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390 Background: 5-fluorouracil and capecitabine are the gold standards in colorectal cancer (CRC) treatment and are often combined with oxaliplatin (FOLFOX and XELOX chemotherapy, respectively). Our purpose was to analyze associations between severe adverse reactions to these regimes and polymorphisms in genes related to these drugs Methods: Retrospective study with 47 adult CRC patients treated with adjuvant FOLFOX/XELOX. 20 polymorphisms in 14 genes were selected: 6 genes [XRCC1 (rs25487), ERCC2 (rs131181), ERCC1 (rs11615), GSTP1 (rs1695), EGFR (rs4559542) and GSTT1 (copy number variation)] related to the pharmacokinetics and dynamics of oxaliplatin and 8 related to fluoropyrimidines [MTHFR (rs1801131 and rs1801133), DPYD (rs2297595 and rs3918290), TYMS (rs34743033 and rs34489327), ABCB1 (rs1128503, rs2032582 and rs1045642), ABCC4 (rs4148551 and rs3742106), ABCC5 (rs3805114), CYP2A6 (rs3742106 ) and CDA (rs2072671)]. Linear by linear association chi-square test (SPSS v.18.0.) was used to study associations. A multivariate analysis including sex and performance status was also conducted. p< 0.05 was considered significant. Results: Mean age was 62 (SD: 12) years and 78.7% male. Univariate analysis: statistically significant associations were obtained between rs11615 (ERCC1), neutropenia and hand-foot syndrome; rs3742106 (CYP2A6) and neutropenia; rs34743033 and rs34489327 (TYMS) and nausea/vomiting; and CNV of GSTT1 and neutropenia. Multivariate analysis: statistically significant associations were obtained between rs3742106 (CYP2A6) and neutropenia (GT vs. TT: OR, 0.042, 95% CI, 0004-0499, p = 0.012); rs2297595 (DPYD) and nausea/vomiting (GA vs AA: OR, 0.051, 95% CI, 0003-0772, p = 0.032); and rs11615 (ERCC1) and neutropenia (CC vs CT / TT: OR, 0.099, 95% CI, 0016-0615, p = 0.013) Conclusions: These results could help oncologists reduce adverse reactions associated to FOLFOX and XELOX chemotherapy by giving patients the best possible option, thus, improving their quality of life. Bigger cohorts are needed to verify the polymorphisms in ERCC1, CYP2A6, TYMS, DPYD and GSTT1 prior application in clinical practice.

Abstract 3270 Introduction: Many of the mutations of the lysosomal acid b-glucosidase (b-glucocerebrosidase) associated with Gaucher disease (GD) translate into enzymes that retain partial catalytic activity in vitro but exhibit impaired cellular trafficking as a consequence of aberrant folding. Current investigational therapeutic strategies for include the development of ligands of the enzyme capable of promoting those conformational changes that are required for efficient folding, restoring trafficking. Although somewhat counter intuitive, competitive inhibitors of this b-glucocerebrosidase, at subinhibitory concentrations, can increase steady-state lysosomal levels of active enzyme through this rescuing mechanism, acting as “pharmacological chaperones”. At the massive lysosomal substrate concentration, the inhibitor would be replaced from the active site of the enzyme and the metabolic activity recovered. However, most of the pharmacological chaperones under study are iminosugars that behave as broad spectrum inhibitors, inhibiting simultaneously several glucosidases, which represents a serious inconvenient for clinical applications. An additional problem is that iminosugars and their derivatives are not active as pharmacological chaperones for glucocerebrosidase mutations located outside the domain containing the active site and are associated with neurological involvement, as the L444P mutation. Aim: The aim of this work is to present molecules with a high binding specificity towards b-glucocerebrosidase, with a high ratio of chaperone versus inhibitor activity and capable of producing an increased in the levels of mutant enzymes associated with Gaucher disease, including mutations located outside the catalytic domain. Methods: Different bicyclic derivatives of L-idonojirimycin were designed and chemically synthesized from D-glucose after in silico structural analysis and identification of the most favorable molecular features to interact with the active site of glucocerebrosidase. The chaperone potential of these compounds was evaluated at different concentrations in vitro using a cell model of GDcarrying the more frequent mutations in Gaucher disease, namely N370S and L444P. (P201230804). Results: The obtained results showed an increase in b-glucocerebrosidase activity at various chaperone concentrations, ranging from 1.96 to 4.98 folds for the L444P mutant and from 2.01 to 3.06 folds for the N370S mutation. Comments: The bicyclic derivatives of L-idonojirimycin could be considered as a therapeutic alternative for GD, mainly in patients with mutations located outside the active site of the enzyme and associated with neurologic involvement. Disclosures: Giraldo: Actelion: Membership on an entity's Board of Directors or advisory committees; Genzyme: Research Funding; Shire: Membership on an entity's Board of Directors or advisory committees, Research Funding.

