Abstracts / Molecular Genetics and Metabolism 120 (2016) S17–S145
osteoclastogenic potential of conditioned media (CM) from CBE
treated osteocytes was evaluated by osteoclast differentiation assays.
RANKL levels were evaluated by immunofluorescence, Cx43 expres-
sion was assessed by qPCR and apoptosis was studied by Annexin-V
and TUNEL staining. To study the mechanism on osteoclast
differentiation, the apoptotic body fraction of the CM and its
supernatant were obtained by centrifugation. Both fractions were
used in osteoclastogenesis assays with or without OPG. CM from CBE
treated osteocytes induced higher levels of osteoclast differentiation
compared to control CM (pb0,01) and higher surface RANKL levels
were observed in treated cells (pb0,05). Cx43 expression diminished
with CBE treatment (pb0,01) and osteocyte apoptosis was increased
(pb0,05). Induction of osteoclast differentiation by apoptotic bodies
and by supernatant was observed in CM from CBE treated cells
(pb0,01). OPG treatment reduced osteoclast levels in both fractions
(pb0,001). In conclusion, we showed that glucocerebrosidase
deficiency in osteocytes leads to an increase in osteoclast differen-
tiation through a mechanism involving osteocyte apoptosis and
soluble molecules. This process could certainly contribute to bone
pathogenesis in Gaucher disease.
doi:10.1016/j.ymgme.2016.11.045
37
Improvement in pulmonary function and serum immunoglobulin
G in long-term enzyme replacement therapy with velmanase
alfa (human recombinant alpha-mannosidase) in
alpha-mannosidosis patients
Line Borgwardta, Allan M Lunda, Linda De Meirleirb, Yasmina
Amraouic, Oluf Andersend, Philippe Dolheme, Mercedes Gil Camposf,
Nathalie Guffong, Bénédicte Héronh, Elisabeth Jamesoni, Simon A
Jonesj, Cecile Larochek, Christoffer Lindbergd, Thorsten Marquardtl,
Karl-Eugene Mengelc, Nicole Muscholm, Johanna MP Van Den Houtn,
Lindsey Wellingo, Frits Wijburgo, Duncan Colep, Ans Van Der Ploegn,
Anna Tylki-Szymanskaq, Jens Foghr, Silvia Geracis, Diego Ardigòs,
Federica Cattaneos, aUniversity Hospital of Copenhagen Rigshospitalet,
Copenhagen, Denmark, bVrije Universiteit Brussel, Brussels, Belgium,
c
Centre for Pediatric and Adolescent Medicine Villa Metabolica, Mainz,
Germany, dSahlgrenska University Hospital, Gothenburg, Sweden,
e
Centre Hospitalier Saint Quentin, Saint Quentin, France, fReina Sofia
University Hospital, Cordoba, Spain, gCentre of Inheroted Metabolic
Disease Edourd Herriot Hospital, Lyon, France, hCentre de Référence des
Maladies Lysosomales CHU Trousseau, Paris, France, iCentral Manches-
ter University Hospitals NHS Foundation Trust, Manchester, United
Kingdom, jWillink Unit St Mary's Hospital, Manchester, United Kingdom,
k
CHU Limoges Hopital de la mere et de l' enfant, Limoges, France,
l
Universitätsklinikum Münster, Münster, Germany, mUniversity Medical
Centre Hamburg-Eppendorf, Hamburg, Germany, nErasmus Medical
Centre, Rotterdam, Netherlands, oAcademic Medical Centre, Amsterdam,
Netherlands, pCentre for Medical Education School of Medicine Cardiff
University, Cardiff, United Kingdom, qThe Children' s Memorial Health
Institute, Warsaw, Poland, rZymenex A/S, Hillerød, Denmark, sChiesi
Farmaceutici S.p.A., Parma, Italy
Velmanase alfa is the first human recombinant alpha-mannosidase
in development for the long-term enzyme replacement therapy of
alpha-mannosidosis, a rare lysosomal disorder in which catabolism of
glycoproteins is impaired. Accumulation of mannose-rich oligosaccha-
rides clinically manifests in a broad range of symptoms, such as motor
function impairment, physical and intellectual disability and respiratory
dysfunction. Immunodeficiency and recurrent infections are important
hallmarks of the disease. Alpha-mannosidase is highly expressed in
