Search

Figure S2. Hazard ratios (black line) and 95% confidence intervals (blue area) from the univariate survival analyses according to the presence of CTCs categorized using different cutoff values (e.g., cutoff 0: 0 CTCs vs. > 0; cutoff 1: 0-1 CTCs vs. > 1; cutoff 2: 0-2 CTCs vs. > 2). Panel A shows breast-cancer-specific survival, and Panel B shows distant disease-free survival.

Purpose: We assessed the impact of donor type in acute myeloid leukemia (AML) patients transplanted with 2 Gy total body irradiation (TBI)-based nonmyeloablative conditioning regimen.Patients and Methods: Data from 1,715 adult patients, with AML in CR1 or CR2 were included in this retrospective survey.Results: Donors consisted either of HLA-matched sibling donors (MSD, n = 701), 10/10 HLA-matched unrelated donors (MUD, n = 611), HLA-haploidentical donors (haplo, n = 112) or single or double umbilical cord bloods (CBT, n = 291). Chronic graft-versus-host disease (GVHD) was less frequent in CBT (28%) and in haplo (30%) patients than in MSD (50%) and MUD (51%) recipients (P < 0.001). Two-year incidence of relapse was 32%, 30%, 34%, and 34% in MSD, MUD, CBT and haplo patients, respectively (P = 0.7). Two-year overall (OS) and GVHD-free relapse-free survival (GRFS) were 59% and 29% in MSD patients, 56% and 39% in CBT recipients, 53% and 23% in MUD recipients, and 43% and 37% in haplo patients, respectively. In multivariate analyses, MUD patients had lower GRFS than MSD patients beyond day 100 (HR 1.3, P = 0.001) while CBT was associated with a better GRFS than MSD beyond day 100 (HR 0.6, P = 0.002).Conclusions: In this large cohort of AML patients transplanted following low-dose TBI-based conditioning, the relapse incidence was not affected by donor type suggesting that the intensity of GVL effects might be comparable with these four transplant approaches. Furthermore, CBT was associated with better GRFS beyond day 100 than MSD while the opposite was observed for MUD. Clin Cancer Res; 24(12); 2794–803. ©2018 AACR.

supplemental figure legend

Figure S1. Flow chart of the data selection process for a pooled analysis of the prognostic relevance of circulating tumor cells in early breast cancer. Patients were ineligible if no valid data on CTC presence at the time of primary diagnosis as assessed using the FDA-cleared CellSearch® System were available (e.g., because of insufficient amount of blood or test failure). * A total of 3,754 patients with early breast cancer were randomized in the SUCCESS A clinical trial; however, blood samples for the enumeration of CTCs with the CellSearch® System were only collected after written informed consent from 1,994 of those patients.

Figure S3. Kaplan-Meier plots of overall survival according to the presence of circulating tumor cells at the time of primary breast cancer diagnosis. Panel A shows the predefined low-risk (T1/N0) patient subgroup, and Panel B shows the high-risk (T234/N123) patient subgroup.

Purpose: Although unequivocal evidence has shown the prognostic relevance of circulating tumor cells (CTC) in the peripheral blood of patients with metastatic breast cancer, less evidence is available for the prognostic relevance of CTCs at the time of primary diagnosis.Experimental Design: We conducted a pooled analysis of individual data from 3,173 patients with nonmetastatic (stage I–III) breast cancer from five breast cancer institutions. The prevalence and numbers of CTCs were assessed at the time of primary diagnosis with the FDA-cleared CellSearch System (Janssen Diagnostics, LLC). Patient outcomes were analyzed using meta-analytic procedures, univariate log-rank tests, and multivariate Cox proportional hazard regression analyses. The median follow-up duration was 62.8 months.Results: One or more CTCs were detected in 20.2% of the patients. CTC-positive patients had larger tumors, increased lymph node involvement, and a higher histologic tumor grade than did CTC-negative patients (all P < 0.002). Multivariate Cox regressions, which included tumor size, nodal status, histologic tumor grade, and hormone receptor and HER2 status, confirmed that the presence of CTCs was an independent prognostic factor for disease-free survival [HR, 1.82; 95% confidence interval (CI), 1.47–2.26], distant disease-free survival (HR, 1.89; 95% CI, 1.49–2.40), breast cancer–specific survival (HR, 2.04; 95% CI, 1.52–2.75), and overall survival (HR, 1.97; 95% CI, 1.51–2.59).Conclusions: In patients with primary breast cancer, the presence of CTCs was an independent predictor of poor disease-free, overall, breast cancer–specific, and distant disease-free survival. Clin Cancer Res; 22(10); 2583–93. ©2016 AACR.

Table S1. Patients and study variables according to study center for the pooled analysis of circulating tumor cells in early breast cancer

Inadequate mobilization of peripheral blood progenitor cells (PBPCs) is a limiting factor to proceeding with autologous hematopoietic cell transplantation (auto-HCT). To assess the impact of clonal hematopoiesis (CH) on mobilization failure of PBPC for auto-HCT, we investigated the characteristics of poor mobilizers (with a total PBPC collection

Patients with poor risk acute myeloid leukemia (AML) have a dismal outcome. We hypothesized that combining decitabine with a standard non-myeloablative (NMA) conditioning regimen prior to allogeneic hematopoietic cell transplantation (allo HCT), might decrease the relapse incidence. We conducted a multicenter prospective phase II study (NCT02252107) with 10-day decitabine (20 mg/m

