Cell Death Dis Research Articles

Retracted] Glutathione peroxidase 2 overexpression promotes malignant progression and cisplatin resistance of KRAS‑mutated lung cancer cells.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that certain of the BrdU incorporation assay data shown in Fig. 7C were strikingly similar to data that had already appeared in another article written by different authors at different research institutes [Lu M, Qin X, Zhou Y, Li G, Liu Z, Geng X and Yue H: Long non‑coding RNA LINC00665 promotes gemcitabine resistance of Cholangiocarcinoma cells via regulating EMT and stemness properties through miR‑424‑5p/BCL9L axis. Cell Death Dis 12: 72, 2021]. Owing to the fact that the contentious data in the above article had already been published prior to its submission to Oncology Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Oncology Reports 48: 207, 2022; DOI: 10.3892/or.2022.8422].

Retracted] MicroRNA‑320a suppresses tumour cell proliferation and invasion of renal cancer cells by targeting FoxM1.

Subsequently to the publication of the above paper, the authors drew to the attention of the Editorial Office that they made a couple of errors in terms of the data assembly in Figs. 2 and 4 in their paper; specifically, the Transwell assay data shown for the 'miR-320a+/FoxM1+' panel in Fig. 5D on p. 1923 also appeared as the 'ACTN/NC' data panel in Fig. 4E on the same page (Fig. 4E contained the erroneously duplicated panel). In addition, data featured in Fig. 2D of the above paper were strikingly similar to data that appeared in Fig. 6e of the following paper, published subsequently to this article, written by different authors (although a Dr Shiyue Zhao worked in the molecular biology laboratory of Harbin Medical University from 2017 to 2018, and the research collaboration was conducted with Dr Chenlong Li's research group): Li C, Zheng H, Hou W, Bao H, Xiong J, Che W, Gu Y, Sun H and Liang P: Long non-coding RNA linc00645 promotes. TGF-β-induced epithelial-mesenchymal transition by regulating miR-205-3p-ZEB1 axis in glioma. Cell Death Dis 10: 17, 2019. Finally, after having conducted an independent investigation of the data in this paper, the Editorial Office noted that one of the Petri dish images in Fig. 2C was also strikingly similar to data that appeared in Fig. 2H of the abovementioned article in the journal Cell Death & Disease. After having considered the authors' request for corrigendum, in view of the problems that were identified with the data, the Editor of Oncology Reports has decided that, owing to a lack of confidence in the presented data, the paper should instead be retracted from the journal. After having informed the authors of this decision, they accepted the decision to retract this paper. The Editor apologizes to the readership for any inconvenience caused. [Oncology Reports 40: 1917‑1926, 2018; DOI: 10.3892/or.2018.6597].

Phase I/II Study to Evaluate the Safety, Feasibility, and Efficacy of FP-1201 (Intravenous Interferon-Beta-1a) to Prevent Toxicities after CD19-Directed CAR T-Cell Therapy: Trial in Progress

BACKGROUND CD19 chimeric antigen receptor (CAR) T-cell therapy has revolutionized the treatment of patients with relapsed/refractory B-cell malignancies. Yet it remains limited by potentially life-threatening toxicities such as CRS and ICANS. The risk of CRS and ICANS restricts the use of CD19 CAR T-cell therapy to large academic centers and leads to high healthcare resource utilization. Current toxicity prevention strategies have shown limited efficacy to date (prophylactic steroids with axi-cel: CRS, 80%; ICANS, 60%; Oluwole, BJH, 2021; early intervention with brexu-cel: CRS, 89%; ICANS, 60%; Shah, The Lancet, 2021). Our group has shown that endothelial activation and dysfunction are associated with the development of CRS and ICANS, suggesting a key role of increased vascular permeability and blood-brain barrier damage in their pathogenesis (Hay, Blood, 2017). Type-I interferons (IFN-alpha and beta) are major regulators of CD73 expression by endothelial cells, an enzyme critical to the maintenance of endothelial integrity. CD73 breaks down extracellular pro-inflammatory ATP into anti-inflammatory adenosine. In addition, extracellular ATP has been shown to be a major and early driver of systemic inflammation upstream from IL-6 production (Cauwels, Cell Death Dis, 2014), which is associated with severe CRS and ICANS. Preclinical models have demonstrated that IFN-beta-1a treatment is associated with reduced capillary and blood-brain barrier permeability via upregulation of CD73 expression (Floris, J Neuroimmunol, 2002; Kraus., Ann Neurol, 2004; Niemela, Eur J Immunol, 2008). In humans, two recombinant IFN-beta-1a therapies are FDA-approved for the treatment of multiple sclerosis. IFN-beta-1a given intravenously (IV) maximizes the drug's bioavailability and its protective effects on the endothelium. In a study of patients with ARDS, IV IFN-beta-1a (FP-1201 and FP-1201-lyo) was safe and induced CD73 expression (Bellingan, Lancet Respir Med 2014). Given the known effects of IFN-beta-1a on preserving endothelial function and blood-brain barrier integrity, we hypothesize that IV IFN-beta-1a (FP-1201) may prevent CRS and ICANS following CD19 CAR T-cell therapy ( Figure 1A). Since higher rates of severe CRS and ICANS have been reported after axi-cel and brexu-cel, which contain CD28-costimulatory domains, we will restrict the inclusion criteria to patients treated with these products to analyze a population with a higher unmet need and a more homogeneous risk of CRS/ICANS. STUDY DESIGN AND METHODS Objectives and endpoints: Primary objective: To evaluate the safety and feasibility of FP-1201 in patients undergoing treatment with axi-cel or brexu-cel with two co-primary endpoints: i) to estimate the incidence of dose-limiting toxicity rates within the first 14 days following the last administration of FP-1201; ii) to study the type, frequency, and severity of adverse events according to the NCI CTCAE v5.0 from the first administration of FP-1201 and until day +28 after CAR T-cell infusion Secondary objectives: i) To decrease the incidence and severity of ICANS; ii) to decrease the incidence and severity of CRS; iii) to decrease corticosteroid usage; iv) to evaluate the effect of FP-1201 on anti-tumor efficacy Exploratory objectives: To characterize the in vivo effects of FP-1201 on endothelial function, the systemic cytokine milieu, and CAR T-cell function Key inclusion criteria: Adults ≥18 years of age with Karnofsky performance status ≥60% and B-cell non-Hodgkin lymphoma eligible for treatment with axi-cel or brexu-cel Key exclusion criteria: Known hypersensitivity to IFN-beta or major organ dysfunction FP-1201 administration: The FDA-approved IFN-beta-1a Rebif® will be formulated for IV administration (FP-1201). Participants will receive FP-1201 at one of four dose levels detailed in Figure 1B. Statistical design and sample size: We will use a Bayesian Optimal Interval Design to guide the FP-1201 dose escalation. We will plan to treat up to 24 participants. SUMMARY This is a phase I/II study of IV IFN-beta-1a in preventing CRS and ICANS following axi-cel and brexu-cel. The aim is to demonstrate safety and efficacy and to investigate the biologic mechanisms of IFN-beta-1a in preventing endothelial dysfunction.Additional preclinical and preliminary clinical data will be presented at the meeting. This study is registered on clinicaltrials.gov as NCT05936229.

