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Mathematical relationships between control group variability and assay quality metrics 对照组变异性与测定质量指标之间的数学关系
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-07-01 DOI: 10.1016/j.slasd.2023.02.003
Andrew Lim

Assay quality metrics have been used in various high-throughput screening (HTS) campaigns to indicate assay quality. Z’-factor has become one of the most widely used metrics, along with other metrics such as standardised mean difference (SSMD). In using these metrics, it is important to understand how these metrics can be impacted by the separation between control groups (indicated by the HZ ratio) and the coefficient of variation (CV) within each control group. In this paper, several mathematical equations have been derived to understand the relationship between assay quality metrics (such as Z’-factor and SSMD) and control group datasets (summarised by CV and HZ). These equations increase our understanding of the factors that improve assay quality metrics, thus providing a quantitative means to visualise how affecting control groups can impact assay quality metrics.

化验质量指标已用于各种高通量筛选(HTS)活动,以指示化验质量。Z’因子与标准化平均差(SSMD)等其他指标一起,已成为最广泛使用的指标之一。在使用这些指标时,重要的是要了解这些指标如何受到对照组之间的分离(由HZ比率表示)和每个对照组内的变异系数(CV)的影响。在本文中,推导了几个数学方程来理解分析质量指标(如Z’-因子和SSMD)与对照组数据集(用CV和HZ总结)之间的关系。这些方程增加了我们对提高测定质量指标的因素的理解,从而提供了一种定量方法来可视化影响对照组如何影响测定质量指标。
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引用次数: 0
Label-free high-throughput screening via acoustic ejection mass spectrometry put into practice 采用声射质谱法进行无标签高通量筛选
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-07-01 DOI: 10.1016/j.slasd.2023.04.001
Martin Winter , Roman P. Simon , Tim T. Häbe, Robert Ries, Yuting Wang, David Kvaskoff, Amaury Fernández-Montalván, Andreas H. Luippold, Frank H. Büttner, Wolfgang Reindl

Acoustic droplet ejection-open port interface-mass spectrometry (ADE-OPI-MS) is a novel label-free analytical technique, promising to become a versatile readout for high-throughput screening (HTS) applications. The recent introduction of ADE-OPI-MS devices to the laboratory equipment market, paired with their compatibility with laboratory automation platforms, should facilitate the adoption of this technology by a broader community. Towards this goal, instrument robustness in the context of HTS campaigns - where up to millions of samples in complex matrices are tested in a short time frame - represents a major challenge, which explains the absence of detailed literature reports on this subject. Here, we present the results of our first fully automated HTS campaign, based on the ADE-OPI-MS technology, aiming to identify inhibitors of a metabolic enzyme in a >1 million compound library. The report encompasses the assay development and validation steps, as well as the adaptation for HTS requirements, where refinement of the capillary cleaning concept was crucial for final success. Altogether, our study unequivocally demonstrates the applicability of the ADE-OPI-MS technology for HTS-based drug discovery.

声液滴喷射开放端口界面质谱(ADE-OPI-MS)是一种新型的无标记分析技术,有望成为高通量筛选(HTS)应用的通用读数。最近将ADE-OPI-MS设备引入实验室设备市场,再加上它们与实验室自动化平台的兼容性,应该有助于更广泛的社区采用这项技术。为了实现这一目标,HTS活动背景下的仪器稳健性——在短时间内测试复杂矩阵中多达数百万个样本——是一个重大挑战,这解释了缺乏关于这一主题的详细文献报告的原因。在这里,我们介绍了我们第一次基于ADE-OPI-MS技术的全自动化HTS活动的结果,旨在鉴定>;100万个复合库。该报告包括分析开发和验证步骤,以及HTS要求的调整,其中毛细管清洁概念的完善对最终成功至关重要。总之,我们的研究明确证明了ADE-OPI-MS技术在基于HTS的药物发现中的适用性。
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引用次数: 2
Fluorescent probe for the identification of potent inhibitors of the macrophage infectivity potentiator (Mip) protein of Burkholderia pseudomallei 伪伯克氏菌巨噬细胞感染增强因子(Mip)蛋白强效抑制剂的荧光探针鉴定
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-07-01 DOI: 10.1016/j.slasd.2023.03.004
Nicolas Julian Scheuplein , Theresa Lohr , Mirella Vivoli Vega , Dyan Ankrett , Florian Seufert , Lukas Kirchner , Nicholas J. Harmer , Ulrike Holzgrabe