The predictive value of measurable residual disease (MRD) for survival in acute myeloid leukemia (AML) has been firmly established in younger patients treated with intensive chemotherapy. The value of MRD after treatment with decitabine in older patients is unknown. This retrospective analysis included patients ≥60 years of age with AML who received an allogeneic hematopoietic cell transplantation (alloHCT) after treatment with decitabine or intensive chemotherapy. Of the 133 consecutively transplanted patients, 109 had available pretransplantation MRD analyses (by flowcytometry [threshold 0.1%]). Forty patients received decitabine treatment (10-day schedule), and 69 patients received intensive chemotherapy (7 + 3 regimen). Patients who received decitabine were older (median 67 versus 64 years) and more often had MRD (70% versus 38%). OS after alloHCT was comparable in both groups. In the chemotherapy group, MRD-positive patients had a significantly higher relapse probability (subdistribution hazard ratio [sHR] 4.81; P= .0031) and risk of death (HR 2.8; P= .02) compared to MRD-negative patients. In the decitabine group there was no significant association between the presence of MRD and relapse (sHR 0.85; P= .83) or death (HR 0.72; P= .60). Pretransplantation MRD in patients receiving decitabine treatment does not have similar predictive value for relapse or survival in older AML patients receiving an alloHCT, compared to patients receiving intensive chemotherapy.

Purpose: Although unequivocal evidence has shown the prognostic relevance of circulating tumor cells (CTC) in the peripheral blood of patients with metastatic breast cancer, less evidence is available for the prognostic relevance of CTCs at the time of primary diagnosis. Experimental Design: We conducted a pooled analysis of individual data from 3,173 patients with nonmetastatic (stage I–III) breast cancer from five breast cancer institutions. The prevalence and numbers of CTCs were assessed at the time of primary diagnosis with the FDA-cleared CellSearch System (Janssen Diagnostics, LLC). Patient outcomes were analyzed using meta-analytic procedures, univariate log-rank tests, and multivariate Cox proportional hazard regression analyses. The median follow-up duration was 62.8 months. Results: One or more CTCs were detected in 20.2% of the patients. CTC-positive patients had larger tumors, increased lymph node involvement, and a higher histologic tumor grade than did CTC-negative patients (all P < 0.002). Multivariate Cox regressions, which included tumor size, nodal status, histologic tumor grade, and hormone receptor and HER2 status, confirmed that the presence of CTCs was an independent prognostic factor for disease-free survival [HR, 1.82; 95% confidence interval (CI), 1.47–2.26], distant disease-free survival (HR, 1.89; 95% CI, 1.49–2.40), breast cancer–specific survival (HR, 2.04; 95% CI, 1.52–2.75), and overall survival (HR, 1.97; 95% CI, 1.51–2.59). Conclusions: In patients with primary breast cancer, the presence of CTCs was an independent predictor of poor disease-free, overall, breast cancer–specific, and distant disease-free survival. Clin Cancer Res; 22(10); 2583–93. ©2016 AACR.

Background Allogeneic hematopoietic stem cell transplantation (alloHSCT) has been established as a powerful treatment modality for patients with hematological malignancies. The graft-versus-leukemia effect, however, is strongly associated with the occurrence of graft-versus-host disease (GVHD) and subsequent transplant-related mortality. The current standard Dutch regimen for prevention of GVHD in T-cell replete (TCR) alloHSCT following reduced intensity conditioning (RIC) consists of mycophenolic acid (MA) and cyclosporine A (CyA) for three and six months post-transplant, respectively. However, as approximately 30% of patients will never develop GVHD, immunosuppressive overtreatment is of concern and might impair outcome. Aims In the present prospective randomized, multicenter, phase III trial we set out to compare time-restricted immunosuppression versus a standard immunosuppressive regimen. The primary objective was to increase the proportion of patients with non-severe GVHD (either acute GVHD grades I-II without gut involvement or chronic GVHD not requiring systemic treatment) within 180 days after transplantation and to reduce the relapse rate, without increasing severe GVHD. Secondary endpoints included time to acute and chronic GVHD, progression-free survival (PFS), GVHD-free/relapse-free survival (GRFS), overall survival (OS), and adverse events. Methods Hematological patients planned to undergo TCR alloHSCT with a related or unrelated 8/8 HLA matched donor were included. The trial randomized patients between three treatment arms. The current analysis includes all patients randomized between arms A and B. Immunosuppression consisted of CyA twice daily aiming for serum trough levels of 250-350 µg/L and MA 16 mg/kg twice daily with a maximum dose of 2160 mg a day. The standard regimen (arm A) prescribed to discontinue MA at day 84 post-transplant and CyA was continued until day +120 followed by tapering until day +180. In those randomized for the time-restricted regimen (arm B) MA was discontinued at day 28 post-transplant and CyA was continued until day +84 followed by tapering. Results A total of 389 patients were randomized 1:1 between arms A and B, of whom 95% (184 in arm A versus 185 in arm B) proceeded to transplant. The median age was 55 (range: 18-71), 57% were male. Fifty-one patients received myeloablative conditioning and 318 (86%) patients RIC. Donors were matched siblings for 135 patients and matched unrelated donor (MUD) for 233 patients. The majority of patients received peripheral blood stem cells, consisting of median 6.46x106/kg CD34+ cells/kg (range: 0.94-26.3) and median 230x106/kg CD3+ T cells (range: 0-936). Baseline patient and transplantation characteristics were equally distributed between the treatment arms. The proportion of patients developing non-severe GVHD within 180 days post-alloHSCT was 24% in both treatment arms, with an odds ratio (OR) of 1.01 (95% confidence interval 0.61-1.67, p 0.98). The cumulative incidence (CI) of grade II-IV and grade III-IV acute GVHD at 6 months post-alloHSCT was not significantly different between the two arms (47% and 15% versus 52% and 18%), nor was the maximum grade or organ involvement of acute GVHD. In addition, no difference was seen in the two-year CI of chronic extensive GVHD between the two treatment arms (51% versus 49%). The three-year estimate of PFS was 51% (44-59%) versus 52% (44-59%), respectively. The three-year CI of progression/relapse was 28% in arm A versus 27% in arm B. OS at three years was 59% (0.51-0.66%) versus 57% (0.50-0.64%). The one-year estimate of the composite endpoint GRFS was 14% (9-19%) in arm A, and 17% (12-22%) in arm B. Incidences and nature of adverse events were comparable in both arms. Conclusion A time-restricted combination of MA and CyA did not increase the proportion of patients with non-severe GVHD within 180 days after transplantation and resulted in similar outcome as compared to standard immunosuppression following alloHSCT using sibling and well-matched unrelated donors. Given the observed high CI of acute grade II-IV and chronic extensive GVHD and low GRFS for both the time-restricted and standard regimen, recipients of TCR RIC alloHSCT should be considered for more intensive immunosuppression. Figure Disclosures Nur: Novartis: Consultancy. Deeren:Alexion, Amgen, Janssen, Roche, Sunesis, Takeda, Sanofi: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding.

