OBJECTIVES/HYPOTHESIS: 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). STUDY DESIGN: 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.
OBJECTIVES/HYPOTHESIS: 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). STUDY DESIGN: 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: Steven E Raper; Marc Yudkoff; Narendra Chirmule; Guang-Ping Gao; Fred Nunes; Ziv J Haskal; Emma E Furth; Kathleen J Propert; Michael B Robinson; Susan Magosin; Heather Simoes; Lisa Speicher; Joseph Hughes; John Tazelaar; Nelson A Wivel; James M Wilson; Mark L Batshaw Journal: Hum Gene Ther Date: 2002-01-01 Impact factor: 5.695
Authors: Salima Hacein-Bey-Abina; Christof von Kalle; Manfred Schmidt; Françoise Le Deist; Nicolas Wulffraat; Elisabeth McIntyre; Isabelle Radford; Jean-Luc Villeval; Christopher C Fraser; Marina Cavazzana-Calvo; Alain Fischer Journal: N Engl J Med Date: 2003-01-16 Impact factor: 91.245
Authors: R Heumann; D Lindholm; C Bandtlow; M Meyer; M J Radeke; T P Misko; E Shooter; H Thoenen Journal: Proc Natl Acad Sci U S A Date: 1987-12 Impact factor: 11.205
Authors: Salima Hacein-Bey-Abina; Françoise Le Deist; Frédérique Carlier; Cécile Bouneaud; Christophe Hue; Jean-Pierre De Villartay; Adrian J Thrasher; Nicolas Wulffraat; Ricardo Sorensen; Sophie Dupuis-Girod; Alain Fischer; E Graham Davies; Wietse Kuis; Lilly Leiva; Marina Cavazzana-Calvo Journal: N Engl J Med Date: 2002-04-18 Impact factor: 91.245
Authors: Steven E Raper; Narendra Chirmule; Frank S Lee; Nelson A Wivel; Adam Bagg; Guang-ping Gao; James M Wilson; Mark L Batshaw Journal: Mol Genet Metab Date: 2003 Sep-Oct Impact factor: 4.797