ALBERT

Description: Abstract 1373 Poster Board I-395 Background: High-dose therapy is the front line treatment of reference in young patients with Multiple Myeloma (MM). Although induction therapy remains a matter of controversy, bortezomib-based therapy is considered more and more as a standard of care. Prior to autologous stem cell transplantation (ASCT), patients undergo PBSC collect, usually starting after cycle 2 to 4 of the induction treatment. Currently, patients receive one transplant at front line, but most of the patients will benefit throughout the MM disease history of a second or a third ASCT procedure. We have noticed that the number of days of collection vary from patient-to-patient following bortezomib-based induction therapy. This increase in collection procedures might increase the cost of PBSC harvest with more patients discomfort and staff unavailability. We have therefore further studied the quality, yields and days of collection in myeloma patients following bortezomib-based therapy as compared to other regimens-based treatment courses. Material and Method: We retrospectively studied 70 patients with myeloma that underwent PBSC harvestsafter mobilization with GCSF following debulking with bortezomib-based therapy (58 days of procedures – 26 patients) versus other agents-based therapy [65 days of procedure – 44 patients; VAD vincristine, adriamycin, dexamethasone). Results: -Yields. CD34 Mobilization is lower following bortezomib-based therapy as compared to other regimen-based therapy. Similarly, more days of collection are also needed to collect the requested yield of PBSC. -Engrafment. No significant differences regarding engraftment was noticed among the 2 groups studied. The days to neutrophil and platelet counts recovery, the number of days with fever and the number of red cell and platelet transfusions were not significantly different between the 2 groups, mobilization following bortezomib-based therapy versus other regimen-based therapy, respectively. Conclusion: In our series, bortezomib-based induction regimen does not increase the number of PBSC harvest failure and the quality of engraftment was identical to other regimen-based induction treatment. However, CD34 mobilization was lower following bortezomib-based therapy, which explains lower daily harvest counts and therefore an increase number of days of collection in bortezomib-based treated patients. Therefore, PBSC harvest procedure following bortezomib-based therapy significantly increases the cost of PBSC collection. These results need to be confirmed in larger studies. New agents in use for mobilization might be considered for future PBSC collection in bortezomib-based treated patients. Disclosures: Leleu: Janssen Cilag: Research Funding.

Description: Abstract 3869 Poster Board III-805 Introduction The outcome of patients with plasma-cell leukemia (PCL) is poor. Avet-Loiseau reported on behalf the IFM, our first experience in PCL patients and showed that the median overall survival (OS) was 8 months (Avet-Loiseau, Blood, 2001). Since 1999, novel agents such as Thalidomide, Bortezomib (Velcade) or Lenalidomide (Revlimid) have been widely used in the treatment of multiple myeloma, both at the time of relapse or part of upfront therapy. Patients and methods In this retrospective analysis, we have looked at the outcome of PCL patients treated within the IFM since 1999 in order to study the impact of novel agents on survival. Results 31 cases, 20 males, 11 females, median age 55 years (34-78) were analyzed. Twenty one patients less than 65 years received high-dose therapy as part of frontline treatment : 19 autologous haematopoietic stem cell transplantation (HSCT) and 5 allogeneic transplantation. Novel agents were used part of induction therapy in 6 cases, at the time of relapse for 9 patients, for both induction and relapse in 16 cases. Thirteen patients received 1 novel agent, 11 received 2 and 7 patients received the 3 novel agents. The median number of lines of therapy was 2 (1 to 4). Bortezomib was used as up front treatment in 15 patients and at relapse for 9 patients. Overall response rate according the IMWG criterias was 70% (17/24) including 11 CR or VGPR (45%). PAD (Bortezomib, Adriamycin and Dexamethasone) and VTD (Bortezomib, Thalidomide, Dexamethasone) regimens provided the best response rates. Lenalidomide was used in 13 patients mostly at relapse. A response was obtained in 53% of patients including 2CR and 2 VGPR (30%). Nineteen patients were treated with Thalidomide-based regimens. Overall response rate was 52% (10/19) including 2 CR and 6 VGPR (31%). Overall, for the whole group of patients, the median progression-free survival was 8 months (0-26) and the median OS was 15 months (6-108). When comparing this survival with that described in our previous experience reported before 1999, we clearly showed that the use of novel agents improved the survival of patients with PCL. Conclusion In this retrospective study, novel agents improved the prognosis of P-PCL. Prospective IFM phase II studies are ongoing to confirm these results. