ALBERT

Description: Background and Aims High dose chemotherapy followed by autologous stem cell transplant (ASCT) is an effective treatment for many patients with haematological malignancies. Adequate stem cell mobilization (SCM) and collection are essential for a successful ASCT. Unfortunately, a small group of patients fail to mobilize sufficient stem cell for transplant. In order to explore factors influencing SCM we conducted a retrospective analysis on a large series of candidates for ASCT in a Portuguese transplantation center. Patients and Methods We analyzed 233 patients who underwent consecutively SCM from 2007 to 2012. Patients with CD34+

Description: BACKGROUND: Despite remarkable therapeutic advances in the last 2 decades and a major improvement in survival, a number of multiple myeloma (MM) patients (pts) present a short-term outcome. AIMS: Our aim was to identify the main factors (baseline characteristics, response to therapy, relapse features) determining early mortality (EM) among a cohort of newly diagnosed symptomatic MM pts treated with novel agents. METHODS: We conducted a national multicenter retrospective study, including a cohort of symptomatic MM pts diagnosed between January/2010 and June/2017, treated with novel agents (bortezomib, thalidomide or lenalidomide) with the maximum age of 75 years-old and living

Description: Several factors influencing disease progression and survival have been identified in multiple myeloma (MM). We analysed a series of 49 consecutive patients with MM that underwent autologous haematopoietic progenitor cell transplant (HPCT) in one center regarding the following variables: use of G-CSF for haematopoietic recovery post-transplant; recovery of normal IgM levels at day +100 post-transplant; levels of lymphocytes namely of the CD4+ and CD8+ subsets also at day +100. Before 2006, all patients had G-CSF starting 24 hours after the cell infusion until neutrophil 〉 500×10^9/L in two consecutive days; in the years 2006–2008, no G-CSF was given to transplanted patients. A historical comparison was done and at the time of this study no significant difference in progression free survival (Kaplan-Meyer analysis), was detected between the two groups, possibly due to the shorter follow-up of the “no G-CSF” (n=19) group; nevertheless median progression free survival (PFS) in the “G-CSF” group was 12 months while median PFS was not attained in the “no G-CSF” group (median follow-up = 7 months). Post transplant IgM levels were also determined in 39 patients. Eighteen patients recovered normal IgM levels at day +100 (46.8 %) and 21 (53.8 %) did not. Comparison of Kaplan-Meyer curves for the two groups did not show any statistically significant difference but there is a sharp difference between median PFS of the “low IgM” (10 months) and the “normal IgM” (27 months) groups. CD4/CD8 ratio was determined in 18 patients at day +100. The ratio varied between 0 and 0.63 (median – 0.305). No correlation was found between post-transplant IgM recovery and CD4/CD8 ratio. In conclusion, in our series of MM patients treated with autologous HPCT we could not find a definite relationship between immunologic recovery and response to treatment although there is a trend to a better outlook of the patients which recover normal IgM levels. It is also uncertain whether use of G-CSF in the post-transplant period would have any effect on disease behaviour.

Description: Background microRNAs (miRs) are small non-coding RNAs that post-transcriptionally regulate gene expression by binding target mRNAs and hampering their translation by translation inhibition or mRNA degradation. miRs have been shown to play an important role in cancer development by controlling several pivotal cellular processes [Lee and Dutta (2009), Annu Rev Pathol 4:199-227]. In particular,miR-128 has also been associated with cancer, namely leukemia and has been shown, , with other miRs, to allow the discrimination between AML and ALL [Mi et al. (2007), PNAS 104(50):19971-6]. Moreover, it is included in a miR signature that associates a subgroup of patients with high-risk molecular features of AML with worse clinical outcome [Marcucci et al. (2008), NEJM 358(18):1919-28]. Nevertheless, all the data associating miR-128 with leukemia derives from expression array analysis and no functional studies have been performed. Therefore, the aim of this study was to understand the role of miR-128 in AML cells and in their response to some chemotherapeutic agents. Methods HL-60 cells were transfected with miR-128 mimic (or control miR mimic) and further treated with etoposide, doxorubicin or their vehicle as control. miR expression was evaluated by RT-qPCR. The effect of miR-128 overexpression in sensitization of HL-60 cells to the effects of doxorubicin or etoposide was analysed by Trypan blue exclusion assay. Cellular proliferation (BrdU assay), cell cycle (flow cytometry following PI labeling), programmed cell death (TUNEL assay) and apoptosis (Annexin V/ PI staining) were analysed. The expression levels of proteins involved in apoptosis (caspase-3, PARP), autophagy (Beclin-1, Vps34 and LC3) and DNA damage (γ-H2AX, 53BP1) were studied (Western Blot). DNA damage was analysed with the Comet assay and by foci formation of γ-H2AX and 53BP1 proteins, visualized by immunofluorescence microscopy. miR-128 expression was analysed in samples from peripheral blood mononuclear cells (PBMCs) of 13 healthy donors and from bone marrow of 11 AML patients by RT-qPCR. Results miR-128 expression was increased upon miR mimic transfection. miR-128 overexpression decreased HL-60 viable cell number to 84.3% and 81.0%, at 24 h and 48 h after transfection respectively, and sensitized HL-60 cells to both doxorubicin and etoposide. Nevertheless, miR-128 overexpression did not affect cell cycle profile, cellular proliferation, apoptosis, or the expression of apoptosis-related or autophagy-related proteins. Interestingly, miR-128 overexpression increased DNA damage analysed by Comet assay (from 3.6% in miR-control transfected cells to 8.1% in miR-128 transfected cells). This increase in DNA damage of miR-128 overexpressing cells was confirmed by verifying an increase in DNA repair foci of γ–H2AX and 53BP1 together with an increase in expression of both those proteins γ–H2AX and 53BP1. Analysis of miR-128 expression in samples from PBMCs of healthy donors and from bone marrow of AML patients showed no statistically significant differences, although the expression levels of miR-128 in the AML samples were higher than in healthy donors. Conclusion miR-128 overexpression per sedecreased HL-60 viable cell number and sensitized cells to doxorubicin and etoposide. miR-128 increased DNA damage, which might justify the increased sensitivity that these cells presented to doxorubicin and etoposide. Concerning patient samples, a slight increase in the expression of miR-128 was found in AML bone marrow samples, when compared to PBMCs from healthy donors, suggesting that these patients maybe more susceptible to DNA damaging agents. Acknowledgments Fundação Calouste Gulbenkian for financial support. FCT for the grants to H. Seca (SFRH/BD/47428/2008) and R. T. Lima (SFRH/BPD/68787/2010). G. M. Almeida was supported by FCT and the European Social Fund. IPATIMUP is an Associate Laboratory of the Portuguese Ministry of Science, Technology and Higher Education and is partially supported by FCT. Disclosures: No relevant conflicts of interest to declare.