Phase I/II Clinical Trial Evaluating Azacitidine + Venetoclax + Donor Lymphocyte Infusion in Post-Transplant Relapse Myelodysplastic Syndromes and Acute Myeloid Leukemia: Preliminary Results of Ventograft, a GFM Study

Background: Azacitidine (AZA) is approved for the treatment of patients with high-risk MDS and in combination to Venetoclax (VEN) in AML. Beyond direct cytotoxic/epigenetic effects of AZA and ability to induce remissions even in patients refractory to chemotherapy, AZA has been shown to increase the immunogenicity of AML blasts by reexpression of key antigens [ Schroeder et al. Leukemia 2013]. Moreover, VEN imparts distinct cell death sensitivity and adaptivity patterns in T cells ( Ludwig et al. Cell Death Dis 2021). DLI have shown efficacy to treat or prevent relapse of AML or MDS following allogeneic stem cell transplantation (allo-SCT) in 20-40% of the cases [ Castagna et al. Transfusion and Apheresis 2016]. A german retrospective study showed an overall response rate (ORR) of 33% (including 27% CR and 6% PR), to AZA and DLI following relapse after allo-SCT in AML and MDS [ Schroeder et al. BBMT 2015], a regimen used since in many centers. We present here preliminary results of a phase I/II evaluating the combination of AZA/VEN/DLI in relapse MDS/AML. Patients and Methods: In this open label multicenter phase I/II study, MDS and AML patients relapsing post allo-SCT, without active graft versus host disease (GVHD) and active infections, are treated with VEN during 14 days in combination with AZA at 75 mg/m²/day for 5 days for each cycle (or 50 mg/m²/day for 5 days each cycle if relapse occurs less than 4 months after allo- SCT), and VEN orally at escalating dose (step 1: VEN 100mg/d; step 2: VEN 200mg/d and step 3: VEN 400mg/d) in the phase I part and at maximal tolerated dose (MTD) on days 1-14 of each cycle in the phase II part. Cycles are administered every 28 day (or delayed until day 42 in case of hematological toxicity). DLI are started at least 8 weeks after immunosuppressive therapy discontinuation, if chimerism in <90% recipient, and administered on day 15 every two cycles (C2, C4, C6, C8) in the absence of GVHD. Patients receive at least 8 cycles of AZA/VEN + DLI unless progression occurs. Response is assessed after 4, 6 and 8 cycles. Patients achieving a response receive 12 cycles of AZA + VEN followed by cycles of AZA alone. In the absence of response after 8 cycles, patients stop treatment. Results: From November 2022 to June 2023, 9 patients were included in part 1 of VENTOGRAFT. M/F was 8/1. Median age 57 years (range 18-71). Two patients had MDS and 7 patients had AML at diagnosis. Five patients had MDS (IPSS-R was low (1), intermediate (2) and very high (2)), and 3 patients had AML (ELN2017 was intermediate (1) and unfavorable (2) and molecular relapse (1)) at the time of relapse. Median interval between allo-SCT and relapse was 44 months (range 2-181). At data cut-off, 3 patients had been treated in step 1, 3 patients in step 2 and 3 patients in step 3 (AZA + VEN 400mg/d D1-14)). Median number of AZA + VEN cycles was 3 (range 1-8) and median number of DLI infused was 1 (range 0-3). One dose limiting toxicity (No ANC recovery on D42 and one patient still neutropenic at D28) was observed in step 3 leading to inclusion 3 more patients on this step. Other adverse events observed were grade 1/2 nausea (2 patients), grade 1 skin lesions (1 patient), ALAT increase grade 3 (1 patient), febrile neutropenia grade 3 (2 patients), grade 3 abdominal pain (1 patient) and grade 3 gastrointestinal bleeding (2 patients). GVHD was seen in 1/9 (11%) patients). CMV and EBV reactivation were observed in 0 and 2 patients, respectively. Overall response rate was 6/9 (67%) patients 67% including 22% complete remission (CR), 22% of CR with incomplete recovery (CRi) and 33% of morphologic leukemia-free state (MLFS). Full donor chimerism was obtained in all patients obtaining a response. Conclusion: The combination of AZA at 75mg/m²/d D1-5 and VEN at 100 to 400mg/d D1-D14 (400mg/d ongoing) was safe, showed a promising ORR of 67%. Final results of the phase 1 and first patients treated in this phase 2 will presented at the ASH meeting.

