Combination Of Romidepsin Research Articles

Acetylation-dependent regulation of core spliceosome modulates hepatocellular carcinoma cassette exons and sensitivity to PARP inhibitors

Despite the importance of spliceosome core components in cellular processes, their roles in cancer development, including hepatocellular carcinoma (HCC), remain poorly understood. In this study, we uncover a critical role for SmD2, a core component of the spliceosome machinery, in modulating DNA damage in HCC through its impact on BRCA1/FANC cassette exons and expression. Our findings reveal that SmD2 depletion sensitizes HCC cells to PARP inhibitors, expanding the potential therapeutic targets. We also demonstrate that SmD2 acetylation by p300 leads to its degradation, while HDAC2-mediated deacetylation stabilizes SmD2. Importantly, we show that the combination of Romidepsin and Olaparib exhibits significant therapeutic potential in multiple HCC models, highlighting the promise of targeting SmD2 acetylation and HDAC2 inhibition alongside PARP inhibitors for HCC treatment.

Romidepsin and Afatinib Abrogate Jak–Signal Transducer and Activator of Transcription Signaling and Elicit Synergistic Antitumor Effects in Cutaneous T-Cell Lymphoma

Cutaneous T-cell lymphomas (CTCL) are mature lymphoid neoplasias resulting from the malignant transformation of skin-resident T cells. A distinctive clinical feature of CTCL is their sensitivity to treatment with histone deacetylase (HDAC) inhibitors. However, responses to HDAC inhibitor therapy are universally transient and non-curative, highlighting the need for effective and durable drug combinations. Here we demonstrate that the combination of romidepsin, a selective class I HDAC inhibitor, with afatinib, an epidermal growth factor receptor (EGFR) family inhibitor, induces strongly synergistic antitumor effects in CTCL models in vitro and in vivo via abrogation of JAK-STAT signalling. These results support a previously unrecognized potential role for HDAC inhibitor plus afatinib combination in the treatment of CTCL.

A multidimensional platform of patient-derived tumors identifies drug susceptibilities for clinical lenvatinib resistance

Lenvatinib, a second-generation multi-receptor tyrosine kinase inhibitor approved by the FDA for first-line treatment of advanced liver cancer, facing limitations due to drug resistance. Here, we applied a multidimensional, high-throughput screening platform comprising patient-derived resistant liver tumor cells (PDCs), organoids (PDOs), and xenografts (PDXs) to identify drug susceptibilities for conquering lenvatinib resistance in clinically relevant settings. Expansion and passaging of PDCs and PDOs from resistant patient liver tumors retained functional fidelity to lenvatinib treatment, expediting drug repurposing screens. Pharmacological screening identified romidepsin, YM155, apitolisib, NVP-TAE684 and dasatinib as potential antitumor agents in lenvatinib-resistant PDC and PDO models. Notably, romidepsin treatment enhanced antitumor response in syngeneic mouse models by triggering immunogenic tumor cell death and blocking the EGFR signaling pathway. A combination of romidepsin and immunotherapy achieved robust and synergistic antitumor effects against lenvatinib resistance in humanized immunocompetent PDX models. Collectively, our findings suggest that patient-derived liver cancer models effectively recapitulate lenvatinib resistance observed in clinical settings and expedite drug discovery for advanced liver cancer, providing a feasible multidimensional platform for personalized medicine.

Multicenter phase 2 study of romidepsin plus lenalidomide for previously untreated peripheral T-cell lymphoma

AbstractPeripheral T-cell lymphomas (PTCLs) are associated with poor prognosis when treated with cytotoxic chemotherapy. We report the findings of a phase 2 study evaluating a chemotherapy-free combination of romidepsin plus lenalidomide as initial treatment for patients with PTCL who were aged >60 years or noncandidates for chemotherapy. Treatment was initiated with romidepsin 10 mg/m2 IV on days 1, 8, and 15 and lenalidomide 25 mg taken orally from days 1 to 21 of 28-day cycle for up to 1 year. The primary objective was overall response rate (ORR). Secondary objectives included safety and survival. The study enrolled 29 patients with a median age of 75 years, including 16 (55%) angioimmunoblastic T-cell lymphoma (AITL), 10 (34%) PTCL– not otherwise specified, 2 ATLL, and 1 EATL. Grade 3 to 4 hematologic toxicities included neutropenia (45%), thrombocytopenia (34%), and anemia (28%). Grade 3 to 4 nonhematologic toxicities included hyponatremia (45%), hypertension (38%), hypoalbuminemia (24%), fatigue (17%), hyperglycemia (14%), hypokalemia (14%), dehydration (10%), and infection (10%). At median follow-up of 15.7 months, 23 patients were evaluable and received a median treatment of 6 cycles. The ORR was 65.2% with complete response (CR) at 26.1%, including 78.6% ORR and 35.7% CR for AITL. Median duration of response was 10.7 months, with 27.1 months for patients achieving CR. The estimated 2-year progression-free survival was 31.5%, and 2-year overall survival was 49.5%. This study provides the first demonstration that the biologic combination of romidepsin and lenalidomide is feasible and effective as initial therapy for PTCL and warrants further evaluation. This trial was registered at www.clinicaltrials.gov as #NCT02232516.