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 2964 Background. Lenalidomide is an oral IMiD®, immunomodulatory compound, approved for use in combination with dexamethasone (Len/Dex) in patients with RRMM who have received one prior therapy. Len/Dex is indicated until evidence of disease progression at the best-tolerated dose of both Len and Dex (Dimopoulos et al. Leukemia 2011). However, the tolerability profile of long term exposure to Len/Dex is not well described, and evidence that long term exposure to Len/Dex would improve on the response rate and survival has yet to be determined. We sought to determine the efficacy and safety profile of long term exposure to Len/Dex in RRMM pts in a multicentre study. Method. We retrospectively reviewed the medical records of 50 RRMM pts treated with Len/Dex and remaining on Len for ≥2 years with a special focus on pts receiving Len for ≥3 years. All pts included had complete follow up records. Results. The median (range min-max) age was 58 years (39–79) with 30% (n=15) 〉 65 years (elderly MM), the sex ratio M/F was 1.2, 49% (n=24) ISS 2 and 3, 12% (n=6) severe renal insufficiency (CrCl 〈 30mL/min), and 8% (n=4) adverse FISH [del17p and/or t(4;14)]. Overall, 25 pts (50%) had Len/Dex at first relapse, 19 pts (38%) at second relapse and 6 (9%) pts in subsequent relapses. Len/Dex was given at first relapse in 10 (66%) elderly patients. The median time from diagnosis to starting Len/Dex was 4.5 years (1–16) for overall cohort and 3 years (1–8) for elderly patients (p=0.05). 28 pts (56%) received Len/Dex for ≥3 years. The median duration on Len/Dex was 3 (2–7) years for the overall cohort, and was 4 (3;7) years for patients exposed to Len ≥3 years. Treatment duration was similar across age categories and across number of previous relapses. With a median follow up of 4 years, 19 patients had stopped Len/Dex. The response rate (ORR, ≥PR) was 96% (n=48), including 37 (74%) patients with ≥VGPR, similar across age categories. Interestingly, the ORR and ≥VGPR were similar irrespective of whether patients have stopped Len/Dex in our study. The ORR was also similar across number of previous relapses, but the ≥VGPR rate was lower in patients at third relapse and beyond, (50%; p=ns). The ORR and ≥VGPR rate was 93% and 77% in patients exposed to Len ≥3 years, similar to the whole cohort. The median time to first response and best response were 2 (1–5) months and 4.5 (2–9) months, respectively. Overall, 9 (18%) patients stopped treatment due to toxicity, 9 (18%) progression of MM, and 1 (0.5%) patient decision. With a median follow up of 4 years, the median (95%CI) TTP was not reached, the estimated 4-yr TTP was 51.5%. There was no imbalance in the incidence of toxicity based on age, number of previous relapses, and patients exposed to Len ≥3 years did not discontinue more often due to toxicity, 14% versus 19% for those receiving Len 〈 3 years. The hematological safety profile was similar across age categories, number of previous relapses, and patients exposed to Len ≥ 3 years; overall, 8 (16%) patients experienced grade 3–4 neutropenia, 6% thrombopenia, and 6% anemia. Ten (20%) patients experienced a thromboembolic event (VTE), all of them of venous type. Two patients had previous history of VTE, but none of them experienced VTE on Len/Dex, likely related to adequate VTE prophylaxis. The median time to first occurrence was 5 (1;28) months, although 4/10 occurred in patients with ≥3 years on Len. All VTE occurred while on VTE prophylaxis except for 1 patient, 5 on aspirin, 2 on prophylactic doses of LMWH, and 2 on VKA (target INR 2–3). The incidence rate of second primary malignancy (SPM) was 3 (6%) (larynx, lung, and MDS). The SPMs occurred at a median time of 4 years from start of Len, while Len was already stopped in 2/3 patients, the latter stopped len at time SPM was diagnosed. Interestingly, none of the patients with more than 3 years exposure on Len had SPM. Conclusions. The current study provides estimates of responses, TTP and safety in a series of MM pts with long-term exposure to Len-based regimen at relapse. 