The presence of circulating tumor cells (CTC) is an independent prognostic factor for progression-free and overall survival for patients with metastatic and newly diagnosed breast cancer. The present study was undertaken to explore whether the presence of CTC before and during follow-up after surgery is associated with recurrence free survival (RFS) and overall survival (OS). In a prospective single center study, CTC were enumerated with the CellSearch system in 30 ml of peripheral blood of 403 stage I-III patients before undergoing surgery for breast cancer (A) and if available 1 week after surgery (B), after adjuvant chemo- and/or radiotherapy or before start of long-term hormonal therapy (C), one (D), two (E) and three (F) years after surgery. Patients were stratified into unfavorable (CTC ≥1) and favorable (CTC=0) prognostic groups. >1 CTC in 30 ml blood was detected in 75/403 (19%) at A, 66/367 (18%) at B, 40/263 (15%) at C, 30/235 (12%) at D, 18/144 (11%) at E and 11/83 (13%) at F. RFS and OS was significantly lower for unfavorable CTC as compared to favorable CTC before surgery (p=0.022 and p=0.006), after adjuvant therapy (p

Familial neurohypophyseal diabetes insipidus (FNDI) is a rare autosomal dominant inherited disease, characterized by serious polyuria and polydipsia, caused by deficient neurosecretion of the antidiuretic hormone, arginine vasopressin (AVP). Vasopressin is a hormone that affects peripherally and centrally regulated functions such as antidiuresis and blood pressure regulation in peripheral tissues and temperature regulation in the central nervous system (1). The antidiuretic function of AVP controls the serum osmolality by stimulating renal water resorption (1)(2). In patients with FNDI, there is insufficient release of AVP, leading to insufficient stimulation of renal water resorption (1)(3)(4)(5). Often, these patients have a constant feeling of thirst that leads to an excessive water intake to compensate for the high urinary output. FNDI is currently diagnosed by means of the patient-unfriendly water dehydration test in which the patient is deprived of water intake for a certain period. In the case of vasopressin deficiency, there will be continuing water loss, producing weight loss and hypertonic dehydration (4), which can be dangerous if the patient is not carefully observed during the test. The onset of FNDI is delayed (early childhood) because of the progressive accumulation of the mutant AVP in the AVP-producing cells. The accumulation of the mutant AVP, as a result of (constant) osmotic stimuli, leads to a slowly progressive degeneration of the AVP-producing cells (2)(6). The molecular background of FNDI is heterogeneous (2)(3)(5)(7)(8), with 33 mutations in the AVP-NP II gene described (Table 1⇓ ). Vasopressin is synthesized in the magnocellular neurons of the hypothalamus as a preprohormone that consists of a signal peptide, vasopressin, neurophysin, and a glycoprotein (3 …

Patients with sickle cell disease (SCD) suffer from frequent and severe episodes of pain that are associated with hospitalizations, impaired quality of life and increased mortality. Current treatment options are scarce. Oxidative stress appears to play a pivotal role in the pathophysiology of SCD. Pilot studies have demonstrated that administration of the antioxidant N-acetylcysteine (NAC) effectively reduces markers of oxidative stress in SCD and may decrease the hospitalization rate for painful crises. NAC is a safe, inexpensive and well-tolerated drug that has been used for years for various indications. The primary aim of this study was to evaluate the effect of NAC on the frequency of daily pain in patients with SCD. We conducted a randomized (1:1), double-blind, placebo-controlled, parallel-group trial at 11 sites across The Netherlands, Belgium and the United Kingdom. Patients were eligible for participation if they were ≥12 years of age, had either HbSS, HbSC, HbSβ⁰- or HbSβ⁺-thalassemia with a history of at least 1 painful crisis per year over the 3 years prior to enrolment. At randomization patients were assigned to receive either oral NAC 600 mg twice daily or placebo for a total duration of 6 months. Patients had monthly checkups during the study. The primary endpoint was the rate of SCD related pain days per patient year, assessed daily with the use of pain diaries. Secondary endpoints included the rate of days with painful crises, admission days, hospitalizations and days with home analgesic use, the severity of pain, the time to first painful crisis and hospitalization, the number of adverse events and the effect on quality of life and various blood markers. The primary intention-to-treat analysis of this study was limited to patients with a minimal completed diary observation time of 110 days. Sensitivity analyses were done in both all randomized patients as well as a subset with ≥80 days of observation time. Lastly, an additional per protocol analysis was performed on patients with adequate adherence (≥80% of tablets used) and ≥110 days of completed diary observation time. A total of 96 patients were randomized of which 67 patients met the minimum observation time of 110 days (27 in the NAC and 40 in the placebo arm). Inclusions were stopped before reaching the estimated sample size of 116 patients due to imposed time restrictions by the main funder of this study. Groups were well balanced for baseline characteristics. The proportion of patients adherent to the assigned study medication regimen was low in both treatment groups (53% in the NAC and 50% in the placebo arm). The rate of SCD related pain days per patient year was 61.4 in the placebo group and 61.6 in the NAC group (rate ratio 0.98; 95% confidence interval [CI] 0.54-1.78; P=.98). Moreover, there were no significant differences in the secondary endpoints between groups. These findings were consistent in the sensitivity analyses. In the total study population, more patients reported gastro-intestinal events in the NAC group (42%) as compared to the placebo group (15%, P In conclusion, treatment with oral NAC did not provide any clinical benefit over placebo in this phase 3 study, possibly due to low compliance rates. Therefore, a potential effect of NAC in SCD cannot fully be excluded based on these results. In an adherent subset of patients we did observe a reduction of days with painful crises and a trend to reduction in other pain related endpoints in the NAC treatment arm. To corroborate these findings additional blood sample analysis of pathophysiological markers is currently being performed. This may further elucidate on the dose-effect relation of NAC in SCD. Disclosures Fijnvandraat: Bayer: Research Funding; CSL Behring: Research Funding.