Abstract CT195: Interim safety results from SEPION/AIO-PAK-0118: A multicenter, Phase I/II study of sequential epigenetic and immune targeting in combination with nab-paclitaxel/gemcitabine in patients with advanced pancreatic ductal adenocarcinoma

Abstract Background: Pancreatic ductal adenocarcinoma (PDAC) remains a hard-to-treat cancer entity with limited therapeutic options. We aim to enhance the anti-tumor and pro-immunogenic effect of standard-of-care nab-paclitaxel/gemcitabine (GnP) by concomitant epigenetic targeting followed by immune-modulating maintenance therapy in patients (pts) with treatment-naive metastatic PDAC. This phase 1/2, open-label, multicenter, dose-escalation/dose-expansion study evaluates the safety and tolerability of azacitidine (AZA) and romidepsin (ROM) in combination with GnP and the potential of immune-modulating consolidation with durvalumab and lenalidomide after three cycles of either GnP or GnP plus epigenetic targeting. Here we report interim results of the dose escalation. Methods: In part 1A, dose escalation of three regimen ROM/GnP (arm A), AZA/GnP (arm B) and ROM/AZA/GnP (arm C) for determination of the recommended dose for expansion (RDE) is performed with a 3 + 3 dose escalation scheme using fixed dose levels. For dose expansion (part 1B), one of the treatment arms (Arm C over B over A) is continued using a Simon two-stage design to a maximum of 35 patients. The efficacy of the sequential programmed death-ligand (PD-L)1 blockade in combination with low-dose lenalidomide is evaluated in study part 2 (consolidation part) for pts with controlled disease (DCR) after 3 cycles of part 1 therapy. Evaluation of the consolidation concept in part 2 requires a sufficient DCR based on the statistical considerations. Thus, in addition to pts in part 1A and 1B, pts treated with GnP alone are additionally recruited (standard arm), so that at least 41 pts with controlled disease after 3 cycles of therapy are available for part 2. Key eligibility includes metastatic PDAC (stage IV), ECOG 0-1 and no prior chemotherapy or radiotherapy therapy for stage IV PDAC. Primary objectives include safety and tolerability of AZA and/or ROM in combination with GnP followed by sequential immune targeting with PD-L1 blockade in combination with low-dose lenalidomide. Results: In October 2022, 76 pts were enrolled, of which 67 pts were treated with at least one dose of study medication. The median age in the treated population was 62.8 years (range: 33 - 83), 32 (47.8 %) were female, 80.6 %/13.4 % were included with ECOG 0/1. In part 1A, an RDE of 2mg/m² for ROM (N=3, dose level L-1) and 30mg/m² for AZA (N=6, dose level L1) was established, respectively. The combined administration of AZA (30 mg/m²), ROM (2 mg/m²) and GnP (arm C) led to severe DLTs, namely neutropenia, non-hematologic toxicities (> grade2) and ROM related non-hematologic toxicities (≥ grade2) in all three patients. Given the best safety profile, Arm B was chosen for the expansion part 1B. Conclusions: ROM in combination with GnP showed considerable toxicity at dose level L1 as did the combination of ROM (dose level L-1) and AZA (dose level L1) with GnP. The combination of 30 mg/m2 AZA with GnP was feasible and chosen for further expansion. Dose expansion (part 1B) and sequential immunotherapy consolidation (part 2) are ongoing. Clinical trial information: NCT04257448. Research Sponsor: GWT-TUD GmbH. Citation Format: Jens T. Siveke, Marianne Sinn, Stefan Boeck, Gabriele Siegler, Thomas Seufferlein, Joerg Trojan, Dirk Waldschmidt, Alexander Koenig, Dorothea Schipp, Ines Winkler, Sven Thorsten Liffers, Volker Kunzmann. Interim safety results from SEPION/AIO-PAK-0118: A multicenter, Phase I/II study of sequential epigenetic and immune targeting in combination with nab-paclitaxel/gemcitabine in patients with advanced pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT195.

Updated Results of an Investigator-Initiated Phase II Study of Pembrolizumab and Romidepsin for Patients with Relapsed or Refractory T-Cell Lymphoma (TCL) with Survival Analysis

