Anti-acute Myeloid Leukemia Research Articles

538P Muscle haptoglobin biomarks cachexia induced by anti-acute myeloid leukemia chemotherapy

538P Muscle haptoglobin biomarks cachexia induced by anti-acute myeloid leukemia chemotherapy

Preparation of a dual-specific antibody targeting human CD123 and exploration of its anti-acute myeloid leukemia effects

Objective: To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) . Methods: Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified. Results: ① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123(+) tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group (P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10(5)/ml to 3.2×10(6)/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8(+) PD-1(+) LAG-3(+) T cells was 10.90%, and the proportion of propidium iodide (PI) (-) Annexin Ⅴ(+) T cells and PI(+) Annexin Ⅴ(+) T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group (P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group (P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group (P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123(+) tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123(+) MV4-11 cells, CD123(+) Molm13 cells, and CD123(+) THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group (P<0.05) . Conclusion: In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123(+) tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Anti-Acute Myeloid Leukemia Activity of CD38-CAR-T Cells with PI3Kδ Downregulation.

We previously constructed a nanobody-based anti-CD38 chimeric antigen receptor T (CD38-CAR-T) cell efficiently against multiple myeloma. As CD38 is also expressed on most tumor cells of acute myeloid leukemia (AML), we wondered about its efficacy in treating AML. In this study, we demonstrated that our CD38-CAR-T cells effectively lysed CD38+ AML cell lines, including NB4, U937, HL-60, THP-1 with an E:T (effector/target cells) ratio of 1:8, and primary AML cells from patients with a low E:T ratio of 1:16. Moreover, recent studies showed that inhibition of PI3Kδ could enhance CAR-T-cell efficacy. We constructed PI3Kδ-downregulated CD38-CAR-T cells with a CD38-CAR lentiviral vector containing short hairpin RNA (shRNA) sequences against PI3Kδ. CD38-CAR-T cells with PI3Kδ downregulation maintained their antileukemia function against both AML cell lines and primary AML cells while reducing the release of IL-2, IFN-γ, and TNF when co-culturing with AML cell lines. Both CD38-CAR-T and PI3Kδ-downregulated CD38-CAR-T-cell therapy significantly improved the survival of AML mice, whereas the latter had an even better effect on survival. In summary, our study demonstrated that CD38-CAR-T cells had promising activity against AML, and PI3Kδ downregulation in CD38-CAR-T cells could reduce some cytokines release without impairing their antileukemia function.

An In Silico Evaluation of Flavonoids in Allium sp. as a Potent Anti-Acute Myeloid Leukemia (AML) Through FLT3 Inhibition

An In Silico Evaluation of Flavonoids in Allium sp. as a Potent Anti-Acute Myeloid Leukemia (AML) Through FLT3 Inhibition

Improving the anti-acute myeloid leukemia activity of CD123-specific engager T cells by MyD88 and CD40 costimulation.

The outcome of patients with acute myeloid leukemia remains poor, and immunotherapy has the potential to improve this. T cells expressing chimeric antigen receptors or bispecific T-cell engagers targeting CD123 are actively being explored in preclinical and/or early phase clinical studies. We have shown that T cells expressing CD123-specific bispecific T-cell engagers (CD123.ENG T cells) have anti-acute myeloid leukemia activity. However, like chimeric antigen receptor T cells, their effector function diminishes rapidly once they are repeatedly exposed to antigen-positive target cells. Here we sought to improve the effector function of CD123.ENG T cells by expressing inducible co-stimulatory molecules consisting of MyD88 and CD40 (iMC), MyD88 (iM), or CD40 (iC), which are activated by a chemical inducer of dimerization. CD123.ENG T cells expressing iMC, iM, or iC maintained their antigen specificity in the presence of a chemical inducer of dimerization, as judged by cytokine production (interferon-γ, interleukin-2) and their cytolytic activity. In repeat stimulation assays, activating iMC and iM, in contrast to iC, enabled CD123.ENG T cells to secrete cytokines, expand, and kill CD123-positive target cells repeatedly. Activating iMC in CD123.ENG T cells consistently improved antitumor activity in an acute myeloid leukemia xenograft model. This translated into a significant survival advantage in comparison to that of mice that received CD123.ENG or CD123.ENG.iC T cells. In contrast, activation of only iM in CD123.ENG T cells resulted in donor-dependent antitumor activity. Our work highlights the need for both toll-like receptor pathway activation via MyD88 and provision of co-stimulation via CD40 to consistently enhance the antitumor activity of CD123.ENG T cells.