Retracted] miR‑216b inhibits glioma cell migration and invasion through suppression of FoxM1.

Following the publication of the above article, a concerned reader reported that certain of the cell invasion assay data shown in Fig. 2D, and the mouse images shown in Fig. 6A, were strikingly similar to data that appeared in the following (subsequently published) article: Li C, Zheng H, Hou W, BaoH, Xiong J, Che W, Gu Y, Sun H and Liang P: Long non-coding RNA linc00645 promotes TGF-β-induced epithelial-mesenchymal transition by regulating miR-205-3p-ZEB1 axis in glioma. Cell Death Dis 10: 717, 2019. The authors regret that the data in question featured in the original Oncology Reports article had been re-used, and accept the Editor's decision to retract this article; furthermore, the authors and the Editor apologize to the readership of the Journal for any inconvenience caused. [Oncology Reports 38: 1751‑1759, 2017; DOI: 10.3892/or.2017.5824].

Retraction statement: Luo L, Liu H and Xi Q (2018) Trastuzumab induces PUMA-dependent apoptosis and inhibits tumor growth in gastric cancer. FEBS Open Bio 8, 1911-1919.

FEBS Open BioEarly View Retraction NoticeOpen Access Retraction statement: Luo L, Liu H and Xi Q (2018) Trastuzumab induces PUMA-dependent apoptosis and inhibits tumor growth in gastric cancer. FEBS Open Bio 8, 1911–1919. This article retracts the following: Retracted: Trastuzumab induces PUMA-dependent apoptosis and inhibits tumor growth in gastric cancer Linghe Luo, Haiyan Liu, Qian Xi, Volume 8Issue 12FEBS Open Bio pages: 1911-1919 First Published online: November 19, 2018 First published: 17 March 2023 https://doi.org/10.1002/2211-5463.13587AboutSectionsPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat https://doi.org/10.1002/2211-5463.12522 The above article, published online on 15 September 2018 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the Editor-in-Chief, FEBS Press, and John Wiley and Sons Ltd. The retraction has been agreed following an investigation into concerns raised by a third party, which revealed inappropriate duplications between this and other published articles [[1-4]]. Thus, the editors consider the conclusions of this manuscript substantially compromised. References 1Wang X, Xue Q, Wu L, Wang B and Liang H (2018) Dasatinib promotes TRAIL-mediated apoptosis by upregulating CHOP-dependent death receptor 5 in gastric cancer. FEBS Open Bio 8, 732– 742. 2Chen G, Chen Z and Zhao H (2020) MicroRNA-155-3p promotes glioma progression and temozolomide resistance by targeting Six1. J Cell Mol Med 24, 5363– 5374. 3Tian T, Guo T, Zhen W, Zou J and Li F (2020) BET degrader inhibits tumor progression and stem-like cell growth via Wnt/β-catenin signaling repression in glioma cells. Cell Death Dis 11, 900. 4Qu H, Song X, Song Z, Jiang X, Gao X, Bai L, Wu J, Na L and Yao Z (2020) Berberine reduces temozolomide resistance by inducing autophagy via the ERK1/2 signaling pathway in glioblastoma. Cancer Cell Int 20, 592. Early ViewOnline Version of Record before inclusion in an issue ReferencesRelatedInformation

Metformin Inhibits STAT3 and MAPK Signaling through AMPK Activation in CSF3RT618I Cells