lungs, and recurrent infections are attributed in part to disease-related
S29
pulmonary abnormalities. Thirty-three patients (14 adults, 19 paediat-
rics) previously enrolled in phase I/II and phase III studies with
velmanase alfa were assessed for efficacy and safety in a comprehen-
sive, open-label, pivotal clinical trial (rhLAMAN-10). Mean (SD)
exposure was 890.5 (461.5) days. Forced Vital Capacity (FVC, % of
predicted) showed a statistically significant and clinically-relevant
improvement of mean absolute and percentage change from baseline to
last observation of 8.1% (p=0.007) and 10.5% (p=0.011), respectively,
in the overall population. A higher level of pulmonary dysfunction
was observed in patients b18 years of age at baseline [mean (SD)
79.6% (16.4%)] compared to adults [mean (SD) 92.5% (19.4%)], with
an improvement of mean (SD) absolute change of 11.6 % (11.7%) at
last observation. Notably, a significant increase in serum immunoglob-
ulin class G (IgG) was observed in all patients with baseline
hypogammaglobulinaemia. Based on the common variable immuno-
deficiency disease categorization (Cassidy, 1974) the number of
patients classified as seriously impaired (n=2) or impaired (n=7) at
baseline decreased to 0 and 3, respectively, at last observation. These
data suggest that weekly infusion of velmanase alfa can significantly
improve pulmonary function and hypogammaglobulinaemia, irrespec-
tive of patient age. The improvement in pulmonary function may result
from a synergistic effect of reduction in tissue accumulation of
oligosaccharides and of improvement in immune system function after
weekly infusion of velmanase alpha.
doi:10.1016/j.ymgme.2016.11.046
38
Improvement in fine and gross motor proficiency after long-term
enzyme replacement therapy with velmanase alfa (human
recombinant alpha mannosidase) in alpha-mannosidosis patients
Line Borgwardta, Allan M Lunda, Yasmine Amraouib, Oluf Andersenc,
Linda De Meirleird, Philippe Dolheme, Mercedes Gil Camposf,
Nathalie Guffong, Bénédicte Héronh, Elisabeth Jamesoni, Simon A
Jonesj, Cecile Larochek, Christoffer Lindbergc, Thorsten Marquardtl,
Karl-Eugene Mengelb, Nicole Muscholm, Anna Tylki-Szymanskan,
Johanna MP Van Den Houto, Ans Van Der Ploego, Lindsey Wellingp,
Frits Wijburgp, Jens Foghq, Silvia Geracir, Diego Ardigor, Federica
Cattaneor, Dawn Phillipss, aUniversity Hospital of Copenhagen
Rigshospitalet, Copenhagen, Denmark, bCentre for Pediatric and Adoles-
cent Medicine Villa Metabolica, Mainz, Germany, cSahlgrenska Univer-
sity Hospital, Gothenburg, Sweden, dVrije Universiteit Hospital, Brussels,
Belgium, eCentre Hospitalier Saint Quentin, Saint Quentin, France, fReina
University Hospital, Cordoba, Spain, gCentre of Inherited Metabolic
Diseases Edouard Herriot, Lyon, France, hCentre de Référence des
Maladies Lysosomales CHU Trousseau, Paris, France, iCentral Manchester
University Hospitals NHS Foundation Trust, Manchester, United Kingdom,
j
Willink Unit St Mary's Hospital, Manchester, United Kingdom, kCHU Limoges
Hopital de la mere et de l' enfant, Limoges, France, lUniversitätsklinikum
Münster, Münster, Germany, mUniversity Medical Center Hamburg-
Eppendorf, Hamburg, Germany, nThe Children' s Memorial Health Institute,
Warsaw, Poland, oErasmus Medical Centre, Rotterdam, Netherlands,
p
Academic Medical Centre, Amsterdam, Netherlands, qZymenex A/S, Hillerød,
Denmark, rChiesi Farmaceutici S.p.A., Parma, Italy, sUniversity of North
Carolina, Chapel Hill, NC, United States
Alpha-mannosidosis is a rare monogenic disorder caused by
deficient activity of alpha-mannosidase. Long-term efficacy of
velmanase alfa, a recombinant form of human alpha-mannosidase,
was investigated in 33 patients (19 paediatrics, 14 adults) treated for