Synergistic efficacy of homoharringtonine and venetoclax on acute myeloid leukemia cells and the underlying mechanisms.

BackgroundTo evaluate whether homoharringtonine (HHT) combined with venetoclax could produce a synergistic anti-acute myeloid leukemia (AML) effect and determine the underlying mechanisms.MethodsThe effect of HHT and venetoclax combination on cell viability, apoptosis, and mitochondrial membrane potential was investigated in vitro using AML cell lines and primary cells. High-throughput mRNA sequencing was used to analyze mRNA level changes after the application of HHT and venetoclax on OCI-AML3 cells. Western blotting was used to verify the changes in protein expression within the mitogen-activated protein kinases/extracellular signal-regulated kinase (MAPK/ERK), phosphatidylinositiol 3-kinase (PI3K)/AKT and p53 pathway. The efficacy of HHT and venetoclax in vivo and their effects on survival time were evaluated in a xenograft model established in severe immunodeficiency (NOD/SCID) mice.ResultsVenetoclax and HHT synergistically inhibited the proliferation of AML cells, decreased the mitochondrial membrane potential, and promoted AML cell apoptosis in a time- and concentration-dependent manner. Venetoclax combined with HHT increased the expression of the caspase-3, Poly (ADP-ribose) polymerase (PARP), and γH2AX proteins. HHT enhanced the proapoptotic effect of venetoclax by reducing the expression of myeloid cell leukemia sequence 1 (Mcl-1). HHT arrested AML cells in G1 phase of the cell cycle. HHT enhanced the proapoptotic effect of venetoclax by inhibiting the activation of the MAPK/ERK and PI3K/AKT pathways and activating the p53 pathway. In vivo experiments confirmed that the combination of HHT and venetoclax could inhibit the growth of tumors in AML xenotransplanted mice and prolong the survival time of tumor-bearing mice.ConclusionsHHT combined with venetoclax synergistically promoted apoptosis in AML cell lines and primary cells by inhibiting the activation of the MAPK/ERK and PI3K/AKT pathways and activating the p53 pathway.

A CD123-specific chimeric antigen receptor augments anti-acute myeloid leukemia activity of Vγ9Vδ2 T cells.

Aim: To investigate whether anti-CD123 chimeric antigen receptor (CAR)-expressing Vγ9Vδ2 T cells could be an alternative for acute myeloid leukemia (AML) treatment. Materials & methods:Ex vivo expanded Vγ9Vδ2 T cells were electroporated with anti-CD123 CAR-encoding mRNA. The effector function and specificity of the modified Vγ9Vδ2 T cells were examined by in vitro cytotoxicity, degranulation and cytokine release level. The in vivo function was analyzed using the xenograft KG1-luc model with NOD-SCID-γc-/- mice. Results: The modified Vγ9Vδ2 T cells exhibited significantly improved effector activities against both AML cell lines and primary AML cells in vitro. In the xenograft mouse model, the modified Vγ9Vδ2 cells displayed an enhanced tumor control potency. Conclusion: Anti-CD123 CAR-expressing Vγ9Vδ2 T cells may serve as an alternative way to target AML.