The macrophage infectivity potentiator (Mip) protein belongs to the immunophilin superfamily. This class of enzymes catalyzes the interconversion between the cis and trans configuration of proline-containing peptide bonds. Mip has been shown to be important for the virulence of a wide range of pathogenic microorganisms, including the Gram-negative bacterium Burkholderia pseudomallei. Small molecules derived from the natural product rapamycin, lacking its immunosuppression-inducing moiety, inhibit Mip's peptidyl-prolyl cis-trans isomerase (PPIase) activity and lead to a reduction in pathogen load in vitro. Here, a fluorescence polarization assay (FPA) to enable the screening and effective development of BpMip inhibitors was established. A fluorescent probe was prepared, derived from previous pipecolic scaffold Mip inhibitors labeled with fluorescein. This probe showed moderate affinity for BpMip and enabled a highly robust FPA suitable for screening large compound libraries with medium- to high-throughput (Z factor ∼ 0.89) to identify potent new inhibitors. The FPA results are consistent with data from the protease-coupled PPIase assay. Analysis of the temperature dependence of the probe's binding highlighted that BpMip's ligand binding is driven by enthalpic rather than entropic effects. This has considerable consequences for the use of low-temperature kinetic assays.

巨噬细胞感染性增强因子(Mip)蛋白属于亲免疫蛋白超家族。这类酶催化含脯氨酸肽键的顺式和反式构型之间的相互转化。Mip已被证明对多种致病微生物的毒力很重要,包括革兰氏阴性细菌假槌伯克霍尔德菌。来源于天然产物雷帕霉素的小分子,缺乏其免疫抑制诱导部分,抑制Mip的肽基脯氨酰顺反异构酶(PPIase)活性,并导致体外病原体负荷减少。在此,建立了一种荧光偏振测定法(FPA),以筛选和有效开发BpMip抑制剂。制备了荧光探针,该探针来源于先前用荧光素标记的哌啶支架Mip抑制剂。该探针显示出对BpMip的中等亲和力,并能够实现高度稳健的FPA,适用于筛选具有中高通量(Z因子~0.89)的大型化合物文库,以鉴定有效的新抑制剂。FPA结果与蛋白酶偶联PPIase测定的数据一致。对探针结合的温度依赖性的分析强调,BpMip的配体结合是由焓效应而非熵效应驱动的。这对低温动力学测定的使用具有相当大的影响。
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引用次数: 1
Evaluating the affinity and kinetics of small molecule glycomimetics for human and mouse galectin-3 using surface plasmon resonance 利用表面等离子体共振评价人和小鼠半乳糖凝集素-3的小分子糖仿制品的亲和力和动力学
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-07-01 DOI: 10.1016/j.slasd.2023.03.005
Henry Kim , Nathalie Weidner , Céline Ronin , Emmanuel Klein , James A. Roper , Barbro Kahl-Knutson , Kristoffer Peterson , Hakon Leffler , Ulf J. Nilsson , Anders Pedersen , Fredrik R. Zetterberg , Robert J. Slack