Background: Peripheral T-cell lymphomas (PTCLs) are a heterogeneous and aggressive group of non-Hodgkin lymphomas. For most subtypes, relapsed/refractory PTCL (r/r TCL) have a poor prognosis and newer therapies have not been approved since 2014. This makes it imperative to develop new, effective therapies for PTCL. Individually, Pembrolizumab and Romidepsin have modest activity in (r/r TCL). Based on their mechanism of action, we hypothesized that their combination may cause synergistic priming of the immune system leading to durable responses. Herein, we report the updated results including survival analysis of the phase I/II study in patients with r/r TCL who had disease progression after at least one prior systemic regimen (NCT03278782). METHODS: Between February 2018-April 2022, 38 patients were enrolled in this study (6 patients in phase I and 32 in phase II portions)We reported the safety and efficacy results in the first 20 patients at the 2020 ASH Annual Meeting. The study schedule was Pembrolizumab at 200 mg on day 1 and Romidepsin 14 mg/m2 on days 1 and 8, in a 21-day cycle. The maximum number of cycles was 35. Adverse events were monitored and graded according to the NCI Common Terminology Criteria for Adverse Events version 4.0 guidelines. The primary endpoint for the phase II part is overall response (OR: CR+PR); the 6 patients enrolled in the phase I part were included in the response assessment. Secondary objectives included progression free survival (PFS) and overall survival (OS). Outcome assessment were performed utilizing Lugano Revised Response Criteria. Kaplan-Meier method was used to estimate PFS and OS. Statistical software SAS 9.4 (SAS, Cary, NC) and S-Plus 8.2 (TIBCO Software Inc., Palo Alto, CA) were used. RESULTS: The number of patients in the intention to treat population (ITT) for the primary endpoint of overall response (Table 1) was 38; the median age was 67 with a male predominance (57.9%). Majority of the patients (86.8%) were >60 years. LDH was elevated in 68.4% and 63.2% of patients had Stage III/IV disease. The median PD-L1 expression was 7.5% (IQR 0-30, max 95). Twenty-eight patients completed more than 1 cycle; among the 10 patients who completed only 1 cycle; 6 died within the first month due to poor PS and PD. The most common ≥ grade 3 adverse events were infections (n=11) and thrombocytopenia (n=10). Three patients stopped therapy due to development of immune related adverse events (iRAEs) - Grade 1 cytokine storm (n=1), Grade 3 gastritis (n=1), Grade 4 colitis (n=1); 2 patients with Grade 2 pneumonitis continued treatment after resolution with steroids. Two patients experienced hyper-progression within the first 10 days of treatment. The interim CR at the end of 3 cycles was 28.9% and final CR at the end of 6 cycles was 34.2% with a final ORR of 39.5%. All 5 patients with iRAEs remain in CR. At a median follow-up of 33.5 months, the median OS was 52.4 months (95% CI: 28 ~ NA months); 23 patients (60.5%) remain alive. The median PFS time was 29.5 months (95% CI: 14.7 ~ NA months); the number of events were 10, 7, 3, 2, 1 in the first 5 years respectively. 2 patients were successfully bridged to allogeneic SCT. There were more responders in other races (57%) compared to whites (34%). CONCLUSIONS: The combination of Romidepsin and Pembrolizumab leads to high response rates and prolonged remissions. The combination improves PFS and OS compared to approved single agents in patients with r/r TCL. Additional studies evaluating the role of the tumor microenvironment will be reported at the Annual Meeting. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Preclinical Validation of EZH2 and HDAC I Dual Inhibition As a Potent Therapy for Refractory Myeloid Leukemia Associated with Down Syndrome

Children with Down Syndrome who are less than 5 years old have approximately 400-fold increased risk of developing myeloid leukemia. Even though children with myeloid leukemia associated with Down syndrome (DS-ML) are more sensitive to chemotherapy than their non-DS-ML counterpart, relapse and treatment-related toxicity needs to be eliminated to achieve better outcome for DS-ML children. Epigenetic alterations including DNA methylation and histone modifications play a key role in the pathogenesis of DS-ML. Enhancer of zeste homolog 2 (EZH2) is one of the epigenetic regulators that harbors high frequency of mutations in DS-ML. EZH2 is a catalytic subunit of PRC2 complex which suppresses gene expression by di- and tri-methylation of lysine27 on histone 3 (H3K27me2/3). EZH2 recruits class I histone deacetylases (HDAC, which remove acetyl groups from histones), on target genes for further suppression of gene expression. Using patient-derived xenograft models of DS-ML, we showed previously that combination of DNA hypomethylating agent azacitidine with HDAC inhibitor panobinostat or BCL2 inhibitor venetoclax showed significant improvement in median survival. Here we aimed to show synergistic effect of EZH2 inhibitor, GSK126, and class I HDAC inhibitor, romidepsin. The viability of CMK, a DS-ML cell line, and NTPL-60 and NTPL-386, in house generated DS-ML PDX lines, was determined by exclusion of propidium iodide-stained cells by flow cytometry. IC50 values for GSK126 and romidepsin were calculated using GraphPad Prism (dose-response curve). The IC50 of GSK126 was 40.5, 18.0, and 17.2 mM, while that of romidepsin was 10.2, 9.2, and 3.0 nM respectively. To determine synergy, the viability of cells exposed to a variety of concentrations of GSK126 and romidepsin alone or in combination was measured. Synergy index was calculated using relative risk ratio (RRR) formula as described previously (Quagliano et al, 56:36, Leuk Res, 2017). All the 3 DS-ML lines showed RRR value of less than 1 indicating synergistic inhibition of cell death by GSK126 and romidepsin treatment in ex vivo culture (Figure 1A). NTPL-386, a refractory DS-ML PDX line, which showed minimal response to chemotherapy (11-day improvement in median survival, Barwe et al., 134:2683, Blood, 2019), was chosen to determine the in vivo efficacy of GSK126 and romidepsin combination. NTPL-386 engrafted mice (n=5 each) were treated with GSK126 (100 mg/Kg) and/or romidepsin (2 mg/Kg) twice weekly for three weeks. Thus, one treatment cycle consisted of six doses. A separate cohort of mice were treated with four cycles of the combination, with two weeks rest between cycles. Disease progression was monitored by percentage of human cells in peripheral blood by flow cytometry. Kaplan-Meier survival estimates were plotted based on the time when mice were euthanized because they met pre-determined experimental endpoints. Compared to untreated mice, GSK126 or romidepsin treatment showed an increase in median survival by 8 days and 11 days respectively (Figure 1B, P <0.005). The combination of these drugs showed a remarkable improvement in survival (41 days). The difference in the median survival of the mice treated with the combination was statistically significant when compared to GSK126 or romidepsin (P <0.005). Further treatment for three additional cycles showed sustained remission upto 150 days post cell injection, when the study was terminated. Prolonged treatment showed a statistically significant improvement in survival compared to one treatment cycle (P <0.05). Further studies to characterize the mechanism of synergy are in progress. In summary, our data show a synergistic effect of EZH2 and HDAC class I inhibition on DS-AML growth and pave the path for clinical evaluation of this combination. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