CAR T cells redirected to cell surface GRP78 display robust anti-acute myeloid leukemia activity and do not target hematopoietic progenitor cells

Developing CAR T cells for acute myeloid leukemia (AML) has been hampered by a paucity of targets that are expressed on AML blasts and not on hematopoietic progenitor cells (HPCs). Here we demonstrate that GRP78 is expressed on the cell surface of primary AML blasts but not HPCs. To target GRP78, we generate T cell expressing a GRP78-specific peptide-based CAR, which show evidence of minimal fratricide post activation/transduction and antigen-dependent T cell differentiation. GRP78-CAR T cells recognize and kill GRP78-positive AML cells without toxicity to HPCs. In vivo, GRP78-CAR T cells have significant anti-AML activity. To prevent antigen-dependent T cell differentiation, we block CAR signaling and GRP78 cell surface expression post activation by using dasatinib during GRP78-CAR T cell manufacturing. This significantly improves their effector function in vitro and in vivo. Thus, targeting cell surface GRP78-positive AML with CAR T cells is feasible, and warrants further active exploration.

Liriopogons (Genera Ophiopogon and Liriope, Asparagaceae): A Critical Review of the Phytochemical and Pharmacological Research.

The closely related genera Liriope and Ophiopogon (Asparagaceae), collectively known in English as liriopogons, have similar therapeutic uses in treating cough, rheumatoid arthritis, and cleaning heat. The main aim of this review is to understand the current phytochemical and pharmacological knowledge including an assessment of the quality of the scientific evidence. A literature search was conducted in line with PRISMA guidelines, by retrieving available information up to 2020 from five online resources. The bioactive metabolites of liriopogons include steroidal saponins, flavonoids, polysaccharides, organic acids, phenols. Cardiovascular protective, anti-inflammatory, anti-diabetic, anti-oxidant, anti-cancer, neuroprotective, anti-viral, anti-acute myeloid leukemia and hepatoprotective effects have been at the center of attention. From a toxicological perspective Ophiopogon japonicus seems to be safe. Some problems with the quality of the pharmacological evidence stand out including the application of excessive dose level and methodological problems in the design. Additionally, a reasonable link between local/traditional uses and pharmacological assessment is often vague or not reflected in the text. Future researches on liriopogons are required to use rigorous scientific approaches in research on evidence-based natural products for the future benefits of patients.

Anti-leukemia properties of cadmium nanoparticles in the in vitro and in vivo condition: A chemobiological study

IntroductionThe present study investigated the anti-acute myeloid leukemia effects of Ziziphora clinopodides Lam leaf aqueous extract conjugated cadmium nanoparticles.Material and methodsTo synthesize CdNPs, Z. clinopodides aqueous extract was mixed with Cd(NO3)2 .4H2O. The characterization of the biosynthesized cadmium nanoparticles was carried out using many various techniques such as UV-Vis. and FT-IR spectroscopy, XRD, FE-SEM, and EDS.ResultsThe uniform spherical morphology of NPs was proved by FE-SEM images with NPs the average size of 26.78cnm. For investigating the antioxidant properties of Cd(NO3)2, Z. clinopodides, CdNPs, and Daunorubicin, the DPPH test was used. The cadmium nanoparticles inhibited half of the DPPH molecules in a concentration of 196 µg/mL. To survey the cytotoxicity and anti-acute myeloid leukemia effects of Cd(NO3)2, Z. clinopodides, CdNPs, and Daunorubicin, MTT assay was used on the human acute myeloid leukemia cell lines i.e., Murine C1498, 32D-FLT3-ITD, and Human HL-60/vcr. The IC50 of the cadmium nanoparticles was 168, 205, and 210 µg/mL against Murine C1498, 32D-FLT3-ITD, and Human HL-60/vcr cell lines, respectively. In the part of in vivo study, DMBA was used for inducing acute myeloid leukemia in mice. CdNPs similar to daunorubicin ameliorated significantly (p≤0.01) the biochemical, inflammatory, RBC, WBC, platelet, stereological, histopathological, and cellular-molecular parameters compared to the other groups.ConclusionsAs mentioned, the cadmium nanoparticles had significant anti-acute myeloid leukemia effects. After approving the above results in the clinical trial studies, these cadmium nanoparticles can be used as a chemotherapeutic drug to treat acute myeloid leukemia in humans.

