Doses Of Etoposide Research Articles

Poly ADP Ribose Polymerase Inhibitor Olaparib Targeting Microhomology End Joining in Retinoblastoma Protein Defective Cancer: Analysis of the Retinoblastoma Cell-Killing Effects by Olaparib after Inducing Double-Strand Breaks.

Retinoblastoma is the most common intraocular cancer in childhood. Loss of function in both copies of the RB1 gene is the causal mutation of retinoblastoma. Current treatment for retinoblastoma includes the use of chemotherapeutic agents, such as the DNA damaging agent etoposide, which is a topoisomerase II poison that mainly generates DNA double-strand breaks (DSBs) and genome instability. Unfaithful repairing of DSBs could lead to secondary cancers and serious side effects. Previously, we found that RB knocked-down mammalian cells depend on a highly mutagenic pathway, the micro-homology mediated end joining (MMEJ) pathway, to repair DSBs. Poly ADP ribose polymerase 1 (PARP1) is a major protein in promoting the MMEJ pathway. In this study, we explored the effects of olaparib, a PARP inhibitor, in killing retinoblastoma cells. Retinoblastoma cell line Y79 and primary retinoblastoma cells expressed the cone-rod homeobox protein (CRX), a photoreceptor-specific marker. No detectable RB expression was found in these cells. The co-treatment of olaparib and etoposide led to enhanced cell death in both the Y79 cells and the primary retinoblastoma cells. Our results demonstrated the killing effects in retinoblastoma cells by PARP inhibitor olaparib after inducing DNA double-strand breaks. The use of olaparib in combination with etoposide could improve the cell-killing effects. Thus, lower dosages of etoposide can be used to treat retinoblastoma, which would potentially lead to a lower level of DSBs and a relatively more stable genome.

Hematopoietic stem cell transplantation for infants with high-risk KMT2A gene–rearranged acute lymphoblastic leukemia

AbstractThe role of allogeneic hematopoietic stem cell transplantation (HSCT) for infants with acute lymphoblastic leukemia (ALL) and KMT2A gene rearrangement (KMT2A-r) is controversial in terms of both its efficacy and potential for acute and late toxicities. In Japanese Pediatric Leukemia/Lymphoma Study Group trial MLL-10, by introducing intensive chemotherapy, indication of HSCT was restricted to patients with high-risk (HR) features only (KMT2A-r and either age <180 days or presence of central nervous system leukemia). Of the 56 HR patients, 49 achieved complete remission. Forty-three patients received HSCT in first remission including 38 patients receiving protocol-specified HSCT with conditioning consisting of individualized targeted doses of busulfan, etoposide, and cyclophosphamide. Three-year event-free survival (EFS) of 56.8% (95% confidence interval [CI], 42.4% to 68.8%) and overall survival of 80.2% (95% CI, 67.1% to 88.5%) were accomplished. Univariable analysis showed that Interfant-HR criteria and flow cytometric minimal residual disease (MRD; ≥0.01%), both at the end of induction and at the end of consolidation (EOC), were significantly associated with poorer EFS. In the multivariable analysis, positive MRD at EOC was solely associated with poor EFS (P < .001). Rapid pretransplant MRD clearance and tailored HSCT strategy in the MLL-10 trial resulted in a favorable outcome for infants with HR KMT2A-r ALL. However, considering the high rate of potentially life-threatening toxicities and the risk of late effects, its indication should be further restricted or even eliminated in the future by introducing more effective therapeutic modalities with minimal toxicities. This trial was registered at the University Hospital Medical Information Network Clinical Trials Registry (UMIN-CTR) as #UMIN000004801.

P-236 Combined squamous cell and neuroendocrine carcinoma of the larynx. A case report

Presented by: Javier Padilla-Cabello (jpadillacabello@gmail.com) Introduction A 67-year-old male patient, smoker of more than a pack a day since childhood, presented to the emergency department with a right cervical tumor and dysphagia of twoweeks' evolution. On examination, cervical palpation revealed an adenopathy in the right area IIa of about 5 cm in diameter, stony and adherent to deep planes. Fibrolaryngoscopy showed a lesion on the right laryngeal surface of the epiglottis and a leukoplakia in the right lateral glossoepiglottic fold. Therefore, it was decided to perform transoral laryngeal microsurgery to take biopsies and request CT for extension study. Materials and Methods The samples taken in the operating room and the CT scan showed that it was an epidermoid carcinoma of the supraglottis stage cT1N2BM0. It was decided to perform a right hemiepiglottectomy with extension to the right lateral glossoepiglottic fold by transoral laser surgery together with bilateral functional cervical emptying. The patient required a tracheotomy due to an episode of severe post-surgical laryngeal edema and the cannula had to be maintained, together with a gastrostomy, because the patient subsequently developed dysphagia with aspiration of secretions and food. Results Histopathological analysis showed that it was a combined tumor: on the one hand a poorly differentiated epiglottic neuroendocrine carcinoma (NEC) (CK-20 +, S100 +, Ki67 90%) with cervical metastasis and on the other hand a supraglottic squamous cell carcinoma in the right glossoepiglottic fold with one cervical metastasis (pT1N1). The case was presented to the oncology committee and it was decided to complete treatment with chemoradiotherapy. Chemotherapy was administered with a reduced dose of carboplatin (AUC = 3) and etoposide. Despite treatment, local cervical tumor progression was observed in the control CT scan and the patient finally died a few weeks later due to COVID19. Conclusions NEC of the larynx are very rare and aggressive, so an accurate histopathologic diagnosis is essential for early treatment. Their most frequent location is the supraglottis and the clinical presentation of the patient is a mild odynophagia or a cervical mass. Very few associated cases of synchronous laryngeal squamous cell carcinoma have been described. Treatment of NEC is based on chemoradiotherapy with a survival rate of less than 5% at 5 years.

