基于NMR代谢组学技术的款冬花生品与蜜炙品化学成分比较

目的探究款冬花蜜炙的科学内涵。方法基于NMR代谢组学技术对款冬花生品和蜜炙品进行分析,采用主成分分析(PCA)、正交偏最小二乘法辨别分析(OPLS-DA)以及单变量分析对款冬花蜜炙前后化学成分进行比较。结果款冬花的代谢指纹图谱共指认出40种代谢物,多元统计结果显示款冬花蜜炙品和生品之间存在显著差异,蜜炙后初级代谢产物1-O-乙基-β-D-葡萄糖苷、β-葡萄糖、蔗糖、α-葡萄糖的量明显升高,缬氨酸、天冬氨酸、苏氨酸的量降低;次级代谢产物款冬酮、芦丁的量升高,绿原酸、咖啡酸等有机酸的量有所降低。结论从整体化学组成上比较了款冬花生品与蜜炙品的差异,为其蜜炙的科学内涵研究奠定了基础。     

Catastrophic ATP loss underlies a metabolic combination therapy tailored for MYCN-amplified neuroblastoma

MYCN-amplified neuroblastoma is a lethal subset of pediatric cancer. MYCN drives numerous effects in the cell, including metabolic changes that are critical for oncogenesis. The understanding that both compensatory pathways and intrinsic redundancy in cell systems exists implies that the use of combination therapies for effective and durable responses is necessary. Additionally, the most effective targeted therapies exploit an “Achilles’ heel” and are tailored to the genetics of the cancer under study. We performed an unbiased screen on select metabolic targeted therapy combinations and correlated sensitivity with over 20 subsets of cancer. We found that MYCN-amplified neuroblastoma is hypersensitive to the combination of an inhibitor of the lactate transporter MCT1, AZD3965, and complex I of the mitochondrion, phenformin. Our data demonstrate that MCT4 is highly correlated with resistance to the combination in the screen and lowly expressed in MYCN-amplified neuroblastoma. Low MCT4 combines with high expression of the MCT2 and MCT1 chaperone CD147 in MYCN-amplified neuroblastoma, altogether conferring sensitivity to the AZD3965 and phenformin combination. The result is simultaneous disruption of glycolysis and oxidative phosphorylation, resulting in dramatic disruption of adenosine triphosphate (ATP) production, endoplasmic reticulum stress, and cell death. In mouse models of MYCN-amplified neuroblastoma, the combination was tolerable at concentrations where it shrank tumors and did not increase white-blood-cell toxicity compared to single drugs. Therefore, we demonstrate that a metabolic combination screen can identify vulnerabilities in subsets of cancer and put forth a metabolic combination therapy tailored for MYCN-amplified neuroblastoma that demonstrates efficacy and tolerability in vivo.