Characterization and anti-acute myeloid leukemia and anti-acute T cell leukemia properties of zinc nanoparticles synthesized by a green approach for bioremediation applications

IntroductionEvery year, many people die due to cancer in all of the world. So, the preparation and formulation of new chemotherapeutic supplements and drugs with remarkable effects to treat cancer are a priority of both developing and developed countries. Recently, zinc nanoparticles have been used as modern chemotherapeutic drugs to treat several cancers such as leukemia, lung cancer, breast cancer, prostate cancer, etc. In this study, zinc nanoparticles (ZnNPs) were synthesized in an aqueous medium using Fumaria officinalis leaf as stabilizing and reducing agents.Material and methodsThe green synthesized ZnNPs@Fumaria officinalis were characterized using different techniques including UV-visible and FT-IR spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive X-ray spectrometry (EDS). The anticancer activity of ZnNPs@Fumaria officinalis was evaluated against acute myeloid leukemia and acute T cell leukemia. According to the XRD analysis, 20.44 nm was measured for the crystal size of ZnNPs@Fumaria officinalis.ResultsSEM images showed a uniform spherical morphology with an average size of 27.96 nm for ZnNPs@Fumaria officinalis. In the cellular and molecular part of the recent study, the cells treated with ZnNPs@Fumaria officinalis were assessed by MTT assay for 48 h regarding the cytotoxicity and anti-human acute leukemia properties on normal (HUVEC), acute myeloid leukemia (32D-FLT3-ITD and Human HL-60/vcr), and acute T cell leukemia (Jurkat, Clone E6-1 and J.RT3-T3.5) cell lines. In the antioxidant test, the IC50 values of ZnNPs@Fumaria officinalis and BHT against DPPH free radicals were 187 and 197 µg/ml, respectively. The IC50 values of ZnNPs@Fumaria officinalis were 227, 200, 250, and 336 µg/ml against 32D-FLT3-ITD, Human HL-60/vcr, Jurkat, Clone E6-1, and J.RT3-T3.5 cell lines, respectively. The viability of the malignant leukemia cell line decreased dose-dependently in the presence of ZnNPs@Fumaria officinalis.ConclusionsIt appears that the anti-human acute leukemia effect of ZnNPs@Fumaria officinalis is due to their antioxidant effects.

Decitabine shows anti-acute myeloid leukemia potential via regulating the miR-212-5p/CCNT2 axis.

Previous research has revealed the involvement of microRNA-212-5p (miR-212-5p) and cyclin T2 (CCNT2) in acute myeloid leukemia (AML). However, whether the miR-212-5p/CCNT2 axis is required for the function of decitabine in AML has not been well elucidated. Quantitative reverse transcription-polymerase chain reaction was used to examine enrichment of miR-212-5p. The relationship between CCNT2 and miR-212-5p was verified by the luciferase reporter assay. Cell apoptosis was evaluated by flow cytometry and western blot. CCK-8 assay was performed to determine cell viability. Decitabine significantly repressed cell viability, while promoted cell apoptosis. Meanwhile, the expression levels of cyclinD1, CDK4, and Bcl-2 were suppressed in cells with decitabine exposure, but Bax and caspase-3 expression levels were upregulated. Besides, miR-212-5p upregulation had the similar function with decitabine in AML cell proliferation and apoptosis. Subsequently, restoration of CCNT2 attenuated miR-212-5p overexpression-induced effects in Kasumi-1 and SKNO-1 cells. In addition, miR-212-5p depletion reversed decitabine-induced CCNT2 downregulation. The miR-212-5p/CCNT2 axis had an implication in the anti-leukemic effect of decitabine in AML.