62% of patients remained on Len beyond 3 years reflecting the efficacy and good safety profile of Len in relapsed MM, irrespective of age and number of prior therapies. Furthermore, no excess of long term side effects, including SPM, was observed with a prolonged long follow-up in this study. Disclosures: Robinson: Celgene: Employment. Miljkovic:Celgene: Employment. Morel:Celgene: Employment. Boccacio:Celgene: Employment. Facon:millenium: Membership on an entity's Board of Directors or advisory committees; janssen: Membership on an entity's Board of Directors or advisory committees; celgene: Membership on an entity's Board of Directors or advisory committees; onyx: Membership on an entity's Board of Directors or advisory committees. Hulin:celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees; janssen: Membership on an entity's Board of Directors or advisory committees. Leleu:Onyx: Honoraria, Speakers Bureau; Sanofi: Honoraria; Amgen: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Janssen: Honoraria, Research Funding, Speakers Bureau; Celgene: Honoraria, Research Funding, Speakers Bureau; LeoPharma: Honoraria, Speakers Bureau.

Description: Abstract 2667 Background. Measurement of serum M-spike is used to determine response to therapy and treatment free survival in Waldenstrom Macroglobulinemia (WM), however, its resolution on serum protein electrophoresis (SPEP) can make accurate measurement difficult. The same applies to total IgM quantification by nephelometry (IgMneph) which inherently includes monoclonal and polyclonal immunoglobulins. IgM levels by either technique do not accurately reflect tumor load or prognosis in WM. There is a need to identify new markers that reflect disease burden and correlate with patients'outcome. Hevylite measures IgMkappa and IgMlambda, separately, and might provide true quantitative measurement of the IgM M-spike. We sought to determine whether Hevylite value can be used as a reliable marker for response to therapy in WM as compared to the M-spike measurement and to assess treatment free and overall survivals. Methods. The study was conducted in a series of 86 WM patients [71 at diagnosis and 15 at relapse], of whom 10 patients with WM were homogeneously treated at front line in the multicentric phase 3 trial, chlorambucil versus fludarabine that included WM. All serum samples were collected prior to treatment and were kept frozen since collection. Responses included partial response (PR) or better, confirmed at 2 consecutive values. Hevylite measurements were made at The Binding Site Ltd, Birmingham, UK. A normal range was produced from normal (blood donor) sera (n=120), median (and 95%ile ranges) were; IgMkappa 0.634g/L (0.29–1.82), IgMlambda 0.42g/L (0.17–0.94), IgMkappa/IgMlambda ratio 1.6 (0.95–2.3). For ease of comparison we have studied the clonal IgM hevylite, expressed as IgMi HL and the IgM hevylite ratios expressed as the involved monoclonal immunoglobulin/uninvolved polyclonal immunoglobulin (IgM HLR). Results. The median age was 66.7 years, Male/Female ratio was 1.77, and 51.2 % patients had WM-ISS score 3. The median (min-max) M-spike level was 20.6 g/L (3.2–90), IgMneph was 19.4 (2.4–87), the median IgM HLR ratio was 100 (2.59–2850) and the median IgMi HL level was 21.9 (1.94–126) g/L. The IgMi HL values correlated well with the M-spike (intra class correlation (icc) coefficient = 0.48 [0.31; 0.60]) shown on the Bland-Altman plot left panel, and with the IgM neph (icc = 0.74 [0.52; 0.86]), right panel of the attached figure. The IgM HLR also correlated to the M-spike (r=0.44, p

Description: Abstract 3970 Background: Measurement of serum M-spike is used to assess response to therapy and treatment-free survival in IgA myeloma. However, its resolution on SPEP and the presence of IgA-dymers can make accurate measurement difficult. The more the M-spike decreases on SPEP, the more imprecise the M-spike measurement is using SPEP. IgA M-spike often migrates in the betaglobines that renders the M-spike measurement often complicated to analyze. Furthermore, quantification of the clonal IgA chain by nephelometry (IgAneph), which inherently includes monoclonal and polyclonal immunoglobulins, does not accurately reflect the tumour burden. Currently, there is a need in myeloma with