Intranasal R-848 administration is an effective treatment for allergic airway disease. It hijacks an otherwise proinflammatory immune process triggered by TLR7 to mediate long-lasting disease suppression. This provides important insight into the efficacy and mode of action of TLR7 ligands in murine models of allergic airway disease and paves the way for their clinical application in humans.

Our studies demonstrate for the first time in vivo the pathogenic consequences of deregulated Activin-A expression in the lung, document novel aspects of Activin-A biology that provide mechanistic explanation for the observed phenotype, link Activin-A to ALI/ARDS pathophysiology, and provide the rationale for therapeutic targeting of Activin-A in these disorders.

Terpos E. Serum Dickkopf-1 is increased and correlates with reduced bone mineral density in patients with thalassemiainduced osteoporosis. Reduction post-zoledronic acid administration. Haematologica 2009; 94:725-728. doi: 10.3324/haematol.2008 This is an open-access paper. ABSTRACTSerum Dickkopf-1 is increased and correlates with reduced bone mineral density in patients with thalassemia-induced osteoporosis. Reduction post-zoledronic acid administration Dkk1 was measured at baseline and after 12 months of therapy, using ELISA methodology (Biomedica Medizinprodukte, No. BI-20412, Gesellschaft GmbH, Wien, Austria; in this ELISA 1 pmol/L=28.68 pg/mL), along with a series of serum bone indices: i) bone resorption markers [C-telopeptide of type-I collagen (CTX), tartrate-resistant acid phosphatase isoform-5b (TRACP5b)], ii) bone formation markers [bone-alkaline phosphatase (bALP), osteocalcin (OC) and C-terminal propeptide of collagen type-I (CICP)], and iii) osteoclast regulators [receptor activator of nuclear factor-κB ligand (RANKL), osteoprotegerin (OPG), and osteopontin], as previously described. 4 Bone mineral density (BMD) of the lumbar spine (L1-L4), femoral neck (FN) and distal radius (R) was determined using Dual-Energy X-ray Absorptiometry (DXA; LUNAR, PRODIGY Version 8.60.006/SYSTEM GE medical system LUNAR USA 726, Heartland Trail, Madison, WI 53717, USA) before and 12 months post-ZOL treatment. 4 The above bone markers were also evaluated in 30, age-and gender-matched, healthy controls (11M/19F, median age 44 years, range: 21-55 years). All controls had BMD measurements (L1-L4/FN/R) to exclude osteopenia/osteoporosis of other etiology, and were examined to ensure that there was no evidence of bone disease (i.e. osteoarthritis) and no receipt of medication that could alter the normal bone turnover during the previous six months. Statistical analysisThe Mann-Whitney test and paired samples t-tests were applied to evaluate the differences between patients and controls while the Wilcoxon signed rank test was used to evaluate differences between baseline and values of the studied parameters at the various time points. Differences between patients of three groups were evaluated using the Mann-Whitney test and the one-way ANOVA. The correlation between changes of various biochemical parameters and BMD was evaluated with the Spearman's (rs) correlation coefficient. Variables found to be statistically significant at the p<0.05 level for the presence of severe osteoporosis (Z-score< -4.0) in at least one studied site were entered into a multivariate model using Cox regression analysis to identify the most statistically significant model. All p values are two sided, the level of significance is <0.05 and confidence intervals refer to 95% boundaries. © F e r r a t a S t o r t i F o u n d a t i o n Results and DiscussionAt baseline, thalassemia patients had increased serum levels of Dkk1 (mean±SD: 39±17.1 pmol/L) compared with controls (27.4±9.7 pmol/L; p<0.0001; Figure 1A Figure 1B) and R-BMD (r=-0.415, p=0.001...