Molecular docking studies, anti-Alzheimer’s disease, antidiabetic, and anti-acute myeloid leukemia potentials of narcissoside

In this research, we explored their capacity for Narcissoside antioxidant and anticholinergic, antidiabetic, and anti-acute myeloid leukaemia. Narcissoside’s antioxidant activities were elucidated by the use of various bioanalytical assays. Narcissoside’s radical scavenging activities were evaluated by DPPH • and ABTS • + scavenging activities. On the other hand, IC50 values were calculated for DPPH • , and ABTS • + scavenging, acetylcholinesterase, and α-glucosidase inhibition effects of narcissoside. IC50 values narcissoside, as 11.54 nM for AChE and 65.58 nM for α-glucosidase were calculated with % Activity-[Inhibitory] graphs. Then, ADME/T analysis of narcissoside molecule was performed to calculate the drug becoming parameters.

Research on the effect of PD-L1 overexpression on CLL-1 CAR-T anti-acute myeloid leukemia

目的探讨程序性死亡受体配体1（PD-L1）对CLL-1 CAR-T细胞抗急性髓系白血病（AML）作用的影响。方法通过构建PD-L1表达载体、制备慢病毒、转导、单克隆筛选技术获得稳定表达PD-L1的THP-1单克隆细胞株（THP1-PDL1），然后以前期制备的CLL-1 CAR-T细胞为效应细胞，以THP-1、THP1-PDL1单克隆细胞株作为靶细胞，分别通过LDH检测、CBA法、CFSE法评价PD-L1过表达对CLL-1 CAR-T细胞裂解功能、炎性因子释放、细胞增殖等功能的影响。结果①成功制备了PD-L1慢病毒，并筛选获得了稳定表达PD-L1的THP1-PDL1单克隆细胞株，流式细胞术及PCR验证成功。②PD-L1过表达抑制了CLL-1 CAR-T细胞裂解THP-1细胞的能力；效靶比为10∶1时，CLL-1CAR-T细胞对THP1-PDL1细胞的杀伤效率明显低于对THP-1细胞的杀伤效率［（15.70±9.90）％对（51.95±2.52）％，P<0.05］。③PD-L1过表达减弱了CLL-1 CAR-T细胞释放细胞因子的能力［与THP1-PDL1细胞共培养时对与THP-1细胞共培养时：IFN-γ（115.66±3.13）pg/ml对（1708.16±26.76）pg/ml，P<0.05；IL-6（17.37±0.72）pg/ml对（124.92±4.26）pg/ml，P<0.05；IL-10（5.69±0.13）pg/ml对（124.12±3.02）pg/ml，P<0.05］；同时抑制了CLL-1 CAR-T细胞的增殖能力。结论成功构建了表达PD-L1的THP1-PDL1单克隆细胞株，同时证实了PD-L1过表达对CLL-1 CAR-T细胞抗AML的不利的影响，为通过PD-1/PD-L1通路调控CLL-1 CAR-T细胞功能提供了一定的理论基础。

Construction of a new anti-CD123 chimeric antigen receptor T cells and effect of anti-acute myeloid leukemia

