In DMD, skeletal and cardiac muscles are affected, leading to wheelchair dependency, respiratory failure and premature death. Recent advances have pointed out a variety of possible therapeutic approaches. A combination of these strategies might enhance the possibility of successful therapy. We isolated CD133+ stem cells from muscle biopsies of GRMD dogs. Five dogs one year old, Joãozinho, Kakà, characterized by a mild clinical phenotype and Marcinho, Danielzihno and Valdihno, characterized by a severe clinical phenotype, were treated with their own trasnduced U7exon 6-8 CD133+ stem cells. Old GRMD dogs are well characterized in term of clinical history and no variation among different dogs in the progression of the disease are observed from this age. All dogs received 2 arterial systemic injections (100 x106 cells each) through a catheter that was introduced in the left femoralis artery and reached the aortic arch at the level of the left subclavia. From a clinical point of view, Kaka had bad performance and was walking with difficulties before injection (at the age of 11 months). Marcihno was in worst condition since unable to walking and full of retraction as well Danielzihno. Valdihno showed the same clinical condition of Marcihno and Danielzhino. In order to test whether morphological and biochemical changes, we performed three different functional measures: claim Stairs (Time), swimming, 6 minute walking test (6MWT), quantitative values. After the injection, all treated dogs had a clinical performance improvement. After the first injection, all dogs had no detectable anti-dystrophin antibodies and its circulating lymphocytes did not react to transduced CD133+ stem cells. All transplanted animals were analysed at different times (6, 12 months after injection); most of the biopsies in all muscles had a morphological amelioration when compared to untreated dogs. Dystrophin expression in the biopsies was variable, ranging from 2 to ≥ 7% in several biopsies of the injected legs. The percentage of dystrophin expressing fibers ranged from 1 up to 7%, in 2 distant sections of 3 different biopsies each of selected muscles from Kaka and Marcihno. Western blot analysis of extracts from different biopsies of the same muscles confirmed the presence of different amount of dystrophin, varying from an undetectable signal to around 6% of a wt canine muscle. Two dogs, Cipo and Ze, received their own muscle-derived CD133+ stem cells Dando, Weiss & Colucci Ltd, Lawrence House, Lower Bristol Road, Bath BA2 9ET, United Kingdom Dando, Weiss & Colucci Limited Cambridge Office P a g e | 4 without lentiviral transduction. No clusters of dystrophin positive myofibers and clinical modifications of the performance. However, they appear stable during the last year of observation. In the group of untreated GRMD dogs, (Valzihno and Caipirihno) died of pneumonia and other complications during the follow up. This is the first demonstration of a clinical effect in old GRMD dogs that regained walking ability after autologous transplantation of engineered stem cells

Muscular dystrophies therapies by engineered stem cells / M. Meregalli, A. Farini, M. Belicchi, D. Parolini, P. Razini, G. Del Fraro, L. Cassinelli, V. Angeloni, S. Maciotta, J.C. Da Silva Bizario, L. Garcia, N. Bresolin, Y. Torrente. ((Intervento presentato al convegno Optistem Endostem Annual Meeting 2012 tenutosi a Sitges, Barcelona, Spain. nel 2012.

Muscular dystrophies therapies by engineered stem cells

M. Meregalli
Primo
;
A. Farini
Secondo
;
M. Belicchi;D. Parolini;V. Angeloni;S. Maciotta;N. Bresolin
Penultimo
;
Y. Torrente
Ultimo
2012

Abstract

In DMD, skeletal and cardiac muscles are affected, leading to wheelchair dependency, respiratory failure and premature death. Recent advances have pointed out a variety of possible therapeutic approaches. A combination of these strategies might enhance the possibility of successful therapy. We isolated CD133+ stem cells from muscle biopsies of GRMD dogs. Five dogs one year old, Joãozinho, Kakà, characterized by a mild clinical phenotype and Marcinho, Danielzihno and Valdihno, characterized by a severe clinical phenotype, were treated with their own trasnduced U7exon 6-8 CD133+ stem cells. Old GRMD dogs are well characterized in term of clinical history and no variation among different dogs in the progression of the disease are observed from this age. All dogs received 2 arterial systemic injections (100 x106 cells each) through a catheter that was introduced in the left femoralis artery and reached the aortic arch at the level of the left subclavia. From a clinical point of view, Kaka had bad performance and was walking with difficulties before injection (at the age of 11 months). Marcihno was in worst condition since unable to walking and full of retraction as well Danielzihno. Valdihno showed the same clinical condition of Marcihno and Danielzhino. In order to test whether morphological and biochemical changes, we performed three different functional measures: claim Stairs (Time), swimming, 6 minute walking test (6MWT), quantitative values. After the injection, all treated dogs had a clinical performance improvement. After the first injection, all dogs had no detectable anti-dystrophin antibodies and its circulating lymphocytes did not react to transduced CD133+ stem cells. All transplanted animals were analysed at different times (6, 12 months after injection); most of the biopsies in all muscles had a morphological amelioration when compared to untreated dogs. Dystrophin expression in the biopsies was variable, ranging from 2 to ≥ 7% in several biopsies of the injected legs. The percentage of dystrophin expressing fibers ranged from 1 up to 7%, in 2 distant sections of 3 different biopsies each of selected muscles from Kaka and Marcihno. Western blot analysis of extracts from different biopsies of the same muscles confirmed the presence of different amount of dystrophin, varying from an undetectable signal to around 6% of a wt canine muscle. Two dogs, Cipo and Ze, received their own muscle-derived CD133+ stem cells Dando, Weiss & Colucci Ltd, Lawrence House, Lower Bristol Road, Bath BA2 9ET, United Kingdom Dando, Weiss & Colucci Limited Cambridge Office P a g e | 4 without lentiviral transduction. No clusters of dystrophin positive myofibers and clinical modifications of the performance. However, they appear stable during the last year of observation. In the group of untreated GRMD dogs, (Valzihno and Caipirihno) died of pneumonia and other complications during the follow up. This is the first demonstration of a clinical effect in old GRMD dogs that regained walking ability after autologous transplantation of engineered stem cells
2-apr-2012
Settore MED/26 - Neurologia
Muscular dystrophies therapies by engineered stem cells / M. Meregalli, A. Farini, M. Belicchi, D. Parolini, P. Razini, G. Del Fraro, L. Cassinelli, V. Angeloni, S. Maciotta, J.C. Da Silva Bizario, L. Garcia, N. Bresolin, Y. Torrente. ((Intervento presentato al convegno Optistem Endostem Annual Meeting 2012 tenutosi a Sitges, Barcelona, Spain. nel 2012.
Conference Object
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/202887
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? ND
social impact