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Dysfunctional autophagy impairs muscle regeneration in lysosomal diseases
S114
Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156
glycosphynsolipid deposits were detected by immunohistochemistry. rh-α-GalA is functional in all the analyzed tissues including the CNS and remains stable after 5 months. Gb3 deposit are less abundant in treated animals compared with not injected knock outs. Currently we are analyzing tissues from adult injected animals of both sexes, which were treated at two doses (15-50x1010vg/Kg) and ages (1 month or 3 months). Preliminary results indicate that rh-αGalA is expressed and functional in all the analyzed tissues (brain, heart, kidney, liver) of the adult mice. These results confirmed that the vector lead the expression of rh-α-GalA when it is injected at early stages (P1) and possibly when injected in presymtomatic (1 month) or symptomatic (3 months) adults. We will analyze LysoGb3 (Mass_Spec), viral genome distribution and antibody production (by Feb19). This project is sponsored by the French patients association Vaincre les Maladie Lisosomales. doi:10.1016/j.ymgme.2018.12.288
273 Qualitative interviews with patients with mucopolysaccharidosis type I (MPS I) and caregivers to evaluate the Mucopolysaccharidosis Health Assessment Questionnaire (MPS-HAQ) Anureet K. Pablaa, Leighann Litcher-Kellyb, Laurence Pollissardc, Andrew Yaworskyb, Sarah Ollisb, Masami Kellyb, Brad Padillab, Alaa Hameda, aSanofi Genzyme, Cambridge, MA, United States, bAdelphi Values, Boston, MA, United States, cSanofi, Chilly-Mazarin, France Mucopolysaccharidosis I (MPS-I) is a rare autosomal recessive genetic disorder that can have physical, social, and emotional impacts. The MPS Health Assessment Questionnaire (MPS-HAQ) was developed to assess these impacts and includes 52 items across 10 domains of activities of daily living (i.e., eating/drinking, dressing, bathing, grooming, tooth brushing, toileting, mobility, walking, stairs, and caregiver assistance). It is intended for patient or caregiver completion. Cognitive debriefing interviews were conducted with patients and/or caregivers to evaluate the readability, comprehensibility, relevance, and comprehensiveness of the MPS-HAQ. Interviews were semi-structured. Participants were asked to “think aloud” about how they chose an answer for each MPS-HAQ item to identify any words or concepts they did not understand and to provide feedback on whether the items assess impacts experienced by individuals with MPS-I. All interviews were audio-recorded, transcribed verbatim, and transcripts were coded for qualitative analysis. Interviews were conducted in the US via telephone representing the experiences of 19 individuals with MPS-I. Among these individuals with MPS-I, ages ranged from 2-37 years (median age of 15 years) and over half were female (n=13, 68.4%). Patients had a diagnosis of Hurler Syndrome (n=14, 73.7%) or Hurler-Scheie (n=5, 26.3%). Preliminary analysis of the interview data indicates that patients and caregivers are able to interpret the content of the MPS-HAQ as intended and that the MPS-HAQ evaluates concepts relevant to the MPS-I experience. Formal qualitative data analyses were conducted on the coded transcripts to evaluate readability, comprehensibility, relevance, and comprehensiveness of the MPSHAQ with the addition of relevant participant quotations. The results from the qualitative interviews support the content validity of the MPS-HAQ for assessing the impacts experienced by patients with MPSI. Sponsor: Sanofi Genzyme
doi:10.1016/j.ymgme.2018.12.289
274 Dysfunctional autophagy impairs muscle regeneration in lysosomal diseases Ron Padilla, Suleiman A. Igdoura, McMaster University, Hamilton, ON, Canada Lysosomal diseases are metabolic diseases which occur because of a deficiency of one of the essential lysosomal enzymes. A mutation in the gene encoding one of these enzymes leads to an accumulation of unwanted substrates, resulting in a variety of clinical manifestations. A common symptom found in lysosomal diseases is skeletal muscle dysfunction, which includes muscle weakness, atrophy and loss of muscle mass. One metabolic pathway which has been shown to be essential for maintenance of skeletal muscles is autophagy. This study sought to investigate the impact of lysosomal diseases on autophagy and how this may potentiate muscle dysfunction. We utilized in-vivo and in-vitro models of sialidosis, Sandhoff disease, and GM1-gangliosidosis to assess autophagy and its impact on myogenic differentiation in skeletal muscles. Our results demonstrated that in lysosomal diseases, autophagy is induced upstream, i.e., ULK1 phosphorylation but is hindered at the autophagosome to lysosome fusion step, i.e., p62 accumulation which led to impairment of myoblast fusion and myogenic differentiation. We conclude that blocking autophagy impairs myogenic differentiation, which potentiates the muscle dysfunction observed in lysosomal diseases. This work highlights autophagy as a new pathway of interest and possible therapeutic target to alleviate muscle dysfunction in lysosomal diseases.