Characteristics and Outcomes of Malignancy-Associated Hemophagocytic Lymphohistiocytosis (HLH) in Adults: A Single-Center Experience

Characteristics and Outcomes of Malignancy-Associated Hemophagocytic Lymphohistiocytosis (HLH) in Adults: A Single-Center Experience

XIAP Knockdown Can Increase the Activity of Caspase 3 and 7, and Restore the Apoptotic Response in Cell Line Models of Rituximab Relapse/Refractory B-Cell Lymphoma

Rituximab-chemotherapy relapse/refractory (R/R) diffuse large B-cell lymphoma (DLBCL) patients have lower response rates to second-line chemotherapy, and have a poor overall prognosis. Pre-clinical models of rituximab resistance demonstrated that the acquirement of resistance to anti-CD20 antibodies leads to concomitant resistance to chemotherapy drugs, suggesting the existence of share pathways of resistance between rituximab and chemotherapy agents. To better understand the mechanisms responsible for rituximab/chemotherapy cross-resistance we developed several rituximab resistance cell lines (RRCL), which also display significant resistance to a wide range of chemotherapy agents. RRCLs exhibit a deregulation in the apoptotic response pathway at several levels. Previous work by our group demonstrated that the X linked inhibitor of apoptosis (XIAP) is a key regulator of chemotherapy resistance in RRCLs. Moreover, we have established that knocking down XIAP in RRCLs can enhance the anti-tumor cytotoxic effect of chemotherapy, but a clear mechanistic role for XIAP in RRCLs has yet to be established. To investigate how XIAP regulates chemotherapy response in resistant lymphoma models we first looked at expression levels of XIAP and its binding partners in RRCLs and rituximab sensitive cell lines (RSCL). Although we observed no change in XIAP expression between the RRCL Raji 4RH and RSCL Raji, we did observe changes in several of the XIAP interacting proteins. Protein levels for both of the primary apoptotic effector caspases (caspase 3 and caspase 7) were substantially decreased in the Raji 4RH cell line compared to Raji cells. In addition, when we exposed Raji and Raji 4RH cells to gemcitabine (50uM) for 28 hours we observed substantially less release of cytochrome C from the mitochondria in Raji 4RH compared to Raji. Cytochrome C is an essential part of the apoptosome, which functions as a primary activator of caspases 3 and 7. Both of these observations support the hypothesis that impaired effector caspase activation and signaling contributes to chemotherapy resistance in RRCLs. To determine if targeting XIAP could restore caspase activation in RRCLs we used a transient siRNA knockdown of XIAP in the Raji 4RH cell line. Cells were treated with either the XIAP targeting siRNA, or a scramble sequence control, and then exposed to chemotherapy for 36 hours. Cellular viability was measured with the Promega CellTiter-Glo ATP assay, while caspase 3 and 7 activity was determined with the Caspase 3/7-Glo assay, also purchased from Promega. XIAP knockdown increased caspase activity by more than 500% compared to controls when cells were exposed to etoposide (20uM). This correlated with a 60% decrease in cellular viability in cells with an XIAP knockdown. In addition, the addition of the caspase inhibitor QVD-OPh along with the RIP kinase inhibitor necrostatin-1 (both at 10uM) was enough to restore cellular viability to the same levels observed in scramble treated controls exposed to the same dose of etoposide. Additional experiments looking at activation of caspases 2 and 6 in Raji 4RH cells following XIAP knockdown did not show any difference compared to scramble controls. In summary, XIAP appears to directly inhibit the caspase dependent apoptotic response in RRCLs. In addition, targeting XIAP can restore the apoptotic response in cell line models of rituximab resistant disease. These results support the hypothesis that selective XIAP inhibitors, currently in development, may be highly active against rituximab rel/ref B-cell lymphoma. DisclosuresNo relevant conflicts of interest to declare.

Predictive Value of Natriuretic Peptide Level Monitoring in Patients with Malignant Lymphoma during High-Dose Chemotherapy in Combination with Autologous Stem Cell Transplantation

