, rs9399137, rs7775698, and 3-bp (TAC) deletion, and SNP rs5006884 in OR51B6 was done by PCR, followed by nucleotide sequencing using the BigDye terminator cycle sequencing kit from Applied BioSystems.HBG1-HBD nucleotide sequencing. Short (500-600 bp) and overlapping fragments of DNA covering the 14.1 kb region were amplified by PCR. All PCR reactions were performed in a total volume of 20 ml. Master mix concentrations, and cycling conditions were optimized based on the region being amplified. Usually, each reaction contained 100-250 ng of DNA, 1X PCR buffer (Applied BioSystems), 2 mM MgCl 2 , 200 mM dNTP, 1 ng of each primer, and 0.5U AmpliTaq polymerase. Cycling conditions consisted of 5 min at 948C, followed by 30 cycles of 40 sec at 948C, 40 sec at 558C, and 3 min at 728C and a 7-min elongation step at 728C.Statistical Analysis. Association between HbF and the minor alleles of each QTL was statistically analyzed by the Fisher exact test performed in R (www.r-project.org). Comparison of hematological parameters and correlation between NOMAP and HbF in both groups was examined using paired T-test. An overall significance level of 0.05 was set for all statistical analyses.

Background: preclinical data suggest that autophagy can serve as an adaptive mechanism facilitating tumor cell survival and resistance to therapy-induced apoptosis. Subsequently, inhibitors of autophagy are thought to enhance the efficacy of therapeutic strategies designed to induce tumor cell apoptosis. Based upon our preclinical results, we hypothesized that the addition of chloroquine, an autophagy inhibitor, to the combination of a proteasome inhibitor, and cyclophosphamide might result in a synergistic effect. Patients and Methods: We designed a phase II trial of bortezomib, cyclophosphamide and chloroquine in patients with refractory myeloma (defined by > 25% increase in M-protein) who progressed on a combination of bortezomib and cyclophosphamide. Treatment consisted of 42 day cycles repeated until disease progression with bortezomib 1.3 mg/m2 IV on days 1, 4, 8, 11, 22, 25, 29 and 32; cyclophosphamide 50 mg orally twice a day; and chloroquine at 500 mg orally daily on days 1-14 and 22-35. The primary endpoint was to estimate clinical benefit (CR+VGPR+PR+SD) after 2 cycles. Results: Between October 2011 and April 2013, 11 patients including 7 men and 4 women were enrolled and treated. The median age was 62 years (range 53-80). All patients were ISS stage II and III. Prior therapies for most patients included transplant, bortezomib, cyclophosphamide and lenalidomide. One patient withdrew after C1D1 and two patients progressed prior to completion of cycle 1. Of the 8 evaluable patients who completed at least 2 cycles, 3/8 achieved a partial response, 1/8 showed stable disease and 4/8 progressed with a clinical benefit rate of 4/10 (40%). Among the 3 patients who achieved a partial response, median duration of response was 4 months (250, 131 and 117 days respectively). The most common non-hematological adverse events (AEs) observed in >25% of patients (any Gr/Gr >3) and felt to be at least possibly related to the study drug were: fatigue (91%/18%); constipation (72%); myalgia, nausea (64% each); cough (55%), diarrhea, neuropathy (45% each); congestion (36%/9%); anorexia and dyspnea (27% each). The most commonly observed hematological AEs were neutropenia (45%/36%) and thrombocytopenia (55%/45%). Data from LC3 and beclin1 assays used to measure correlations of clinical response with in vitro autophagy inhibition will be presented. Conclusions: this study suggests that the addition of chloroquine to bortezomib and cyclophosphamide is effective in overcoming proteasome inhibitor resistance in a significant fraction of heavily pretreated patients, with an acceptable toxicity profile. Disclosures No relevant conflicts of interest to declare.