Jimma, Thitina

Abstract Abstract 5158 Background: Vitamin D insufficiency is commonly observed in clinical practice. Observational studies have suggested an association of vitamin D insufficiency with increased risk of cancer development. Further, vitamin D insufficiency has been associated with poorer prognosis in solid tumors as well as hematologic malignancies, including non-Hodgkin lymphoma and chronic lymphocytic leukemia. In this study, we examined the clinical and prognostic correlates of low plasma levels of 25-hydroxyvitamin D (25[OH]D), which reflects whole-body vitamin D stores, in a cohort of myeloproliferative neoplasm (MPN) and myelodysplastic syndrome (MDS) patients. Methods: We studied a total of 409 patients: 247 (60%) had primary myelofibrosis (PMF), 74 (18%) de novo MDS, 63 (15%) polycythemia vera (PV) and 25 (6%) essential thrombocythemia (ET). Inclusion criteria required availability of archived plasma, bone marrow aspirate and biopsy and cytogenetic information at the time of first referral to our institution. Vitamin D measurements were performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). For the purposes of this study, plasma 25(OH)D levels <10 ng/mL indicated severe deficiency, <25 ng/mL insufficiency, and 25–80 ng/mL optimal levels. Results: Patient characteristics were as follows: (i) PMF: median age=63 years, >65 years=41%, males=68% and serum creatinine above the upper normal limit (UNL)=15%; (ii) MDS: median age=72 years, >65 years=77%, males=69% and creatinine >UNL=12%; (iii) PV median age=61 years, >65 years=37% and males=50%; and (iv) ET: median age=48 years, >65 years=10% and males=28%. Median (range) plasma 25(OH)D levels (ng/mL) were as follows: PMF 25 (0–60), PV 27 (6.8–51), ET 31 (6–50) and MDS 30 (7.7–72) (p=0.005). The proportion of patients with 25(OH)D insufficiency were: PMF 48%, PV 43%, ET 28% and MDS 28% (p=0.01). Similarly, the proportion of patients with severe 25(OH)D deficiency were: PMF 9%, PV 3%, ET 12% and MDS 1% (p=0.05). In PMF and MDS, 25(OH)D insufficiency was not significantly correlated with age (≤65 vs. >65 years), gender, red cell transfusion need, DIPSS-plus/IPSS category, hemoglobin (<10 vs. ≥10 g/dL), thrombocytopenia (<100 vs. ≥100 × 109/L), circulating blasts, unfavorable karyotype, serum creatinine >UNL, or JAK2V617F (PMF only) (p>0.05). 25(OH)D insufficiency was not predictive of inferior overall survival (OS) in PMF, MDS, PV, or ET (p>0.05). Similarly, in PMF and MDS, 25(OH)D insufficiency was not predictive of inferior leukemia-free survival (LFS) (p>0.05). Severe 25(OH)D deficiency was infrequently observed in MDS or PV patients; in PMF or ET patients, there was no prognostic impact of severe 25(OH)D deficiency on OS or LFS (p>0.05). Conclusions: Vitamin D insufficiency is relatively common in PMF, PV, ET and de novo MDS however this finding had no prognostic value for OS in the aforementioned patients. In PMF and MDS patients, low 25(OH)D levels had no clearly identifiable clinical correlates and further did not have prognostic value for LFS. Disclosures: No relevant conflicts of interest to declare.