Introduction. The natriuretic peptides are neurohormones that act to relieve the symptoms associated with volume expansion and pressure overload by promoting natriuresis and diuresis, vasodilatation, and suppression of the Renin Angiotensin Aldosterone system. Serum N-terminal prohormone of brain natriuretic peptide (NT-proBN) levels in the blood have been used for screening and diagnosis of congestive heart failure (CHF) in cardiology practice. However, there is no data to support the predictive value of its levels in patients with malignant lymphomas (ML) during the high-dose chemotherapy (HDC) with autologous hematopoietic stem cells transplantation (auto-HSCT).The goal of the study was to evaluate the cardiotoxic effect of HDC with auto-HSCT in patients with malignant lymphomas using NT-proBN at different time before and after the treatment; to evaluate dependence of increasing levels of this biochemical marker with age and different conditioning regimen.Material and metods. The study included 127 patients (67 - Hodgkin's lymphoma, 28 - non-Hodgkin's lymphomas, and 32 - multiple myeloma) The median age was 37 year (range: 19 to 68 years); 80 females, 47 males. The level of NT-proBNP was monitored before starting the HDC (point1), after the HDC (point 2), and on day 12 after the auto-HSCT (point 3).Results; Levels of NT-proBNP at time of admission to the hospital (point 1) was significantly lower than at the end of the HDC (point 2) and on D + 12 (point 3).There were no significant correlations between the age of patients and the level of NT-proBNP before HDC (point 1) and level on D + 12 (point 3); however, in point 2, after HDC, a significant positive Spearmen correlation coefficient, R = 0.20, was obtained. So the older the patient, the higher the probability of an increase in the concentration of NT-proBNP after HDC.Analysis of the dependence of changes in the level of NT-proBNP on the sex of patients at different periods of the study revealed significant differences in marker levels in men and women, in women its concentration is higher than in men before HDC (poin 1). After completion of conditioning and on day 12, the level of NT-proBNP in women still significantly exceeded the corresponding value in men.Analisis of the level of the marker, depending on different conditioning regimens, showed that before the start of conditioning there were no significant differences in marker levels in patients who received the CBV regimen and high doses of melphalan. After the end of conditioning, a significant increase in the concentration of the marker was established in patients undergoing CBV, the median of NT-proBNP concentration was 526 pg/ml, whereas the patients receiving melphalan received 245 pg/ml. However, in point 3 the revealed differences were leveled off.Comparison of the concentrations of NT-proBNP in the groups of patients receiving BEAM and melphalan did not reveal significant differences. At the same time, the assessment of the marker levels at the poin after HDC showed significant differences. Median NT-proBNP concentrations in this period were 152.9 pg/ml in patients treated with BEAM, 293 pg/ml in patients treated with CBV (Figure 1)The results of the analysis of the relationship between the level of NT-proBNP and the total dose of cyclophosphamide showed the presence of a moderate significant correlation of the cyclophosphamide dose and the concentration of the marker in at the end of HDC (point 2), R = 0.40 (Table 1). At the same time, an assessment of the relationship between the level of NT-proBNP and the total dose of etoposide did not reveal the presence of significant correlation coefficients at all times of the study.Conclusion: The results obtained can serve as a basis for considering NT-proBNP as a marker of cardiotoxicity of HDC and auto-HSCT. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Survival Benefit and Efficiency of Low Dose Decitabine With CEG Regimen Compared to Decitabine Alone in the Elderly MDS – A Multicenter, Retrospective Study

BackgroundDecitabine are used in the treatment of myelodysplastic syndrome (MDS), but none trials reported overall survival improvement. MethodsHigh-risk MDS and MDS transformed AML (sAML) patients (IPSS-R > 4.5, age above 60 years) in 6 medical centers of China were treated and compared a new regimen (decitabine with CEG) consisted of low dose decitabine (15 mg/m2, days 1-3), low dose etoposide (30 mg/m2, days 4,6,8,10,12), cytarabine (10 mg/m2 per day, days 4-12) and granulocyte colony-stimulating factor (G-CSF, 5ug/kg, adjusted by patients’ WBC level, 12 hours prior to decitabine administration) with decitabine alone. The endpoints were death and disease progression. ResultsThe baseline characteristics of these 2 groups were equivalent and none patients received prior chemotherapy. The treatment response rate (P= .048) and progression free survival (PFS, P = .030) all demonstrated significant improvement compared with decitabine alone. Decitabine with CEG regimen had attained a CR rate of 45.7%, a median OS of 36 (19-53) months and a median PFS of 34 (16.7-51.3) months in high-risk MDS patients, a CR rate of 40% in sAML. While decitabine alone only attained a median OS of 26 (24.5-27.5) months and a CR rate of 18.2% as well as a median progression free survival of 20 (17.6-22.4) months in MDS patients. Treatment response to CR or PR and TP53 mutation were 2 prognostic factor for OS and PFS in decitabine with CEG regimen. ConclusionDecitabine with CEG regimen showed some promising advantage in elderly, high-risk MDS.

Lysosome Fe2+ release is responsible for etoposide- and cisplatin-induced stemness of small cell lung cancer cells.

Iron metabolism has been shown to hand over cancer stem cell, which is regarded as the root of tumor progression, recurrence and chemoresistance. This study aims to explore whether iron metabolism is involved in etoposide- and cisplatin-induced stemness in small cell lung cancer (SCLC) cells. Here, analysis on tumor-sphere formation and stemness marker expression is performed to determine whether etoposide and cisplatin can induce SCLC cell stemness. Online dataset analysis is constructed to determine the correlation between iron transportation and the survival of lung cancer patients. Chromatin immunoprecipitation combined with rescuing experiments are carried out to reveal the underlying mechanisms. Additionally, the non-lethal doses of etoposide and cisplatin can induce SCLC cell stemness in a concentration-dependent manner and reduce the lysosome iron concentration dependent on Ferritin expression, which is positively regulated by HIF-1α/β. Moreover, HIF-1α/β can directly bind to Ferritin promoter region. This HIF/Ferritin axis is responsible for etoposide- and cisplatin-induced iron reduction in lysosomes and stemness of SCLC cells. This work demonstrates that iron in lysosomes is essential for etoposide and cisplatin-induced stemness of SCLC cells, which is regulated by the HIF/Ferritin axis.

Efficacy of Moderately Dosed Etoposide in Macrophage Activation Syndrome-Hemophagocytic Lymphohistiocytosis.