HDAC inhibitor Chidamide, but not romidepsin, synergizes with humanized PD1 antibody to enhance T-cell chemokine expression and augment IFN-gamma response in NK-T-cell related lymphoma: A preclinical study.

e14568 Background: The incidence of NK-T cell lymphoma (NKTL) in Asia is significantly higher than that in European and American countries. The median survival time of EKTL is only 8-12 months, and the 5-year overall survival rate is still less than 30%. New therapies or combinations are warranted. Methods: The murine NKTL cell line EL4 and RMA were used to establish cell line xenograft models in transgenic humanized PD1 mice. Different dosages or sequential HDAC inhibitors were combined with humanized PD1 antibody in vivo to test whether the synergistic effect is existing. RNA-seq, qPCR, and IHC were performed to illustrate the potential mechanisms. Three NTKL patients who were treated with hPD1 and Chidamide in clinical were recorded in this study. Results: Romidepsin combined with hPD1 antibody is superior to hPD1 antibody alone in the treatment of two NKTL CDX models (EL4 and RMA), but it is similar with romidepsin single agent, either in different dosages or sequence. However, Chidamide combined with hPD1 antibody significantly inhibits the tumor growth compared to single agents in these two pre-clinical models. RNA-seq reveals Chidamide synergizes with humanized PD1 antibody to enhance T-cell chemokine expression, augment IFN-gamma response and increase CD8 T cell infiltration. String PPI analysis and ingenuity pathway analysis suggest IFN-gamma is the hub upstream regulator in the combination. On the contrary, the romidepsin combinations decrease the activity of T cell immune responses and the function of immune cell migration. The expression of IFN-gamma gene set signatures is decreased in the romidepsin combination. Three NKTL patients who received PD1 antibody and Chidamide are showed promising efficacy, and the progression-free survival is 26+ months, 8 months, and 9+ months respectively. Conclusions: Different HDAC inhibitor shows different combination effects with immunotherapy. Chidamide combination shows a synergistic effect with immunotherapy by enhancing INF-gamma response, but romidepsin combination antagonizes this effect significantly.

Azacytidine and Decitabine Exhibit Differential Effects on Cytotoxicity and Methylation and Exhibit Class Synergy with Hdaci in Models of Ptcl

Introduction: The PTCL represent a challenging group of lymphoid malignancies with a poor outcome. The PTCL appear to be the prototypical ‘epigenetic disease’, based on the following observations: (1) it is the one disease for which HDAC inhibitors have been shown to have universal activity; (2) multiple lines of evidence have begun to establish that the PTCL are characterized by recurrent mutations in important genes governing genome wide methylation, including TET2, IDH1/2, and DNMT3. While commonly invoked as a reason to explain sensitivity to HDAC inhibitors, the fact is only hypomethylating agents and not HDAC inhibitors, would be expected to affect this underlying biology.Methods: We have compared Azacitidine (AZA) and Decitabine (DEC) activities in a panel of seven cell lymphoma (TCL) cell lines (H9, HH, P12, PF382, C5MJ, TLMo1 and MT-2) with regard to their dose response effects on cell viability, apoptosis, DNMT protein expression and DNA methylation. We have also evaluated the preclinical merits of combining the hypomethylating (HoMe) agents and HDAC inhibitors (Romidepsin and Belinostat) using cytotoxicity assay (CellTiter Glo). We have performed RNASeq analysis on TCL cell lines to identify unique gene sets perturbed by the AZA-Romidepsin combination.Results: AZA and DEC demonstrated different effects on cell viability, apoptosis, DNMT levels and DNA methylation. Significant heterogeneity in DEC sensitivity was seen among the lines studies, which was not noted with AZA exposure. AZA was consistently more potent than DEC at high drug concentration (> 1μM) but DEC was active at lower drug concentration (< 0.1μM). The difference in activity was mirrored by the dose dependent effects of AZA and DEC on markers of apoptosis. A strong correlation between DNMT depletion and DNA methylation was noted for both drugs, though the concentration:effect relationships were different. Although decitabine was more effective at lower concentration a more sustained depleting effect on the DNMT1 expression was observed in azacitidine treated cells when compared to the decitabine treated cells. Among the synergistic interactions between the two classes of drugs (HoMe and HDACi), the combination of romidepsin or belinostat plus decitabine was more synergistic than the combination of romidepsin or belinostat plus azacitidine. Not all cell lines exhibit the same degree of toxicity with the combination, with synergy being most notable in HH and H9, while MT-2 appears no better than additively cytotoxic. Moreover, not all cell line lines exhibit the same degree of toxicity upon exposure to romidepsin in combination with repetitive addition of azacitidine over a 96 hour time period. However > 50% hypometylation was observed in all seven cell lines at concentration of 250 nM of azacitidine indicating the synergy was mainly dependent on HDACi and not HoMe agent concentrations. Gene expression profile analysis of untreated and combination treated TCL cell lines estimated that 38, 409 and 1215 genes were differentially expressed when untreated cells were compared to romidepsin, azacitidine and combination treated cells respectively. Statistically over-representation of down-regulated genes related to the cholesterol (8.77 fold) and steroid (6.68 fold) metabolic process and upregulated genes related to gamete generation (5.96 fold) was identified in the combination gene signature. In particular, the expression of 39 genes coding for cancer testis antigens was found upregulated.Conclusions: These data suggest that HoMe based combinations are active and synergistic in models of TCL. Concentrations that induce DNMT loss may be a valuable biomarker in identifying optimal doses for clinical development. These data established important differences between the HDAC inhibitors and hypomethylating agents. Assumptions these drugs within a class are identical is inaccurate, though clinically they have been hard to differentiate. GEP reveals they affect vast areas of cellular biology, and is likely no singular mechanism will explain the basis for synergy. However, key information with regard to the emergence of cancer testes antigen expression may suggest future rational combinations. DisclosuresO'Connor:Trillium Therapeutics: Research Funding; Celgene: Honoraria, Research Funding.