Galectin-3 is a beta-galactoside-binding mammalian lectin that is one of a 15-member galectin family that can bind several cell surface glycoproteins via its carbohydrate recognition domain (CRD). As a result, it can influence a range of cellular processes including cell activation, adhesion and apoptosis. Galectin-3 has been implicated in various diseases, including fibrotic disorders and cancer, and is now being therapeutically targeted by both small and large molecules. Historically, the screening and triaging of small molecule glycomimetics that bind to the galectin-3 CRD has been completed in fluorescence polarisation (FP) assays to determine KD values. Surface plasmon resonance (SPR) has not been widely used for compound screening and in this study it was used to compare human and mouse galectin-3 affinity measures between FP and SPR, as well as investigate compound kinetics. The KD estimates for a set of compounds selected from mono- and di-saccharides with affinities across a 550-fold range, correlated well between FP and SPR assay formats for both human and mouse galectin-3. Increases in affinity for compounds binding to human galectin-3 were driven by changes in both kon and koff whilst for mouse galectin-3 this was primarily due to kon. The reduction in affinity observed between human to mouse galectin-3 was also comparable between assay formats. SPR has been shown to be a viable alternative to FP for early drug discovery screening and determining KD values. In addition, it can also provide early kinetic characterisation of small molecule galectin-3 glycomimetics with robust kon and koff values generated in a high throughput manner.

半乳糖凝集素-3是一种β-半乳糖苷结合哺乳动物凝集素,是半乳糖凝集素家族的15个成员之一,可以通过其碳水化合物识别结构域(CRD)结合几种细胞表面糖蛋白。因此,它可以影响一系列细胞过程,包括细胞活化、粘附和凋亡。半乳糖凝集素-3与多种疾病有关,包括纤维化疾病和癌症,目前正被小分子和大分子靶向治疗。从历史上看,与半乳糖凝集素-3 CRD结合的小分子糖模拟物的筛选和试验已经在荧光偏振(FP)测定中完成,以确定KD值。表面等离子体共振(SPR)尚未广泛用于化合物筛选,在本研究中,它被用于比较FP和SPR之间的人和小鼠半乳糖凝集素-3亲和力测量,以及研究化合物动力学。从亲和力在550倍范围内的单糖和二糖中选择的一组化合物的KD估计值在人和小鼠半乳糖凝集素-3的FP和SPR测定形式之间良好相关。对与人半乳糖凝集素-3结合的化合物的亲和力的增加是由kon和koff的变化驱动的,而对小鼠半乳糖凝集素3,这主要是由于kon。在人与小鼠半乳糖凝集素-3之间观察到的亲和力降低在不同的测定形式之间也是可比较的。SPR已被证明是FP的一种可行的替代品,用于早期药物发现筛选和确定KD值。此外,它还可以提供以高通量方式产生的具有强大kon和koff值的小分子半乳糖凝集素-3糖模拟物的早期动力学表征。
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引用次数: 1
A novel fluorogenic reporter substrate for 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-2 (PLCγ2): Application to high-throughput screening for activators to treat Alzheimer's disease 一种新的1-磷脂酰肌醇4,5-二磷酸磷酸二酯酶γ -2 (plc - γ2)荧光报告底物:用于阿尔茨海默病激活剂的高通量筛选
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.03.003
Ramya Visvanathan , Tadanobu Utsuki , Daniel E. Beck , Emma Lendy , Kuai-lin Sun , Yinghui Liu , Kirk W. Hering , Andrew Mesecar , Zhong-Yin Zhang , Karson S. Putt

A rare coding variant in PLCγ2 (P522R) expressed in microglia induces a mild activation of enzymatic activity when compared to wild-type. This mutation is reported to be protective against the cognitive decline associated with late-onset Alzheimer's disease (LOAD) and therefore, activation of wild-type PLCγ2 has been suggested as a potential therapeutic target for the prevention and treatment of LOAD. Additionally, PLCγ2 has been associated with other diseases such as cancer and some autoimmune disorders where mutations with much greater increases in PLCγ2 activity have been identified. Here, pharmacological inhibition may provide a therapeutic effect. In order to facilitate our investigation of the activity of PLCγ2, we developed an optimized fluorogenic substrate to monitor enzymatic activity in aqueous solution. This was accomplished by first exploring the spectral properties of various “turn-on” fluorophores. The most promising turn-on fluorophore was incorporated into a water-soluble PLCγ2 reporter substrate, which we named C8CF3-coumarin. The ability of PLCγ2 to enzymatically process C8CF3-coumarin was confirmed, and the kinetics of the reaction were determined. Reaction conditions were optimized to identify small molecule activators, and a pilot screen of the Library of Pharmacologically Active Compounds 1280 (LOPAC1280) was performed with the goal of identifying small molecule activators of PLCγ2. The optimized screening conditions allowed identification of potential PLCγ2 activators and inhibitors, thus demonstrating the feasibility of this approach for high-throughput screening.