Macrophage activation syndrome (MAS) constitutes 1 subtype of the hyperinflammatory syndrome hemophagocytic lymphohistiocytosis (HLH), and the term MAS-HLH was recently proposed for HLH with underlying autoimmune/autoinflammatory conditions. The mortality of MAS-HLH has been estimated at 5-10%. Here we report our experiences with moderately dosed etoposide in severe MAS-HLH; the objective was to effectively reduce severe hyperinflammatory activity with limited side effects. In addition to conventional antiinflammatory treatment, moderately dosed etoposide was administered to 7 children affected by rapidly progressing MAS-HLH with central nervous system (n = 5) and/or pulmonary (n = 5) involvement. Three had underlying systemic juvenile idiopathic arthritis (sJIA), 2 had atypical sJIA (no arthritis at diagnosis), and 2 had systemic lupus erythematosus. We performed lymphocyte cytotoxicity analyses in all 7 and genetic analyses in 6. All children promptly responded to moderately dosed etoposide (50-100 mg/m2 once weekly), added to conventional MAS-HLH treatment that was considered insufficient. The mean accumulated etoposide dose was 671 mg/m2 (range 300-1050 mg/m2) as compared to 1500 mg/m2 recommended in the first 8 weeks of the HLH-94/HLH-2004 protocols. One child developed neutropenic fever and another neutropenic sepsis (neutrophils 0.3 × 109/L at therapy onset). Five of 7 children had low percentages (< 5%) of circulating natural killer (NK) cells prior to or in association with diagnosis; NK cell activity was pathologically low in 2 of 5 children studied. Disease-causing variants in HLH-associated genes were not found. All children were alive at latest follow-up (2-9 yrs after onset); neurological symptoms had normalized in 4 of 5 affected children. Moderately dosed etoposide may be beneficial in severe and/or refractory MAS-HLH.

The benefits of etoposide capsules as maintenance therapy for patients with extensive-stage small cell lung cancer: a prospective two-stage, two-center study

BackgroundDue to the high incidence and mortality of lung cancer, and etoposide is the standard first-line chemotherapy for small cell lung cancer, to evaluate the efficacy and safety of etoposide capsules at different doses as maintenance therapy for patients with extensive-stage small cell lung cancer (ES-SCLC) who show a response to etoposide plus platinum.MethodsThe study was divided into two stages: stage I, a single-center, one-arm prospective study, and stage II, a multicenter, controlled non-randomized prospective study (patients were chosen from ClinicalTrials.gov Identifier: NCT02179528). All patients received six cycles of etoposide plus platinum. Patients who were evaluated as complete remission (CR) or partial remission (PR) entered the maintenance treatment (MT) (etoposide capsule, once a day for 20 days, every 28 days as a cycle, until disease progression). In stage I, the dose of etoposide was 25 mg; in stage II, patients were non-randomized into etoposide capsule (25 mg/50 mg) and observation groups. In this study, the primary endpoints were progression-free survival (PFS) and safety; the secondary endpoint was overall survival (OS). Toxicity was graded according to the Common Terminology Criteria for Adverse Events v3.0.ResultsNinety-two patients were enrolled. In stage I, the median PFS was 6.700 months (95% CI: 6.408–6.992). In stage II, the median PFS of the MT group was better than that in the NMT group (8.930 vs. 5.900 months, P=0.002). In the pooled analysis, the overall median PFS of the MT group was better than that of the NMT group (7.870 vs. 5.900 months, P=0.003). However, there was no significant difference in OS between the groups (15.030 vs. 14.330 months, P=0.813). Multivariate Cox regression analysis showed that maintenance therapy was an independent protective factor for PFS in patients with ES-SCLC.ConclusionsEtoposide capsules as maintenance therapy significantly prolonged the PFS of patients with ES-SCLC who responded to etoposide plus platinum, with acceptable tolerability.

DDEL-03. LONG-TERM INTRAVENTRICULAR THERAPY ALTERNATING ETOPOSIDE AND LIPOSOMAL CYTARABINE: EXPERIENCE IN 75 CHILDREN AND ADOLESCENTS WITH MALIGNANT BRAIN TUMORS

BACKGROUNDMalignant brain tumors of childhood carry a high risk for leptomeningeal dissemination and tumor cells floating in the CSF are often not amenable to systemic and/or antiangiogenic chemotherapy. We report on our experience with an intraventricular therapy consisting of alternating cycles of liposomal cytarabine and etoposide.PATIENTS AND METHODSBetween 2004 and 2017, 75 patients aged 0.6 to 22 years (median 11) with various malignant brain tumors received intraventricular etoposide 0.25mg (<1year) - 0.5mg on five consecutive days alternating with liposomal cytarabine at a dose of 25mg (<3 years) - 50mg via an Ommaya reservoir.RESULTS5533 doses of etoposide (5–277/patient, median 141) corresponding to 1–56 five-day-cycles/patient alternating with 534 doses of liposomal cytarabine (1–21/patient, median 11) were administered. Treatment was given over a period of 1 – 146 months (median 73.5). Toxicities did occur but were infrequent and mostly mild. Since all patients received some sort of concurrent anti-cancer therapy, the efficacy of intrathecal therapy cannot be assessed independently. However, 29/75 patients are still alive, and none of the patients had tumor cells in the CSF at their last evaluation.CONCLUSIONIn conclusion, alternating intraventricular liposomal cytarabine and etoposide produced responses and proved to be an important adjunct for patients receiving drugs with a low penetrance into the CSF. Since production of liposomal cytarabine was discontinued in 2017 it remains to be determined whether substitution of the slow release formulation by aqueous cytarabine on days 1, 4, 8, and 11 may produce similar results.