2018 Poster Board I-1040 Angiogenesis is a crucial process in the pathogenesis of several inflammatory, autoimmune and malignant diseases. Endothelial damage and inflammation make a significant contribution to the pathophysiology of sickle cell disease (SCD) and the beta-thalassemia syndromes. However, there are very limited data in the literature for the role of angiogenic cytokines in the pathogenesis of thalassemia major (TM) and of double heterozygocity of SCD and beta-thalassemia (HbS/beta-thal). Deferasirox (Exjade®) is a once-daily orally administered iron chelator approved for the treatment of transfusional iron overload in patients with transfusion-dependent anemia which seems to reduce markers of inflammation in TM. The aim of this prospective study was to evaluate the levels of angiogenic and inflammatory cytokines in patients with TM and HbS/beta-thal with iron overload who received chelation therapy with deferasirox. Forty-five patients (16M/29F) with TM and 20 patients (7M/13F) with HbS/beta-thal were evaluated. Deferasirox was administered at a dose between 10-30 mg/kg/day based on the number of blood transfusions received before the initiation of treatment. After 3 months, dose adjustments (increases) were allowed in increments of 5 mg/kg/day every 3 months as required to reduce markers of iron overload. Deferasirox was given for a period of 12 months. Serum levels of angiogenic cytokines, such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), angiogenin (Ang), angiopoietin (Angp)-1 and -2 and of inflammatory cytokines including interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-á), IL-1á, IL-1β, IL-4, IL-10 and transforming growth factor (TGF)-β1 and β2, were measured at baseline (day 1 of deferasirox administration) and then after 12 months post-deferasirox therapy, using ELISA methodology (R&D Systems, Minneapolis, MN, USA, for angiogenin cytokines and ILs and Diaclone, Bensancon, France for TNF-á, TGF-β1 and TGF-β2). Twenty healthy blood donors of similar age and gender were also evaluated as control group. Standard hematology and biochemistry was evaluated monthly in all patients. Patients with both TM and HbS/beta-thal had increased levels of all studied angiogenic cytokines compared with healthy controls but reduced levels of Angp-1/Angp-2 ratio (p<0.01 for all comparisons). Patients with HbS/beta-thal had increased levels of Angp-2 (p=0.002) and reduced levels of Angp-1/Angp-2 ratio (p<0.001) compared to TM patients. Both TM and HbS/beta-thal patients had also increased levels of IL-6 (p=0.008), IL-1á (p=0.01), TGF-β2 (p=0.01), IL-10 (p=0.02), IL-4 (p=0.03), and TNF-á (p=0.01) compared to controls. There were no differences between TM and HbS/beta-thal patients in terms of inflammatory cytokines. In TM patients, VEGF strongly correlated with TGF-β1 (r=0.652, p<0.001), platelet count (r=0.503, p=0.001) and white blood cell (WBC) count (r=0.484, p=0.002), while Angp-2 strongly correlated with TNF-á (r=0.536, p=0.001), WBC count (r=0.430, p=0.008) and platelet count (r=0.330, p=0.01). In HbS/beta-thal patients Angp-1/Angp-2 ratio strongly correlated with ALT (r=-0.611, p=0.007) and Hb (r=0.498, p=0.036). Twelve months post-deferasirox administration, there was a dramatic reduction of ferritin, SGOT and SGPT compared with baseline values in both patient groups (p<0.0001). TM patients showed a continuous increase of both VEGF (mean±SD: from 680±489 ng/ml prior to deferasirox to 801±547 ng/ml post-deferasirox; p=0.002) and bFGF (from 16.2±17.1 ng/ml prior to deferasirox to 33.1±28.8 ng/ml post-deferasirox; p=0.001) after 12 months of deferasirox therapy. No other changes were observed in TM group regarding angiogenic molecules. Patients of HbS/beta-thal showed no alterations in terms of angiogenic cytokines that continued to be elevated compared to controls. In terms of inflammatory cytokines deferasirox produced a reduction of TNF-á in both studied groups (p<0.01 for all comparisons). Our study suggests that angiogenic and inflammatory cytokines are increased in patients with TM and HbS/beta-thal and thus they have a role in the pathogenesis of these disorders. Angiopoietins seem to be strongly implicated in the biology mainly of HbS/beta-thal. However, further studies are needed to fully elucidate this role as well as the effect of deferasirox, if any, on inflammation. Deferasirox seems to have no impact on angiogenic cytokines. Disclosures: No relevant conflicts of interest to declare.