与野生型相比,小胶质细胞中表达的PLCγ2(P522R)的一种罕见编码变体诱导酶活性的轻度激活。据报道,这种突变对与迟发性阿尔茨海默病(LOAD)相关的认知能力下降具有保护作用,因此,野生型PLCγ2的激活已被认为是预防和治疗LOAD的潜在治疗靶点。此外,PLCγ2与其他疾病有关,如癌症和一些自身免疫性疾病,其中已发现PLCγ2活性增加更大的突变。在这里,药理学抑制可以提供治疗效果。为了便于我们对PLCγ2活性的研究,我们开发了一种优化的荧光底物来监测水溶液中的酶活性。这是通过首先探索各种“开启”荧光团的光谱特性来实现的。最有前途的开启荧光团被掺入水溶性PLCγ2报告底物中,我们将其命名为C8CF3香豆素。确认了PLCγ2对C8CF3香豆素的酶促反应能力,并测定了反应动力学。优化反应条件以鉴定小分子活化剂,并对药理学活性化合物库1280(LOPAC1280)进行中试筛选,目的是鉴定PLCγ2的小分子激活剂。优化的筛选条件允许鉴定潜在的PLCγ2激活剂和抑制剂,从而证明了这种方法用于高通量筛选的可行性。
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引用次数: 0
Development of a high-throughput TR-FRET screening assay for LAG-3/FGL1 interaction 高通量TR-FRET筛选LAG-3/FGL1相互作用试验的开发
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.04.003
Somaya A. Abdel-Rahman , Longfei Zhang , Moustafa T. Gabr

Lymphocyte activation gene 3 (LAG-3) is a negative immune checkpoint and a key regulator of immune homeostasis with multiple biological activities related to T-cell functions. Fibrinogen-like protein 1 (FGL1) is a major LAG-3 functional ligand that is upregulated in various human cancers. LAG-3 positive T cells bind FGL1 expressed by cancer cells, which inhibits T-cell activation and cytokine secretion via indirect blocking of T cell receptor (TCR) signaling. High expression of LAG-3 and FGL1 in patients with solid tumors is associated with drug resistance and decreased survival in response to FDA-approved immune checkpoint inhibitors. Therefore, targeting the LAG-3/FGL1 pathway represents a promising therapeutic strategy to maximize the number of patients benefiting from checkpoint blockade therapy. However, there are no small molecules in existence that target LAG-3/FGL1 interaction. Herein, we report a time-resolved fluorescence resonance energy transfer (TR-FRET) assay to evaluate the ability of small molecules to inhibit LAG-3/FGL1 interaction. We further demonstrate the implementation of the developed assay in screening chemical libraries of small molecules from the NCI Diversity Set VII, FDA-approved drugs, and a focused library of NF-κB modulators. This work will pave the way for drug discovery efforts focused on therapeutic targeting of LAG-3/FGL1 interaction using small molecules.