Introduction: In the light of the discovery of the CSF3R mutation in chronic neutrophilic leukemia (CNL) patients, the T618I mutation demonstrated to be a molecular marker and a possible target for new therapies due to the JAK/STAT signaling pathway activation [N Engl J Med (2013) 368(19):1781-90]. Since no curative treatment options are available for patients diagnosed with CNL, it is of interest the study new therapeutical approaches for these patients. In this regard, the treatment of JAK2V617F murine models with metformin, an FDA-approved drug for type 2 diabetes, reduced splenomegaly, extramedullary hematopoiesis, and colony formation of HSPC [Cell Death Dis. 2018;9(3):311]. Hence, we characterized the preclinical cellular and molecular effects of metformin in CSF3RT618I models. Methods: The in vitro experiments were performed in Ba/F3 cells transduced retrovirally with CSF3RT618I overexpression (OE) model. Cells were treated with metformin (1-20 mM) to access cell viability and survival. Protein expression was evaluated by western blot. For in vivo evaluation, eight-week-old female NSG mice were transplanted with 2x106 Ba/F3 CSF3RT618I cells, resuspended in Matrigel: PBS (1:1 ratio). After the tumor formation (2 days), mice were treated for 10 days by intraperitoneal injection of vehicle (PBS) or metformin (125 mg/kg, n=10/group). Tumor volume was measured using the formula (V = W2 × L × 0.52), where W and L are the smallest and largest diameters, respectively. Ex vivo primary cells from a patient diagnosed with CNL harboring the CSF3RT618I mutation were used to assess the effects of metformin treatment. The primary bone marrow mononuclear cells were evaluated regarding cell survival and clonogenic capacity upon metformin treatment. All experiments were approved by the Animal and Human Ethics Committee of the Institution. Statistical analyses were performed by Mann-Whitney or ANOVA test and Bonferroni post-test. A p-value < 0.05 was considered. Results: After 24 and 48 hours, Ba/F3 CSF3RT618I treated with 1, 5, 10 and 20 mM of metformin presented a reduction in cell viability in a dose- and time-dependent manner (p < 0.01). The IC50 for metformin treatment for 24, 48, and 72 hours was 17.44, 1.3, and 3.2 mM, respectively. Increased frequency of apoptotic cells was observed at 5, 10, and 20 mM of metformin at times of 24, 48, and 72 hours (p < 0.01). Molecularly, CSF3RT618I cells displayed reduced STAT3 and ERK phosphorylation and increased AKT/mTOR activation, upon metformin 5 mM treatment for 24 hours. AMPK activation was observed in cells treated with 1-20 mM of metformin compared to vehicle. In vivo, metformin treatment decreased the tumor burden of CSF3RT618I transplanted mice after 9 days of treatment (vehicle versus metformin mean ± SEM: 731.9±90.14 vs. 481.6±38.14; p = 0.035). At the protein level, we observed inhibition in Cyclin D1 and the presence of caspase 3 cleavage in the isolated tumor from the metformin-treated mice. Ex vivo metformin treatment of the primary cells isolated from the CNL patient harboring CSF3RT618I resulted in increased apoptosis, with 51.94% (p < 0.05), 52.40% (p < 0.05), and 63.1% (p < 0.001), of apoptosis induction upon treatment with 5, 10 and 20 mM. Metformin ex vivo treatment strongly reduced hematopoietic progenitor cells colony formation on primary CNL CSF3RT618I peripheral blood (83.5% at 10 mM and 100% at 20 mM) and bone marrow mononuclear cells (100% at 10 mM and 98% at 20 mM). Conclusions: We provide evidence that metformin exerts anticancer activity in CSF3RT816I cells, by downregulation of STAT3 and MAPK signaling through AMPK activation. These results support the idea that metformin can be repurposed in the CNL clinics, as a therapeutical option.

Efficacy and Safety of Lenalidomide, Anti-PD-1 Antibody Combined with Orelabrutinib or Rituximab in the Treatment of Patients with Relapsed/Refractory Diffuse Large B-Cell Lymphoma

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma worldwide. Although most patients with DLBCL respond to first-line chemoimmunotherapy with R-CHOP, approximately 40% of patients will have relapsed or refractory disease. Currently, the second-line therapeutic regimen is chemoimmunotherapy followed by consolidative autologous hematopoietic stem cell transplantation (AHCT) for relapsed/refractory DLBCL (R/R DLBCL). However, the prognosis has remained poor. (Sawalha. J Pers Med. 2021). Thus, discovering effective treatment regimens for patients with R/R DLBCL has become extremely crucial. Recently, a combination regimen of lenalidomide, anti-programmed cell death protein-1 (PD-1) antibody, and rituximab demonstrated a good efficacy with an overall response rate (ORR) of 40% in R/R DLBCL (Sethi et al. Blood. 2019). Activation of the Bruton's tyrosine kinase (BTK)-mediated B-cell receptor (BCR) signaling pathway is a hallmark of DLBCL (Kapoor et al. Cell Death Dis. 2019). Orelabrutinib, a novel and potent irreversible BTK inhibitor, has shown encouraging efficacy with an ORR of 61.5% in combination with a new generation anti-CD20 antibody MIL62 in R/R DLBCL (Xu et al. Blood. 2020; Thus, we assessed the efficacy and safety of lenalidomide, anti-PD-1 antibody combined with orelabrutinib or rituximab in R/R DLBCL. Methods: This was a prospective, multicenter, single-arm, open-label, phase II study (ChiCTR2200056256). Patients aged 18 to 75 years were eligible if they had histologically confirmed R/R DLBCL, an Eastern Cooperative Oncology Group performance status of 0-2, adequate organ and bone marrow function, and survival time ≥ 6 months. Eligible patients received a combination regimen (21-day/cycle) of lenalidomide (10-25 mg, days 1-10, QD, oral), Tislelizumab (200 mg, day 1, intravenous), and orelabrutinib (150 mg, QD, oral) or rituximab (375 mg/m2, day 1, intravenous). The primary endpoint was ORR. Secondary endpoints included complete response (CR), progression-free survival (PFS), duration of response (DOR), time to response (TTR) and adverse events (AEs). Efficacy was assessed using Lymphoma Response to Immunomodulatory therapy Criteria (LYRIC, 2016). AEs were assessed according to the Common Terminology Criteria for Adverse Events (CTCAE) v 4.0. Results: Between August 2021 and March 2022, 9 patients (6 males; median age, 73 years, range 65-76) were enrolled. Among them, 8 patients were non-germinal center B-cell-like and 1 was unknown. Most patients had comorbidities (n=8, 88.9%), an Ann Arbor stage of III or IV (n=4, 44.4%), International Prognostic Index (IPI) ≥ 3 (n=4, 44.4%). All patients received lenalidomide, anti-PD-1 antibody combined with orelabrutinib regimen, and the median duration of treatment was 5 cycles (range 3-6). Among them, 4 patients (44.4%) received a second-line therapy, and 5 patients (55.5%) received ≥ third-line therapy. Before enrollment, 3 patients attained a CR as their best response during the previous therapies. Eight patients were included in the efficacy analysis. The ORR, CR and median TTR were 87.5%, 25.0%, and 2.45 months (range 1.97-2.73), respectively. Among them, 3 patients completed 6 cycles of therapy with ORR of 100% (Figure 1). The PFS and DOR were not completely mature due to the short follow-up period at the end of follow-up. Eight (80%) patients reported AEs. The most common hematological AEs were anemia (77.8%), platelet count reduced (55.6%), neutrophil count decreased (33.3%) and white blood cell count decreased (22.2%). Three patients (33.3%) discontinued treatment due to AEs (atrioventricular block, n=1; rash, n=2). In addition, the most common AEs were grade 1 or 2, and no grade 4 AEs. One patient died due to electrolyte disturbances and depression. Conclusion: Lenalidomide, anti-PD-1 antibody combined with orelabrutinib or rituximab regimen is effective and well-tolerated for patients with R/R DLBCL in this phase II study. This study provides a potential novel strategy for the treatment of R/R DLBCL. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Retraction statement: Lymphocyte‐specific protein 1 inhibits the growth of hepatocellular carcinoma by suppressing ERK1/2 phosphorylation