XPO1 Relieves MYC-Induced Replication Stress Limiting the Immunogenicity of DLBCL Cells

The soluble nuclear pore component XPO1 is a therapeutic target in DLBCL and its selective inhibitor selinexor received FDA approval in relapsed/refractory DLBCL. XPO1 inhibitors block the nuclear export of a subset of cargo proteins. Since the spectrum of XPO1 cargos varies among cell types, the mechanism underlying the anti-lymphoma effect of selinexor has not been fully elucidated. Lack of this knowledge hampers the clinical use of XPO1 inhibitors in terms of patient selection and development of rational therapeutic combinations. We reported that XPO1 is amplified and overexpressed in DLBCL. At the molecular level, we described that XPO1 overexpression functionally remodels the nuclear pore to sustain the expression of DNA damage repair proteins increasing lymphoma cells' tolerance to genotoxic stress. Here, we investigated whether this XPO1 role is relevant for the anti-lymphoma effect of selinexor. Aberrant MYC expression induces DNA damage resulting from replication stress; thus, we speculated that selinexor kills MYC expressing (MYC+) DLBCL cells by limiting their ability to tolerate replication stress. Using B-cells expressing MYC in an inducible manner, we demonstrated that exposure to selinexor (6 h) caused G2-M cell cycle arrest upon MYC expression (p&amp;lt;0.0001, vs. vehicle-treated cells). Cell cycle arrest was associated with increased levels of DNA damage in S-phase as measured by yH2AX (p&amp;lt;0.0001). Furthermore, although MYC increased the expression of key replication stress repair proteins (i.e. RAD51, WEE and BRCA1), this effect was blunted by selinexor. To determine the clinical relevance of this mechanism, we interrogated publicly available datasets of DLBCLs and found that XPO1 amplification and overexpression was associated with MYC translocation (p&amp;lt;0.005), double-hit lymphoma signature (p&amp;lt;0.005), and elevated levels of chromosomal instability (p&amp;lt;0.005). Overall, these observations suggest that MYC-driven lymphomas sustaining high levels of genomic instability may critically depend on XPO1. To therapeutically capitalized this mechanism, we evaluated the effect of selinexor on MYC+ DLBCLs. Clinically achievable doses of selinexor induced cell cycle arrest and apoptosis in 83% (5/6) of cell lines investigated in vitro. In a patient-derived xenograft (PDX) MYC+ DLBCL model, a human-equivalent dose of selinexor impaired tumor growth (p&amp;lt; 0.05, vs. vehicle) without significant toxicities. Reduced tumor growth was secondary to apoptosis as determined by TUNEL (p&amp;lt;0.05). To characterize the biological effects of XPO1 inhibition, we compared the transcriptome of PDX from the previous experiment. Pathway analysis of differentially expressed genes revealed that selinexor induced pathways regulating innate immune signaling. The upregulation of key genes from this pathway (i.e. DDX58, DDX60, IRF7 and STAT1) was validated by qRT-PCR. Since unrepaired DNA damage elicits an innate immune response, we hypothesized that these effects could be mechanistically linked. To test this notion, we exposed MYC+ DLBCL cells to sub-lethal doses of the DNA damaging agent etoposide, selinexor, or their combination. Expression of innate immune genes was significantly higher in cells exposed to the combination compared to either drug alone (p&amp;lt;0.001, for both comparisons), indicating that selinexor increases the immunogenicity of MYC+ DLBCL cells and this effect is enhanced by concomitant replication stress. Furthermore, by analyzing primary DLBCLs we found that poorly immunogenic lymphomas, characterized by an immune deserted microenvironment, expressed higher levels of XPO1 and MYC compared to DLBCL with an immune rich microenvironment (P&amp;lt;0.001, for both genes). To determine a potential effect of selinexor in the function of immune effector cells, we performed single cells secretomic analysis (Isolight) of circulating CD8+ T cells obtained from two DLBCL patients before and after selinexor treatment and found that selinexor increases the secretory activity of CD8+ T cells in both patients. Overall, our study reveals a key oncogenic role for XPO1 in enabling DLBCL cells to tolerate MYC-induced replication stress while limiting their immunogenicity. This suggest that patients with MYC+ DLBCL and elevated genomic instability may benefit from therapeutic approaches based on XPO1 inhibitors in combination with immunomodulatory drugs, an approach we will test prospectively. Disclosures Rutherford: AstraZeneca: Consultancy; Celgene: Consultancy; Dova: Consultancy; Genentech/Roche: Research Funding; Heron: Consultancy; Juno: Consultancy; Karyopharm: Consultancy, Research Funding; Kite: Consultancy; LAM Therapeutics: Research Funding; Regeneron: Research Funding; Seattle Genetics: Consultancy. Martin:Regeneron: Consultancy; Cellectar: Consultancy; Janssen: Consultancy; Karyopharm: Consultancy, Research Funding; Teneobio: Consultancy; Celgene: Consultancy; Bayer: Consultancy; Beigene: Consultancy; Sandoz: Consultancy; I-MAB: Consultancy; Morphosys: Consultancy; Kite: Consultancy; Incyte: Consultancy. Leonard:Miltenyi: Consultancy; Sutro: Consultancy; Roche/Genentech: Consultancy; BMS/Celgene: Consultancy; Regeneron: Consultancy; ADC Therapeutics: Consultancy; MEI Pharma: Consultancy; Bayer: Consultancy; Gilead/Kite: Consultancy; Karyopharm: Consultancy; GenMab: Consultancy; Epizyme: Consultancy; AstraZeneca: Consultancy. Cerchietti:Cellgene: Research Funding; Bristol Myers Squibb: Research Funding.