up to 48 months in phase I-III studies and long-term follow-up. The
Bruininks-Oseretsky test (BOT-2) was a secondary efficacy endpoint
S30 Abstracts / Molecular Genetics and Metabolism 120 (2016) S17–S145
used to capture fine and gross motor proficiency and key deficits
such as decreased visual motor coordination and ataxia. The
integrated analysis indicated a severe impairment at baseline with
mean age-equivalent (AE) and scale scores (SS) well below
functional expectations for chronological age. Across all treated
patients, the mean BOT-2 total point score improved at month 12
with an absolute change of +7.5 (p b0.05), 10.6 % (pb0.01) and at
last assessment of +5.1 (p=0.230), 13.0% (p=0.035). Analysis by
age showed a trend for greater change in the paediatric than adult
patients. Subtest scores showed some improvement trends at last
observation with AE and SS increases in paediatrics for the Fine
Motor Precision (FMP; AE 12.9%, SS 10.2%), Manual Dexterity (MD;
AE 9.3%, SS 27.4%), Upper Limb Co-ordination (ULC; AE 10.7%, SS
34.2%), and Bilateral Co-ordination (BC; AE 8.5%, SS 15.9%). FMP, MD
and ULC values provide an overall measure of visual motor
coordination, MD values represent the level of speed and accuracy
during goal-directed activities that correspond to eating, dressing
and recreation and BC items require simultaneous coordination of
the upper and lower limbs and and provide a measure of ataxia.
Patients with Alpha-Mannosidosis require assistance for many
activities of daily living and subtest improvements may contribute to
increasing independence, reducing caregiver burden and preventing
further decline. Overall, these data provide evidence of the positive
clinical effect of velmanase alfa.
doi:10.1016/j.ymgme.2016.11.047
39
Long-term enzyme replacement therapy with velmanase alfa
(human recombinant alpha-mannosidase) slows disease progression
in adult patients suffering from alpha-mannosidosis
Line Borgwardta, Allan M Lunda, Yasmina Amraouib, Oluf Andersenc,
Linda De Meirleird, Philippe Dolheme, Mercedes Gil Camposf,
Nathalie Guffong, Bénédicte Héronh, Cecile Larochei, Christoffer
Lindbergc, Thorsten Marquardtj, Karl-Eugene Mengelb, Nicole
Muscholk, Anna Tylki-Szymanskal, Johanna MP Van Den Houtm, Ans
Van Der Ploegm, Lindsey Wellingn, Frits Wijburgn, Duncan Coleo, Jens
Foghp, Silvia Geraciq, Diego Ardigòq, Federica Cattaneoq, aUniversity
Hospital of Copenhagen Rigshospitalet, Copenhagen, Denmark, bCentre
for Pediatric and Adolescent Medicine Villa Metabolica, Mainz,
Germany, cSahlgrenska University Hospital, Gothenburg, Sweden, dVrije
Universiteit Brussel, Brussels, Belgium, eCentre Hospitalier Saint Quentin,
Saint Quentin, France, fReina Sofia University Hospital, Cordoba, Spain,
g
Centre of Inherited Metabolic Diseases Edouard Herriott Hospital, Lyon,
France, hCentre de Référence des Maladies Lysosomales CHU Trousseau,
Paris, France, iCHU Limoges Hopital de la mere et de l'enfant, Limoges,
France, jUniversitätsklinikum Münster, Münster, Germany, kUniversity
Medical Center Hamburg-Eppendorf, Hamburg, Germany, lThe Children’s
Memorial Health Institute, Warsaw, Poland, mErasmus Medical Centre,
Rotterdam, Netherlands, nAcademic Medical Centre, Amsterdam,
Netherlands, oCentre for Medical Education in the School of Medicine
Cardiff University, Cardiff, United Kingdom, pZymenex A/S, Hillerød,
Denmark, qChiesi Farmaceutici S.p.A., Parma, Italy
Velmanase alfa is an enzyme replacement therapy in develop-
ment for the long-term treatment of alpha-mannosidosis, a lyso-
somal disorder caused by defective activity of the enzyme alpha-
mannosidase and leading to progressive skeletal deformities and
impairment in motor function, immune system, and life quality.