FEBS Open BioVolume 12, Issue 12 p. 2255-2255 Retraction NoticeOpen Access Retraction statement: Lymphocyte-specific protein 1 inhibits the growth of hepatocellular carcinoma by suppressing ERK1/2 phosphorylation This article retracts the following: Retracted: Lymphocyte-specific protein 1 inhibits the growth of hepatocellular carcinoma by suppressing ERK1/2 phosphorylation Hongyong Zhang, Yufeng Wang, Zhikui Liu, Bowen Yao, Changwei Dou, Meng Xu, Qing Li, Yuli Jia, Shengli Wu, Kangsheng Tu, Qingguang Liu, Volume 6Issue 12FEBS Open Bio pages: 1227-1237 First Published online: November 7, 2016 First published: 15 November 2022 https://doi.org/10.1002/2211-5463.13509AboutReferencesRelatedInformationPDFPDF ToolsExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessClose modalShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat https://doi.org/10.1002/2211-5463.12139 The above article published online on 06 October 2016 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the Editor-in-Chief and John Wiley and Sons Ltd. The retraction has been agreed following an investigation based on allegations raised by a third party, which revealed inappropriate and unexplained duplications between this and another article [[1]]. Thus, the editors consider the conclusions of this manuscript substantially compromised. References 1Yang N, Chen J, Zhang H, Wang X, Yao H, Peng Y, et al. LncRNA OIP5-AS1 loss-induced microRNA-410 accumulation regulates cell proliferation and apoptosis by targeting KLF10 via activating PTEN/PI3K/AKT pathway in multiple myeloma. Cell Death Dis. 2017; 8:e2975. https://doi.org/10.1038/cddis.2017.358 Volume12, Issue12December 2022Pages 2255-2255 ReferencesRelatedInformation

Sting Agonist GSK3745417 Induces Apoptosis, Antiproliferation, and Cell Death in a Panel of Human AML Cell Lines and Patient Samples

GSK3745417 is currently in phase 1 development targeting solid tumor indications. Recent literature has suggested a role for STING (Stimulator of Interferon Genes) activation induced cytotoxicity in AML cells (Baba et al, Cell Death Dis, 2021; 12(4):322.), indicating potential for therapeutic benefit in AML/MDS. To determine the implications of STING activation on tumor cells directly, GSK3745417 treatment was assessed in a screen against a panel of 93 tumor cell lines from various tumor origin. GSK3745417 treatment resulted in cytokine production across cell lines but cell growth inhibition in only 3 of the cell lines, including the only 2 AML cell lines in the panel. GSK3745417 was subsequently tested in an expanded panel of 13 AML cell lines. The data confirmed the cell growth inhibitory effect of GSK3745417 in 11/13 AML cell lines. Overall, growth inhibition was more potent than IFNβ induction, indicating a distinct growth inhibition/cytotoxicity mechanism in AML cells upon STING activation in addition to the potential to induce anti-AML immune response through type 1 IFN induction (Curran et al, Cell Rep, 2016;15(11):2357-66.). Further study demonstrated caspase induction in 7 out of 11 responding cell lines upon GSK3745417 treatment which was blocked by apoptosis inhibitor QVD. This supports the hypothesis that GSK3745417 induced cytotoxicity in AML cells is driven through an apoptosis mechanism. GSK3745417 also blocked colony formation in primary AML samples from 5 donors. GSK3745417 is able to induce growth inhibition with transient compound incubation in vitro in as little as 30 minutes to 1 hour. Taken together, this data suggests a role for GSK3745417 as a therapeutic in AML/MDS indications through dual mechanisms of direct tumor cell killing and immune activation at a clinically relevant dose. The human biological samples were sourced ethically and their research use was in accord with the terms of the informed consents under an IRB/EC approved protocol.