IgA isotype (approximately 30–40% of patients) to identify new markers that better reflect the disease burden and the response to treatment, and correlate to patients' outcome. Hevylite® measures IgAkappa and IgAlambda and might provide precise quantitative measurement of the IgA M-spike. We sought to determine whether Hevylite® can be used as a reliable marker for diagnosis and response to therapy in IgA myeloma as compared to the M-spike measurement on SPEP and nephelometry. Methods: We conducted a retrospective analysis on 113, smoldering or symptomatic, IgA myeloma patients at diagnosis referred to our department from 1997 to 2011. All serum samples were collected prior to treatment or at relapse (for sequential data) and were kept frozen since collection. Hevylite® measurements were made at The Binding Site Ltd, Birmingham, UK. A normal range was produced from normal (blood donor) sera (n=138), and were for IgA kappa 0.48–2.82g/L, IgA lambda 0.36–1.98g/L and IgA K/L ratio 0.80–2.04. For ease of comparison we have studied the IgA hevylite ratios expressed as IgA K/L ratio. Results: The median age at diagnosis was 65 years (range: 33–93) and the M/F sex ratio was 0,91. Forty-eight percent of patients had an ISS greater than 2 (n=86). On nephelometry the median IgA level was 22g/L (range min-max, 0–100). Fifty patients had M-spikes migrating among the betablobulines, 27 among the gammaglobulines, 10 migrated in both and 23 unspecified. Fourteen patients had oligosecretory disease (M-component 〈 10g/L). Forty-two patients had an IgA kappa clonal chain, 70 others an IgA lambda and for one patient the data was not available. Among IgA kappa patients the mean HCLratio was 616.6 (median 117.7 [0.021–4323.7]) whereas it was 0.61 (0.21 [0.004–0.455]) among the IgA lambda patients. Across the entire population, 58 patients were identified by SPEP and HCL ratios. Among the 55 patients whose M-component was not quantified on SPEP, HCL ratios were abnormal in 53/55 (96%) cases. In the subgroup of patients whose M-protein migrates in the beta-region 29 out of 50 are identified by HCL ratios and SPEP. Another 19 patients (38%) had abnormal HCL ratios while unquantified on SPEP. The same was seen in patients whose M-protein migrates among the gammaglobulins with 16/27 and 11/27 (40%), respectively. In the last subgroup of patients with beta and gamma migrating M-proteins, all (10/10) were identified by HLC and SPEP. More interestingly, among the oligosecretory MM patients identified by an M-component 〈 10g/L, all (14 patients) had an abnormal HCL ratio. In our series of IgA myeloma, 51% of patients were accurately quantified on SPEP. When using HCLratios, an extra 47% of patients became measurable, with an abnormal HLC ratios allowing IgA myeloma in up to 98% of cases. When considering IgA measurement using nephelometry, 102 patients had both high IgA levels (90%) and abnormal HCL ratios. Interestingly, 7 patients had normal IgA levels with IgAneph and abnormal HCL ratio offering a useful diagnostic tool for 96% of patients as compared to IgAneph. In our series, IgA myeloma was neither associated with a poor outcome (median [range] OS: 119 months [0,5–604]) nor a poor response to therapy (median [range] TTP: 16 months (1–92). HCL involved chain ROC analysis identified a 37g/L cut-off as prognostic in IgA myeloma(p=0.039). Conclusion: Hevylite® is a new and reliable marker for diagnostic and monitoring of IgA myeloma. It enables to quantify accurately up to 98% of IgA-MM patients. These preliminary data need confirmation in further prospective trials in order to monitor further the impact of this marker in IgA myeloma patients before it becomes the gold standard to monitor the IgA M-protein in years to come. Disclosures: Boyle: Chugai: Consultancy, Honoraria. Combat:The Binding Site: Employment. Pietrantuono:The Binding Site: Employment. Facon:onyx: Membership on an entity's Board of Directors or advisory committees; celgene: Membership on an entity's Board of Directors or advisory committees; janssen: Membership on an entity's Board of Directors or advisory committees; millenium: Membership on an entity's Board of Directors or advisory committees. Harding:The Binging Site: Employment. Leleu:Celgene: Honoraria, Research Funding, Speakers Bureau; Janssen: Honoraria, Research Funding, Speakers Bureau; Novartis: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Sanofi: Honoraria; Onyx: Honoraria, Speakers Bureau; LeoPharma: Honoraria, Speakers Bureau.