Despite their relative rarity compared to blood cancers, solid-tumor pediatric cancers are now the leading cause of pediatric cancer-related deaths. Among the most deadly is high-risk neuroblastoma (NB): amplification of MYCN confers high risk and is the clear driver of NB in these cancers (1). As such, MYCN remains the most important drug target in NB and one of the most important in pediatric cancer. Unfortunately, direct chemical targeting of MYCN has not yet been successful, and despite advancements in anti-GD2 immunotherapy (2), alternate ways of targeting MYCN-amplified NB may be needed to successfully treat this cancer.One approach is to find tumor-specific vulnerabilities, which are exploitable pharmacologically. Many efforts, including ours (3), have exhaustively looked for kinase inhibitors with particular efficacy in MYCN-amplified NBs. However, the emerging picture is a lack of kinase inhibitor efficacy in MYCN-amplified NB. Other vulnerabilities may be classified under the broad category of drugs targeting epigenetic modifiers. For example, using a CRISPR/Cas9 screen, Stegmaier and colleagues demonstrated that MYCN-amplified NB may be susceptible to targeting the H3K27me methylase EZH2 (4); in a different study, they demonstrated the susceptibility of MYCN-amplified NB to the combination of BRD4 inhibitors with CDK7 inhibitors (5). In addition, Thiele and colleagues (6) demonstrated high-risk NBs were susceptible to inhibition of the lysine methyltransferase SETD8. As promising as these data are, it remains unknown whether tolerability and/or clinical activity in MYCN-amplified NB will occur and SETD8, BRD4, and CDK7 inhibitors so far are not in the pediatric clinic. Cell death inducers constitute a third category. To this point, we recently uncovered a susceptibility of MYCN-amplified NB to the BCL-2 inhibitor venetoclax (3), confirmed by others (7). There, MYCN-driven NOXA expression sensitizes cells to venetoclax (3). Venetoclax is now in early phase trials in pediatric patients including those with NB ({"type":"clinical-trial","attrs":{"text":"NCT03236857","term_id":"NCT03236857"}}NCT03236857). It remains to be seen whether or not it will elicit responses in NB patients as a single agent.A fourth distinct category of therapeutic strategies to indirectly target oncogenes is through metabolism targeting, involving the growing coterie of drugs targeting the pathways fulfilling the high-energy demands of cancer cells. A major energy currency in cells is adenosine triphosphate (ATP). The Warburg effect describes the propensity of cancer cells (and highly proliferating normal cells) to produce ATP in the presence of oxygen with the less efficient, extramitochondrial glycolysis, as opposed to the more efficient mitochondria-based oxidative phosphorylation occurring in most noncancerous cells (8). The mechanistic explanation of the Warburg effect and how it might benefit cancer cells has been revised dramatically over the years. It was originally proposed that mitochondria from cancer cells were defective and lacked oxidative phosphorylation capabilities (9); on the contrary, emerging data show that many cancers rely on oxidative phosphorylation to facilitate the generation of ATP (8, 10). Interestingly, while amplified MYCN directly regulates the expression of many of the key glycolytic enzymes and as such contributes to the Warburg effect (11, 12), a study utilizing a Seahorse respirator demonstrated that a MYCN-amplified NB cell line favored oxidative phosphorylation over glycolysis for the metabolic needs, while the reverse was true for a MYCN wild-type NB cell line (13). In an independent study, MYCN was associated with higher glycolytic flux and oxidative phosphorylation and conferred sensitivity to fatty acid oxidation disruption (12). Overall, since c-MYC, which shares ∼40% binding homology to DNA-binding sites throughout the genome with MYCN, has been extensively characterized as a metabolic master regulator (14, 15), it is likely there are other MYCN-driven metabolic processes that may represent significant drug targets.Monocarboxylate transporters (MCTs) consist of four members (MCT1–4) in mammalian cells. Among their most critical substrates are lactate and pyruvate; MCT1 and MCT4 are responsible for lactate export across the plasma membrane to the extracellular space (16). AZD3965 (17) (AstraZeneca) is the first in-class–specific MCT1/2 dual inhibitor and is currently in early phase trials for diverse cancers; however, other inhibitors from different companies have recently been developed as well (18). Of note, AZD3965 has demonstrated good tolerability in diverse patients (clinical trial number {"type":"clinical-trial","attrs":{"text":"NCT01791595","term_id":"NCT01791595"}}NCT01791595). Although rare (65 cases/100,000 person-years), lactic acidosis led to the market retrieval of phenformin in America (19), yet phenformin remains in use as a type II antidiabetic drug in Europe, functioning centrally as a mitochondrial complex I electron transport chain (ETC) inhibitor. Phenformin reduces both glycolytic intermediates and pyruvate, increases shunting of glucose-derived carbon (increasing total lactate production), and markedly reduces tricarboxylic acid cycle intermediates (20). Indeed, there has been a recent resurgence in interest in the use of phenformin to treat cancer. For example, in BRAF mutant melanoma, phenformin sensitized cells to BRAF inhibitor through cooperative suppression of the metabolic sensor pathway mTORC1 (21). These preclinical data have led to a clinical trial of phenformin in combination with BRAF inhibitor in BRAF mutant melanoma ({"type":"clinical-trial","attrs":{"text":"NCT03026517","term_id":"NCT03026517"}}NCT03026517). Overall, while targeting individual metabolic pathways has demonstrated some preclinical success in different cancer models, it is limited with significant redundancy in pathways to generate ATP and regenerate NAD+ (22). We therefore assessed potential combination therapies involving metabolic targeting drugs to identify a strategy for MYCN-amplified NB.     

