and retinoic acid. 
     
     
         32 . The pharmaceutical formulation of  claim 25 , wherein the sustained delivery composition releases from about 1 ng/ml to about 20 ng/ml Shh for at least 7 days.