A Phase 1 Trial of Alisertib and Romidepsin for Relapsed/Refractory Aggressive B-Cell and T-Cell Lymphomas

A Phase 1 Trial of Alisertib and Romidepsin for Relapsed/Refractory Aggressive B-Cell and T-Cell Lymphomas

A Multicenter Phase I Dose-Finding and Preliminary Efficacy Study of the Histone Deacetylase Inhibitor Romidepsin in Combination with Pegylated Liposomal Doxorubicin for the Treatment of Adults with Relapsed or Refractory Cutaneous and Peripheral T-Cell Lymphomas

IntroductionLiposomal doxorubicin and romidepsin have single agent activity in PTCL and CTCL. In pre-clinical studies, we demonstrated that romidepsin had synergistic effects in combination with doxorubicin. The aim of this study is to determine the safety and feasibility of the combination of romidepsin and liposomal doxorubicin in patients with relapsed/refractory CTCL and PTCL.Materials and Methods:This is a multi-center, single arm, open-label phase I dose-escalation study. Patients with relapsed/refractory CTCL (stage IIA-IV), or PTCL of any stage or subtype who received at least one systemic therapy are eligible. The primary endpoint is to identify the maximum tolerated dose (MTD) of romidepsin in combination with liposomal doxorubicin and the secondary endpoint is to determine the overall response rate (ORR) of this combination. Patients were treated with liposomal doxorubicin 20 mg/m2 on day 1 and romidepsin at four dose levels on days 1, 8 and 15, every 28 days, until 2 cycles beyond the best response, 8 cycles, disease progression (PD), or intolerability; whichever comes first. Dose levels of romidepsin include 8, 10, 12 and 14 mg/m2. Dose limiting toxicity (DLT) is defined as any adverse event during therapy including: grade 5 toxicity, any grade 4 hematologic toxicity lasting >7 days (except neutropenia), or any grade 4 non-hematologic toxicity according to CTCAE version 4.0. The MTD was defined as the highest tested dose level where two or more participants experienced a DLT utilizing a standard 3+3 design. Response was evaluated using the Cheson criteria in PTCL and patients with nodal disease and the mSWAT was utilized in CTCL. Pre-specified in the protocol, after determination of the MTD of romidepsin, up to 12 patients will be treated in a dose-expansion cohort if efficacy is noted.Results:Fourteen patients (5 PTCL, 9 CTCL) were treated at four dose-levels with a median 4 cycles (range 1-8) of therapy. Clinical data regarding these patients is included in Figure 1. Grade 3/4 non-hematologic adverse events occurred in 4 patients including 3 infection, 1 fatigue and 1 atrial fibrillation. Four patients experienced grade 3/4 hematologic toxicities: 1 neutropenia, 2 thrombocytopenia and 3 anemia. The MTD of romidepsin was determined to be 12 mg/m2 and the dose limiting toxicity was prolonged (>7 days) grade 4 thrombocytopenia. The ORR was 57%, including 2 complete responses and 6 partial responses.Given these results, three additional patients have been treated in a pre-specified dose-expansion cohort. For the 17 total patients (including 3 additional patients treated at the MTD), the ORR was 52%. In CTCL the ORR was 78% (1 CR, 6 PR, 2 PD) and among PTCL patients the ORR was 25% (1 CR, 1 PR, 1 PD).Conclusion:Here we report the results of the dose-escalation study of the combination of romidepsin and liposomal doxorubicin with preliminary data on the ORR. This combination has an acceptable toxicity profile at an MTD of 12 mg/m2 for romidepsin and has promising efficacy in both relapsed/refractory PTCL and CTCL. An additional eight patients will be included in a dose-expansion cohort. [Display omitted] DisclosuresAi:Celgene: Research Funding. Porcu:Kiowa: Research Funding; Miragen: Research Funding; Tetralogic: Research Funding; Celgene: Research Funding; Innate Pharma: Research Funding; Cell Medica: Research Funding; Galderma: Research Funding; Kura: Research Funding. Wieduwilt:Sigma-Tau: Research Funding; Reata Pharmaceuticals: Equity Ownership. Brammer:Celgene: Research Funding.