Chronic delta-9-tetrahydrocannabinol (THC) treatment counteracts SIV-induced modulation of proinflammatory microRNA cargo in basal ganglia-derived extracellular vesicles

BackgroundEarly invasion of the central nervous system (CNS) by human immunodeficiency virus (HIV) (Gray et al. in Brain Pathol 6:1–15, 1996; An et al. in Ann Neurol 40:611–6172, 1996), results in neuroinflammation, potentially through extracellular vesicles (EVs) and their micro RNAs (miRNA) cargoes (Sharma et al. in FASEB J 32:5174–5185, 2018; Hu et al. in Cell Death Dis 3:e381, 2012). Although the basal ganglia (BG) is a major target and reservoir of HIV in the CNS (Chaganti et al. in Aids 33:1843–1852, 2019; Mintzopoulos et al. in Magn Reson Med 81:2896–2904, 2019), whether BG produces EVs and the effect of HIV and/or the phytocannabinoid–delta-9-tetrahydrocannabinol (THC) on BG-EVs and HIV neuropathogenesis remain unknown.MethodsWe used the simian immunodeficiency virus (SIV) model of HIV and THC treatment in rhesus macaques (Molina et al. in AIDS Res Hum Retroviruses 27:585–592, 2011) to demonstrate for the first time that BG contains EVs (BG-EVs), and that BG-EVs cargo and function are modulated by SIV and THC. We also used primary astrocytes from the brains of wild type (WT) and CX3CR1+/GFP mice to investigate the significance of BG-EVs in CNS cells.ResultsSignificant changes in BG-EV-associated miRNA specific to SIV infection and THC treatment were observed. BG-EVs from SIV-infected rhesus macaques (SIV EVs) contained 11 significantly downregulated miRNAs. Remarkably, intervention with THC led to significant upregulation of 37 miRNAs in BG-EVs (SIV–THC EVs). Most of these miRNAs are predicted to regulate pathways related to inflammation/immune regulation, TLR signaling, Neurotrophin TRK receptor signaling, and cell death/response. BG-EVs activated WT and CX3CR1+/GFP astrocytes and altered the expression of CD40, TNFα, MMP-2, and MMP-2 gene products in primary mouse astrocytes in an EV and CX3CR1 dependent manners.ConclusionsOur findings reveal a role for BG-EVs as a vehicle with potential to disseminate HIV- and THC-induced changes within the CNS.

Development of highly potent and selective GRK5 inhibitors

G protein‐coupled receptor kinases (GRKs) regulate cell signaling by triggering receptor desensitization via phosphorylation on G protein‐coupled receptors (GPCRs). (1) The seven human GRKs (GRK1−GRK7) are classified into three subfamilies: GRK1 (GRK1 and GRK7), GRK2 (GRK2 and GRK3), and GRK4 (GRK4, GRK5, and GRK6). GRK2 and GRK5 are the most abundant in cardiovascular tissue, where they are potential targets for treatment of cardiovascular disease. GRK5 also undergoes Ca2+/calmodulin‐dependent nuclear localization, where it phosphorylates histone deacetylase 5 (HDAC5), inducing an increase in transcription of cardiac hypertrophy‐related genes. In GRK5‐knockdown mice, cardiomyocytes are protected from hypertrophy; however, the specific roles of GRK5 in heart failure and hypertrophic cardiomyopathy are still unclear. Furthermore, GRK5 is required for cancer progression in various cancer types. Depletion of GRK5 has been shown to suppress prostate cancer, breast cancer, and non‐small‐cell lung cancer. (2) Therefore, targeting GRK5 can also be a chemotherapeutic strategy. Here we are testing a series of inhibitors for GRK5/6 derived from the indolinone scaffold, utilizing Cys474 residue unique in GRK5/6 to enhance selectivity by covalent capture. An immediate goal is to facilitate the development of more selective inhibitors via rational design by determining x‐ray crystal structures of inhibitor complexes with GRK5. Recently, we have crystallized a hyper‐phosphorylated GRK5, and the active site structure suggested the possibility for compound soaking. Overall, discovery of the inhibitors can significantly facilitate understanding and treatment of cardiovascular diseases and cancer.1. Z. Yang et al., Phosphorylation of G Protein‐Coupled Receptors: From the Barcode Hypothesis to the Flute Model. Molecular Pharmacology 92, 201‐210 (2017).2. L.‐P. Jiang et al., GRK5 functions as an oncogenic factor in non‐small‐cell lung cancer. Cell Death Dis 9, 295 (2018).