Abstract Abstract 3850 Background: We have recently shown that primary myelofibrosis (PMF) was associated with increased plasma levels of IL-1b/IL-1RA/IL-2R/IL-6/IL-8/IL-10/IL-12/IL-13/IL-15/TNF-α/G-CSF/INF-α/MIP-1α/MIP-1b/HGF/IP-10/MIG/MCP-1/VEGF in PMF (J Clin Oncol 2011; 29: 1356). In addition, increased levels of IL-8, IL-2R, IL-12, IL-15 and IP-10 were independently associated with inferior survival. In the current study, we focused on the plasma cytokine profile in polycythemia vera (PV) and examined how it compares with PMF or in a cohort of PV patients whose samples were collected during follow-up. We also looked for phenotypic and prognostic correlates. Methods: Study inclusion for patients with PV required availability of archived plasma within one year of diagnosis. Samples from PV patients seen at different points post-diagnosis were also collected, in order to compare their cytokine profile with that of patients whose samples were collected within one year of diagnosis. Standard procedures were followed to centrifuge peripheral blood samples at 4°C and store aliquots at −80°C. Concentrations of 30 plasma cytokines/chemokines were analyzed in duplicates using Multiplex Bead-based Luminex Technology (Invitrogen, Carlsbad, CA, USA): IL-1β, IL-1RA, IL-2, IL-2R, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-15, IL-17, EGF, eotaxin, FGF-b, GM-CSF, G-CSF, HGF, IFN-α, IFN-γ, IP-10, MCP-1, MIG, MIP-1α, MIP-1β, RANTES, TNF-α, and VEGF. Measurements were performed on a Luminex 200 analyzer (Luminex Corporation, Austin, TX, USA) and resulting data were evaluated using STarStation Software Version 2.3 (Luminex Corporation, Austin, TX, USA). Results I: Plasma cytokines in PV (n=36 for samples collected within one year of diagnosis and n=40 for samples collected beyond that time point) vs. PMF (n=92) vs. normal controls (n=35) There was no significant difference between the three groups in the plasma levels of the following cytokines: IL-1β, IL-2, IL-4, EGF, and IL-17. PV and normal controls had similar levels of IL-2R, IL-5, FGF-b, and TNF-α; all four cytokines were elevated in PMF. The levels of the following cytokines were equally increased in PV and PMF: IL-1RA, IL-7, HGF, MIG, and VEGF. The following cytokines were also elevated in both PMF and PV but predominantly so in PV (MIP-1α) or PMF (IL-6, IL-8, IL-12, IL-13, IL-15, MIP-1β, IP-10, and MCP-1). Eotaxin and GM-CSF were elevated in PV but not PMF. Compared to normal controls, significantly lower levels were noted for IL-10 in PV and IFN-α and IFN-γ for PMF. IFN-α and IFN-γ levels were both elevated in PV. Comparison of PV samples collected within one year of diagnosis (n =36) vs. those collected beyond that time point (n =40) showed significantly increased levels of IL-8 (p=0.0005) and borderline significant increases in IP-10 (p=0.09) and IL-2R (p=0.09). Results II: Clinical correlates of plasma cytokines in PV Among the above-listed cytokines that were abnormally increased or decreased in PV, significant correlations were seen with sex (IP-10 and GM-CSF), age (IP-10, eotaxin), platelet count (MIG), hemoglobin level (IL-6), arterial thrombosis (IFN-γ, IL-1RA, VEGF, MIP-1α), and abnormal karyotype (IL-13, MIP-1α). None of the cytokines correlated with leukocyte count, venous thrombosis, pruritus or microvascular symptoms. The following, treated as continuous variables, were associated with inferior survival: IL-7, HGF, IFN-α, MCP-1, GM-CSF and IL-10 (p<0.05 for all). The number of events and sample size were too small to perform multivariable analysis. Conclusions: Plasma cytokine levels in PV are markedly abnormal and show both similarities and differences with PMF; the similarities became more pronounced in patients whose samples were examined at later stages of their disease. The current study suggests prognostic implications that warrant validation in a larger number of patients. Disclosures: No relevant conflicts of interest to declare.