doi:10.1016/j.ymgme.2018.12.290
275 Platform technology for treatment of the brain in lysosomal diseases: Application to NCL1 Batten disease William M. Pardridge, Eric K.-W. Hui, Jeff Z. Lu, Huilan Lin, Ruben J. Boado, ArmaGen Inc., Calabasas, CA, United States Batten disease has primary brain manifestations. Progress in the treatment has primarily been limited to the fact that recombinant enzymes are large molecules that do not cross the blood-brain barrier (BBB). BBB-penetration of enzyme therapeutics is enabled by re-engineering the recombinant enzyme as a bi-functional IgGenzyme fusion protein, wherein the IgG domain is a monoclonal antibody (MAb) that targets a specific endogenous receptormediated transporter within the human BBB, such as the human insulin receptor (HIR). A proof of concept phase II clinical trial in pediatric subjects with severe Hurler MPSI with a BBB penetrating HIRMAb-iduronidase fusion protein (AGT-181, valanafusp alpha) produced stabilization of cognition, and reduced hepatosplenomegaly. The aim of the present investigation was to determine if a similar BBB penetrating IgG-fusion protein can be engineered for the treatment of neuronal ceroid lipofuscinosis type 1 (NCL1), which is caused by mutations in the CLN1 gene encoding for palmitoyl-protein thioesterase type 1 (PPT1). Genes encoding the heavy chain and the light chain of a HIRMAb-PPT1 fusion protein were engineered followed by stable transfection of Chinese hamster ovary (CHO) cells. The identity of the CHO derived HIRMAb-PPT fusion protein (AGT-194) was confirmed by hIgG1 and PPT1 Western blot analyses and the purity by SDS-PAGE and SEC-HPLC. Fusion
Abstracts / Molecular Genetics and Metabolism 126 (2019) S17–S156
glycosphynsolipid deposits were detected by immunohistochemistry. rh-α-GalA is functional in all the analyzed tissues including the CNS and remains stable after 5 months. Gb3 deposit are less abundant in treated animals compared with not injected knock outs. Currently we are analyzing tissues from adult injected animals of both sexes, which were treated at two doses (15-50x1010vg/Kg) and ages (1 month or 3 months). Preliminary results indicate that rh-αGalA is expressed and functional in all the analyzed tissues (brain, heart, kidney, liver) of the adult mice. These results confirmed that the vector lead the expression of rh-α-GalA when it is injected at early stages (P1) and possibly when injected in presymtomatic (1 month) or symptomatic (3 months) adults. We will analyze LysoGb3 (Mass_Spec), viral genome distribution and antibody production (by Feb19). This project is sponsored by the French patients association Vaincre les Maladie Lisosomales. doi:10.1016/j.ymgme.2018.12.288
273 Qualitative interviews with patients with mucopolysaccharidosis type I (MPS I) and caregivers to evaluate the Mucopolysaccharidosis Health Assessment Questionnaire (MPS-HAQ) Anureet K. Pablaa, Leighann Litcher-Kellyb, Laurence Pollissardc, Andrew Yaworskyb, Sarah Ollisb, Masami Kellyb, Brad Padillab, Alaa Hameda, aSanofi Genzyme, Cambridge, MA, United States, bAdelphi Values, Boston, MA, United States, cSanofi, Chilly-Mazarin, France Mucopolysaccharidosis I (MPS-I) is a rare autosomal recessive genetic disorder that can have physical, social, and emotional impacts. The MPS Health Assessment Questionnaire (MPS-HAQ) was developed to assess these impacts and includes 52 items across 10 domains of activities of daily living (i.e., eating/drinking, dressing, bathing, grooming, tooth brushing, toileting, mobility, walking, stairs, and caregiver assistance). It is intended for patient or caregiver completion. Cognitive debriefing interviews were conducted with patients and/or caregivers to evaluate the readability, comprehensibility, relevance, and comprehensiveness of the MPS-HAQ. Interviews were semi-structured. Participants were asked to “think aloud” about how they chose an answer for each MPS-HAQ item to identify any words or concepts they did not understand and to provide feedback on whether the items assess impacts experienced by individuals with MPS-I. All interviews were audio-recorded, transcribed verbatim, and transcripts were coded for