Simvastatin-romidepsin combination kills bladder cancer cells synergistically

The HMG-CoA reductase inhibitor simvastatin activates AMP-activated protein kinase (AMPK) and thereby induces histone acetylation. We postulated that combining simvastatin with the histone deacetylase (HDAC) inhibitor romidepsin would kill bladder cancer cells by inducing histone acetylation cooperatively. The combination of romidepsin and simvastatin induced robust apoptosis and killed bladder cancer cells synergistically. In murine subcutaneous tumor models using MBT-2 cells, a 15-day treatment with 0.5 mg/kg romidepsin and 15 mg/kg simvastatin was well tolerated and inhibited tumor growth significantly. Mechanistically, the combination induced histone acetylation by activating AMPK. The combination also decreased the expression of HDACs, thus further promoting histone acetylation. This AMPK activation was essential for the combination's action because compound C, an AMPK inhibitor, suppressed the combination-induced histone acetylation and the combination's ability to induce apoptosis. We also found that the combination increased the expression of peroxisome proliferator-activated receptor (PPAR) γ, leading to reactive oxygen species production. Furthermore, the combination induced endoplasmic reticulum (ER) stress and this ER stress was shown to be associated with increased AMPK expression and histone acetylation, thus playing an important role in the combination's action. Our study also suggests there is a positive feedback cycle between ER stress induction and PPARγ expression.

Multicenter phase II study of romidepsin plus lenalidomide for patients with previously untreated peripheral T-cell lymphoma (PTCL).

7514 Background: PTCL are aggressive malignancies associated with poor prognosis when treated with cytotoxic chemotherapy. Novel agents, such as HDAC inhibitor romidepsin and immunomodulatory agent lenalidomide, have shown clinical activities as single agents and in combination in R/R PTCL. We hypothesize that upfront treatment with these agents is an effective and well-tolerated option to defer chemotherapy, particularly in patients who are not candidates for intensive approach. We report the findings of the first chemo-free combination of romidepsin plus lenalidomide as initial treatment for PTCL (ClinicalTrials.gov-NCT02232516). Methods: Patients with untreated PTCL who were over 60 or noncandidates for chemotherapy based on comorbidity CIRS score were eligible. Treatment was initiated with romidepsin 10 mg/m2 IV on d 1, 8, 15, and lenalidomide 25 mg PO on d 1-21 of 28-day cycle for up to 1 year, unless discontinued prior due to POD, toxicities, or withdrawal of consent. The primary objective was to evaluate ORR per Cheson criteria. Secondary objectives included safety, PFS, OS, DOR, and delay to chemotherapy. The sample size was 20 evaluable patients, which allows to estimate the underlying true response rate with the margin of error of an approximate 95% confidence interval equal to 0.22, assuming the true ORR = 0.5. Results: The study enrolled 29 subjects at 3 US centers, including 16 (55%) AITL, 11 (38%) PTCL-NOS, 1 ATLL and 1 EATCL. The median age was 75 (range 49-84), and M:F ratio was 1:1. Nineteen (66%) had stage III/IV disease, 23 (79%) had elevated LDH, and 9 (31%) had IPI 3-5. Treatment was well tolerated with expected side effects. Grade 3-4 hematologic toxicities included neutropenia (45%), thrombocytopenia (34%) and anemia (28%). Grade 3-4 non-hematologic toxicities included hyponatremia (45%), hypertension (38%), hypoalbuminemia (24%), fatigue (17%), hyperglycemia (14%), hypokalemia (14%), dehydration (10%), lung infection (10%) and sepsis (10%). At a median follow-up of 8 months, 20 subjects were evaluable with at least one response assessment, and received a median treatment of 6 cycles. The ORR was 75% (95%CI: 50.9%, 91.3%) with CR at 30% (11.9%, 54.3%). For AITL, the ORR was 85% (54.6%, 98.1%) with CR at 38% (13.9%, 68.4%). Median DOR was 4.2 months for all responders, and 14.3 months for CR patients. The estimated 1-yr PFS was 54.3% with 3-yr PFS at 36.2%, and the estimated 1-yr OS was 76.0% with 3-yr OS at 51.3%. Two subjects moved onto consolidative ASCT in remission, and 4 received additional cytotoxic chemotherapy after progression. Conclusions: This study provides the first demonstration that chemo-free biologic combination of romidepsin and lenalidomide is feasible and effective as initial therapy for PTCL patients who are not candidates for cytotoxic chemotherapy. These data justify further evaluation of such novel agents as a frontline strategy. Clinical trial information: NCT02232516.

Romidepsin (HDACi) plus cisplatin and nivolumab triplet combination in patients with metastatic triple negative breast cancer (mTNBC).