e14058 Background: Patients with colorectal cancer (CRC) and liver metastases have a poor prognosis, only a small proportion has initially resectable metastases. Bevacizumab (BEV) added to chemotherapy (CT) is active for metastatic CRC. This study aimed to assess efficacy and safety of BEV as neoadjuvant or conversion treatment in patients with CRC liver metastases. Methods: Observational retrospective review in CRC patients with liver metastases who have undergone neoadjuvant or conversion CT with BEV as first or second line treatment. Results: 34 patients analyzed: 53% male; median age 55; 77% had metastases at diagnosis; ECOG 0-1 (38-62%); primary tumor location: 52% colon, 37% rectum, 11% colon/rectum; 84% had synchronic metastases; 16% initially resectable metastases 9% irresectable and 75% potentially resectable. 77% and 24% received first and second line BEV prior surgery respectively. Complete resection was achieved in 85%. All patients with initially unresectable metastases become resectable. 44% suffered post-operative complications; most frequent being abdominal and wound infection (12-9%). Complete pathologic response of liver metastases was seen in 7% and 61% reached major response. Median OS after surgery was 46 months. Disease free survival (DFS) since surgery was 8.8 months without significant differences between those receiving BEV as first or second line treatment. 88% of patients did not expeirence grade 3-4 toxicity. KRAS mutation was detected in 10 of 26 patients with available KRAS status (39%). Median DFS was significantly longer in wild type wt-KRAS patients compared with those mutated m-KRAS (12 vs 7 months p

e14671 Background: Analysis of the mechanisms of action of somatostatin analogs (SSAs) and mTOR inhibitors or tyrosine kinase inhibitors suggest that they may provide synergistic effects when used in combination for the treatment of pts with NETs. Methods: This is a Spanish multicenter cohort of pts with NETs treated with the SSAs lanreotide (LAN) and MTTs at 35 Spanish referral centers. Data of 159 combined treatments (133 pts) was retrospectively collected in order to evaluate the efficacy and safety of such combinations. Results: 133 pts (52,6% M) with a median age of 59,9 years; ECOG 0/1/2/3: 34%/49%/16%/1%; Lung/Pancreas/Gastrointestinal//Unknown(UK) origin: 9%/48% /32%/11%; G1/G2/G3/UK: 41%/32%/1%/26%; Non Functioning/ Functioning (69%/31%) received treatment with MTT + LAN for synergistic antiproliferative purpose in 85% of the cases, hormonal control (13,5%) or both objectives (1,5%). 115 pts received one combination treatment and 18 pts received between 2 (12 pts) and 5 (1 pt). LAN was administrated in combination with sunitinib (61), everolimus (73), bevazucimab (9), sorafenib (8) and pazopanib (8). The reported toxicity was determined by the MTT profile with no significant additional severe adverse events related to the combination with LAN. The probability of progression-free survival (PFS) at 6, 12 and 18months was 89%, 73% and 67% respectively for those pts who received LAN and sunitinib (n=50) and 78%, 69% and 57%, respectively for those pts who received LAN and everolimus (n=56). Median PFS was not reached at the time of analysis. Conclusions: The combination of LAN and MTT, mainly everolimus and sunitinib, is widely used in clinical practice without unexpected toxicities. Median PFS will be higher than data observed in phase III studies with sunitinib and everolimus, raising the hypothesis of enhanced efficacy combining SSAs and MTT that should be confirmed in randomized prospective clinical trials. [Table: see text]

368 Background: VTE is considered a main cause of mortality and morbidity in cancer pts and is commonly underestimated by oncologists. Pancreatic cancer is associated with the highest risk of VTE and the true incidence remains uncertain. Chemotherapy has been identified as an independent risk factor for VTE. The aim of this study is to analyze the incidence of cancer-associated thrombosis in ambulatory pts with pancreatic cancer receiving chemotherapy. Methods: We performed a retrospective review to determine the incidence of VTE in the gastrointestinal cancer unit of our centre. Between 2008 and 2010, 64 consecutives pts were identified and included in the analysis. Pancreatic neuroendocrine tumors were excluded. Results: Clinical characteristics ( table 1). Twenty pts (31.2%) experienced VTE including 7 pulmonary emboli (PE), 8 deep-vein thromboses (DVT) and 8 visceral vein thromboses (VVT). PE:25%. DVT:25%. VVT:35%. PE+DVT:10%. DVT+VVT:5%. Arterial thromboembolism (ATE): 2 pts (3.1%). 56% of the events were diagnosed during the first six months after diagnosis. Khorana’s predictive model for chemotherapy-associated thrombosis (risk category): intermediate risk (IR) 17 pts (27.9%) and high risk (HR) 44 pts (72.1%). VTE in HR and IR category: 14/44 pts (31.8%) and 6/17 pts (35.3%). Conclusions: The high incidence of VTE observed in this study is consistent with prior reports. Primary antithrombotic prophylaxis should be considered in this setting. Specific predictive model for chemotherapy-associated thrombosis in pancreas cancer must be investigated. [Table: see text]

144 Background: Neoadjuvant intraperitoneal and systemic chemotherapy (NIPS) is a new treatment modality in gastric cancer with peritoneal seeding. Several phase I-II clinical trials with NIPS and cytoreductive surgery have shown an increase in overall survival in Japanese patients (pts) with acceptable toxicity. We report the toxicity related to neutropenia and granulocyte colony stimulating factor (G-CSF) use of the first experience in Spain with NIPS (DCF-like) in gastric cancer with peritoneal carcinomatosis. Methods: Ten consecutive pts have been enrolled in this protocol in a compassionate use program in our center from 2009 to 2011. Chemotherapy (DCF-like) was delivered through an implantable peritoneal catheter. Primary prophylaxis with GCS-F was not used as described in the phase II studies, except in one pt. Clinical characteristics (Table). Results: The incidence of grade 3–4 neutropenia was 70% and any grade neutropenia 80%. Febrile neutropenia was described in 50% of the pts. All pts received secondary prophylaxis according to ASCO guidelines. No treatment-related deaths were observed. One pt discontinued chemotherapy due to febrile neutropenia and diarrhoea. There were no complications due to the infection of peritoneal catheter. Late or unexpected toxicities have not been observed. Conclusions: The first results suggest febrile neutropenia rate of this treatment modality exceeds the 20% ASCO threshold. Primary prophylaxis with G-CSF should be considered in this setting. [Table: see text]