Investigating β‐Lapachone Mediated Metabolic Disruption Using Stable Isotope Tracers

β‐lapachone has been found to be a highly potent anti‐cancer agent in phase 1 clinical trials, but has dose limiting toxicity in red blood cells [Gerber et. al Br J Cancer. 2018 Oct;119(8):928‐936]. Identifying a safe and quantitative method of monitoring treatment efficacy will be highly beneficial to establishing an adaptive dosing platform. Adaptative dosing can allow personalized treatments that significantly reduce off target effects while maintaining an effective therapeutic regimen. In this study we present the use of a safe and noninvasive tracer strategy utilizing both [2H7]glucose and [U‐13C]glucose tracers to assess the metabolic effects caused by β‐lapachone treatment in vitro. Treating cancers with β‐lapachone bioactivates NAD(P)H: quinone oxidoreductase 1 (NQO1) to produce a highly unstable hydroquinone intermediate that reestablishes itself through an aggressive futile cycle, generating reactive oxygen species (ROS) in the form of hydrogen peroxide. Accelerated accumulation of ROS causes large amounts of DNA damage, triggers poly‐ADP‐ribose polymerase‐I hyperactivation, induces global depletion of NAD+ and ATP, and hinders overall glycolytic flux [Moore et. al Cell Death Dis. 2015 Jan; 6(1): e159]. Cancer cells that overexpress NQO1 subsequently die through NAD+‐keresis. Measuring changes in glycolysis and downstream metabolism caused by the bioactivation of NQO1 would establish a platform for assessing the efficacy of treatment, potentially allowing the reduction of chemotherapeutic dosage thereby diminishing off‐target toxicities. Changes in central carbon metabolism caused by NQO1 bioactivation were detected and assessed in various cancer cells using both tracer strategies. Deuterated lactate and HDO (deuterated water) were easily quantified, and the production of these metabolic products were linearly correlated with [2H7]glucose consumption. Mass isotopologue distribution analysis by gas chromatography‐mass spectrometry demonstrated downregulated energy metabolism with a reduction in 2H and 13C labeling in NQO1+ cancer cells treated with β‐lapachone. These methods are highly beneficial for measuring rates of glycolytic metabolism, identifying potential therapeutic synergies, the efficacy of chemotherapeutic agents that hamper central carbon metabolism, and can be potentially translated to in vivo models.

Assessing PfGARP-Mediated Apoptosis of Blood-Stage Plasmodium falciparum Parasites.

Apoptosis is conventionally regarded as an evolutionarily conserved and genetically controlled process of programmed cell death confined to metazoan organisms. However, recently, conserved features of apoptosis have also been demonstrated in unicellular eukaryotes (Holzmuller et al. Parasitology 132:S19-S32, 2006; Le Chat et al. Mol Biochem Parasitol 153:41-47, 2007; Madeo et al. Curr Opin Microbiol 7:655-660, 2004; Welburn et al. Parasitology 132:S7-S18, 2006; Jensen et al. Science 216:1230-1233, 1982) including malaria parasites (Al-Olayan et al. Int J Parasitol 32:1133-1143, 2002; Ch'ng et al. Cell Death Dis 1:e26, 2010; Meslin et al. J Infect Dis 195:1852-1859, 2007; Picot et al. Trans R Soc Trop Med Hyg 91:590-591, 1997; Raj et al. Nature 582:104-108, 2020). P. falciparum glutamic-acid-rich protein (PfGARP) is an antigen of 80kDa that is uniquely expressed on the exofacial surface of red blood cells (RBCs) infected by early-to-late-trophozoite-stage P. falciparum parasites (Raj et al. Nature 582:104-108, 2020). We have recently demonstrated that antibodies against PfGARP bind to the PfGARP displayed on the surface of P. falciparum trophozoite-infected RBCs and trigger apoptosis in the intracellular parasites (Raj et al. Nature 582:104-108, 2020). This is the first demonstration of antibody-induced apoptosis in blood-stage malaria parasites and is characterized by several conserved features such as crisis form morphology, loss of mitochondrial membrane potential, loss of integrity of food vacuole, activation of caspase-like cysteine proteases, and fragmentation of chromosomal DNA. Here we describe the assays used to detect these features of apoptosis in the mature blood stage of malaria parasites.

BMP6 Mediates BMP2-Increased Human Trophoblast Invasion

TGF-β superfamily proteins play divergent roles in regulating human extravillous trophoblast (EVT) invasion and their coordinated effects are essential for adequate placentation during pregnancy 1. Bone morphogenetic protein 2 (BMP2), which belongs to the BMP subfamily of TGF-β superfamily, has been shown to promote human EVT invasion and the acquisition of endothelial-like phenotype 2,3. It has been reported that BMP2 promotes EVT invasion by up-regulating Activin A, a growth factor which also belongs to TGF-β superfamily. However, whether BMP6 mediates the pro-invasive effect of BMP2 has yet to be determined. Herein, we firstly treated immortalized trophoblast cells (HTR8/SVneo) with recombinant BMP2 protein for 6 and 24 hrs, and our bulk-RNA sequencing results demonstrated significantly increased BMP6 mRNA levels after BMP2 treatment. Furthermore, we confirmed the up-regulatory effects of BMP2 on BMP6 mRNA and protein levels in both HTR8/SVneo and primary EVTs isolated from first-trimester villi. Notably, siRNA-mediated down-regulation of BMP6 significantly attenuated both basal and BMP2-induced cell invasion in HTR8/SVneo cells as measured by Matrigel-coated transwell invasion assay. In summary, our results firstly demonstrated the up-regulatory effect of BMP2 on BMP6 expression in human trophoblasts and identified the mediation role of BMP6 in BMP2-promoted EVT invasion, suggesting the interplay between BMP subfamily members during EVT invasion regulation. Our ongoing research focusing the underlying molecular mechanisms and signaling pathways could further benefit the advancement of diagnostic and therapeutic strategies for EVT invasion dysregulation-related pregnancy disorders, e.g., pre-eclampsia. Reference: (1) Li Yan et al., Trends Endocrinol Metab 2021 18: S1043-2760(20)30266-6. (2) Hong-Jin Zhao et al., FASEB J 2020;34(2):3151-3164. (3) Hong-Jin Zhao et al., Cell Death Dis 2018;9(2):174.