qualitative analysis. Interviews were conducted in the US via telephone representing the experiences of 19 individuals with MPS-I. Among these individuals with MPS-I, ages ranged from 2-37 years (median age of 15 years) and over half were female (n=13, 68.4%). Patients had a diagnosis of Hurler Syndrome (n=14, 73.7%) or Hurler-Scheie (n=5, 26.3%). Preliminary analysis of the interview data indicates that patients and caregivers are able to interpret the content of the MPS-HAQ as intended and that the MPS-HAQ evaluates concepts relevant to the MPS-I experience. Formal qualitative data analyses were conducted on the coded transcripts to evaluate readability, comprehensibility, relevance, and comprehensiveness of the MPSHAQ with the addition of relevant participant quotations. The results from the qualitative interviews support the content validity of the MPS-HAQ for assessing the impacts experienced by patients with MPSI. Sponsor: Sanofi Genzyme
doi:10.1016/j.ymgme.2018.12.289
274 Dysfunctional autophagy impairs muscle regeneration in lysosomal diseases Ron Padilla, Suleiman A. Igdoura, McMaster University, Hamilton, ON, Canada Lysosomal diseases are metabolic diseases which occur because of a deficiency of one of the essential lysosomal enzymes. A mutation in the gene encoding one of these enzymes leads to an accumulation of unwanted substrates, resulting in a variety of clinical manifestations. A common symptom found in lysosomal diseases is skeletal muscle dysfunction, which includes muscle weakness, atrophy and loss of muscle mass. One metabolic pathway which has been shown to be essential for maintenance of skeletal muscles is autophagy. This study sought to investigate the impact of lysosomal diseases on autophagy and how this may potentiate muscle dysfunction. We utilized in-vivo and in-vitro models of sialidosis, Sandhoff disease, and GM1-gangliosidosis to assess autophagy and its impact on myogenic differentiation in skeletal muscles. Our results demonstrated that in lysosomal diseases, autophagy is induced upstream, i.e., ULK1 phosphorylation but is hindered at the autophagosome to lysosome fusion step, i.e., p62 accumulation which led to impairment of myoblast fusion and myogenic differentiation. We conclude that blocking autophagy impairs myogenic differentiation, which potentiates the muscle dysfunction observed in lysosomal diseases. This work highlights autophagy as a new pathway of interest and possible therapeutic target to alleviate muscle dysfunction in lysosomal diseases.
doi:10.1016/j.ymgme.2018.12.290
275 Platform technology for treatment of the brain in lysosomal diseases: Application to NCL1 Batten disease William M. Pardridge, Eric K.-W. Hui, Jeff Z. Lu, Huilan Lin, Ruben J. Boado, ArmaGen Inc., Calabasas, CA, United States Batten disease has primary brain manifestations. Progress in the treatment has primarily been limited to the fact that recombinant enzymes are large molecules that do not cross the blood-brain barrier (BBB). BBB-penetration of enzyme therapeutics is enabled by re-engineering the recombinant enzyme as a bi-functional IgGenzyme fusion protein, wherein the IgG domain is a monoclonal antibody (MAb) that targets a specific endogenous receptormediated transporter within the human BBB, such as the human insulin receptor (HIR). A proof of concept phase II clinical trial in pediatric subjects with severe Hurler MPSI with a BBB penetrating HIRMAb-iduronidase fusion protein (AGT-181, valanafusp alpha) produced stabilization of cognition, and reduced hepatosplenomegaly. The aim of the present investigation was to determine if a similar BBB penetrating IgG-fusion protein can be engineered for the treatment of neuronal ceroid lipofuscinosis type 1 (NCL1), which is caused by mutations in the CLN1 gene encoding for palmitoyl-protein thioesterase type 1 (PPT1). Genes encoding the heavy chain and the light chain of a HIRMAb-PPT1 fusion protein were engineered followed by stable transfection of Chinese hamster ovary (CHO) cells. The identity of the CHO derived HIRMAb-PPT fusion protein (AGT-194) was confirmed by hIgG1 and PPT1 Western blot analyses and the purity by SDS-PAGE and SEC-HPLC. Fusion