淋巴细胞活化基因3(LAG-3)是一种阴性免疫检查点,是免疫稳态的关键调节因子,具有与T细胞功能相关的多种生物活性。纤维蛋白原样蛋白1(FGL1)是一种主要的LAG-3功能配体,在各种人类癌症中上调。LAG-3阳性T细胞结合癌症细胞表达的FGL1,其通过间接阻断T细胞受体(TCR)信号传导来抑制T细胞活化和细胞因子分泌。LAG-3和FGL1在实体瘤患者中的高表达与对FDA批准的免疫检查点抑制剂的耐药性和生存率降低有关。因此,靶向LAG-3/FGL1途径是一种很有前途的治疗策略,可以最大限度地增加受益于检查点阻断治疗的患者数量。然而,目前还没有靶向LAG-3/FGL1相互作用的小分子。在此,我们报道了一种时间分辨荧光共振能量转移(TR-FRET)测定法,以评估小分子抑制LAG-3/FGL1相互作用的能力。我们进一步证明了所开发的检测方法在筛选来自NCI Diversity Set VII、FDA批准的药物和NF-κB调节剂的集中库的小分子化学文库中的应用。这项工作将为利用小分子靶向LAG-3/FGL1相互作用的药物发现工作铺平道路。
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引用次数: 2
Comparison of two supporting matrices for patient-derived cancer cells in 3D drug sensitivity and resistance testing assay (3D-DSRT) 两种患者源性癌细胞支撑基质在3D- dsrt药物敏感性和耐药试验中的比较
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.03.002
Michaela Feodoroff , Piia Mikkonen , Laura Turunen , Antti Hassinen , Lauri Paasonen , Lassi Paavolainen , Swapnil Potdar , Astrid Murumägi , Olli Kallioniemi , Vilja Pietiäinen

Central to the success of functional precision medicine of solid tumors is to perform drug testing of patient-derived cancer cells (PDCs) in tumor-mimicking ex vivo conditions. While high throughput (HT) drug screening methods have been well-established for cells cultured in two-dimensional (2D) format, this approach may have limited value in predicting clinical responses. Here, we describe the results of the optimization of drug sensitivity and resistance testing (DSRT) in three-dimensional (3D) growth supporting matrices in a HT mode (3D-DSRT) using the hepatocyte cell line (HepG2) as an example. Supporting matrices included widely used animal-derived Matrigel and cellulose-based hydrogel, GrowDex, which has earlier been shown to support 3D growth of cell lines and stem cells. Further, the sensitivity of ovarian cancer PDCs, from two patients included in the functional precision medicine study, was tested for 52 drugs in 5 different concentrations using 3D-DSRT.

Shortly, in the optimized protocol, the PDCs are embedded with matrices and seeded to 384-well plates to allow the formation of the spheroids prior to the addition of drugs in nanoliter volumes with acoustic dispenser. The sensitivity of spheroids to drug treatments is measured with cell viability readout (here, 72 h after addition of drugs). The quality control and data analysis are performed with openly available Breeze software. We show the usability of both matrices in established 3D-DSRT, and report 2D vs 3D growth condition dependent differences in sensitivities of ovarian cancer PDCs to MEK-inhibitors and cytotoxic drugs. This study provides a proof-of-concept for robust and fast screening of drug sensitivities of PDCs in 3D-DSRT, which is important not only for drug discovery but also for personalized ex vivo drug testing in functional precision medicine studies. These findings suggest that comparing results of 2D- and 3D-DSRT is essential for understanding drug mechanisms and for selecting the most effective treatment for the patient.

实体瘤功能精准医学成功的核心是在模拟肿瘤的体外条件下对患者来源的癌症细胞(PDC)进行药物测试。虽然高通量(HT)药物筛选方法已被公认用于以二维(2D)形式培养的细胞,但这种方法在预测临床反应方面的价值可能有限。在此,我们以肝细胞系(HepG2)为例,描述了HT模式下三维(3D)生长支持基质(3D-DSRT)中药物敏感性和耐药性测试(DSRT)的优化结果。支持基质包括广泛使用的动物来源的Matrigel和基于纤维素的水凝胶GrowDex,该水凝胶早些时候已被证明支持细胞系和干细胞的3D生长。此外,使用3D-DSRT对5种不同浓度的52种药物测试了功能精准医学研究中包括的两名患者的卵巢癌症PDCs的敏感性。很快,在优化方案中,PDC嵌入基质并接种到384孔板上,以允许在使用声学分配器添加纳升体积的药物之前形成球体。球体对药物处理的敏感性通过细胞活力读数(此处为添加药物后72小时)来测量。质量控制和数据分析使用公开的Breeze软件进行。我们展示了两种基质在已建立的3D-DSRT中的可用性,并报告了卵巢癌症PDC对MEK抑制剂和细胞毒性药物敏感性的2D与3D生长条件依赖性差异。本研究为在3D-DSRT中稳健快速筛选PDC的药物敏感性提供了概念证明,这不仅对药物发现很重要,而且对功能精准医学研究中的个性化离体药物测试也很重要。这些发现表明,比较2D和3D-DSRT的结果对于了解药物机制和为患者选择最有效的治疗方法至关重要。
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引用次数: 0
Application of human iPSC-derived macrophages in a miniaturized high-content-imaging-based efferocytosis assay 人ipsc衍生巨噬细胞在小型化高含量成像的efferocytosis实验中的应用
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.04.002
Sarah Bitzer , Mozhgan Dehghan Harati , Karim C. El Kasmi , Daniela Schloesser , Julia Sauer , Heiko Olbrich , Michael Schuler , Florian Gantner , Ralf Heilker