Background Gaucher disease (GD) is due to deficiency of the glucocerebrosidase enzyme. It is panethnic, but its presentation reveals ethnicity-specific characteristics. Methods We evaluated the distribution, and clinical and genetic characteristics of GD patients in the Iberian Peninsula (IP). We analysed geographical distribution, demographic, genetic and clinical data, age at diagnosis, type, and years of therapy in 436 GD patients from the IP. Results The prevalence of GD was 1/149,000 inhabitants; 88.3% were type 1, 6.7% type 2, and 5.0% type 3. The mean age at diagnosis in type 1 was 28.7 years. A total of 72.7% were classified as having mild forms, 25.5% moderate, and 1.7% severe. Anemia and thrombocytopenia were present in 56% and 55%, respectively. Bone disease and hepatomegaly were reported in 62% and 68%, respectively, and were more likely in asplenic than in non-splenectomized patients. Sixty-nine mutant alleles were identified, and five mutations accounted for 75% of the GBA alleles. Several patients described in our series had interesting phenotypes. A total of 58.7% of patients had received enzyme replacement therapy and 12.6% were treated with miglustat. Conclusions A broad spectrum of GBA mutations is present in the IP, with 98.2% of type 1 GD being mild and 23.0% never treated. These data highlight genetic and phenotypic heterogeneities among geographic populations.

Abstract 4729 Background and Objective: The actual population of Iberian Peninsula (IP) is about 55 millions of inhabitants with a combination of various ethnic groups There are several reports that have analyzed GBA mutations in specific IP population. The aim of this study is to describe the distribution and main clinical characteristics of GD patients in IP. Patients and Methods: Data from the National Spanish Gaucher Disease Registry, Genetic Department of Barcelona University and Genetic Medical Institute of Porto since 1970 were jointed. Statistical analysis of demographic data, geographical distribution of GBA mutations and allelic frequency of GBA mutations in IP were analyzed. As well as identified the existence of clusters and the distribution of different types, the type and years on therapy. In addition we have determined the age at diagnosis (aDx) and SSI. Results: The prevalence of GD in IP had been 436 diagnosed patients, 421 of them born in IP. The mean aDx was 26.3±19.88 y, there are not differences between aDx before and after the last decade (24.8 vs 26.9 years respectively). Gender: female: 47.74 %, male: 47.98%, unknown: 4.28% type 1: 375 (89.1%), type2: 27(6.4%), type3: 19(4.5%). Mean SSI in type1 GD was 7.7 (range 1–30), and the distribution was 72.7% mild, 25.5% moderate and 1.7% severe. We have identified 70 alleles and 76 different mutations. The most prevalent allelic frequencies were: N370S: 48.8%, L444P: 18.3%, D409H: 3.1%, G3877S: 2.9%, double mutation: L444P+E326K: 2.0%, which account the 75% of total alleles, in 3.3% of alleles the mutation has not been yet identified and the remaining alleles corresponding to private mutations. Nevertheless the most frequent IP genotypes were N370S/L444P, 27.8%, N370S/N370S, 15%, N370S/? 5.2%, G377S/D409H, N370S/G202R, N370S/c.84insG, 2.4% respectively and homozygous for L444P 2.1%. Both Spain and Portugal the most prevalent mutations were N370S and L444P, however N370S is more frequent in Portugal than in Spain (57.1% vs 46.5%) by contrast L444P is more frequent in Spain than Portugal (19.4% vs 14.3%). 247 patients (58.7%) have received ERT for a mean of 10.2±3.89 years and 53 (12.6%) has been treated with miglustat for a mean of 2.1±1.9 years. The 90.4% of type1 and 31.0% of type 3 are alive and all type 2 are dead. Conclusions: There are broad spectrums of GBA mutations in GD patients from IP. Five different mutations account for 75% of GBA mutant alleles including N370S, L444P, D409H and G377S. No differences in gender distribution and tendency of aDx were observed. The 98.2% of type1 GD has a mild or moderate grade of severity and 23.0% has never received ERT or SRT. Disclosures: No relevant conflicts of interest to declare.

Abstract 4725 In the last twenty years enzyme replacement therapy (ERT) with imiglucerase has been a clinically effective for Gaucher disease (GD). The recombinant glucocerebrosidase administered intravenously - usually at biweekly intervals by lifelong has improved the quality of life of patients, avoided spleen removal and bone complications. In the last months an acute shortage of imiglucerase manufactured by the Genzyme Corporation (MA, USA) has occurred as a result of viral contamination firstly and other deficiencies in the production facility. In September 2009 a position statement based on the findings of the European Working Group for Gaucher Disease and European Gaucher Alliance, established a set of key recommendations about identification and monitoring of at-risk patients threatened. In Spain the follow-up of patients and the strict complementation of rules of therapy have permitted to obtain a profile of the situation in a group of patients with restricted ERT. Patients and Methods: A total of 50 GD1 patients have been analyzed before and after 6 and 12 months of imiglucerase shortage. Have been excluded for analysis children in order to dose reduction has been minimal as well as patients who have switched to another ERT or miglustat therapy. Results: Gender: 25 males/25 females. Mean age of group: 45.3±15.3 (range:18-84) SSI at diagnosis(Dx): 8.7±3.8 (range:3-19) Chitotriosidase (CT) activity at Dx:13,383±12,783 nM/mL.h; CCL18/PARC at Dx: 767±1,198 ng/mL. 20% of patients were splenectomized and 78% had bone disease at Dx. During shortage 23 patients (46%) discontinued therapy, in this period only one patient suffered a bone crisis and other anaemia as complications. Mean reduction of haemoglobin level: 2.7% (NS), platelet counts: 5.4% (NS). CT activity was increased 135% (p Disclosures: Off Label Use: Lenalidomide is not approved for the treatment of smoldering multiple myeloma.