Description: Extracellular vesicles (EVs) mediate intercellular signaling and communication, allowing the intercellular exchange of proteins, lipids, and genetic material. Their recognized role in the maintenance of the physiological balance and homeostasis seems to be severely disturbed throughout the carcinogenesis process. Indeed, the modus operandi of cancer implies the highjack of the EV signaling network to support tumor progression in many (if not all) human tumor malignancies. We have reviewed the current evidence for the role of EVs in affecting cancer hallmark traits by: (i) promoting cell proliferation and escape from apoptosis, (ii) sustaining angiogenesis, (iii) contributing to cancer cell invasion and metastasis, (iv) reprogramming energy metabolism, (v) transferring mutations, and (vi) modulating the tumor microenvironment (TME) by evading immune response and promoting inflammation. Special emphasis was given to the role of EVs in the transfer of drug resistant traits and to the EV cargo responsible for this transfer, both between cancer cells or between the microenvironment and tumor cells. Finally, we reviewed evidence for the increased release of EVs by drug resistant cells. A timely and comprehensive understanding of how tumor EVs facilitate tumor initiation, progression, metastasis and drug resistance is instrumental for the development of innovative EV-based therapeutic approaches for cancer.

Description: Introduction: Systemic light chain (AL) amyloidosis is a plasma cell dyscrasia in which a toxic plasma cell clone causes devastating multiorgan complications and death, and substantially affects the quality of life of patients (pts). Current therapeutic options are not regulatory approved and are typically based on modified treatments that are used for multiple myeloma. The aim of EMN23 study is to thoroughly describe patterns of AL amyloidosis management in a large-scale real-world evidence (RWE) setting throughout Europe, to understand different treatment strategies and their evolution, and evaluate resource utilization. Methods: EMN23 is an ongoing, retrospective, observational, multicenter study aiming to enroll 5000 pts in 10 countries (13 Sites) across Europe; Austria, Czech Republic, France, Germany, Greece, Italy, the Netherlands, Portugal, Spain, and UK. Pts must have documented systemic AL amyloidosis, and a first-line treatment start date between 2004 and 2018. Results are presented in 2 chronological cohorts, 2004-2010, and 2011-2018, with 2010 being an important landmark, since bortezomib, cyclophosphamide, dexamethasone (CyBorD) regimen was introduced in clinical practice. Pts who received treatment in the context of an interventional clinical trial were not analyzed for treatment and efficacy results. The current analysis presents baseline disease characteristics, treatment patterns and efficacy outcomes. Informed consent or a non-opposition letter was collected for all alive pts, depending on the regulatory environment of each country, and a waiver of consent was obtained for the deceased. Results: In total, 2031 pts from Austria (1.9%), Czech Republic (0.9%), France (9.2%), Germany (3.6%), Greece (12.6%), Italy (58.7%), the Netherlands (7.2%), Portugal (1.0%), and Spain (4.9%) have been analyzed; 67.9% of pts started treatment after 2010. Median age at diagnosis was 66 years and 54.5% were males, while 48.2% had ECOG performance status ≤1. Heart (71.9%), kidney (67.2%), soft tissue (19.7%) and nervous system (18.5%) were most often involved at diagnosis, and 17.2%, 36.2%, 21.1% and 15.2% of pts were at cardiac stage I, II, IIIA and IIIB, according to Mayo staging system, respectively, while for 10.3% of pts staging was not reported. There was a slight increase of pts with stage III in the more recent era. In the period 2004-2010, 52.8% of the pts had at least 2 lines of therapy, which dropped to 35.2% in the period 2011-2018. A 6.4% of pts received autologous stem cell transplant at first line. The prevailing first-line regimen in 2004-2010 was melphalan with dexamethasone (MDex; 50.6% of the pts), replaced by CyBorD (46.1%) in the post-2010 period. The predominant first-line regimens were different in various countries in 2004-2010, but in general CyBorD was the most extensively used