Novel serum metabolomics‐based approach by gas chromatography/triple quadrupole mass spectrometry for detection of human skin cancers: Candidate biomarkers

Skin cancer incidence rates are continuing to rise; however, if detected at an early stage, they can be cured with minimally invasive treatment. Therefore, the identification of novel and robust biomarkers for the early detection of skin cancer is required to improve the quality of life of the patient after treatment. In the present study, we aimed to identify novel biomarkers of skin cancers. We carried out serum metabolomics using gas chromatography/triple quadrupole mass spectrometry for two types of skin cancer: squamous cell carcinoma and melanoma. The changes in the expression of metabolites compared with healthy volunteers were analyzed by principal component analysis. Among all 118 metabolites, 27 in patients with squamous cell carcinoma and 33 in patients with melanoma showed significant changes in comparison with healthy volunteers. Principal component analysis showed that both skin cancer groups could be distinguished from the healthy volunteers group. We further investigated the specific metabolites most useful for these distinctions. In the squamous cell carcinoma group, these metabolites were glycerol, 4‐hydroxybenzoic acid, sebacic acid, fucose and suberic acid. In the melanoma group, these metabolites were glutamic acid, sebacic acid, suberic acid, 4‐hydroxybenzoic acid and phenylalanine. The present study identified several metabolites that were distinct for certain skin cancer types, which could potentially be used as diagnostic biomarkers leading to novel clinical management strategies.     

Understanding the metabolome – challenges for metabolomics

Metabolomics is a comprehensive method for metabolite assessment, measuring the overall metabolic signature of biological samples. This approach opens up many possibilities in areas such as new biomarker discovery and hypothesis generation. The application of metabolomics is growing rapidly but many challenges must first be addressed before it can reach its true potential. Metabolomics organisations are currently working towards guidelines for commonality in metabolomics experiments as development of optimal methodologies and study designs are needed. Blood and urine appear to be the most useful biofluids for nutrition research, but an array of biofluids, cells and tissues can be used. The key steps required for the successful understanding of metabolomics data are compound identification and biological interpretation. Many databases of compounds are available but are still under construction with much information remaining to be populated. An understanding of the effects of normal physiological variation on metabolic profiles is essential for accurate interpretation of profile changes, particularly in human studies, because of diversity in lifestyle and environmental factors. The effects of factors such as ethnicity, gender, age, body composition, health, dietary intake, physical activity, gut microflora and stress need to be further explored in order to advance the understanding of the human metabolome and therefore improve data interpretation.     

代谢组学在中医药领域的应用与展望

代谢组学作为系统生物学的重要组成部分，与中医药基础理论的“整体观”具有一致性，通过现代分析技术手段检测生物体系内代谢产物的变化，更准确、直接地反映生物体系的终端和表型信息，揭示中医药治疗复杂疾病的作用机制。将中医药研究与代谢组学方法相互结合是中医药现代化进程的重要之举，为探求中医药研究新思路和新方法奠定重要基础。通过对近10年相关研究成果进行归纳，概述代谢组学在中医证候、中药作用机制、中药安全性评价和中药质量控制等方面的应用和研究进展，分析和总结代谢组学在中医药领域中的研究方法，并提出代谢组学在中医药研究中的思考和展望。     

高血压病与中医证型的代谢组学研究探析

高血压病是以体循环动脉压升高、周围小动脉阻力增高,同时伴有不同程度代谢障碍为主要表现的临床综合征,是一种危害极大的世界范围内的疾病。虽然中医学与代谢组学在不同时代背景产生和发展,但两者具有诸多相同之处。两者均强调整体观,且具有相似的思维模式。代谢组学技术与中医证的研究相结合,可把证的研究再次推向了科学发展的前沿,成为中医现代化最重要的研究方向和有效突破口之一。     

代谢组学在眼部疾病中的研究进展

眼部疾病是全球的重要健康问题。代谢组学是对一个系统中的代谢物进行识别和定量的新兴研究技术，可用于阐明疾病机制及识别新的生物标志物，为深入探究眼部疾病的发生和发展提供了新的策略。本文简介了代谢组学的基本技术，汇总分析了代谢组学在糖尿病视网膜病变、年龄相关性黄斑变性、青光眼和干眼四种眼部疾病中的最新研究进展，总结了潜在的生物标志物和代谢途径，并对代谢组学在未来眼部疾病管理和治疗中的应用前景进行了展望。