Background: Bone disease is the most important source of disability in type 1 Gaucher disease (GD1). High percentage of patients present generalized osteopenia and osteoporosis with reduced bone mass. It is important to know the risk of bone mass lost in order to avoid the bone resorption. The standard technique to evaluate bone disease is Dual-Energy X-ray Absorptiometry (DEXA) that measures bone density in the axial skeleton. Nevertheless other procedure to exam bone density using ultrasound (US) can measure the amount of mechanical energy transmitted by the bone, as well as their speed of transmission (BUA). Aims: In order to evaluate bone disease indicators in GD1, we have planned a comparative analysis in GD1 by both techniques to exam individual variability in bone mineral density and to study the response to therapy. Patients and Methods: 24 adults GD1 patients diagnosed in last 5 years and 15 healthy controls stratified by age and gender were analyzed. Bone marrow changes have been evaluated by a semi quantitative score (S-MRI, Roca et al 2006 Eur J Radiol). To analyse the bone density we have used the Z-score quantification in L5 and femur head, calcaneus US was performed simultaneously determined BUA and bone density, in addition GBA genotype, clinical characteristics and subrogate biomarkers profile included chitotriosidase, CCL-18, MIP- 1b were analyzed. The advantages/disadvantages of each procedure and the correlation of results with the clinical characteristics, S-MRI, response to therapy, have been analyzed. Results: Osteopenia in GD is present in 85% of cases, it progresses with age and has been correlated with the severity of the disease and response to therapy. BMD evaluation by DEXA in GD1 is well documented but the special physio-pathological characteristics of GD make the use of US feasible and BUA results provide more information about the risk of skeletal complications. Comments: More experience is needed in order to precisely define the role of bone US exam in the management of GD patients. This work has been partially sponsored by FIS 07/90938 and CIBERER U-752

Background and objectives: Several reports describe the use of miglustat (Zavesca®), an oral inhibitor of glucosylceramide synthase, for type 1 Gaucher disease (GD1) in clinical trials. There are few data on the use of miglustat for GD1 in the real-world clinical experience. Here, we assess the long-term efficacy, tolerability and safety of miglustat in a prospective, open-label, observational study of GD1 patients attending routine clinic visits. Design and Methods: 48 patients with mild-to-moderate GD1 received miglustat 100 mg t.i.d. over 36 months. Patients were analysed in two groups: therapy-naïve patients (n = 11) and those switched from previous treatment with enzyme replacement therapy (ERT; n = 37). Assessments of clinical status, haematological parameters, and organomegaly were performed before treatment and after 6, 12, 24 and 36 months of therapy. Data were compared with a historical reference cohort 29 GD1 patients treated for 36 months with ERT. Results: Nine treatment-naïve patients completed 36 months of miglustat therapy. Sustained improvements in haemoglobin were seen in all patients who had anaemia before treatment. Platelet counts improved in patients with initial thrombocytopenia, and were maintained in patients with normal counts at baseline. Plasma chitotriosidase activity decreased in treatment-naïve patients during the first year on therapy and remained stable thereafter. S-MRI findings indicated improved bone status at 36 months. In patients switched from ERT, 15 completed 36 months of miglustat therapy, and displayed maintained or improved clinical and haematological parameters, and surrogate disease severity markers. No statistically significant differences were observed at 36 months between treatmentnaïve patients with similar baseline characteristics treated with either miglustat or ERT. Overall, miglustat was well tolerated. Interpretation and conclusions: Miglustat provided appreciable therapeutic responses on clinical, haematological, biomarker and S-MRI parameters at 36 months in patients with GD1, and was well tolerated. Therapeutic responses with miglustat were comparable with those seen with ERT.

Background: Gaucher disease (GD), caused by a deficiency of the glucocerebrosidase (GC) enzyme, is characterized by the accumulation of glucosylceramide (GlcCer) within lysosomes, resulting in cellular dysfunction and damage. GlcCer is catalyzed by the glucosylceramide synthase (GCS), also known as UDP-glucose ceramide glucosyltransferase, (UGCG), EC 2.4.1.80, first enzyme in glycosphingolipids biosynthesis. Mutations in the glucocerebrosidase (GBA) gene are required to cause GD, but other factors play an important role. Aims: GCS gene is a logical candidate to hypothesize that its variability could be involved in clinical severity. Patients and methods: We have analyzed GCS variants (g.(−295)C>T, g.(−222)ins10, g.148A>G, g.166A>T, g.25510C>G, g.29312A>G and g.34991A>G) in a group of 80 [N370S]+[L444P] heterozygous and 23 N370S homozygous patients. Results: were related with severity score index (SSI). g.(−295)C>T and g.166A>T SNPs were in complete linkage disequilibrium. In the N370S homozygous, carriers of g.(−222)ins10 have higher SSI than non carriers (6.6 vs 2.4, pG had higher SSI than the G-allele carriers (6.7 vs 1.5, pg variant at intron 1 changes the affinity for the transcription factor AP-2. GlcCer were elevated in patient carriers of one mutated allele of these two variants. Conclusion: our results suggest that the genetic GCS variants could influence in the development and progression of GD severity.