Abstract Abstract 1751 Because IDH mutations are frequent in blast-phase myeloproliferative neoplasms (MPN), they might contribute to leukemic transformation. We examined this possibility in 301 consecutive patients with chronic-phase primary myelofibrosis (PMF). All study patients were fully characterized for karyotype, JAK2 and MPL mutational status, and Dynamic International Prognostic Scoring System-plus (DIPSS-plus) risk status. DNA from bone marrow or peripheral blood was used to screen for IDH1 and IDH2 mutations, by direct sequencing and/or high resolution melting (HRM) assay. Mutant IDH was detected in 12 patients (4%): seven IDH2 (five R140Q, one R140W and one R172G) and five IDH1 (three R132S and two R132C). MPL exon 10 was mutated in 18 patients (6.3%) and constituted W515L in 14 patients, W515K in 3 and a frameshift mutation in 1 patient. JAK2V617F was detected in 169 (56%) patients. Six patients displayed both JAK2V617F and IDH mutations (IDH2R140Q in 2 patients, IDH2R140W in 1 and IDH1R132S in 3); JAK2V617F allele burden was 1%, 7%, 22%, 27%, 30% and 96%, respectively. One patient displayed both IDHR140Q and MPLW515R. One-hundred and seven (36%) patients were negative for all three mutations. The 12 IDH-mutated patients were clinically compared to patients belonging to the three other molecular subgroups: mutated for JAK2 only (n=164), mutated for MPL only (n=18) and unmutated for all three (n=107). The four molecular subgroups were remarkably similar in their phenotype with few exceptions; IDH-mutated patients were significantly older than those with no mutations (p=0.04) whereas age distribution was similar between patients with mutant IDH, MPL or JAK2. In univariate analysis, overall survival (OS) for IDH-mutated patients was significantly shorter than those for JAK2-mutated (p=0.03), MPL-mutated (p=0.047) or unmutated (p=0.0009) patients. IDH-mutated patients also showed significantly shorter leukemia-free survival (LFS), compared to those with mutant JAK2 (p=0.0008), mutant MPL (p=0.02) or no mutations (p=0.001). After accounting for age, the presence of mutant IDH remained a significant disadvantage for both OS (p=0.04) and LFS (p=0.005). Multivariable analysis of OS that included risk categorization per DIPSS-plus confirmed the independent prognostic relevance of mutant IDH (p=0.03): HR for patients with no mutations =0.39, 95% CI 0.2–0.75; HR for JAK2-mutated patients =0.50, 95% CI 0.27–0.95; HR for MPL-mutated patients =0.53, 95% CI 0.23–1.2. A similar analysis for LFS that included risk factors for LT (i.e. unfavorable karyotype and platelet count <100 × 109/L) as covariates also confirmed the prognostic relevance of mutant IDH (p=0.003): HR for patients with no mutations =0.16; 95% CI 0.06–0.46; HR for JAK2-mutated patients =0.18; 95% CI 0.06–0.48; HR for MPL-mutated patients =0.09; 95% CI 0.01–0.76). Further analysis disclosed that the negative OS and LFS effect of mutant IDH was most pronounced in the presence (p=0.0002 and <0.0001, respectively) as opposed to the absence (p=0.34 and 0.64, respectively) of concomitant JAK2V617F expression. Analysis of paired samples obtained during the chronic and blast phases of the disease was possible in 2 IDH-mutated patients and showed the presence of both IDH and JAK2 mutations at both time points. Our observations suggest that IDH mutations in PMF are independent predictors of leukemic transformation and raise the possibility of leukemogenic collaboration with JAK2V617F. Disclosures: No relevant conflicts of interest to declare.

Abstract Among spliceosome component mutations, those involving SF3B1 are most frequent in myelodysplastic syndromes with ring sideroblasts (MDS-RS; ∼ 75% incidence) and SRSF2 in chronic myelomonocytic leukemia (∼ 28% incidence). We recently reported on the lack of prognostic significance for SF3B1 mutations in both MDS-RS and primary myelofibrosis (PMF). In the current study, we examined the prevalence and prognostic relevance of SRSF2 mutations in PMF. Among 187 patients screened, 32 (17%) harbored SRSF2 monoallelic mutations affecting residue P95. Significant associations were demonstrated between SRSF2 mutations and advanced age (P < .01), IDH mutations (P < .01), and higher DIPSS-plus risk category (P = .03). SRSF2 mutations were associated with shortened overall (P < .01) and leukemia-free (P < .01) survival; the adverse effect on survival was independent of DIPSS-plus (P = .01; HR = 1.9; 95% CI, 1.1-3.0) and IDH mutations (P < .01; HR = 2.3; 95% CI, 1.4-3.8). In conclusion, SRSF2 mutations are relatively common in PMF, cluster with IDH mutations, and are independently predictive of poor outcome.

Abstract DIPSS-plus (the Dynamic International Prognostic Scoring System-plus) includes 8 risk factors for survival in primary myelofibrosis. In the present study of 884 karyotypically annotated patients with primary myelofibrosis, we sought to identify 1 or 2 parameters that can reliably predict death in the first 2 years of disease. After a median of 8.2 years from time of referral to the Mayo Clinic, 564 deaths (64% of patients in the study) had been recorded. Risk factors associated with > 80% 2-year mortality included monosomal karyotype, inv(3)/i(17q) abnormalities, or any 2 of the following: circulating blasts > 9%, leukocytes ≥ 40 × 109/L, or other unfavorable karyotype. Patients with any 1 of these risk profiles (n = 52) displayed significantly shorter overall survival than those otherwise belonging to a high-risk category per DIPSS-plus (n = 298); respective median survivals were 9 and 23 months (hazard ratio 2.2, 95% confidence interval 1.6-3.1; P < .01). The present information complements DIPSS-plus in the selection of primary myelofibrosis patients for high-risk treatment approaches.