Macrophages play a pivotal role in drug discovery due to their key regulatory functions in health and disease. Overcoming the limited availability and donor variability of human monocyte-derived macrophages (MDMs), human induced pluripotent stem cell (iPSC)-derived macrophages (IDMs) could provide a promising tool for both disease modeling and drug discovery. To access large numbers of model cells for medium- to high-throughput application purposes, an upscaled protocol was established for differentiation of iPSCs into progenitor cells and subsequent maturation into functional macrophages. These IDM cells resembled MDMs both with respect to surface marker expression and phago- as well as efferocytotic function. A statistically robust high-content-imaging assay was developed to quantify the efferocytosis rate of IDMs and MDMs allowing for measurements both in the 384- and 1536-well microplate format. Validating the applicability of the assay, inhibitors of spleen tyrosine kinase (Syk) were shown to modulate efferocytosis in IDMs and MDMs with comparable pharmacology. The miniaturized cellular assay with the upscaled provision of macrophages opens new routes to pharmaceutical drug discovery in the context of efferocytosis-modulating substances.

巨噬细胞由于其在健康和疾病中的关键调节功能,在药物发现中发挥着关键作用。克服人类单核细胞衍生巨噬细胞(MDMs)的有限可用性和供体变异性,人类诱导多能干细胞(iPSC)衍生巨噬细胞(IDMs)可以为疾病建模和药物发现提供一种有前途的工具。为了获得大量的模型细胞用于中高通量应用,建立了一种将iPSC分化为祖细胞并随后成熟为功能性巨噬细胞的扩大方案。这些IDM细胞在表面标记物表达、吞噬和泡腾功能方面都类似于MDM。开发了一种统计上稳健的高含量成像测定法,以量化IDM和MDM的泡腾细胞增多率,从而允许以384孔和1536孔微孔板形式进行测量。为了验证该测定的适用性,脾脏酪氨酸激酶(Syk)抑制剂被证明可以通过类似的药理学调节IDMs和MDM的泡腾细胞增多症。在泡腾调节物质的背景下,巨噬细胞的小型化细胞测定为药物发现开辟了新的途径。
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引用次数: 0
High-throughput approaches to uncover synergistic drug combinations in leukemia 高通量方法揭示白血病的协同药物组合
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.04.004
Emma J. Chory , Meng Wang , Michele Ceribelli , Aleksandra M Michalowska , Stefan Golas , Erin Beck , Carleen Klumpp-Thomas , Lu Chen , Crystal McKnight , Zina Itkin , Kelli M. Wilson , David Holland , Sanjay Divakaran , James Bradner , Javed Khan , Berkley E. Gryder , Craig J. Thomas , Benjamin Z. Stanton

We report a comprehensive drug synergy study in acute myeloid leukemia (AML). In this work, we investigate a panel of cell lines spanning both MLL-rearranged and non-rearranged subtypes. The work comprises a resource for the community, with many synergistic drug combinations that could not have been predicted a priori, and open source code for automation and analyses. We base our definitions of drug synergy on the Chou-Talalay method, which is useful for visualizations of synergy experiments in isobolograms, and median-effects plots, among other representations. Our key findings include drug synergies affecting the chromatin state, specifically in the context of regulation of the modification state of histone H3 lysine-27. We report open source high throughput methodology such that multidimensional drug screening can be accomplished with equipment that is accessible to most laboratories. This study will enable preclinical investigation of new drug combinations in a lethal blood cancer, with data analysis and automation workflows freely available to the community.