Description: Abstract 3921 Background. Osteolytic disease is a major complication of multiple myeloma (MM) that may lead to devastating skeletal-related events (SREs). 70% of patients have osteolytic lesions at diagnosis, and up to 90% develop lytic lesions during the course of their disease. Furthermore, almost 15% of patients present with diffuse osteopenia at diagnosis. Various imaging techniques have been performed for the diagnosis and follow-up of bone disease in MM, however, conventional radiography (CR) remains the gold standard. CR has several limitations. One is that CR reveals lytic disease when more than 30% of the trabecular bone has been lost. Thus, some patients may have a suboptimal assessment of generalized osteopenia. Another limitation is that CR cannot be used for the assessment of response to therapy because the lytic bone lesions seldom show evidence of healing. On the other hand new vertebral fractures do not always indicate disease progression and may occur due to ongoing bone loss or reduction of tumor mass that supports the bony cortex. Other limitations include lack of accurate visualization of some areas, observer dependency, lengthy period on the examination table, and poor tolerance by patients with severe pain and extended lytic disease. The EOS System is a new 2D and 3D imaging system for musculo-skeletal physiology and pathology assessment with low radiation dose and standing position. We hypothesized that EOS would not be inferior to CR in terms of routine imaging and diagnosis of bone lytic lesions of patients with MM, but would improve on the quality of life during the procedure of the imaging for the frail patients with MM. Methods and Materials. Fifty six consecutive patients with symptomatic MM (at diagnostic and at first relapse) were included in this prospective study. Each patient provided informed consent. All patients underwent an EOS® examination (frontal and lateral views from skull to knees) and radiographs (axial skeleton: skull, spine, pelvis, femurs, humeri, ribs, as per International Myeloma Working Group guidelines) the same day, prior to start any treatment. Each imaging modality was read in random order by 2 reviewers independently for the detection of bone lesions (osteolytic lesions, vertebral collapses). Whole-body MRI was performed in case of disagreement between the 2 imaging modalities. Radiation dose and technical comfort were also assessed. The length of time of either exam was measured and the patients had to fill in a quality of life questionnaire aimed at comparing the roughness of the 2 techniques. Our study received prior approval from our Ethics Committee. Results. The median age was 62 (range, 32–90), gender ratio was 30 male / 26 female. CR and EOS® diagnosed 467 and 451 bone lytic lesions, respectively. There was no significant difference between the 2 imaging techniques, as 445 out of 473 bone lesions were detected by both the EOS®system and the CR. The median length of time to perform the CR exam was 6 to 8 times longer than EOS® technique. The average radiation dose with the EOS®system was 7.8 times less than with CR. The majority of the patients found the EOS®system examination to be more comfortable than the multiple radiographic incidences. The main limitation to EOS® technique was in patients with high BMI greater than 30, in whom CR remains the most sensitive technique. Furthermore, EOS® system was not able to differentiate old versus novel lytic lesions, as expected with standard radiographs. Finally, EOS presented with the same difficulty to count the number of lytic lesions per patient, as for the CR technique. Conclusion. EOS® system is a new low-dose radiation device which allows a quicker scan of the whole body. In this preliminary study performed in patients with MM, this technique allowed detection of bone lesions with better comfort for the patients as compared to CR. This is of paramount importance in patients with MM that often presented with altered health status and bone pain that hampered the ability to perform CR with optimal conditions. Disclosures: Facon: Celgene: Consultancy, Honoraria; Janssen-Cilag: Consultancy, Honoraria.