1076 Background: Histone deacetylase inhibitors (HDACi) upregulate genes involved in antigen presentation machinery and increase expression of natural killer group 2, member D ligands (NKG2DL), thus resulting in enhanced tumor cell recognition and response to PD-1/CTLA-4 blockade. Cisplatin and HDACi combination synergistically induces cytotoxicity, apoptosis, and DNA damage. This phase I-II trial investigated combination of romidepsin (HDACi) plus cisplatin and nivolumab (PD-1 inhibitor) in mTNBC. Patients and Methods: Eligible patients had mTNBC with any number of prior chemotherapies. Phase I was 3+3 dose-escalation design with three dose levels of romidepsin (8, 10, 12mg/m2, D2, 9) plus cisplatin 75mg/m2 D 1 every 21 days. Phase II treatment included romidepsin plus cisplatin plus nivolumab 360mg every 21 days and was designed according to Simon’s two stage minimax design. Primary endpoints were recommended phase 2 dose (RP2D) and objective response rate (ORR). Additional endpoints included safety, PFS, and pharmacokinetics. Results: 51 patients were enrolled (N=13 phase I, N=38 phase II) between 2015-2020. 69% had received ≥1 prior metastatic chemotherapy, 47% had prior platinum, 53% had liver metastasis, 12% had BRCA1/2 mutation, and 11% had PD-L1 positive disease. There were no dose limiting toxicities in phase I. The RP2D was romidepsin 12mg/m2 D2,9 + cisplatin 75mg/m2 D1 + nivolumab 360mg D1 every 21 days. Thrombocytopenia (G3:27%, G4:0%), neutropenia (G3:25%, G4:0%), anemia (G3:22%, G4:0%), nausea (G3:22%, G4:0%), and vomiting (G3:20%, G4:0%) were the most common grade 3/4 adverse events. 21% of patients had immune AEs (G3-4:8%). Among 34 evaluable phase II patients, ORR was 44% (Table), median PFS was 4.4 months, and 1-year PFS was 23%. Median OS was 10.3 months and 1-year OS was 43%. No pharmacokinetic interactions were detected with co-administration of romidepsin-cisplatin-nivolumab. Conclusions: The triplet combination of romidepsin plus cisplatin and nivolumab was well tolerated and shows encouraging efficacy in pretreated mTNBC, including in patients with PD-L1 negative disease and in those with liver metastasis. Correlative biomarker work is ongoing. This combination warrants further evaluation in larger studies. Clinical trial information: NCT02393794 .[Table: see text]

Tamoxifen enhances romidepsin-induced apoptosis in T-cell malignant cells via activation of FOXO1 signaling pathway

Although romidepsin as monotherapy appears to be useful for treating T-cell lymphoma, combined chemotherapy with other therapeutic agents is required for improvement of the treatment outcome. To establish safer and more effective regimens, systematic screening was conducted to identify suitable drugs to be used in combination with romidepsin for T-cell malignancies, and the underlying molecular mechanisms were examined. The most effective agent was tamoxifen. The combination of romidepsin and tamoxifen had a significant synergistic effect in inducing apoptosis. The growth-inhibitory effects of the combined treatment were reversed by α-tocopherol. FOXO1 expression was greatly upregulated in MOLT-4 cells treated with romidepsin plus tamoxifen. Knockdown of FOXO1 expression by siRNA significantly reduced the cell death induced by romidepsin plus tamoxifen. The combination of romidepsin and tamoxifen might be considered for the treatment of T-cell lymphoma patients.

A Phase I Trial Assessing the Feasibility of Romidepsin Combined with Brentuximab Vedotin for Patients Requiring Systemic Therapy for Cutaneous T-Cell Lymphoma

A Phase I Trial Assessing the Feasibility of Romidepsin Combined with Brentuximab Vedotin for Patients Requiring Systemic Therapy for Cutaneous T-Cell Lymphoma

A Phase II Study of Pembrolizumab in Combination with Romidepsin Demonstrates Durable Responses in Relapsed or Refractory T-Cell Lymphoma (TCL)

A Phase II Study of Pembrolizumab in Combination with Romidepsin Demonstrates Durable Responses in Relapsed or Refractory T-Cell Lymphoma (TCL)

Protein-Driven Mechanism of Multiorgan Damage in COVID-19

We propose a new plausible mechanism by mean of which SARS-CoV-2 produces extrapulmonary damages in severe COVID-19 patients. The mechanism consist on the existence of vulnerable proteins (VPs), which are (i) mainly expressed outside the lungs; (ii) their perturbations is known to produce human diseases; and (iii) can be perturbed directly or indirectly by SARS-CoV-2 proteins. These VPs are perturbed by other proteins, which are: (i) mainly expressed in the lungs, (ii) are targeted directly by SARS-CoV-2 proteins, (iii) can navigate outside the lungs as cargo of extracellular vesicles (EVs); and (iv) can activate VPs via subdiffusive processes inside the target organ. Using bioinformatic tools and mathematical modeling we identifies 26 VPs and their 38 perturbators, which predict extracellular damages in the immunologic endocrine, cardiovascular, circulatory, lymphatic, musculoskeletal, neurologic, dermatologic, hepatic, gastrointestinal, and metabolic systems, as well as in the eyes. The identification of these VPs and their perturbators allow us to identify 27 existing drugs which are candidates to be repurposed for treating extrapulmonary damage in severe COVID-19 patients. After removal of drugs having undesirable drug-drug interactions we select 7 drugs and one natural product: apabetalone, romidepsin, silmitasertib, ozanezumab, procaine, azacitidine, amlexanox, volociximab, and ellagic acid, whose combinations can palliate the organs and systems found to be damaged by COVID-19. We found that at least 4 drugs are needed to treat all the multiorgan damages, for instance: the combination of romidepsin, silmitasertib, apabetalone and azacitidine.