Introduction: Anti-thymocyte globulin (ATG) prophylaxis was shown to be effective in reducing both acute and chronic graft-versus-host disease (GVHD) following allogeneic stem cell transplantation (allo-SCT). In previously published studies, the addition of ATG to the pre-allo-SCT conditioning regimen resulted in substantial reduction in GVHD incidence, but theoretically ATG could be associated with increased relapse rate, especially if administered at high doses (> 6 mg/kg of ATG Thymoglobulin or >15mg/kg of ATG Fresenius) to patients with high-risk acute myeloid leukemia (AML). To further address this question, we assessed transplantation outcome in adult AML patients who underwent reduced intensity conditioning (RIC) allo-SCT from a matched sibling donor (MSD) while in complete remission (CR1), with or without ATG. We used the registry data of the Acute Leukemia Working Party of the European Society forBlood and Marrow Transplantation (EBMT). Patients and Methods: Inclusion criteria were adult patients with AML (de novo or secondary) who underwent RIC allo-SCT from an MSD in CR1 between the years 2000 and 2014. RIC conditioning was defined as per the EBMT criteria. Only patients presenting with intermediate- or poor-risk cytogenetics or secondary AML were included in the analysis. Patients who received any kind of T cell depletion including Campath were excluded. Patients were considered to receive high-dose ATG if they received > 6mg/kg of ATG-Thymoglobulin or >15mg/kg of ATG-Fresenius. All other patients were considered as a control. Results: Two hundred and five of the 1750 patients, included in the analysis, received high-dose ATG. The median patient age at diagnosis was 56 years and the median donor age was 55 years for both groups. Fifty four percent of the patients were males. The graft properties were similar in the high-dose ATG and control groups, with female-to-male transplants in 48/205 (23.4%) and 378/1545 (24.6%) patients, respectively. Peripheral blood was the source of grafts in 193/205 (94.1%) and 1413/1545 (91.5%) patients, respectively (p=NS). Poor cytogenetics was reported in 28/205 (13.7%) patients who were given high-dose ATG and in 247/1545 (16%) patients from the control group. Secondary AML was more prevalent among patients from the high-dose ATG group compared to the control group [84/205 (41%) vs 425/1545 (27.5%; p=0.0003]. The median follow-up was 44.6 (0-177) months. The 3-year overall survival (OS) and leukemia-free survival (LFS) were almost identical in both groups, amounting to 54.9 and 46.3 months, respectively for patients who received high-dose ATG and to 54 and 49.5 months, respectively for patients in the control group (p=0.978 and 0.376 ). Differences in relapse incidence (RI) (40.9% vs 34.4%; p=0.1) and non-relapse mortality (NRM) (11.7% vs 16%; p=0.152) were insignificant. The composite endpoint of GVHD-free/relapse-free survival (GRFS) was similar in the groups after one and three years of follow-up (42.9 vs 49.7 and 31.1 vs 36 months, respectively; p=0.152 and p=0.124). In multivariate analysis, the use of ATG prophylaxis was not found to be associated with any of the evaluated parameters. In contrast, OS, LFS, RI, NRM and GRFS were all affected by patient age and presence of secondary AML. Poor cytogenetics correlated with inferior OS, LFS, RI and GRFS. Donor CMV positivity was associated with lower OS, LFS and GRFS and a higher rate of NRM. Conclusions: In AML patients presenting with intermediate- or poor-risk cytogenetics who are referred to RIC allo-SCT from an MSD in CR1, the addition of ATG to conditioning regimens does not influence the outcome. The composite GRFS endpoint is mainly affected by AML risk factors (hazard ratio of 1.43 for secondary AML and 1.37 for poor-risk cytogenetics; p=0.0001). The role of ATG prophylaxis in AML patients undergoing RIC allo-SCT should be prospectively evaluated. Disclosures Maertens: Gilead: Consultancy, Honoraria, Speakers Bureau; Amgen: Consultancy; Astellas: Consultancy, Speakers Bureau; Merck Sharp & Dohme: Consultancy, Honoraria, Research Funding, Speakers Bureau; Pfizer: Consultancy, Honoraria, Research Funding, Speakers Bureau.