Gaucher disease (GD) is caused by a deficiency of the lysosomal enzyme acid β-glucosidase (GC) leading to accumulation of glucosylceramide within the macrophages of the reticuloendothelial systems. Plasma of GD patients contains low levels of total cholesterol, LDL-cholesterol and HDL-cholesterol. Enzyme replacement therapy (ERT) with imiglucerase and Substrate Reduction Therapy (SRT) with miglustat restores the haemoglobin levels, platelet counts and organomegalies. Previously we have demonstrated that ERT had significant effects on the concentration and metabolism of plasma lipoproteins. However, the effects of SRT on plasma lipid and other atherogenic profiles of GD patients have not been explored. We report the results of long term (36 months) SRT on plasma lipoprotein concentrations in 26 (11 men and 15 women) GD patients. Mean age of patients was 51 (SD ±20.7; range 22–74) years. Ten patients were therapy-naïve to SRT and 16 switched from previous enzyme replacement therapy (ERT). All patients received 100 mg of Miglustat t.i.d with recommendations of a low hydrocarbonate diet in the first weeks. Total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-c), high density lipoprotein cholesterol (HDL-C), Apolipoproteins (Apo AI, ApoB and Lp(a)), CRP, MPI1b, CCL18 concentrations and chitotriosidase activity were measured before (baseline) and after 12, 24 and 36 months of follow-up. Treatment resulted in a significant increases in HDL-c concentration in naïve patients during the first 24 months (p

Aims: Gaucher disease (GD) is characterized by a wide spectrum of manifestations. Several reports have indicated that GD relatives could developed neurological abnormalities. In order to detect neurological symptoms in GD patients and their relatives we have designed a specific prospective inquiry in Spanish GD families. Patients and Methods: the Spanish GD Registry is an independent registry working since 1993, sponsored by the Spanish Gaucher Foundation (FEETEG). From January to December 2006 we have perform a postal survey contacting 42 physicians and 92 families to evaluate neurological symptoms and correlated with genetic characteristics. The data were included in a SPSS 12.0 data base to perform statistical analysis using descriptive parameters, ANOVA, t-test and correlation study including Pearson coefficient. Results: We have obtained information from 72 families (78.3% responses) that included 92 type 1 and 7 type 3 GD patients and 266 relatives. Thirty (32.6%) of type 1 GD reported any neurological problem: tremor 8 (8.7%), uncordinated movements 9 (13.8%), concentration defects 11 (11.9%), strabismus 7 (7.6%), deafness 8 (8.7%), Parkinson disease (PD) 7 (7.6%), peripheral neuropathy 10 (10.9%). Thirty six (13.5%) first or second degrees relatives from 72 families presented neurological manifestations: PD 14 (5.3%), epilepsia 8 (3.0%), tremor 7 (2.6%), deafness 2 (0.7%), others 5 (1.9%). Patients with PD had mutations: S364R, D409H, L444P, [IVS2-2A>G]+ [c.(− 203)A>G], c.500insT, L336P. In relatives with PD a wide spectrum of mutations were observed: L444P, N370S, V398I, G202R, c.1439–1445del7, [E326K]+[N188S] and c.953delT, in others neurological manifestations as epilepsia, tremor or peripheral neuropathy predominant mutations were D409H, G195W, R120W; R147X, L336P and G377S. Conclusions: We have found a higher incidence than expected of PD and other neurological symptoms in GD1 patients and relatives. These manifestations appear frequently in carriers of L444P or rare mutations. It is important to perform a systematic neurological exam in GD1 patients and carriers with risk mutations. This work is partially supported by the grant FIS: 06/1253.

Background: Semiquantitative MRI is a very useful procedure to evaluate the bone marrow burden in Gaucher disease (GD). Score systems have been applied to obtain a parameter to evaluate the severity of bone disease. Our purpose was applied a simple, reproducible and accurate score to evaluate completely bone marrow involvement in GD patients Patients and Methods: MRI was performed in spine, pelvis and femora at diagnosis in 54 adult consecutively diagnosed as GD1, 61.1% females. Three MRI patterns and punctuation in each location were defined: normal: 0; non-homogeneous infiltration subtypes reticular: mottled: diffuse: and homogeneous infiltration: This score was called as Spanish-MRI (S-MRI): Two independent observers applied the S-MRI and Bone Marrow Burden (BMB) score and to compare the differences between both systems by using non parametric Mann-Whitney test. Correlation rank test was calculated. Results: In 46 patients (85.2%) bone involvement was observed, 39 (72.3%) has spine affected, 35 (64.8%) pelvis and 33 (61.2%) femorae, 14 patients had bone infarcts, 14 avascular necrosis, 2 vertebral fracture and 2 bone crisis. Correlation analysis between S-MRI and BMB was significant (r2:.675; p= .0001). No evidence of correlation was observed between CT activity and S-MRI and neither between CT activity and BMB. We have analysed the relationship between the patient genotype and bone infiltration according S-MRI, the location and complications. Patients with genotype N370S/rare allele (n=22), have more bone complications (90.1%) Infarcts: 9; AVN: 8, Bone crisis: 2; and vertebral fracture: 1. The mean S-MRI in this group was 7.9. Patients N370S/L444P (n=16) were the higher group with spine involvement (87.5%) mean S-MRI: 8.7. By contrast all patients homozygous for N370S (n=8), have femorae involvement, but a low S-MRI 7.5. The remaining 5 patients have complex genotypes (rare/rare), nevertheless they are not significant data(p=.335, p=.795) Conclusions: S-MRI is a simple method that provides useful information to evaluate bone infiltration and detect silent complications. Our results were correlated with the BMB score with higher sensitivity and specificity and provide accuracy to classify the degrees of the bone disease This work has been performed with the collaboration of the Spanish Group on Gaucher Disease and partially sponsored by a grant from FEETEG.