目的构建一种新的靶向CD123的嵌合抗原受体T细胞(CAR-T细胞)，为CD123阳性白血病的免疫治疗提供实验参考。方法通过单克隆筛选技术获得能稳定分泌CD123抗体的杂交瘤细胞株6E11，将杂交瘤细胞扩增后腹腔注射至Balb/c小鼠腹腔内，收集腹水并处理、纯化得到单克隆抗体，测定抗体效价并对其特异性进行验证；RT-PCR法获得轻链和重链可变区序列，并以此为基础利用分子克隆技术构建一种新的靶向CD123嵌合抗原受体，包装病毒后感染T细胞制备CD123CAR-T细胞，通过功能实验初步探讨6E11CAR-T细胞体外抗白血病的能力。结果①获得1株稳定分泌抗人CD123抗体的杂交瘤细胞株6E11并获得其可变区序列。②6E11单克隆抗体对CD123蛋白亲和性高，解离常数(Kd值)为2.10nmol/L，特异性识别CD123阳性细胞且与CD123阴性细胞无交叉反应。 ③成功构建了CD123CAR慢病毒载体，感染T细胞后获得了靶向CD123的CAR-T细胞(6E11CAR-T)，感染效率大于60％。 ④6E11CAR-T能明显杀伤CD123阳性靶细胞MV4-11，效靶比1∶1时6E11CAR-T细胞对MV4-11细胞的杀伤比例明显高于 Vecor-T细胞[(98.60±1.20)％对(20.28±6.74)％，P<0.001]，但对CD123阴性靶细胞K562没有明显杀伤作用。 ⑤MV4-11细胞可以显著激活6E11CAR-T，但对Vecor-T细胞无明显激活作用[(26.33±3.30)％对(1.17±0.06)％，P<0.001]。⑥6E11CAR-T与MV4-11细胞共培养上清中细胞因子水平均显著高于Vecor-T组[IL-2：(92.90±1.51)pg/ml对(6.05±3.41)pg/ml，P<0.001；TNF-α：(1 407.20±91.95)pg/ml对(7.86±0.85)pg/ml，P<0.001；IFN-γ：(5 614.60±170.17)pg/ml对(8.42±2.70)pg/ml，P<0.001]但与 K562细胞共培养后，两组各细胞因子水平差异无统计学意义。 ⑦6E11CAR-T在与CD123阳性髓系白血病(AML)原代细胞共培养过程中被显著激活，且能有效杀伤原代 AML细胞。结论杂交瘤细胞株6E11能稳定分泌高效特异的抗人CD123单克隆抗体，可用于检测表达人CD123的细胞，也能应用在靶向人CD123蛋白的肿瘤免疫治疗中，以 6E11Ig可变区序列为抗原识别区的CD123CAR-T细胞，具有明确的体外抗白血病活性，为进一步的临床研究奠定了基础。

Novel green synthesis of Boswellia serrata leaf aqueous extract conjugated gold nanoparticles with excellent anti‐acute myeloid leukemia property in comparison to mitoxantrone in a leukemic mice model: Introducing a new chemotherapeutic drug

The present research confirms the capacity of aqueous extract of Boswellia serrata grown under in vitro condition for the green synthesis of gold nanoparticles (AuNPs). Also, we showed the cytotoxicity, antioxidant, and anti‐acute myeloid leukemia properties of AuNPs compared to mitoxantrone in a leukemic mouse model. The synthesized AuNPs were characterized using several techniques including XRD, TEM, FE‐SEM, UV–Vis, and FT‐IR. From the XRD pattern, four distinct diffraction peaks at 38.2°, 44.2°, 64.7° and 77.4° are indexed as (111), (200), (220) and (311) planes of FCC metallic gold. TEM and FE‐SEM images revealed an average diameters of 15–30 nm for the nanoparticles. FT‐IR findings offered antioxidant compounds in the nanoparticles were the sources of reducing power, reducing gold ions to AuNPs. UV–Vis revealed an absorption band at 536 nm that is related to the surface plasmon resonance of AuNPs. In vivo design, induction of acute myeloid leukemia was done by DMBA in 75 mice. Then, the mice were randomly divided into six subgroups, including untreated, control, HAuCl4, B. serrata, AuNPs, and mitoxantrone. AuNPs (In the dose of 1 mg/kg body weight) similar to mitoxantrone, significantly (p ≤ 0.05) increased the platelet, lymphocyte, and RBC parameters and the anti‐inflammatory cytokines (IL4, IL5, IL10, IL13, and IFNα) and reduced the weights and volumes of liver and spleen and their sub‐compartment, the total WBC, blast, monocyte, neutrophil, eosinophil, and basophil counts, and the pro‐inflammatory cytokines (IL1, IL6, IL12, IL18, IFNY, and TNFα) as compared to the untreated mice. By quantitative Real‐Time PCR, S1PR1 and S1PR5 mRNA expression in lymphocytes were significantly (p ≤ 0.05) increased by treating the leukemic mice with the AuNPs and mitoxantrone. In vitro design, AuNPs similar to mitoxantrone had low cell viability dose‐dependently against Human HL‐60/vcr, 32D‐FLT3‐ITD, and Murine C1498 cell lines without any cytotoxicity on HUVEC cell line. Besides, the DPPH assay showed similar antioxidant potentials for AuNPs and mitoxantrone. In conclusion, the results of this research indicated the excellent capacity of synthesized gold nanoparticles using B. serrata leaf aqueous extract in the treatment of acute myeloid leukemia in leukemic mice.