BACKGROUND: The treatment of recurrent transitional cell carcinoma (TCC) remains an unmet clinical need. This study assessed lapatinib, a dual tyrosine kinase inhibitor of epidermal growth factor receptor (EGFR) and HER-2, as second-line therapy in patients with locally advanced or metastatic TCC. METHODS: This was a single-arm, multicenter, open-label, prospective phase 2 study. Patients with TCC whose disease progressed on prior platinum-based chemotherapy received lapatinib until disease progression or unacceptable toxicity, with evaluations for response by Response Evaluation Criteria In Solid Tumors criteria performed every 8 weeks. The primary endpoint of the current study was objective tumor response rate. Secondary endpoints included safety, time to disease progression, and overall survival. RESULTS: Fifty-nine patients were enrolled in the study, 25 of whom (42%) could not be evaluated for response. The primary endpoint of an objective response rate (ORR) >10% was observed in 1.7% (95% confidence interval [95% CI], 0.0%-9.1%) of patients: however, 18 (31%; 95% Cl, 19%-44%) patients achieved stable disease (SD). The median time to disease progression and overall survival (OS) were 8.6 weeks (95% Cl, 8.0 weeks-11.3 weeks) and 17.9 weeks (95% Cl, 13.1 weeks-30.3 weeks), respectively. Clinical benefit (ORR and SD) was found to be correlated with EGFR overexpression (P = .029), and, to some extent, HER-2 overexpression. The median OS was significantly prolonged in patients with tumors that overexpressed EGFR and/or HER2 (P = .0001). Lapatinib was well tolerated. CONCLUSIONS: The study was considered to be negative because it did not meet its primary endpoint; however, further analysis demonstrated an improvement in OS in a subset of patients with tumors overexpressing EGFR and/or HER-2, which is encouraging and warrants further investigation. Cancer 2009;115:2881-90. (C) 2009 American Cancer Society

Introduction: Platelet support in patients who are pancytopenic after autologous stem-cell transplantation (auSCT) is difficult in the presence of human leukocyte antigen antibodies (HLA-AB) class I. Multiple transfusions and pregnancy are risk factors for development of HLA-AB. In renal transplantation rituximab (R-mab) is used to reduce HLA-AB levels in HLA-incompatible kidney transplantation. We evaluated whether the incidence of HLA-AB was lower in lymphoma patients who were treated with R-mab during salvage therapy prior to auSCT. Methods: Single-centre retrospective evaluation of patients treated with DHAP-VIM-DHAP salvage (with or without R-mab) and BEAM plus auSCT for B-cell non-Hodgkin lymphoma (B-NHL) or Hodgkin's lymphoma (HL). After becoming available R-mab was given to patients with CD20-positive lymphoma; R-mab was not used before it's availability and in CD20-negative lymphoma. HLA-AB were measured in case of insufficient platelet yield after transfusion. Results: Between January 1st 1999 and December 31st 2013, 164 patients with a B-NHL or HL underwent auSCT (Table). Age at auSCT was median 52.5 (17.8 – 70.2) years, 73 (45%) patients were female and 89 (54%) were treated with R-mab. Type of lymphoma was diffuse large B-NHL (DLBCL) in 82 (50%) patients, HL in 46 (28%) patients, follicular lymphoma (FL) in 30 (18%) patients and other in 6 (4%) patients. All transfused cellular blood products were leucoreduced. Of R-mab treated patients, 42 (47%) were female versus 31 (41%) non R-mab treated patients, p=0.529 (Table). R-mab treated patients were older at auSCT than non R-mab treated patients: median 57.4 (18.3 – 70.2) years versus 48.1 (17.8 – 64.6) years, p All 3 patients who developed HLA-AB were female (100%) versus 44% (n=70) of patients without HLA-AB, p=0.086. All 3 women (100%) with HLA-AB have been pregnant versus 51 (73%) women without HLA-AB (p=0.704); in 8 women without HLA-AB their history of pregnancy was unknown. None (0%) of 3 patients with HLA-AB were treated with R-mab versus 89 of 161 (55%) patients without HLA-AB, p=0.094. Age at auSCT was median 44.2 (39.1 – 52.4) years in patients with HLA-AB and median 52.6 (17.8 – 70.2) years in patients without HLA-AB, p=0.246. Number of rPC transfused was median 7 (4 – 14) in patients with HLA-AB and median 6 (1 – 54) in patients without HLA-AB, p=0.666. There was no significant difference seen in type of lymphoma between patients with and without HLA-AB. Conclusion: The incidence of HLA-AB in our cohort of lymphoma patients treated with DHAP-VIM-DHAP salvage with or without rituximab followed by BEAM plus auSCT for B-NHL or HL is very low. A trend was seen towards treatment without R-mab and female gender, not towards a history of pregnancy. TableOverallRituximabNo Rituximabpn16489 (54)75 (46)Female (n)73 (45)42 (47)31 (41)0.529Age auto-Tx (years)52.5 (17.8 – 70.2)57.4 (18.3 – 70.2)48.1 (17.8 – 64.6)< 0.001HLA-antibodies (n)3 (2)03 (4)0.094Transfused rPC (n)6 (1 – 54)6 (1- 53)5 (1 – 54)0.529Type of lymphomaDLBCL (n)82 (50)61 (69)21 (28)< 0.001Hodgkin (n)46 (28)2 (2)44 (59)< 0.001FL (n)30 (18)23 (26)7 (9)0.008Other (n)6 (4)3 (3)3 (4)1.000 Denoted as median (range), categorical variables as number (%). Disclosures No relevant conflicts of interest to declare.