A Multi-Center Analysis of the Impact of Dose Level of R-EPOCH on Outcomes of Patients with Double/Triple-Hit B-Cell Lymphoma

A Multi-Center Analysis of the Impact of Dose Level of R-EPOCH on Outcomes of Patients with Double/Triple-Hit B-Cell Lymphoma

Adding Etoposide to R-CHOP (R-CHOEP) Does Not Significantly Increase the Risk of Secondary Neoplasms in Patients with Aggressive B-Cell Lymphoma - Results from Randomized Phase 3 Trials of the German Lymphoma Alliance (GLA)

Introduction:Considering the increasing numbers of lymphoma patients (pts) surviving long-term, late effects of current treatment strategies such as secondary (sec) malignancies gain increasing importance. Many treatment regimens used in pts with lymphoma (R-CHOEP, DA-EPOCH-R, BEACOPP) include etoposide, a cytotoxic agent reported to increase sec leukemias. In order to further investigate the role of etoposide in inducing sec malignancies in pts with aggressive lymphoma we analyzed the R-MegaCHOEP trial (Schmitz et al., Lancet Oncology 2012) where young, high-risk pts with aggressive B-cell lymphomas had received R-CHOEP or R-MegaCHOEP, a regimen containing very high doses of etoposide (4g/m2). We compared rates of secondary tumors to incidences found in young patients treated with R-CHOP only. Methods:We analyzed 1536 pts aged 18-60 years with aggressive B-cell lymphoma treated in the prospective phase 3 trials FLYER (NCT00278421; n=588, median observation time (OT)=66 months), UNFOLDER (NCT00278408; n=695, median OT=72 months) and MegaCHOEP (NCT00129090; n=253, median OT=112 months) to compare the cumulative incidences of sec neoplasms. We performed a competing risk analysis for time from randomization to occurrence of sec malignancy (myelodysplastic syndrome/acute myeloid leukemia (MDS/AML) or other) according to first-line therapy with R-CHOP (n=1283) vs. R-CHOEP (n=127) vs. R-MegaCHOEP (n=126). We used a cause-specific hazard model adjusted for gender, age (&amp;gt;50 vs. &amp;lt;= 50 years) and radiotherapy. Results:Adjusting for competing events, we found sec MDS/AML in 0.3% of pts treated with R-CHOP, in 1% of pts treated with R-CHOEP and in 2% of pts following treatment with R-MegaCHOEP. Cause-specific hazard ratios showed a trend for increased risk for sec AML following R-MegaCHOEP in comparison to R-CHOP (HR 5.2, 95%CI (1.0; 27.5), p=0.052) while R-CHOEP did not significantly increase the incidence of sec AML (HR 1.4, 95%CI (0.1; 14.0), p=0.777). Male gender and age &amp;gt;50 years showed a trend for increasing sec AML. We found very similar incidences of sec solid tumors (not MDS/AML) affecting 4% of pts treated with R-CHOP or R-MegaCHOEP and 6% of pts following R-CHOEP regimen. The only factor significantly increasing the risk of sec solid tumors was age &amp;gt;50 years (HR 2.6, 95%CI (1.5; 4.3), p&amp;lt;0.001). Conclusions:This analysis shows that sec malignancies represent rare events in younger patients with aggressive B-cell lymphoma, even after extreme dose-escalation of etoposide. Compared to standard R-CHOP, 8 cycles of R-CHOEP were not associated with increased risk of sec AML or solid tumors thus remaining an attractive first-line treatment for young high-risk pts with DLBCL. Disclosures Haenel: Amgen, Novartis, Roche, Celgene, Takeda, Bayer:Honoraria.Truemper:Janssen:Consultancy;Mundipharma:Research Funding;Nordic Nanovector:Consultancy;Roche:Research Funding;Seattle Genetics:Research Funding;Takeda Europe:Consultancy, Research Funding.Held:MSD:Consultancy;Acrotech, Spectrum:Research Funding;Amgen:Research Funding;BMS:Consultancy, Other: Travel Grants, Research Funding;Roche:Consultancy, Other: travel grants, Research Funding.Borchmann:Bristol Myers Squibb:Research Funding;Takeda:Research Funding.Dreyling:Celgene:Consultancy, Research Funding, Speakers Bureau;Roche:Consultancy, Research Funding, Speakers Bureau;Beigene:Consultancy;Janssen:Consultancy, Research Funding, Speakers Bureau;Novartis:Consultancy;Abbvie:Research Funding;Astra Zeneca:Consultancy;Bayer:Consultancy, Speakers Bureau;Gilead:Consultancy, Research Funding, Speakers Bureau.Viardot:Roche:Honoraria, Other: advisory board;Kite/Gilead:Honoraria, Other: advisory board;Novartis:Honoraria, Other: advisory board;Amgen:Honoraria, Other: advisory board.Kroschinsky:Riemsser:Research Funding;Roche:Consultancy, Honoraria, Other: Support of parent study and funding of editorial support;Gilead:Consultancy;BMS/Celgene:Consultancy, Honoraria;Sandoz:Research Funding.Ott:NIH:Consultancy, Other: Co-inventor on a patent related to the MCL35 assay filed at the National Institutes of Health, United States of America..Lenz:Bayer:Consultancy, Honoraria, Research Funding, Speakers Bureau;Janssen:Consultancy, Honoraria, Research Funding, Speakers Bureau;Novartis:Consultancy;Gilead:Consultancy, Honoraria, Research Funding, Speakers Bureau;AQUINOX:Research Funding;AstraZeneca:Consultancy, Honoraria, Research Funding;Celgene:Consultancy, Honoraria, Speakers Bureau;Verastem:Research Funding;Morphosys:Consultancy, Honoraria, Research Funding;Roche:Consultancy, Honoraria, Research Funding, Speakers Bureau;Agios:Research Funding;BMS:Consultancy.Schmitz:Riemser:Honoraria;Takeda:Honoraria;Janssen:Research Funding;Bristol-Myers Squibb:Current equity holder in publicly-traded company.