我们报道了一项针对急性髓细胞白血病(AML)的综合药物协同作用研究。在这项工作中,我们研究了一组横跨MLL重排和非重排亚型的细胞系。这项工作包括一个社区资源,包括许多无法先验预测的协同药物组合,以及用于自动化和分析的开源代码。我们将药物协同作用的定义建立在Chou-Talalay方法的基础上,该方法有助于在等辐射图和中值效应图等表示中可视化协同作用实验。我们的主要发现包括影响染色质状态的药物协同作用,特别是在组蛋白H3赖氨酸-27修饰状态的调节方面。我们报告了开源高通量方法,这样就可以使用大多数实验室都可以使用的设备来完成多维药物筛选。这项研究将使人们能够对致命的癌症中的新药组合进行临床前研究,并向社区免费提供数据分析和自动化工作流程。
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引用次数: 0
In Vitro three-dimensional (3D) cell culture tools for spheroid and organoid models 体外三维(3D)细胞培养工具,用于球体和类器官模型
IF 3.1 4区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS Pub Date : 2023-06-01 DOI: 10.1016/j.slasd.2023.03.006
Sang-Yun Lee , In-Seong Koo , Hyun Ju Hwang , Dong Woo Lee

Three-dimensional (3D) cell culture technology has been steadily studied since the 1990′s due to its superior biocompatibility compared to the conventional two-dimensional (2D) cell culture technology, and has recently developed into an organoid culture technology that further improved biocompatibility. Since the 3D culture of human cell lines in artificial scaffolds was demonstrated in the early 90′s, 3D cell culture technology has been actively developed owing to various needs in the areas of disease research, precision medicine, new drug development, and some of these technologies have been commercialized. In particular, 3D cell culture technology is actively being applied and utilized in drug development and cancer-related precision medicine research. Drug development is a long and expensive process that involves multiple steps—from target identification to lead discovery and optimization, preclinical studies, and clinical trials for approval for clinical use. Cancer ranks first among life-threatening diseases owing to intra-tumoral heterogeneity associated with metastasis, recurrence, and treatment resistance, ultimately contributing to treatment failure and adverse prognoses. Therefore, there is an urgent need for the development of efficient drugs using 3D cell culture techniques that can closely mimic in vivo cellular environments and customized tumor models that faithfully represent the tumor heterogeneity of individual patients. This review discusses 3D cell culture technology focusing on research trends, commercialization status, and expected effects developed until recently. We aim to summarize the great potential of 3D cell culture technology and contribute to expanding the base of this technology.

与传统的二维细胞培养技术相比,三维细胞培养技术由于其优越的生物相容性,自20世纪90年代以来一直在稳步研究,最近已发展成为一种进一步提高生物相容性的类器官培养技术。自90年代初在人工支架中进行人体细胞系的3D培养以来,由于疾病研究、精准医学、新药开发等领域的各种需求,3D细胞培养技术得到了积极发展,其中一些技术已经商业化。特别是3D细胞培养技术在药物开发和癌症相关精准医学研究中得到了积极的应用和利用。药物开发是一个漫长而昂贵的过程,涉及多个步骤——从靶点识别到先导发现和优化、临床前研究,以及临床使用批准的临床试验。癌症在危及生命的疾病中排名第一,因为肿瘤内的异质性与转移、复发和治疗耐药性相关,最终导致治疗失败和不良预后。因此,迫切需要使用3D细胞培养技术开发有效的药物,该技术可以密切模拟体内细胞环境,并定制肿瘤模型,忠实地代表个体患者的肿瘤异质性。这篇综述讨论了3D细胞培养技术,重点是研究趋势、商业化现状和最近开发的预期效果。我们旨在总结3D细胞培养技术的巨大潜力,并为扩大该技术的基础做出贡献。
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引用次数: 5
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SLAS Discovery
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