Description: Abstract 4930 Purpose AL amyloidosis is a monoclonal disorder responsible for secretion of a monoclonal free light chain which will deposit as aggregated fibrils and cause organ dysfunction. Prognosis without treatment is poor with median survival around 1 year. Since our multicentric randomized trial comparing M-dex (oral Melphalan and Dexamethasone combination) and high dose treatment with stem cell support (Jaccard et al, NEJM 2007), M-Dex is our reference front line therapy in AL patients, whatever the risk group. Median survival with this strategy has dramatically improved, reaching 5 years in our study as well as in the Italian one (Palladini et al, Blood 2007). Nevertheless survival of refractory patients was poor in the absence of valuable rescue treatment between 2000 and 2005(Figure 1). New drugs as Thalidomide, Lenalidomide or Bortezomib, whose efficacy has been proven in multiple myeloma, has been reported to be effective and tolerable in AL patients. We performed a retrospective multicentric study to determine outcome of M-Dex refractory patients in the era of these new drugs. Patients and methods Patients with biopsy proven AL amyloidosis, treated with M-Dex, front line, since June 2006 were included if they were considered as refractory by there referent physician in 10 centres belonging to the French network for AL amyloidosis. We recorded the hematological response with second line treatment. Survival was analyzed from the first treatment date using Kaplan Meier model. Results We included 29 patients with a median age of 60 years (32-76), 16 patients had cardiac involvement, 19 renal involvement. The median number of organ involvement was 2 (1-5). Isotype of monoclonal light chain was kappa in 38% of cases, and lambda in 62%. Median abnormal free light chain level was 158 mg/L (25.9-2100). Twenty patients (69 %) were considered as non responders because they did not reached a 50% decrease in free light chain serum level and 9 patients (31%), who achieved a partial hematological response, because they did not have a clinical response. The median time between the first M-Dex cycle and the second line treatment was five months (1-17). Second line consisted in thalidomide in 5 patients, lenalidomide in 7 patients, and Bortezomib in 17 patients, in combination with sequential Dexamethasone. Hematological response occurred in 69% of the whole series, with 27% complete response. Depending on treatment, partial hematological response was obtained in 4/5 patients with thalidomide, 2/7 patients with lenalidomide, and 14/17 patients with bortezomib responded with 8 complete responses. With a median follow-up of 21 months (0-32) 69% of patients are alive (Figure 2). Conclusion As expected introduction of new drugs for treatment of refractory AL patients gives a high level of hematological response leading to a better survival. Bortezomib seems to be particularly attractive with hematological response rate of 82%, and 47% complete response. The combination of M-Dex and bortezomib will be compared soon with M-Dex in a prospective international multicentric study. Disclosures No relevant conflicts of interest to declare.

