Lysosomal acid lipase deficiency high-risk screening of patients with fatty liver and dyslipidemia using DBS in Japan

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Abstracts / Molecular Genetics and Metabolism 120 (2016) S17–S145

in untreated adults with GD1. In the 9-month primary analysis, all visceral and hematologic efficacy parameters improved significantly in eliglustat-treated but not placebo-treated patients (Mistry, JAMA. 2015;313:695); 39/40 patients entered the open-label extension phase, during which all received eliglustat. Long-term data are analyzed with respect to time on eliglustat, which varied from 2.5 to ≥4.5 years depending on when randomization occurred, initial randomization group, and protocol-mandated switches to commercial eliglustat when it became available. Overall, seven patients were switched to commercial eliglustat, two discontinued due to pregnancy, and four chose to withdraw. Among patients with 4.5 years of data, mean spleen and liver volumes decreased by 66% and 23%, respectively, and mean hemoglobin level and platelet count increased by 1.4 g/dL and 87%, respectively. With regard to long-term therapeutic goals for these four parameters (Pastores, Semin Hematol. 2004;4:4), among the 33 patients with ≥2.5 years of data, the majority met all 4 goals, and 91% met at least 3 goals. Mean spine T-score increased by a mean of 21%, and median levels of the biomarkers chitotriosidase, GL-1, and MIP-1β decreased by 82%, 79%, and 71%, respectively. Eliglustat was generally well-tolerated; 99% of all 559 adverse events (AEs) and all 7 serious AEs were mild or moderate, 83% were considered unrelated to treatment, and no patients discontinued due to AEs. In summary, 4.5-year results of the ENGAGE trial demonstrated continuation and maintenance of improvements in hematologic, visceral, biomarker, and bone measures, consistent with the 4-year results of the Phase 2 trial in treatment-naïve patients (Lukina, Blood Cells Mol Dis. 2014;53:274). doi:10.1016/j.ymgme.2016.11.243

235 Lysosomal acid lipase deficiency high-risk screening of patients with fatty liver and dyslipidemia using DBS in Japan Takashi Miyajimaa,b, Keiko Akiyamaa, Junko Igarashia,b, Mohammad Arif Hossaina, Hiroko Yanagisawaa, Yoshikatsu Etoa,c, aAdvanced Clinical Research Center, Kawasaki, Japan, bAnGes MG Co, Kawasaki, Japan, cTokyo Jikei University School of Medicine, Tokyo, Japan Lysosomal acid lipase deficiency (LALD) is characterized by the accumulation of triglyceride and cholesterol ester in various organs including liver and spleen. The cholesteryl ester storage disease (CESD) is generally late onset type of LALD and patients clinically present wit fatty liver, liver dysfunction and dyslipidemia associated with atherosclerosis and liver cirrhosis in late stage. Therefore, it is essentially necessary to treat these patients by enzyme replacement therapy, since recently Sebelipase alfa (Kanuma, Alexion Pham.) has been produced by genetically engineered construct of chicken egg to produce rhLAL. We reported here high risk screening of LALD for more than 1000 adult patients with fatty liver and dyslipidemia, using DBS. The LAL activity was measured by 4 methylumbelliferyl palmitate, following the method described by Dairaku et al (2014). Among the 1000 adult fatty liver patients, we could not find any positive patient with a complete deficiency of LAL activity. However, we found a 11 years old Japanese boy and two more cases from Korea and Thailand with complete LAL deficiency in our high risk patient screening. In Asia, there are few cases with Wolman disease and CESD. 12 cases with Wolman disease and 9 cases with CESD are reported in Japan. We summarized all these cases with LALD and the significance of high risk screening of adult patients with fatty liver in Japan.

doi:10.1016/j.ymgme.2016.11.244

236 Scale-up processing of recombinant human glucosamine (n-acetyl)-6-sulfatase for the treatment of mucopolysaccharidosis type IIID Derek R Moena,b, Mengqing Chenb, Shih-hsin Kanb, Le Q Stevenb, Jill Wooda, Sean Ekinsa, Tsui-Fen Chouc, Patricia I Dicksonc, aPhoenix Nest, Inc., Brooklyn, NY, United States, bDavid Geffen School of Medicine at UCLA / LA BioMed, Torrance, CA, United States, cPhoenix Nest, Inc., Torrance, CA, United States Sanfilippo syndrome (mucopolysaccharidosis type III; MPS III) is a devastating pediatric neurodegenerative disorder with no cure or effective treatment available. The fundamental cause of MPS III is an inherited mutation in an enzyme required to catabolize heparan sulfate (HS), a glycosaminoglycan, which plays important structural and functional roles in the brain and elsewhere. The symptoms of MPS IIID are largely localized to the brain, therefore, our strategy proposes to deliver recombinant human glucosamine (N-acetyl)-6sulfatase (rhGNS) intrathecally in order to effectively treat the underlying cause. We have previously purified ~200 ug of myctagged secreted rhGNS to a specific activity N13,000 nmol/h/mg and demonstrated stability over one month at 4°C in artificial cerebrospinal fluid. Additionally, we observed intracellular enzymatic activity of rhGNS in MPS IIID fibroblasts when rhGNS is added to the media and confirmed radiolabelled HS is diminished (33-65%) to WT levels in MPS IIID fibroblasts treated with rhGNS. We have now begun the scale-up process in preparation for proof of concept studies in the MPS IIID knock-out mouse utilizing our newly established quality control guidelines for enzymatic activity, glycosylation pattern, and intracellular uptake. We are also developing a native rhGNS product using chromatographic methods to expeditiously bring our ERT to the clinic.

doi:10.1016/j.ymgme.2016.11.245

237 Pathogenic mutation at IDS gene on a newborn girl with clinical symptoms of Hunter syndrome (MPS II) Lina J Moreno, Adalberto Sanchez, Jose M Satizabal, Universidad del Valle, Cali, Colombia Hunter syndrome, or mucopolysaccharidosis type II, MPS II, is a lysosomal disease that is heterogeneous, chronic, and progressive. It is caused by deficiency of iduronate-2-sulfatase enzyme. The disease interferes with the body's ability to metabolize, and recycle mucopolysaccharides or glycosaminoglycans-GAG- in the lysosome, resulting in a multisystem organ dysfunction. The clinical spectrum ranges from mild forms-attenuated- to severe forms that differ by the presence or absence of progressive brain involvement. It is inherited X-linked, the causative gene is the IDS,locusXq28, covering more than 300 mutations described and affect 1 in every 162,000 live births. Although this rare disorder is X-linked, therefore occurring almost exclusively in males, Hunter syndrome has also been reported in a small group of female patients, manifesting with equal severity. The most common mechanism for disease expression in female patients is thought to involving the process of X-chromosome inactivation. Female patient, 11 months old with hemihypertrophy on left side of the body, globose abdomen with splenomegaly, dysmorphic facial features, no perinatal or family history of importance, karyotype 46XX,and radiography long bones signs of disostosis. The result of sequencing on IDS gene showed heterozygous mutation IDS gene in