regimen after 2010. Overall, the most frequent second-line regimen was BorD (29.8%), which was also the treatment of choice for 15.7% of the pts who had frontline therapy with MDex in the pre-2010 period; and lenalidomide dexamethasone (RD, 25.5%) which was also favored following frontline treatment with CyBorD for 10.6% of the pts in the post-2010 period. In the 2004-2010 era, 31.7% of the pts achieved a hematologic VGPR or CR at first line, which was slightly improved to 37.2% in the post-2010 period. Notably, 29.9% and 23% of pts did not achieve a hematologic response (no response or disease progression) in the two periods, respectively (p=0.001). Mortality in the first months of treatment remained roughly at the same level, with the 3-month overall survival (OS) rate at 82.3% for both cohorts, whereas the median OS increased from 43.5 months in the pre-2010 period to 50.1 months in the post-2010 period (p=0.525). There was a trend towards an improvement in the median OS of stage IIIA pts from 9.5 to 25.9 months, pre- and post-2010, respectively, however, no change was observed for stage IIIB pts (2.7 to 3.5 months). Conclusions: Therapeutic options for AL amyloidosis have changed considerably over time. After 2010 CyBorD became the prominent first-line treatment; the proportion of pts not achieving a response was reduced by 23% and a trend towards improvement of OS for stage IIIA disease was observed, but the prognosis of stage IIIB pts remained dismal and early mortality remained unchanged. Early diagnosis is still an unmet need, and improved therapies are essential. Figure 1 Disclosures Palladini: Celgene: Other: Travel support; Jannsen Cilag: Honoraria, Other. Schönland:Janssen, Prothena, Takeda: Honoraria, Other: travel support to meetings, Research Funding. Milani:Celgene: Other: Travel support; Pfizer: Other: Speaker honoraria; Janssen: Other: Speaker honoraria. Jaccard:Janssen: Consultancy, Honoraria, Other: A.J. has served in a consulting or advisory role for Janssen and has received honoraria from, received research funding from, and had travel, accommodations, or other expenses paid for or reimbursed by Janssen., Research Funding; Celgene: Honoraria, Other: A.J. has served in a consulting or advisory role for Janssen and has received honoraria from, received research funding from, and had travel, accommodations, or other expenses paid for or reimbursed by Celgene., Research Funding. Bridoux:Celgene: Honoraria; Baxter: Consultancy; Janssen: Honoraria. Cibeira:Amgen: Honoraria, Other: Educational lectures; Celgene: Honoraria, Other: Educational lectures; Akcea Therapeutics: Honoraria, Membership on an entity's Board of Directors or advisory committees; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Educational lectures. Agis:Janssen: Honoraria, Research Funding; Amgen: Honoraria; BMS: Honoraria; Celgene: Honoraria; Takeda: Honoraria. Minnema:Amgen: Honoraria; Servier: Honoraria; Janssen Cilag: Honoraria; Celgene Corporation: Honoraria, Research Funding; Gilead: Honoraria. Bergantim:AMGEN, Celgene, Janssen, Takeda: Membership on an entity's Board of Directors or advisory committees, Other: Speaker honoraria; Celgene, AMGEN/SPH/APCL: Other: Grants, Research Funding. Hajek:Oncopeptides: Consultancy; Janssen: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Consultancy, Research Funding; BMS: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; PharmaMar: Consultancy, Honoraria; Abbvie: Consultancy, Honoraria. João:Takeda: Consultancy, Research Funding; Amgen: Consultancy; Janssen: Consultancy; BMS: Consultancy. Wechalekar:Celgene: Honoraria; Janssen: Honoraria, Other: Advisory; Caelum: Other: Advisory; Takeda: Honoraria, Other: Travel. Sonneveld:Karyopharm: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Sanofi: Consultancy; Skyline Dx: Honoraria, Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria, Research Funding; Amgen: Consultancy, Honoraria, Research Funding. Kastritis:Amgen: Consultancy, Honoraria, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Genesis Pharma: Consultancy, Honoraria; Takeda: Consultancy, Honoraria; Pfizer: Consultancy, Honoraria. OffLabel Disclosure: This is a RWE study on AL amyloidosis, and currently there are no regulatory approved therapeutic options for the disease.