Description: Abstract 2493 Poster Board II-470 Background. The Prognostic Inflammatory and Nutritional Index (PINI) is a simple scoring system which aggregates two blood markers of inflammatory (C reactive protein and orosomucoid) and of nutritional (albumin and prealbumin) states. This marker is predictive of nearest lethality and chronic institutionalization in geriatric routine practice for hospitalized elderly patients. MM is a malignant plasma cell disorder with a median age around 70 years old at diagnosis, a population characterized with several comorbid conditions that indirectly determine the prognosis and the treatment decision. The current most powerful adverse prognostic markers in MM are the adverse karyotypic abnormalities, such as 17p deletion and (4;14) translocation, and the high serum beta-2 microglobulin (b2m) that corresponds to the high International Staging System (ISS) group in combination with albumin. However, approximately 70% of MM patients have no know adverse karyotypic abnormalities, and high b2m and ISS are essentially informative for a minority of patients. We have sought to evaluate the prognostic value of PINI in MM in a large retrospective study. Method. The PINI score was determined in 226 previously untreated MM patients from sera collected at diagnosis before treatment, of whom 112 pts were 65 or older. The Serum albumin, prealbumin, orosomucoid (human α1-acid glycoprotein) and hsCRP were measured by immunonephelometry. PINI is calculated with the following formula: PINI=[orosomucoid (mg/L) × CRP (mg/L)]/[albumin (g/L) × prealbumin (mg/L)]. To avoid inadequate value of CRP, patients with fever and documented infection at diagnosis were excluded from the study. Results. Main patients characteristics at sample collection for the overall population were as follows: median age=64, ISS 2 and 3 of 29% and 21%, respectively. 35% of pts had chromosome 13 deletion (del13q). These characteristics were similar in pts older than 65. With a median follow-up of 57 months, median (+/-se) overall survival (OS) were 57 (+/-7) and 40 (+/-5) months in the overall and elderly population, respectively. The median (min-max) value of PINI were 0,29 (0,01-123) and 0,47 (0,01-123), respectively. The most informative PINI cut-off was determined at 4 and corresponded to its best impact on survival; 14% and 12.5% of pts had PINI ≥ 4, in the overall and elderly population, respectively. PINI correlated with markers of tumour burden such as high b2m level, low hemoglobin level and, high creatinin value. PINI ≥ 4 was also related to high ISS score (II + III), in 46% and 32% of pts, respectively. PINI did not correlate to del13q. In univariate analysis, median OS (+/-se) was significantly lower in patients with PINI ≥ 4 (number of death/number of pts in the group (O/N)=24/31) than pts with PINI

Description: Background: Acute myeloid leukemia (AML) is a heterogeneous disease even within the same genetic subgroup, e.g. within the NPM1 mutated subgroup, some patients have additional mutations in FLT3, IDH1/2, DNMT3A or TET2. Recent reports have shown that minimal residual disease (MRD) in AML, during or after treatment has prognostic impact. However, the clinical assessment of MRD in AML faces many questions. Firstly, which of the potential molecular and/or cellular markers should be included? Secondly, what type of biological sample should be analyzed? Thirdly, what is the sensitivity threshold to set and what are the relevant time frames for the MRD evaluation. The current technology development next-generation sequencing (NGS), opens new perspectives for monitoring MRD in AML, with the opportunity to expand and multiplex various markers. In this context, we tested the NGS technology for MRD monitoring IDH1/2 and DNMT3A in AML cases with mutated NPM1. We validated our results against MRD evaluated by RTqPCR. Patients and methods: In this study, we included 94 patients (pts) from the ALFA 0701 trial. NPM1 monitoring by RTqPCR was performed as previously described (Lambert et al, Oncotarget july 2014). IDH1/2 and DNMT3A monitoring was done by NGS using the Ion Torrent Proton instrument. In order to obtain very high coverage (approximately 2 Millions reads by sample), 24 samples were analyzed per run. Bioinformatic analysis was performed according to our previous work (Boyer et al, Am JHematol 2013). Results. 94 samples from 31 NPM1 mutated pts were analyzed. (17pts were positive for IDH1/2 and 15 were positive for DNMT3A). Sequencing data showed an excellent depth with a median of 2012459 reads (range 102,657 to 5,160,118 reads) and 966,298 reads (range 565,152 to 2,700,349 reads) for IDH1/2 and DNMT3A, respectively. Unfortunately, despite of this excellent coverage, a median of 520 reads were found positive in the negative controls due to a multi steps cross contaminations (OT2, Clonal amplification, purification). Thus, limiting the sensitivity to 0.07% (0.001-0.097%, p