Background: The presence of circulating tumor cells (CTC) is associated with poor prognosis in patients with metastatic colorectal cancer (CRC). This study was conducted to determine if the presence of CTC prior to surgery and during follow-up in patients with newly diagnosed non-metastatic CRC identifies patients who are at risk for disease recurrence. Methods: In a prospective single center study 183 patients with newly diagnosed non-disseminated CRC scheduled for surgery were enrolled from 2003 till 2008 and followed up for a median of 5.1 years. CTC were enumerated with the CellSearch System in 4 aliquots of 7.5 ml of peripheral blood before and after surgery (1-26 weeks), after adjuvant therapy and 1, 2, 3 and 4 years after surgery. Findings: ≥1 CTC/ 30ml of blood were detected in 44 (24%) patients before surgery. CTC frequency did not change significantly at the time points after surgery. Patients with CTC before surgery had a significant decrease in Recurrence Free Survival (RFS, logrank test p=0.014) and Colon Cancer Related Survival (CCRS, p=0.002). Five year RFS dropped from 75% to 61% and five year CCRS from 83% to 69% for patients with CTC before surgery. In a multivariate analysis of CTC, T-Stage and N-stage, the presence of CTC and N-stage remained as significant factors for RFS and CCRS. Surprisingly the presence of CTC after surgery was not significantly associated with RFS and CCRS whereas CTC 2-3 years after surgery was again significantly associated with RFS and CCRS. Interpretation: The presence of CTC in patients with stage I-III CRC before surgery is associated with a significant reduction RFS and CCRS. Although similar amounts of CTC were detected within 3 months after surgery they were not associated with RFS or CCRS. In contrast CTC were again highly significant for RFS and CCRS 2-3 years after surgery. These findings suggest a role of CTC detection, to assess which patients need adjuvant treatment. To implement CTC detection in the non-metastatic setting a validated CTC detection technology is needed with increased sensitivity and specificity. Citation Format: Guus van Dalum, Marco R. de Groot, Gerrit-Jan Stam, Loes F.A. Scholten, Walter J.B. Mastboom, Istvan Vermes, Arjan G.J. Tibbe, Leon W.M.M. Terstappen. Importance of circulating tumor cells in newly diagnosed colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3064. doi:10.1158/1538-7445.AM2014-3064

Patients with suspected pulmonary embolism often receive heparin therapy for hours to days before ventilation/perfusion scintigraphy is completed. We assessed to what extent the lung scan classification and pulmonary vascular perfusion changed over a period of 2-4 days of full anticoagulant therapy. In 312 consecutive patients with at least one segmental perfusion defect on the initial perfusion scan, classification of both the initial and the final lung scan allowed us to study alteration in scan classification. Changes in pulmonary perfusion were assessed scintigraphically in a subgroup of 64 patients with proven pulmonary embolism. Among 79 patients with an initial high-probability lung scan, the final scan remained high probability in 77 whereas it became non-diagnostic and normal in one patient each. The lung scan classification did not change in any of the 233 patients who initially had a non-diagnostic scan. Thus, a different lung scan category was observed in only 2 out of 312 patients (0.6%; 95% CI 0.1%-2.3%). The mean pulmonary perfusion at baseline in the subgroup of 64 patients with pulmonary embolism was 62% (SD +/-17%; range 26%-89%). A mean absolute improvement in pulmonary perfusion of only 4%+/-11% (NS) was observed in the 2-4 days of observation. It is concluded that intravenous heparin therapy for a period of 2-4 days has only a minimal influence on the diagnostic lung scan classification and induces only minor changes in pulmonary vascular obstruction in the majority of patients.

Background: While there is unequivocal evidence regarding the prognostic relevance of circulating tumor cells (CTCs) in peripheral blood of patients (pts.) with metastatic breast cancer (BC), less data is available for the prognostic relevance at time of primary diagnosis. Methods: We conducted a pooled analysis with individual data of 3172 pts with non-metastatic (Stage I-III) BC from five academic institutions. Prevalence and number of CTCs in the peripheral blood were assessed at time of primary diagnosis using the FDA-approved CellSearch System (Veridex, USA). Patient outcomes were analyzed using univariate log-rank tests and multivariate Cox regressions. The median follow-up time was 61 months. Results: At least one CTC was detected in 20.2% of the pts. The presence of CTCs was associated with larger tumors, more axillary lymph node metastases, and higher histological tumor grade (all p < 0.005). The presence of CTCs was significantly related to poor progression-free (PFS, log-rank test, p < 0.001, hazard ratio [HR] 2.0) and overall survival (OS, p < 0.001, HR 2.6). Multivariate Cox regressions, including tumor size, nodal status, histological tumor grade, hormone receptor status and HER2 status, and CTC prevalence confirmed that the presence of CTCs was an independent prognostic factor for both poor PFS (HR 1.8) and OS (HR 2.1, both p < 0.001). In the univariate subgroup analysis concerning OS, the HR for pts. with positive HRS was 3.1 (p< 0.0001), and 2.0 (p = 0.001) for pts. with a negative HRS. The prognostic relevance of CTC status was similar for patients with HER2 negative and HER2 positive tumors (HR 2.6 and 2.5, respectively). The HR in CTC-positive pts. with HRS positive/HER2 positive tumors was 3.4 (p = 0.003). For patients with more than 3 CTCs, the hazard ratio for death was increased to 5.2 (1.5 - 2.8) in univariate and to 4.0 (2.6 - 6.2) in multivariate analysis. Cox Regression Analysis for Overall SurvivalVariableHazard ratio95% CIP-valueCTCs pos vs. neg2.1081.594 - 2.788 Conclusions: In patients with early breast cancer, the presence of CTCs in peripheral blood is an independent predictor of poor progression-free and overall survival. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr PD6-6.

BackgroundPresence of circulating tumor cells (CTC) in patients with metastatic breast, colorectal and prostate cancer is indicative for poor prognosis. An automated CTC (aCTC) algorithm developed previously to eliminate the variability in manual counting of CTC (mCTC) was used to extract morphological features. Here we validated the aCTC algorithm on CTC images from prostate, breast and colorectal cancer patients and investigated the role of quantitative morphological parameters.MethodologyStored images of samples from patients with prostate, breast and colorectal cancer, healthy controls, benign breast and colorectal tumors were obtained using the CellSearch system. Images were analyzed for the presence of aCTC and their morphological parameters measured and correlated with survival.ResultsOverall survival hazard ratio was not significantly different for aCTC and mCTC. The number of CTC correlated strongest with survival, whereas CTC size, roundness and apoptosis features reached significance in univariate analysis, but not in multivariate analysis. One aCTC/7.5 ml of blood was found in 7 of 204 healthy controls and 9 of 694 benign tumors. In one patient with benign tumor 2 and another 9 aCTC were detected.Significance of the studyCTC can be identified and morphological features extracted by an algorithm on images stored by the CellSearch system and strongly correlate with clinical outcome in metastatic breast, colorectal and prostate cancer.

