First test of a ‘‘high-density injection’’ protocol for myogenic cell transplantation throughout large volumes of muscles in a Duchenne muscular dystrophy patient: eighteen months follow-up Daniel Skuk a , Marlyne Goulet a , Brigitte Roy a , Vincent Piette b , Claude H. Co ˆte ´ c , Pierre Chapdelaine a , Jean-Yves Hogrel e , Martin Paradis a , Jean-Pierre Bouchard f , Michel Sylvain d , Jean-Guy Lachance g , Jacques P. Tremblay a, * a Unite ´ de recherche en Ge ´ne ´tique humaine, Centre Hospitalier de l’Universite ´ Laval, 2705 boulevard Laurier, Que., Canada GIV 4G2 b Service d’ergothe ´rapie et physiothe ´rapie, Centre Hospitalier de l’Universite ´ Laval, Que., Canada c Unite ´ CRML, Centre Hospitalier de l’Universite ´ Laval, Que., Canada d De ´partement de neurologie pe ´diatrique, Centre Hospitalier de l’Universite ´ Laval, Que., Canada e Institut de Myologie, Paris, France f De ´partement de neurologie, Ho ˆpital de l’Enfant-Je ´sus, Que., Canada g Service de niphrologie, Uniti de Greffe rinale of the Httel-Dieu, Que., Canada Received 8 June 2006; received in revised form 18 September 2006; accepted 11 October 2006 Abstract A 26-years old Duchenne muscular dystrophy (DMD) patient received normal muscle-precursor cells, proliferated in vitro and implanted in a thenar eminence, biceps brachii, and in a portion of a gastrocnemius by injections placed 1 mm from each other or less. Saline was injected in the contralateral gastrocnemius. The patient was immunosuppressed with tacrolimus. The protocol of cell transplantation was well tolerated and did not cause permanent sequels. Some injected sites were biopsied at 1, 14 and 18 months post-transplantation. Muscles were replaced by fat and fibrosis. In the cell-grafted site of the gastrocnemius, 27.5% of the myofiber profiles expressed donor-derived dystrophin 1 month post-transplantation and 34.5% 18 months post-transplantation. The contra- lateral gastrocnemius was dystrophin-negative. Myofibers were virtually absent in the biceps brachii, where only two dystrophin-posi- tive myofibers were observed. In conclusion, a ‘‘high-density injection’’ protocol was feasible for intramuscular cell-transplantation in a DMD patient and long-term expression of donor-derived dystrophin was observed. Ó 2006 Elsevier B.V. All rights reserved. Keywords: Cell transplantation; Clinical trial; Duchenne muscular dystrophy; Dystrophin; Muscle-precursor cell; Tacrolimus 1. Introduction Available treatments for Duchenne muscular dystro- phy (DMD) only slow down muscle degeneration for a few years (corticotherapy [1]), control some of its conse- quences (orthopedic surgery of contractures and spinal deformities [2]), or prolong life expectancy (mechanical ventilation assistance [3]). More efficient strategies are needed for the treatment of this disease, aiming to stop skeletal muscle degeneration and, if possible, to restore muscle mass. Since DMD is produced by a severe defi- ciency of dystrophin, a protein that has a protective role for the sarcolemma, the main objective of most experi- mental therapeutic strategies to stop muscle degeneration is to induce its expression in DMD skeletal muscles. The 0960-8966/$ - see front matter Ó 2006 Elsevier B.V. All rights reserved. doi:10.1016/j.nmd.2006.10.003 * Corresponding author. Tel.: +1 418 654 2186; fax: +1 418 654 2207. E-mail address: Jacques-P.Tremblay@crchul.ulaval.ca (J.P. Tremblay). www.elsevier.com/locate/nmd Neuromuscular Disorders 17 (2007) 38–46