PET-Guided Strategy Improves the Safety of Beacopp-Based Treatment in Advanced Hodgkin Lymphoma: Prolonged Follow-up of the Lysa Ahl 2011 Phase 3 Study

PET-Guided Strategy Improves the Safety of Beacopp-Based Treatment in Advanced Hodgkin Lymphoma: Prolonged Follow-up of the Lysa Ahl 2011 Phase 3 Study

All-oral metronomic DEVEC schedule in elderly patients with peripheral T cell lymphoma.

PurposePeripheral T cell lymphomas (PTCLs) have an overall poor prognosis. Indeed, registry data in elderly patients show that the median progression-free survival (mPFS) following first- and second-line therapies are only 6.7 and 3.1 months, respectively. The aim of the study is to show the activity of metronomic chemotherapy, a regular administration of low chemotherapeutic drug doses allowing a favourable toxicity profile, on elderly PTCL patients.MethodsWe report a series of 17 PTCL patients, treated with the all-oral metronomic schedule DEVEC (prednisolone–etoposide–vinorelbine–cyclophosphamide) in four Italian centres. Patients 5/17 (29.4%) were treatment-naïve (naïve) and 12/17 (70.6%) were relapsed-refractory (RR), respectively. The median age was 83 years (range 71–87) and 71.5 years (range 56–85) for naïve and RR, respectively. In vitro activity of metronomic vinorelbine (VNR), etoposide (ETO) and their concomitant combination on HH, a PTCL cell line, was also assessed.ResultsHistology: PTCL-not-otherwise-specified = 12; angioimmunoblastic = 2; NK/T nasal type = 1; adult-type leukaemia lymphoma = 1, transformed Mycosis Fungoides = 1. The overall response rate was 80 and 58% in naïve and RR, respectively; whereas the PFS was 20 in naïve (95% CI 0–43) and 11 months (95% CI 4.2–17.8) in RR. The occurrence of relevant adverse events was 23.5%, which was managed with ETO dose reduction. In vitro experiments showed that both metronomic VNR and ETO caused a significant inhibitory activity on HH cells and a strong synergism when administered concomitantly.ConclusionAll-oral DEVEC showed an encouraging activity and acceptable toxicity. This schedule deserves further studies in elderly PTCL also for assessing combinations with targeted drugs.Electronic supplementary materialThe online version of this article (10.1007/s00280-020-04172-3) contains supplementary material, which is available to authorized users.

Abstract 5260: Preclinical studies with ONC201 in combination with clinically approved chemotherapy as a novel treatment strategy against Small Cell Lung Cancer (SCLC)

Abstract According to the American Cancer Society, about 10-15 percent of all lung cancer diagnoses are small cell lung cancer (SCLC). The median survival rate of limited-stage small cell lung cancer is about 12 to 16 months. Several clinically approved chemotherapy drugs have been used to treat SCLC, with several more currently under clinical trials. However, a novel combination is still needed to increase patient life expectancy. ONC201 is an impridone, currently in clinical trials, that has proven to have proapoptotic effects in multiple tumor types. We hypothesized that treating SCLC cell lines with ONC201 in conjunction with clinically approved chemotherapy drugs, such as etoposide and carboplatin, will have a synergistic effect and serve as a potential novel combination treatment plan. SCLC cell lines H1048 and H1105 were treated for 72 hours with ONC201 and etoposide, or ONC201 and carboplatin. CellTiter-Glo cell viability assay and Western blotting of PARP cleavage and ATF4 were used to check synergy in the combination treatments. The combination of ONC201 and clinically approved chemotherapy drugs, etoposide and carboplatin, revealed that there was an upregulation in cleaved PARP and a synergistic effect on the overall cell samples. When looking at the combination index of the cell proliferation assays, the best index for each combination was 0.867 with an ONC201 dosage of 4 μM and an etoposide dosage of 0.05 μg/mL in H1048, 0.43191 at ONC201 dosage 0.3125 μM and etoposide dosage 2.5 μg/mL in H1105, and 0.20278 at ONC201 dosage 2.5 μM and carboplatin dosage 3.125 μg/mL in H1105. We are currently testing drug combinations including a triple drug combination of ONC201 and two clinically approved drugs and studying the mechanism underlying the synergies, with the potential to translate this novel combination treatment plan of ONC201 and clinically approved drugs into a clinical regimen to improve the outcome of SCLC patients. Citation Format: Young S. Lee, Lanlan Zhou, Christopher Azzoli, Wafik S. El-Deiry. Preclinical studies with ONC201 in combination with clinically approved chemotherapy as a novel treatment strategy against Small Cell Lung Cancer (SCLC) [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5260.

Abstract 575: Disulfiram plus copper synergizes with etoposide for the treatment of Merkel cell carcinoma