Analysis of the Mechanisms of Action of Naphthoquinone-Based Anti-Acute Myeloid Leukemia Chemotherapeutics.

Acute myeloid leukemia (AML) is a neoplastic disorder resulting from clonal proliferation of poorly differentiated immature myeloid cells. Distinct genetic and epigenetic aberrations are key features of AML that account for its variable response to standard therapy. Irrespective of their oncogenic mutations, AML cells produce elevated levels of reactive oxygen species (ROS). They also alter expression and activity of antioxidant enzymes to promote cell proliferation and survival. Subsequently, selective targeting of redox homeostasis in a molecularly heterogeneous disease, such as AML, has been an appealing approach in the development of novel anti-leukemic chemotherapeutics. Naphthoquinones are able to undergo redox cycling and generate ROS in cancer cells, which have made them excellent candidates for testing against AML cells. In addition to inducing oxidative imbalance in AML cells, depending on their structure, naphthoquinones negatively affect other cellular apparatus causing neoplastic cell death. Here we provide an overview of the anti-AML activities of naphthoquinone derivatives, as well as analysis of their mechanism of action, including induction of reduction-oxidation imbalance, alteration in mitochondrial transmembrane potential, Bcl-2 modulation, initiation of DNA damage, and modulation of MAPK and STAT3 activity, alterations in the unfolded protein response and translocation of FOX-related transcription factors to the nucleus.

Chimeric antigen receptor T-cell therapy for acute myeloid leukemia: how close to reality?

The approval of the anti-CD19 chimeric antigen receptor (CAR) T-cell product tisagenlecleucel (Kymriah®) by the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for relapsed pediatric B-lineage acute lymphoblastic leukemia (B-ALL) was a landmark event in acute leukemia therapy. The approval was based on data from a phase II global trial in which 75 pediatric and young adult B-ALL patients received tisagenlecleucel, demonstrating safety, feasibility and biological response, with complete remissions (CR) at three months in 81% of patients, and event-free survival rates of 73% and 50% at six and 12 months, respectively.1 A detailed summary of the design and basic biology of CAR T cells was recently published, providing an excellent summary of the history and the current state of the field of CAR T-cell therapy for the treatment of malignant diseases.2 Unfortunately, the successes of CAR T cells in treating B-ALL have not yet been translated to the treatment of acute myeloid leukemia (AML), where progress has been delayed by the lack of a suitable targetable surface antigen. In BALL and other B-cell malignancies, elimination of malignant B cells occurs alongside that of normal B cells/B-cell progenitors. B-cell depletion has been clinically tolerated for years, since the ensuing hypogammaglobulinemia is easily corrected. In stark contrast, elimination of normal myeloid cells/progenitors is unlikely to be tolerated for long, as the targeted AML antigen is frequently co-expressed on healthy hematopoietic stem/progenitor cells (HSPC), leading to ablation of all myeloid progeny. Creative solutions are being sought to overcome these obstacles in order to make CART therapy a viable option for patients with AML. The current state of play: anti-acute myeloid leukemia chimeric antigen receptor T cells in the clinic Thirteen CART trials for patients with AML are currently enrolling patients (Table 1), though none have yet yielded mature published data. The first trial demonstrating biological activity for CART in AML was published in 2013, evaluating a second-generation (CD28 co-stimulatory domain) retrovirally transduced anti-Lewis Y CAR T-cell.3 Four of five enrolled patients with relapsed/refractory (RR) AML received a mean 4.46×106/kg CAR-positive T cells after lymphodepleting chemotherapy. The best responses achieved for each patient were: stable disease in two patients (for 49 days and 23 months respectively), reduction in blasts in one patient, and a cytogenetic remission in a patient with abnormal cytogenetics as the sole abnormality (i.e. blast count not elevated). Although all patients eventually progressed, this study was important as it demonstrated biological activity of CAR T cells against AML without significant hematopoietic toxicity, and the possibility of targeting a non-protein antigen.4 Table 1. Chimeric antigen receptor (CAR) T-cell therapy trials for patients with acute myeloid leukemia open for enrollment.