Bone Morphogenetic Protein 2 Increases Human Trophoblast Invasion by Up-Regulating Integrin Beta3

The adequate invasion of extravillous trophoblast cells (EVTs) is indispensable for the implantation of embryos and subsequent remodeling of uterine spiral arteries in early human gestation. Bone morphogenetic protein 2 (BMP2), which is abundantly expressed at the maternal-fetal interface, has been shown to promote the human EVT invasion process (1). Integrin switching (i.e., a switch from α6β4 to αvβ3) plays essential roles in cell-extracellular matrix adhesion and has been reported to influence EVT migration and invasion (2). Moreover, integrin β3 has been found to promote the adhesion, invasion, and migration abilities of embryonic trophoblasts (3). However, whether integrin β3 participates in BMP2 signaling and mediates BMP2-increased-human trophoblast invasion remains unknown. The purpose of our study was to explore the effects of BMP2 on integrin αvβ3 expression and the possible mediation role of integrin β3 in BMP2-regulated human trophoblast invasion. We used immortalized human trophoblast cell line (HTR8/SVneo) and primary human extravillous trophoblast cells (EVTs) isolated from first-trimester villi as study models. RT-qPCR and Western blot assay were respectively utilized to detect the messenger RNA and protein levels of intergrin αv and β3. The function of the target protein was studied by siRNA knockdown, and the trophoblast invasion ability was checked by Matrigel-coated transwell invasion assays. Our results demonstrated that the mRNA and protein levels of integrin β3, rather than integrin αv, were up-regulated after BMP2 treatment in HTR8/SVneo and primary EVT cells. Importantly, siRNA-mediated down-regulation of integrin β3 significantly inhibited basal and BMP2-induced HTR8/SVneo cell invasionas measured by transwell invasion assay. In conclusion, we findings support that BMP2 promotes human trophoblast cell invasion by up-regulating integrin β3 expression, benefiting the in-depth understanding of the pro-invasive effect of BMP2 on human trophoblasts during early pregnancy. Reference: (1) Hong-Jin Zhao et al., Cell Death Dis 2018;9:174. (2) Damsky, C.H. et al, Development 1994; 120, 3657-3666. (3) Dong-Mei He et al., Reproduction 2019;157:423-430.

Identification of ILC2 in the Lung Using Flow Cytometry.

Group 2 innate lymphoid cells (ILC2), despite their scarcity, are the dominant innate lymphoid cell population in the lung, orchestrating innate immunity and adaptive immunity (Germain and Huang. Curr Opin Immunol 56:76-81, 2019; Krabbendam et al. Immunol Rev 286:74-85, 2018; Mindt et al. Front Immunol 9:840, 2018) . Recent studies reveal that ILC2 play critical roles in inflammation-associated lung injury during sepsis (Lai et al. Cell Death Dis 9:369, 2018; Xu et al. Immunol Cell Biol 96:935-947). Therefore, studies aiming to understand the pathobiology of ILC2 may reveal new therapeutic strategies for sepsis. However, the identification of ILC2 requires multiple surface and intracellular markers. This makes the detection of ILC2 in the lung challenging. Here we describe a method to detect ILC2 in the mouse lung using flow cytometry.

A Phase 1/2 Study of Umbralisib, Ublituximab, and Venetoclax (U2-Ven) in Patients with Relapsed or Refractory Chronic Lymphocytic Leukemia (CLL)

A Phase 1/2 Study of Umbralisib, Ublituximab, and Venetoclax (U2-Ven) in Patients with Relapsed or Refractory Chronic Lymphocytic Leukemia (CLL)

Mitochondrial Potentiation Ameliorates Age-Related Heterogeneity in Hematopoietic Stem Cell Function.

Aging is associated with reduced fitness and increased myeloid bias of the hematopoietic stem cell (HSC) compartment, causing increased risk of immune compromise, anemia, and malignancy. We show that mitochondrial membrane potential (MMP) can be used to prospectively isolate chronologically old HSCs with transcriptional features and functional attributes characteristic of young HSCs, including a high rate of transcription and balanced lineage-affiliated programs. Strikingly, MMP is a stronger determinant of the quantitative and qualitative transcriptional state of HSCs than chronological age, and transcriptional consequences of manipulation of MMP in HSCs within their native niche suggest a causal relationship. Accordingly, we show that pharmacological enhancement of MMP in old HSCs invivo increases engraftment potential upon transplantation and reverses myeloid-biased peripheral blood output at steady state. Our results demonstrate that MMP is a source of heterogeneity in old HSCs, and its pharmacological manipulation can alter transcriptional programs with beneficial consequences for function.

Type 3 IP3 receptors driving oncogenesis

Type 3 Inositol 1,4,5-trisphosphate (IP3) receptors (IP3R3s) have been identified as anti-oncogenic channels by propelling pro-apoptotic Ca2+ signals to mitochondria. Yet, recent studies (Rezuchova et al, Cell Death Dis, 2019; Ueasilamongkol et al, Hepathology, 2019; Guerra et al, Gut, 2019) revealed that IP3R3 upregulation drives oncogenesis via ER-mitochondrial Ca2+ crosstalk, adding complexity to IP3R3’s role in cancer.