Abstract Merkel cell carcinoma (MCC) is an aggressive neuroendocrine skin cancer with limited treatment options. Currently two immunotherapies are approved for the treatment of MCC, the PD-L1 inhibitor avelumab and the PD-1 inhibitor pembrolizumab. Unfortunately, these treatments only have durable benefit for less than half of patients that can be treated with immunotherapy. Therefore, new and effective treatments are needed for MCC. To address this, we conducted a high-throughput drug screening of ~4,000 small molecules. Our drug screen identified disulfiram (DSF), an aldehyde dehydrogenase inhibitor used in the treatment of alcoholism, as an agent that selectively reduced MCC cell viability. Recent studies suggest that DSF complexed with copper can have anti-cancer effects. Consistent with this, the addition copper at physiological levels increased the potency of DSF in MCC cells lines. Next, we demonstrated that disulfiram combined with copper synergized with the topoisomerase II inhibitor etoposide, lowering the effective dose of etoposide required to reduce MCC cell viability. Mechanistically, combining DSF, copper and etoposide was cytotoxic but did not induce apoptosis in MCC cell lines. This combination increased the formation of double stranded DNA breaks while disulfiram plus copper induced autophagy, which could mechanistically be an indication of immunogenic cell death signaling. Taken together, our data suggest that disulfiram combined with copper and etoposide could be repurposed for the effective treatment of advanced MCC. One strong advantage of this combination is that all agents can be given orally and at dosages with minimal adverse effects. Citation Format: Natasha Tremayne Hill, Tara Gelb, Dan Urban, Tyler Kellenberger, Matthew Hall, Isaac Brownell. Disulfiram plus copper synergizes with etoposide for the treatment of Merkel cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 575.

Damage-associated molecular patterns (DAMPs) related to immunogenic cell death are differentially triggered by clinically relevant chemotherapeutics in lung adenocarcinoma cells

BackgroundChemotherapeutics can stimulate immune antitumor response by inducing immunogenic cell death (ICD), which is activated by Damage-Associated Molecular Patterns (DAMPs) like the exposure of calreticulin (CRT) on the cell surface, the release of ATP and the secretion of High Mobility Group Box 1 (HMGB1).MethodsHere, we investigated the levels of ICD-associated DAMPs induced by chemotherapeutics commonly used in the clinical practice of non-small cell lung cancer (NSCLC) and the association of these DAMPs with apoptosis and autophagy. A549 human lung adenocarcinoma cells were treated with clinically relevant doses of cisplatin, carboplatin, etoposide, paclitaxel and gemcitabine. We assessed ICD-associated DAMPs, cell viability, apoptosis and autophagy in an integrated way.ResultsCisplatin and its combination with etoposide induced the highest levels of apoptosis, while etoposide was the less pro-apoptotic treatment. Cisplatin also induced the highest levels of ICD-associated DAMPs, which was not incremented by co-treatments. Etoposide induced the lower levels of ICD and the highest levels of autophagy, suggesting that the cytoprotective role of autophagy is dominant in relation to its pro-ICD role. High levels of CRT were associated with better prognosis in TCGA databank. In an integrative analysis we found a strong positive correlation between DAMPs and apoptosis, and a negative correlation between cell number and ICD-associated DAMPs as well as between autophagy and apoptosis markers. We also purpose a mathematical integration of ICD-associated DAMPs in an index (IndImunnog) that may represent with greater biological relevance this process. Cisplatin-treated cells showed the highest IndImmunog, while etoposide was the less immunogenic and the more pro-autophagic treatment.ConclusionsCisplatin alone induced the highest levels of ICD-associated DAMPs, so that its combination with immunotherapy may be a promising therapeutic strategy in NSCLC.

Manipulating Senescence of Cardiac Fibroblasts In Vivo and In Vitro to Investigate Arrhythmogenic Effect of Senescence

Cellular senescence is a stress response characterized by irreversible cell cycle arrest, secretion of inflammatory cytokines and growth factors termed the “senescence associated secretory phenotype” (SASP), and resistance to apoptosis. Senescence plays beneficial roles in tumor suppression, wound healing, and development in young animals, but is thought to underlie aging‐associated pathology in numerous tissues, mainly via the SASP. Our previous work in the aged rabbit myocardial infarction (MI) model demonstrated an increase and persistence of senescence, which we hypothesize to drive increased risk of arrhythmogenesis with age post‐MI. Here, we investigate the molecular mechanisms by which we believe senescence drives arrhythmogenesis. Specifically, we hypothesize that inflammatory cytokines in the SASP locally produced by senescent myofibroblasts acts on border zone myocytes to disrupt cellular‐level electrophysiology. Concordantly, we hypothesize that genetically or pharmacologically eliminating senescent cells post‐MI will mitigate arrhythmogenesis in vivo and in vitro, and that genetically increasing senescence post‐MI in a mouse Cre‐Lox model will increase arrhythmogenesis.Here, we show that commercially‐available aged human cardiac fibroblasts and primary adult rabbit cardiac fibroblasts can be induced into senescence via treatment with low doses of Etoposide in a time‐dependent manner. Additionally, via a senolytic drug screening, we found that high doses of Dasatinib, Quercetin, and Navitoclax effectively eliminate senescent cells but not non‐senescent cells. However, we found combinations of Dasatinib and Quercetin or Dasatinib and Venetoclax did not perform better than either drug in solo. Additionally, we verify that calcium linescan imaging as well as patch clamp measurements of the IKr current can be measured in primary 3‐week‐old rabbit cardiomyocytes.Using genetically altered mice to conditionally knock out the pro‐senescence p53 gene in myofibroblasts under control of the Periostin gene (p53 CKO mice), we find that p53 CKO mice have significantly less senescence post‐MI compared to littermate controls, and the effect is stronger in females compared to males. We also report that fibroblast‐specific knockout of the MDM2 gene in the same condition is embryonic lethal, as Periostin is expressed in the developing valve leaflets and endocardial cushion cells, where senescence plays a vital role in pruning transient fetal structures.In conclusion, we demonstrate methods to genetically and pharmacologically manipulate senescence in vivo and in vitro, which will allow us to investigate the molecular mechanisms through which senescence might play a role in promoting arrhythmogenesis with age post‐MI.Support or Funding InformationThe authors would like to acknowledge our supporting NIH funding sources, R21 grant 1R21AG049608‐01 and R01 grant R01HL139467