Self-nanoemulsifying system improves oral absorption and enhances anti-acute myeloid leukemia activity of berberine

BackgroundRecently, we found that berberine (BBR) exerts anti-acute myeloid leukemia activity, particularly toward high-risk and relapsed/refractory acute myeloid leukemia MV4-11 cells in vitro. However, the poor water solubility and low bioavailability observed with oral BBR administration has limited its clinical use. Therefore, we design and develop a novel oil-in-water self-nanoemulsifying system for BBR (BBR SNE) to improve oral bioavailability and enhance BBR efficacy against acute myeloid leukemia by greatly improving its solubility.ResultsThis system (size 23.50 ± 1.67 nm, zeta potential − 3.35 ± 0.03 mV) was prepared with RH40 (surfactant), 1,2-propanediol (co-surfactant), squalene (oil) and BBR using low-energy emulsification methods. The system loaded BBR successfully according to thermal gravimetric, differential scanning calorimetry, and Fourier transform infrared spectroscopy analyses. The release profile results showed that BBR SNE released BBR more slowly than BBR solution. The relative oral bioavailability of this novel system in rabbits was significantly enhanced by 3.41-fold over that of BBR. Furthermore, Caco-2 cell monolayer transport studies showed that this system could help enhance permeation and prevent efflux of BBR. Importantly, mice with BBR SNE treatment had significantly longer survival time than BBR-treated mice (P < 0.001) in an MV4-11 engrafted leukemia murine model.ConclusionsThese studies confirmed that BBR SNE is a promising therapy for acute myeloid leukemia.

Anti-acute myeloid leukemia activity of 2-chloro-3-alkyl-1,4-naphthoquinone derivatives through inducing mtDNA damage and GSH depletion

2-Chloro-3-alkyl-1,4-naphthoquinone derivatives were synthesized and tested as the anti-acute myeloid leukaemia agents. The compound 9b (2-chloro-3-ethyl-5,6,7-trimethoxy-1,4-naphthoquinone) was the most potent toward HL-60 leukaemia cells. In mechanistic study for 9b, the protein levels of mtDNA-specific DNA polymerase γ (poly-γ) and mtDNA transcription factor A (mt-TFA) were decreased after the 24 h treatment, showing the occurrence of mtDNA damage. And 9b triggered cell cycle arrest at S phase accompanied by a secondary block in G2/M phase which had a direct link to the process of mtDNA damage. The dissipations of mitochondrial membrane potential and ATP also proceeded. On the other hand, 9b promoted the generation of ROS and resulted in the oxidation of intracellular GSH to GSSG. This process was coupled to the formation of adduct between 9b and GSH, detected by the UV–Vis spectrum and HRMS analysis. Depletion of GSH by buthionine sulfoximine enhanced ROS level and produced higher cytotoxicity, suggesting GSH was involved in the anti-leukemic mechanism of 9b. Together, our results provide new insights on the molecular mechanism of the derivatives of 2-chloro-1,4-naphthoquinone and 9b might be useful for the further development into an anti-leukemia agent.