title: Neurotrophic factor-secreting autologous muscle stem cell therapy for the treatment of laryngeal denervation injury.
abstract: OBJECTIVE
  To determine if the spontaneous reinnervation that characteristically ensues after recurrent laryngeal nerve (RLN) injury could be selectively promoted and directed to certain laryngeal muscles with the use of neurotrophic factor (NF)-secreting muscle stem cell (MSC) vectors while antagonistic reinnervation is inhibited with vincristine (VNC).
  
  
  METHODS
  Basic science investigation involving primary cell cultures, gene cloning/transfer, and animal experiments.
  
  
  METHODS
  MSC survival assays were used to test multiple individual NFs in vitro. Motoneuron outgrowth assays assessed the trophic effects of identified NF on cranial nerve X (CNX)-derived motoneurons in vitro. Therapeutic NF was cloned into a lentiviral vector, and MSCs were transduced to secrete NF. Sixty rats underwent left RLN transection injury, and at 3 weeks received injections of either MSCs (n = 24), MSCs secreting NF (n = 24), or saline (n = 12) into the left thyroarytenoid muscle complex; half of the animals in the MSC groups simultaneously received left posterior cricoarytenoid injections of VNC, whereas half of the animals received saline.
  
  
  RESULTS
  Ciliary neurotrophic factor (CNTF) had the greatest survival-promoting effect on MSCs in culture. The addition of CNTF (50 ng/mL) to CNX motoneuron cultures resulted in enhanced neurite outgrowth and branching. In the animal model, the injected MSCs fused with the denervated myofibers, immunohistochemistry demonstrated enhanced reinnervation based on motor endplate to nerve contact, and reverse transcriptase-polymerase chain reaction confirmed stable CNTF expression at longest follow-up (4 months) in the CNTF-secreting MSC treated groups.
  
  
  CONCLUSIONS
  MSC therapy may have a future role in selectively promoting and directing laryngeal reinnervation after RLN injury.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA. shalum@iupui.edu
authors.forename: Stacey L
authors.initials: SL
authors.lastname: Halum
authors.forename: Bryan
authors.initials: B
authors.lastname: McRae
authors.forename: Khadijeh
authors.initials: K
authors.lastname: Bijangi-Vishehsaraei
authors.forename: Kelly
authors.initials: K
authors.lastname: Hiatt
mesh_terms: D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D020934:Ciliary Neurotrophic Factor / Q000494:pharmacology; D004195:Disease Models, Animal; D004576:Electromyography; D005822:Genetic Vectors; D007150:Immunohistochemistry; D007828:Laryngoscopy; D016086:Lentivirus; D032342:Muscle Cells / Q000637:transplantation*; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009416:Nerve Regeneration / Q000187:drug effects*; D010957:Plasmids; D051381:Rats; D011916:Rats, Inbred F344; D061226:Recurrent Laryngeal Nerve Injuries / Q000628:therapy*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D014182:Transplantation, Autologous; D014750:Vincristine / Q000008:administration & dosage / Q000494:pharmacology
chemical_list: D020934:Ciliary Neurotrophic Factor; D009414:Nerve Growth Factors; D014750:Vincristine