Osteoporosis represents an important cause of morbidity in adult patients with thalassemia. Its pathogenesis is multifactorial, and includes mainly bone marrow expansion, endocrine dysfunction and iron overload. Bone metabolism is altered in thalassemia. Osteoclast function is elevated, while osteoblast activity seems to be reduced and thus the balance of bone remodeling is in favor of bone loss. The exact mechanisms of osteoblast dysfunction have not been fully clarified to-date. Wingless-type (Wnt) signaling is an important pathway for osteoblast differentiation. Dickkopf-1 (Dkk-1) protein is an inhibitor of Wnt pathway and is implicated in the pathogenesis of several bone disorders. Collagen type-I is the main structural protein of the bone. The collagen type-I alpha (COLIA)-1 specific protein (Sp)-1 polymorphism has been related to osteoporosis in thalassemia. The aim of this study was to evaluate the serum levels of Dkk-1 in patients with thalassemia-induced osteoporosis who receive therapy with zoledronic acid (ZOL) and evaluate possible correlations with clinical and laboratory data, including the COLIA-1 Sp1 polymorphism. Sixty-six patients (21M/45F; median age 35.5 years) with thalassemia and osteoporosis were studied. Patients were blindly randomized to receive ZOL at a dose of 4 mg, iv, in 15 min infusion, every 6 months (group A, n=23) or every 3 months (group B, n=21), or to receive placebo every 3 months (group C, n=22) for a period of one year. All patients received oral calcium (500 mg) during the treatment period. Dkk-1 was measured at baseline and after 12 months of therapy using ELISA methodology (Biomedica Medizinprodukte, Wien, Austria) along with a series of serum bone remodeling indices: bone resorption markers [C-telopeptide of type-I collagen (CTX), tartrate-resistant acid phosphatase isoform-5b (TRACP-5b)], bone formation markers [bone-specific alkaline phosphatase (bALP), osteocalcin, and C-terminal propeptide of collagen type-I (CICP)], and osteoclast regulators [receptor activator of nuclear factor-kappa B ligand (RANKL), osteoprotegerin (OPG), and osteopontin]. The above bone markers were also evaluated in 30, age- and gender-matched, healthy controls. The G-->T mutation at base 1 of intron 1 at the binding site of the Sp1 transcription factor of the COLIA-1 gene was detected by polymerase chain reaction using mutagenesis primers followed by restriction enzyme analysis in all patients. BMD of the lumbar spine (L1-L4), femoral neck (FN) and wrist (W) was determined using DEXA, before and 12 months after treatment. At baseline, all patients had increased serum levels of Dkk-1 (mean±SD: 39±17.1 pmol/L) compared to controls (27.4±9.7 pmol/L; p<0.0001). Furthermore, thalassemia patients had increased values of CTX (p<0.0001), bALP (p<0.001), CICP (p=0.003), sRANKL (p=0.02), and OPG (p=0.001) compared to controls. Results for the COLIA-1 Sp1 polymorphism were available for 53 patients. Seventeen patients (32%) were G/T heterozygotes at the polymorphic Sp1 site (Ss), while 3 (5.6%) were T/T homozygotes (ss). Dkk-1 serum levels correlated with L1-L4 BMD (r=−0.290, p=0.022) and W-BMD (r=−0.415. p=0.001), but also with TRACP-5b (r=0.310, p=0.011) and bALP levels (r=−0.289, p=0.018). Ss and ss patients tended to have lower L1-L4 BMD compared with SS patients (p=0.09). No significant correlations were observed between Ss and ss patients with the measured bone markers or the response to ZOL. As reported previously, patients of group B experienced an increase of L1-L4 BMD, while no other alterations in BMD were observed in the 3 studied groups after 12 months of ZOL administration. Interestingly, patients of groups A+B showed a strong reduction of Dkk-1 after 12 months of ZOL (from 39.6±16.6 to 28.9±16.3 pmol/L; p=0.004); indeed they almost normalized Dkk-1 levels (no difference from control values). In contrast, patients of group C showed a borderline increase of Dkk-1 (from 33.1±16.8 to 40.1±23.2 pmol/L, p=0.08). These results show for the first time in the literature that Dkk-1 is increased in the serum of patients with thalassemia and osteoporosis, correlates with their BMD and is reduced post-ZOL therapy. This Dkk-1 elevation may be at least partially responsible for osteoblast dysfunction in thalassemia and reveal a novel possible target for the development of new agents for the management of bone loss in thalassemia patients.