14512 Background: The presence of CTCs is associated with poor prognosis in patients with metastatic carcinomas. The significance of CTCs at the time of diagnosis and after therapy is not known. A prospective clinical trial is conducted to determine if monitoring CTC levels in pts diagnosed with breast and colorectal cancer identify pts at risk for recurrence. CTCs are correlated with pathological stage as follow-up is too short to assess recurrence risk. Methods: Currently 213 pts with newly diagnosed breast and 119 pts with colorectal cancer before surgery with curative intend have been enrolled as well as a control group of 120 patients in whom diagnosis of breast or colorectal cancer was excluded. Thirty ml of blood was screened for the presence of CTCs at baseline before surgery and every following year using the CellSearch™ System. Results: Presence of CTCs in 30mL of blood in breast, colorectal cancer and the control group is provided in the table . In colorectal cancer the average number of CTCs /30 mL at baseline for Dukes A, B and C was 0.2, 0.7 and 1.1 respectively, for stage 0, I, II, III breast cancer 0.3, 0.5, 0.6 and 1.8 respectively and for the control group 0.24 CTCs /30 mL. Conclusions: CTC can be detected in a substantial proportion of pts before and after definitive surgery for breast and colorectal cancer. The number of CTCs before surgery correlated with the stage of the disease. Specificity of the CellSearch™ System is being increased to improve the discrimination of patients with benign and malignant disease. Follow-up must clarify whether the presence CTCs is an identifier for the risk of recurrence. [Table: see text] No significant financial relationships to disclose.

395 Background: The presence of circulating tumor cells (CTC) is associated with poor prognosis in patients with metastatic colorectal cancer (CRC). The current study was conducted to determine if the presence of CTC prior to surgery in patients with newly diagnosed non-metastatic CRC can identify those patients at risk for disease recurrence. Methods: In a prospective single center study 180 patients with newly diagnosed non-disseminated CRC scheduled for surgery were enrolled and followed up for a median of 41 months. CTC were enumerated with the CellSearch System in 4 aliquots of 7.5 ml of peripheral blood before undergoing surgery. The control group consisted of sixty patients with benign tumors after surgery or colonoscopy. Results: ≥1 CTC/ 30ml of blood were detected in 9 (15%) persons of the control group, and in 41 (23%) CRC patients. From the 180 CRC patients, 44 developed recurrent disease (24%) and 38 patients (21%) died during follow up. Presence of CTC correlated with disease recurrence (p= 0.040) and death (p=0.006). Patients with CTC had a significantly decreased three year Disease Free Survival (DFS) and overall survival (OS) versus those without CTC (DFS 57%vs. 78%, p=0.018 and OS 70% vs. 80%, p=0.003%). Conclusions: Patients with stage I-III CRC with CTC before surgery have a significantly lower 3 years DFS and OS. These findings warrant further development of technology to increased the sensitivity and specificity of CTC detection for incorporation as prognostic marker along side Duke Stage to assess which patients need adjuvant treatment.

Background: Patients (pts) relapsing with CML after allogeneic hematopoietic stem cell transplantation (alloHSCT) may be treated with tyrosine kinase inhibitors (TKI) and/or donor lymphocyte infusions (DLI). As nowadays the majority of CML patients would have received at least imatinib prior to transplantation, we were interested in analizing a) the type of TKI used after alloHSCT, b) the indication for TKI treatment, c) the outcome of this treatment and d) the temporal relationship with DLI if given. Patients and methods: 435 pts received TKI after first allogeneic HSCT for CML for different reasons. Transplants had been performed in first chronic phase (CP1, n=194), accelerated phase (AP, n= 60) or for more advanced disease (blast crisis (BC)/> CP1, n=177) from HLA identical siblings (n=231) or unrelated donors (n=204) between 2000 and 2013. TKI given prior to transplant was imatinib (n=268), dasatinib (n=162), nilotinib (n=88), bosutinib (n=4) and ponatinib (n=7). Median age at transplant was 44 (18.5-68) years, 274 pts (63%) were male. TKI post alloHSCT were given between 2000 and 2015. 1st TKI given was either imatinib (n=223), dasatinib (n=131), nilotinib (n=67), bosutinib (n=2) or ponatinib (12). The indications for TKI therapy were the same as for transplantation (n=262), for relapse/progression/persistent disease (n=124), for prophylaxis/pre-emptive (n=32), planned (n=5), others (n=8) and missing (n=4). Results: Median follow-up from start of TKI was 55 (1-171) months. The median time interval from transplant to TKI was 6 (0.2-165) months. It was longer for TKI given for relapse/progression with 15 (1-89) months and shorter for TKI given for prophylaxis/pre-emptive with 1.6 (0.2-43) months. It was longer for imatinib with 11 (0.2-121) months vs 3.8 (0.2-165) months for other TKI. Imatinib as 1st TKI was mainly given for relapse/progression/persistent disease (48%) and the other TKI for the same reason as for transplantation (83%). 103/223 (46%) of pts with imatinib, 99/131 (76%) with dasatinib, 55/67 (82%) with nilotinib and 11/14 (79%) with bosutinib/ponatinib post-transplantation had been treated with imatinib prior to transplantation. In total, 196 (45%) patients received DLI after alloHSCT, of which 63/435 (14.5%) had DLI prior to TKI post-alloHSCT, 19/435 (4.4%) had DLI at the same time of TKI and 114/435 (26%) had DLI post-TKI. Best response after TKI was complete molecular remission in 17.7%, cytogenetic remission in 4.4%, hematological remission in 20.2% and no response/progression/relapse in 57.7% of pts. 50% of pts treated with imatinib had a response (molecular/cytogenetic/hematological) vs 34% with nilotinib, 33% with dasatinib and 33% with bosutinib/ponatinib, p=0.014. OS was 60% (55-65%) at 5 years. It was 66% (60-73%) with imatinib vs 51% (42-60%) with other TKI, p=0.0024. 5 years RFS was 47% (42-53%). It was 53% (46-60%) with imatinib vs 40% (32-48%) with other TKI, p=0.0102. 5 years RI was 25% (21-30%). It was 21% (16-27%) with imatinib vs 31% (24-38%) with other TKI, p=0.0454. 5 years NRM was 27% (23-32%). It was 26% (20-31%) with imatinib vs 29% (22-36%) with other TKI, p=0.365. In multivariate analysis for OS, imatinib vs other TKI post-transplant did not show anymore an effect, HR 1.19 (0.85-1.67), p=0.317. Factors influencing OS were time from diagnosis to transplant, HR 1.01 (1.00-1.01), p=0.009, AP vs CP1, HR 1.80 (1.11-2.91), p=0.017 and BC/>CP1 vs CP1, HR 2.3 (1.58-3.33), p In multivariate analysis for RFS as for OS, imatinib vs other TKI did not have an effect, HR 1.11 (0.83-1.48), p=0.496. Other factors having a tendency or influencing RFS were time from diagnosis to transplant, HR 1.00 (1.00-1.01), p=0.054, AP vs CP1, HR 1.52 (1.00-2.31), p=0.050 and BC/>CP1 vs CP1, HR 2.11 (1.55-2.88), p Conclusion: These data suggest that TKI after alloHSCT induce a response in about 42% of pts regardless of the type of TKI used and that time from diagnosis to transplantation as well as the phase of disease at transplant remain the main factors influencing the outcome of CML patients relapsing after alloHSCT. Disclosures Kröger: Riemser: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; Neovii: Honoraria, Research Funding; Sanofi: Honoraria, Research Funding.