Background: The Spanish Gaucher Foundation (FEETEG) is an independent and non-profit organization that keeps the Spanish Gaucher Registry (SGR) established in 1993. This SGR contains demographic, genetic, clinical, analytical and image data from a cohort of 306 Gaucher Disease (GD) patients (269 GD1, 22 GD2, 16 GD3), and 751 relatives. The SGR is a useful tool to perform epidemiological-genetic studies and analyse associated comorbilities. Patients and Methods: From January to July 2006 an epidemiological survey to physicians and to the patients through GD Associations was conducted to ascertain the incidence of neurological symptoms in patients and their relatives as well as analyse genotype-phenotype relationship. The statistical analysis was performed in a SPSS 12.0 database using descriptive parameters, ANOVA, t-test and correlation study including Pearson coefficient. Results: Twenty-four (38.7%) out of 62 GD1 patient respondents from 56 families have neurological symptoms, 8 (12.5%) tremor, 8 (12.5%) uncoordinated movements, 11 (17.7%) concentration defects, 3 (4.8%) strabismus and 8 (12.5%) deafness, 3 (4.8%) Parkinson Disease (PD). Twenty-one out of 56 families have one or more relatives with neurological manifestations. There were 29(8.6%) out of 336 relatives with neurological symptoms:13 (3.8%) PD, 5 (1.5%) epilepsy, 6 (1.8%) essential tremor, 5 (1.5%) others. Subjects with PD were carriers of mutations:: S364R, G202R, V398I, R47X, L336P, L444P, G195W, recNci and insertion alleles. In families with epilepsy the predominant carrier mutations were: L444P, G195W, R130W and in essential tremor: L444P. Comments: There is a high incidence of neurological symptoms between GD1 patients and carriers. These manifestations appear frequently in carriers of rare mutations. It is very recommendable to perform a systematic neurological exam in GD1 patients and in carriers with risk mutations.

Introduction: Chitotriosidase protein (CT) is the most important biochemical marker described for Gaucher disease (GD). CT activity is increased several hundred-fold in plasma of GD patients and shows a strong positive correlation with the severity of the disease. However, a recessively inherited enzyme deficiency, with an incidence of about 6% in the Caucasian population, means that not all patients with GD can be monitored by measuring CT activity. Materials and Methods: This study was performed with plasma samples from 22 type 1 GD patients and 23 healthy subjects. All patients are from Spanish GD Registry. Applying two dimensional gel electrophoresis (2-DE) we compared the plasma proteome profile of GD patients with that of healthy subjects. Differential proteins were excised and prepared for MALDI-TOF analysis. Results were verify with 2-DE western blot. Results: In this study we have described five CT isoforms in a 2-DE gel from GD plasma samples. We show and compare the 2-DE images of each CT spot in plasma protein samples from healthy controls, GD patients with wild-type CT protein, and GD patients either homozygous or heterozygous for the 24 bp duplication. We propose a connection between the quantitative image analysis of each CT spot and the enzymatic activity of CT in each patient. Lastly, our results verifies that post-translational glycosylation explains at least some of differences between CT isoforms. Conclusion: The possibilities of proteomic technology in the investigation of biological biomarkers of GD and the nature of the CT protein are demonstrated, as well as in detecting new biochemical markers useful for the diagnosis of the disease and for monitoring patient response to therapy.

Background: Classically, type 1 Gaucher disease (GD) is characterized by organomegaly, haematological manifestations as anaemia and thrombocytopenia and frequent skeletal complications in absence of neurological manifestations. However, there is growing evidence for an association between GD and Parkinson’s disease and tremors, peripheral neuronopathy and others. These manifestations recently have been described in type 1 GD patients. The aim of our study has been to evaluate systematically a group of type 1 GD patients in order to assess about the presence of neurological manifestations. Design and methods: In this study, we have included 21 type 1 GD Spanish patients in follow-up for five years or more (male 7/female 14; mean age at diagnosis 24±16.88 years; SSI 6.5±3.2; genotype N370S or G377S heterozygous) 7 patient under enzyme replacement therapy (ERT) every two weeks at mean dose of 45 U/kg for ten years or more, 5 received therapy since four years ago and 4 under ERT since three years ago, all of them with satisfactory response (reduction in visceral size and normalization of hematological parameters). Five patients had not received any pharmacological therapy. All patients accept to participate in the study and the neurological exam and neurophysiological tests were performed by the same specialists. The protocol included a detailed inquiry about symptoms and concomitant medications and a neurological exam by a modified total neuropathy score to determine neurological signs. A superficial electroneurogram in sural and peroneal nerve was performed. As a cognoscenti test we used the memory impairment screen for dementia. Results: From a total of 21 patients, 8 patients (38.09%) showed some neurological deficits: a 8 years old child (GBA genotype G377S/R463C) develop ocular alteration and psychomotor delayed and he was reclassified as type 3 GD patient; two females siblings of 64 and 61 years old (GBA genotype N370S/IVS4-2A>G +c.( −203)A>G) has evident neurological symptoms. One of them presented parkinsonism since 40 years old and she did not response favourably to treatment with L-Dopa. The other sister carrier a joint hip replacement, has been under ERT since 1994 and she has developed dementia and Parkinson’s disease at 68 years old. Another splenectomized patient (age at diagnosis: 15 years, genotype N370S/L444P) was classified with mild or moderate and stable disease without ERT the neurological exam showed saccades ocular movements. One male 25 years old (genotype N370S/N370S) has been under ERT three years ago and presents abnormalities of eye movement. A 32 years old woman, (genotype N370S/L444P) had peripheral neuronopathy in arms. A 17 years old woman (genotype N370S heterozygous) had clonic movements in upper extremities. A child 3.5 years old (genotype N370S heterozygous) had psycomotor incordination. Comments: Due to the high incidence of neurological complaints in patients with GD type 1 it is necessary a precise and detailed clinical study to discard concurrent and medical problems and/or side effects from concomitant medications or true neurological problems related to GD.