Abstract CT160: The integration of PD1 blockade with epigenetic therapy is highly active and safe in heavily treated patients with T-cell lymphoma

Abstract Our group has demonstrated that combinations of epigenetic modifiers produce potent synergy in pre-clinical models of PTCL, and has confirmed this data in early phase clinical studies. We showed that the combination of romidepsin and hypomethylating agents, decitabine or azacytidine, uniquely induces the expression of genes, including cancer testis antigens, gamma interferon and endogenous retrovirus. We are conducting a phase 1 study of pembrolizumab combined with pralatrexate alone (Arm A), with pralatrexate + decitabine (Arm B), or decitabine alone (Arm C) in patients with peripheral T-cell lymphoma (PTCL) and cutaneous T-cell lymphoma. In addition, we are conducting a Phase 1/2A study of durvalumab combined with oral 5-azacitidine + romidepsin (Arm A), pralatrexate + romidepsin (Arm B), romidepsin (Arm C), or oral 5-azacitidine (Arm D) in patients with PTCL. Patient characteristics are listed in Table 1. We have treated a total of 12 patients across the 2 trials and 4 different Arms including pralatrexate plus pembrolizumab (3 patients), pralatrexate, decitabine and pembrolizumab (3 patients), decitabine and pembrolizumab (3 patients) and azacytidine, romidepsin and durvalumab (3 patients). Twelve patients out of 12 received at least one dose of drug and were evaluable for toxicity. There was a dose limiting toxicity (DLT) for each arm including prolonged grade 3 thrombocytopenia, febrile neutropenia, grade 3 hyponatremia and rash, and cytokine release syndrome, respectively. There were no treatment-related deaths. Eight patients out of 12 are evaluable for response at the time of this analysis. Three out of 8 patients achieved a complete remission, 3 out of 8 patients had a partial remission, and 2 out of 8 patients experienced progression of disease. Interestingly, all of the responses were seen in the triple combination of pralatrexate, decitabine, pembrolizumab and azacytidine, romidepsin and durvalumab. These preliminary clinical data suggest that the addition of immune-checkpoint inhibitors on an epigenetic backbone is safe and demonstrates encouraging responses in patient with PTCL and CTCL. These trials are registered at www.clinicaltrials.gov (NCT03240211, NCT03161223). Patient characteristics (n=12)Median age (range)68 (26-77)Sex, n (%) Male Female6 (50) 6 (50)Race, n (%)White/Non-Hispanic White/Hispanic Black Asian Unknown5 (42) 1 (8) 3 (25) 1 (8) 2 (17)Histology, n (%) PTCL, NOS AITL Mycosis Fungoides PTCL, NOS TFH ATLL Sezary Syndrome ALCL, ALK+2 (17) 2 (17) 2 (17) 1 (8) 1 (8) 1 (8) 1 (8)Stage at diagnosis IB II III IV Unknown1 (8) 2 (17) 3 (25) 4 (33) 2 (17)Median number of prior therapies (range) **2 unknown3 (0-5) Citation Format: Enrica Marchi, Helen Ma, Francesca Montanari, Ahmed Sawas, Jennifer K. Lue, Changchun Deng, Kareema T. Whitfield, Sandra Klein, Matthew V. Matthew, Freedy J. Loffredo, Luigi Scotto, Hyejung Lee, Hye A. Kim, Alice T. Jacobs, Aishling M. Rada, Karen A. Khan, Salvia Salvia, John Lister, Nora N. Benanni, Mark A. Francescone, Pier Luigi Zinzani, Wonseog Kim, Owen A. O'Connor. The integration of PD1 blockade with epigenetic therapy is highly active and safe in heavily treated patients with T-cell lymphoma [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 CT160.

Romidepsin (FK228) regulates the expression of the immune checkpoint ligand PD-L1 and suppresses cellular immune functions in colon cancer

Romidepsin (FK228), a histone deacetylase inhibitor (HDACi), has anti-tumor effects against several types of solid tumors. Studies have suggested that HDACi could upregulate PD-L1 expression in tumor cells and change the state of anti-tumor immune responses in vivo. However, the influence of enhanced PD-L1 expression in tumor cells induced by romidepsin on anti-tumor immune responses is still under debate. So, the purpose of this study was to explore the anti-tumor effects and influence on immune responses of romidepsin in colon cancer. The results indicated that romidepsin inhibited proliferation, induced G0/G1 cell cycle arrest and increased apoptosis in CT26 and MC38 cells. Romidepsin treatment increased PD-L1 expression in vivo and in vitro via increasing the acetylation levels of histones H3 and H4 and regulating the transcription factor BRD4. In subcutaneous transplant tumor mice and colitis-associated cancer (CAC) mice, romidepsin increased the percentage of FOXP3+ regulatory T cells (Tregs), decreased the ratio of Th1/Th2 cells and the percentage of IFN-γ+ CD8+ T cells in the peripheral blood and the tumor microenvironment. Upon combination with an anti-PD-1 antibody, the anti-tumor effects of romidepsin were enhanced and the influence on CD4+ and CD8+ T cells was partially reversed. Therefore, the combination of romidepsin and anti-PD-1 immunotherapy provides a more potential treatment for colon cancer.