Fourteen adult patients [mean (SD) age 24.6 (5.3) years at baseline]
participated in the clinical development program (phase III study
and long-term follow-up). Last observation corresponds to either
18 or 36 months of exposure. Biological activity of velmanase alfa
was demonstrated by statistically significant reduction of serum
oligosaccharides (alpha-mannosidase substrate) with mean (SD) %
change from baseline of -57.6% (30.5) at last observation. Increased
serum immunoglobulin G (IgG) levels with correction of hyp-
ogammaglobulinemia when present was observed, suggesting that
the clearance of oligosaccharides reverted the impaired function of
plasma cells [-38.6% (21.6) change at last observation]. The 3-Minute
Stair Climb Test (3MSCT) results suggest a stabilization of the disease
[baseline 53.0 steps/min (11.8), last observation 53.6 (14.6)]; 3 out
of 5 patients that required aids or help for walking at baseline,
improved to the extent of not needing aids at last observation. In
particular one adult forced to use the wheelchair for long distances at
baseline discontinued the use at last observation. Clinically-relevant
improvements were also observed in the Childhood Health Assess-
ment Questionnaire (CHAQ) Pain Index [mean change (SD) from
baseline -35.3 (54.3)], with an average improvement exceeding the
minimal clinically important difference (Dhanani 2002). The preser-
vation of respiratory function was also observed in the long-term
follow-up. Albeit starting enzyme replacement in early childhood
has proven to be the most effective strategy to prevent functional
deterioration in several lysosomal disorders, initiating treatment
with velmanase alfa in adulthood has proven to be effective in
improving key disease biomarkers and in slowing the disease pro-
gression in multiple domains.
doi:10.1016/j.ymgme.2016.11.048
40
Ophthalmologic findings in Fabry disease patients: relation to
disease severity and to enzyme replacement therapy in a
Mexican family
Araceli Borja-Borjaa, Gabriela Salas-Péreza, Jorge Gutiérrez-Poncea,
Antonio Sánchez-Acostaa, Pablo Radillo-Díazb, Fátima Sierrac, Juan C
García-Pérezd, aHospital de Especialidades de Puebla, Puebla, Mexico,
b
Centro Médico Coyoacán, Mexico city, Mexico, cHospital de la Mujer,
SSA, Puebla, Puebla, Mexico, dHospital Universidad Popular del Estado
de Puebla, Puebla, Mexico
Fabry disease is an X linked disorder of lysosomal glyco-
sphyngolipid metabolism which affects multiple organ systems
leading to their progressive deterioration. Target organs are the
kidney, heart and nervous system; however, other organs have
demonstrated accumulation and symptom and sign appearance at
some point in the disease. Ophthalmologic manifestations are
frequent and have been described extensively; we present our
analysis of ophthalmologic findings in a Mexican family. Material and
methods. We elaborated a clinical and ophthalmologic review and its
correlation with kidney, heart and nervous system damage in 10
members of a family with a c.260delA (P.Glu87Glyfs*34) mutation in
exon 2 of the GLA gene. Results. Nine females and one male were
evaluated. Mean age was 16.5 years. Ophthalmologic findings
included cornea verticillata, conjunctival telangiectasias, and retinal
capillary tortuosity. Findings were more evident in patients with
more evidence of renal and/or cardiovascular damage; all females
exhibited a high incidence of ophthalmologic anomalies. Interest-
ingly, the patient with more ophthalmologic findings also exhibited
more cutaneous anomalies (generalized xerosis, folliculitis,
dermatofibromas). Conclusions. Ophthalmologic findings in FD are
frequent and seem to be associated more frequently with cardiovas-
cular and renal manifestations of the disease. The potential of these
anomalies as markers of disease progression as well as of enzyme