Gaucher’s disease (GD) is an autosomal recessive lysosomal storage disorder, characterized by accumulation of glycosphingolipid in so-called Gaucher cells. The clinical manifestations of Gaucher disease are highly variable, and although certain genotypes are often associated with mild or severe symtomps, a defined correlation between genotype and phenotype does not exist. Identification of serum biochemical markers characteristic of disease may be useful in the diagnosis and monitoring of GD, nevertheless about 6% of the population does not express the chitotriosidase (CT) gene. In this study, we analyzed using currently available enzyme analysis the relationship among two known surrogate markers: CT, which utility in initiation and optimization of costly therapeutic interventions has been highly demonstrated, and a newly-described chemokine, pulmonary and activation-regulated chemokine (CCL18/PARC) as associated to Gaucher disease. Patients and methods: We have analysed 21 control samples, 149 samples of GD patients on enzyme replacement therapy (ERT), and 52 samples of GD patients on substrate reduction therapy (SRT), 4 samples of GD on combined therapy (ERT+SRT) and 7 samples of GD patients without therapy. The samples were stored at −80°C in the biobank of Biochemistry and Molecular and Cellular Biology Department of Zaragoza University. The CT activity and CCL18/PARC quantification were performed simultaneously in the samples obtained at baseline and yearly during 7 years under ERT. Results: our results concluded that both markers levels similarly fell with time and their variations correlated strongly each other, mainly in patients under ERT. The poor correlation of both variables in the case of SRT might be due to small number of samples. These findings demonstrated that CCL18 is a good biomarker of monitoring GD, comparable to CT and very useful in patients without expression of CT gene. Conclusion: The availability of sensitive plasma surrogate markers may be of great value for monitoring the efficacy of treatment, especially in cases of deficiencies of some marker.

Background and objective Miglustat (Zavesca®), an inhibitor of the enzyme glucosylceramide synthase, is an orally active molecule that has been shown to be a safe and effective therapy for type I GD. Here we report on our experience of miglustat monotherapy for the treatment of type 1 Gaucher disease (GD) patients in Spain. Design and Methods: We designed a prospective, open-label study of 21 patients with mild-to- moderate type 1 GD, to investigate the efficacy and tolerability of miglustat over a 12-month period in therapy-naïve patients and in patients who have previously received enzyme replacement therapy (ERT). Clinical assessments, including haematological parameters and organomegaly, were carried out prior to and after 6 and 12 months of miglustat therapy. In addition, the results were compared with data analysed retrospectively from 40 patients with type 1 GD who had been treated for 6 months with ERT. The data following 6 months of miglustat therapy are presented here. Results At present, 16 patients have completed 6 months of miglustat treatment. All patients with anaemia had improved haemoglobin concentrations, and in the treatment-naive group a mean increase of 1 g/dl was observed. Platelet counts improved in patients with thrombocytopenia and were maintained in patients with counts within normal limits at baseline. Chitotriosidase activity was maintained in switched patients and decreased in naïve patients. Miglustat was well tolerated, and the efficacy after 6 months therapy was comparable to that observed in the clinical trials and in patients treated with ERT for 6 months. Interpretation and conclusions In our experience, properly selected patients with mild to moderate type I GD had a satisfactory clinical and biochemical response to 6 months of miglustat therapy. The therapy was well tolerated, and clinical benefits were comparable to those obtained with ERT.