MYH9 -related disease ( MYH9 -RD) is an autosomal dominant thrombocytopenia with giant platelets variably associated with young-adult onset of progressive sensorineural hearing loss, presenile cataract, and renal damage. MYH9 -RD is caused by mutations of MYH9 , the gene encoding for non-muscle heavy-chain myosin-9. Wild-type and mutant myosin-9 aggregate as cytoplasmic inclusions in patients’ leukocytes, the identification of which by immunofluorescence has been proposed as a suitable tool for the diagnosis of MYH9 -RD. Since the predictive value of this assay, in terms of sensitivity and specificity, is unknown, we investigated 118 consecutive unrelated patients with a clinical presentation strongly consistent with MYH9 -RD. All patients prospectively underwent both the immunofluorescence assay for myosin-9 aggregate detection and molecular genetic analysis of the MYH9 gene. Myosin-9 aggregates were identified in 82 patients, 80 of which (98%) had also a MYH9 mutation. In the remaining 36 patients neither myosin-9 aggregates nor MYH9 mutations were found. Sensitivity and specificity of the immunofluorescence assay was evaluated to be 100% and 95%, respectively. Except for the presence of aggregates, we did not find any other significant difference between patients with or without aggregates, demonstrating that the myosin-9 inclusions in neutrophils are a pathognomonic sign of the disease. However, the identification of the specific MYH9 mutation is still of importance for prognostic aspects of MYH9 -RD. Fil: Savoia, Anna. Universita Degli Studi Di Trieste; Italia. Italian Registry for MYH9-Related Disease; Italia Fil: de Rocco, Daniela. Universita Degli Studi Di Trieste; Italia. Italian Registry for MYH9-Related Disease; Italia Fil: Panza, Emanuele. Italian Registry for MYH9-Related Disease; Italia. Universita Di Bologna; Italia Fil: Bozzi, Valeria. Italian Registry for MYH9-Related Disease; Italia. University of Pavia; Italia Fil: Scandellari, Raffaella. Italian Registry for MYH9-Related Disease; Italia. Universita Di Padova; Italia Fil: Loffredo, Giuseppe. Italian Registry for MYH9-Related Disease; Italia. Pausilipon Hospital; Italia Fil: Mumford, Andrew. University of Bristol; Reino Unido Fil: Heller, Paula Graciela. Italian Registry for MYH9-Related Disease; Italia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina Fil: Noris, Patrizia. Italian Registry for MYH9-Related Disease; Italia Fil: de Groot, Marco R.. University of Bristol; Reino Unido Fil: Giani, Marisa. Italian Registry for MYH9-Related Disease; Italia. Istituto di Ricovero e Cura a Carattere Scientifico; Italia Fil: Freddi, Paolo. Italian Registry for MYH9-Related Disease; Italia Fil: Scognamiglio, Francesca. Italian Registry for MYH9-Related Disease; Italia Fil: Riondino, Silvia. Italian Registry for MYH9-Related Disease; Italia. Istituto di Ricovero e Cura a Carattere Scientifico; Italia Fil: Pujol Moix, Núria. Universitat Autonoma de Barcelona; España Fil: Fabris, Fabrizio. Italian Registry for MYH9-Related Disease; Italia. Universita Di Padova; Italia Fil: Seri, Marco. Italian Registry for MYH9-Related Disease; Italia. Universita Di Bologna; Italia Fil: Balduini, Carlo L.. Italian Registry for MYH9-Related Disease; Italia. University of Pavia; Italia Fil: Pecci, Alessandro. Italian Registry for MYH9-Related Disease; Italia. University of Pavia; Italia