Background: Extreme elevations of plasma chitotriosidase (CT) are observed in Gaucher’s disease (GD) patients. This enzyme has been considered as a response marker to enzyme replacement therapy (ERT). It has been previously described the normalization of enzymatic activity only in patients who have been treated by bone marrow transplantation (Young E et al, 1997). It is widely accepted that GD subjects receiving ERT show improvements in clinical symptoms and regression of signs of disease, such as disappearance or reduction of visceral enlargement, normalized haematological parameters and skeletal improvement and also reduce the CT activity values, but these do not actually reach a normal range. Surprisingly, in the follow-up of patients included in the Spanish Gaucher Registry, some cases showed normalization of their CT activity after several years under therapy. The purpose of this study is to analyze the clinical, analytical and genotype characteristics of type 1 GD patients that have normalized their CT activity and to compare with the rest of patients under ERT for same period of follow-up. Patients and Methods: We have studied a cohort of 64 type 1 GD patients receiving ERT. We observed 20 GD patients who reach a normal CT activity range (CT activity < 200 nmol/mL.h) under therapy 2–7 years (group A) and 44 GD patients whose CT activity maintained increased under therapy 2–10 years (group B). Clinical and analytical data have been obtained from Spanish GD registry. Assessment of response included serial measurement of haemoglobin (Hb), platelet count, liver and spleen sizes and CT activity. Plasma CT activity was measured with the fluorogenic substrate 4-methylumbelliferyl-β-D-N, N′, N″ triacetylchitotrioside. Determination of the 24-bp duplication in the CT gene was performed by PCR followed by electrophoresis of the amplified fragments. For statistical analysis we used the StatView database (version 4.5). Results: At baseline characteristics of both groups were as indicated in tables. Patients of group B had worse indicators of disease severity at baseline. SSI, percentages of bone disease and spleen removal were higher in group B than group A. In addition we found twice more heterozygous patients for 24 bp duplication of the CT gene in group A compared to group B Nevertheless, as the table shows, patients of group B received higher doses of ERT and for a longer time. Conclusion: The normalization of CT activity is infrequent in GD patients under ERT. It is influenced by CT genotype and probably by severity of disease. Clinical data Age at diagnosis Male/Female SSI Spleen removal(%) Bone disease (%) N370S Homozygous/N370S Heterozygous N: number, SSI: severity score system Analytical data Group A (n=20) 32 ± 15.0 7/13 7.6 ± 2.55 5 55 0/20 Group B (n=44) 29 ± 15.9 19/25 8.4 ± 3.15 27 66 5/39 Analytical data Hb (g/dL) Platelets x 109/L CT activity(nmol/mL.h) Heterozygous CT genotype(%) ERT Dose (Units/2 weeks) N: number; Hb: Haemoglobin; CT: chitotriosidase; ERT: Enzyme replacement theraphy Group A (n=20) 12.3 ± 1.85 72.8 ± 31.27 7,489 ± 3,751 65 34.3 ± 10.33 Group B (n=44) 11.9 ± 1.84 107.5 ± 83.85 8,459 ± 3,563 32 41.4 ± 15.08

Introduction: AM3(Inmunoferon®), is a glycoconjugate of natural origin with immunomodulatory properties indicated in secondary immnunodeficiencies as well as coadjuvant treatment in neoplastic diseases with cellular immunity deficiency. Aim: To evaluate the influence of AM3 in the biological recovery of immune system in patients undergoing hematopoietic stem cells transplantation (HSCT)(autologous/allogenic). As well as to register the clinical incidences as intercurrent infections and mucositis from day of infusion hematopoietic cells to 90 days postransplantion in patients with and without AM3. Patients and methods: Group A (cases): Inclusion of 19 consecutive patients undergoing HSCT. April 2004 - June 2005. Inclusion criteria: age ≥ 18 years, first HSCT. and signed consent form. AM3 dose: 20mg/8h orally since infusion day to 90 days postransplantation. Group B (controls): 19 patients consecutively undergoing HSCT. from February 2003- March 2004. Biological evaluation of the immune system recovery: BCC, immunoproteins quantification: IgG, IgA, IgM, C3, C4, lymphoid populations distribution and activity of monocyte biomarkers (chitotriosidase, CCL18/PARC) on days −7, 0,+7,+14,+21,+28,+56,+90. Clinical evaluation and comparing cases/controls of incidence of mucositis severity evaluated according to WHO classification and microbiological documented infectious diseases. Results: The addition of AM3 as coadjuvant therapy in patients undergoing HSCT reduce significantly the percentage of oral mucositis (63.1% vs 89.5%) (p= 0.025) being those mucositis of less complexity in the first 2 weeks postransplantation. The number of infectious diseases was lower in patients under AM3 therapy (31.5% vs 47.4%) (p= 0.254) (Detailed in table). The recovery of the immune system evaluated by blood cells counts and immunobiomarkers showed a initial recuperation at day +14 in autologous HSCT and at day 21 for allogenic HSCT. The tolerance was satisfactory and only two patients needed discontinue therapy because of digestive intolerance. Wide studies must be performed in order to evaluate the benefit of this coadjuvant therapy. The inclusion of AM3 as a coadjuvant therapy in patients undergoing HSCT is associated with a significant minor percentage of oral mucositis. This work has been partially sponsored by a grant from FEHHA. Group A Group B Mean age (range) 44 (18–67) 48 (18–68) Females (%) 6 (31%) 7 (36,8%) Allogenic HSCT (M/F) 4 (21%) (3M/1F) 6 (31,5%) (5M/1F) Autologous HSCT(M/F) 15 (78,9%) (10H/5M) 13(68,4%) (7M/6F) Diagnosis Acute leukemia: 7 (37%), MM 7 (37%), MDS 2 (10%), Hodgkin disease 2nd remission 2 (11%), lymphoblastic lymphoma 1 (5%) Acute leukemia 4 (21%), MM 9 (47%), Hodgkin disease 2nd remission 2 (11%), NHL 4 (21%) Mucositis 12 (63%). Grade I: 1 patient (9%); grade II: 4 (33%); grade III: 4 (33%) 17 (89.5%). Grade II: 1 patient (6%); grade III: 6 (35%); grade IV: 9 (53%) Infectious diseases 6 (31,5%): S. Epidermidis 4 (21%), S. hominis 1 (5%), CMV 2 (10,5%), C. Albicans 1 (5%) 8 (42%): S. coagulase negative 7 (36,8%), S. viridans 1 (5.2%), S. epidermidis 1 (5.2%), Acinetobacter junii 1 (5.2%), Ochrobactrum anthropi 1 (5.2%), P. aeruginosa 1 (5.2%), C. difficile 1 (5.2%), ADN Herpes virus 6: 2 (10.5%), CMV 3 (15.7%), C. albicans 2 (10.5%), C. Krusei 1 (5.2%), P. carinii 1 (5.2%)