BACKGROUND & AIMS: BACKGROUND:Sperm epigenetics is an emerging area of study supported by observations reporting that abnormal sperm DNA methylation patterns are associated with infertility. Here, we explore cytosine-guanine dinucleotides (CpGs) methylation in high (HM) and low motile (LM) Bos taurus sperm populations separated by Percoll gradient. HM and LM methylation patterns were investigated by bisulfite sequencing. RESULTS:Comparison between HM and LM sperm populations revealed that methylation variation affects genes involved in chromatin organization. CpG Islands (CGIs), were highly remodelled. A high proportion of CGIs was found to be methylated at low/intermediate level (20-60%) and associated to the repetitive element BTSAT4 satellite. The low/intermediate level of methylation in BTSAT4 was stably maintained in pericentric regions of chromosomes. BTSAT4 was hypomethylated in HM sperm populations. CONCLUSIONS:The characterization of the epigenome in HM and LM Bos taurus sperm populations provides a first step towards the understanding of the effect of methylation on sperm fertility. Methylation variation observed in HM and LM populations in genes associated to DNA structure remodelling as well as in a repetitive element in pericentric regions suggests that maintenance of chromosome structure through epigenetic regulation is probably crucial for correct sperm functionality.

背景与目标： 背景：精子表观遗传学是一个新兴的研究领域，其观察结果表明，精子DNA甲基化异常与不育有关。在这里，我们探索由Percoll梯度分离的高（HM）和低运动（LM）的金牛座精子群体中的胞嘧啶-鸟嘌呤二核苷酸（CpGs）甲基化。亚硫酸氢盐测序研究了HM和LM甲基化模式。
结果：HM和LM精子种群之间的比较表明，甲基化变异影响染色质组织所涉及的基因。 CpG岛（CGI）进行了高度重塑。发现高比例的CGI在低/中水平（20-60％）处被甲基化，并与重复元素BTSAT4卫星相关。 BTSAT4的甲基化水平处于低/中级水平，在染色体的外周中心区域保持稳定。 BTSAT4在HM精子群体中被低甲基化。
结论：HM和LM金牛座的精子群体的表观基因组的表征为迈向了解甲基化对精子繁殖力的影响迈出了第一步。在HM和LM群体中与DNA结构重塑相关的基因以及周围中心区域的重复元件中观察到的甲基化变化表明，通过表观遗传调控维持染色体结构可能对正确的精子功能至关重要。

BACKGROUND & AIMS: OBJECTIVES:Famine exposure in utero can 'programme' an individual towards type 2 diabetes and obesity in later life. We sought to identify, (1) whether Bangladeshis exposed to famine during developmental life are programmed towards diabetes and obesity, (2) whether this programming was specific to gestational or postnatal exposure windows and (3) whether epigenetic differences were associated with famine exposure. DESIGN:A historical cohort study was performed as part of a wider cross-sectional survey. Exposure to famine was defined through birth date and historical records and participants were selected according to: (A) exposure to famine in postnatal life, (B) exposure to famine during gestation and (C) unexposed. SETTING:Matlab, a rural area in the Chittagong division of Bangladesh. PARTICIPANTS:Young adult men and women (n=190) recruited to a historical cohort study with a randomised subsample included in an epigenetic study (n=143). OUTCOME MEASURES:Primary outcome measures of weight, body mass index and oral glucose tolerance tests (0 and 120 min glucose). Secondary outcome measures included DNA methylation using genome-wide and targeted analysis of metastable epialleles sensitive to maternal nutrition. RESULTS:More young adults exposed to famine in gestation were underweight than those postnatally exposed or unexposed. In contrast, more young adults exposed to famine postnatally were overweight compared to those gestationally exposed or unexposed. Underweight adults exposed to famine in gestation in utero were hyperglycaemic following a glucose tolerance test, and those exposed postnatally had elevated fasting glucose, compared to those unexposed. Significant differences in DNA methylation at seven metastable epialleles (VTRNA2-1, PAX8, PRDM-9, near ZFP57, near BOLA, EXD3) known to vary with gestational famine exposure were identified. CONCLUSIONS:Famine exposure in developmental life programmed Bangladeshi offspring towards diabetes and obesity in adulthood but gestational and postnatal windows of exposure had variable effects on phenotype. DNA methylation differences were replicated at previously identified metastable epialleles sensitive to periconceptual famine exposure.

背景与目标： 目的：子宫内的饥荒暴露可以使个体“编程”走向以后的2型糖尿病和肥胖症。我们试图确定：（1）孟加拉国在发育过程中是否暴露于饥荒是否针对糖尿病和肥胖症；（2）该计划是否特定于妊娠或出生后暴露窗口；（3）表观遗传差异是否与饥荒暴露相关。
设计：进行了一项历史队列研究，作为更广泛的横断面调查的一部分。通过出生日期和历史记录定义饥荒的暴露，并根据以下条件选择参与者：（A）出生后生活中的饥荒；（B）孕期中的饥荒暴露；（C）未暴露。
地点：Matlab，孟加拉国吉大港地区的农村地区。
参与者：参加历史队列研究的年轻成年男性和女性（n = 190），其表观遗传学研究（n = 143）中包含随机分组的子样本。
观察指标：体重，体重指数和口服葡萄糖耐量试验（0和120µmin葡萄糖）的主要观察指标。次要结果测量包括使用全基因组的DNA甲基化以及对孕产妇营养敏感的亚稳态表位等位基因的靶向分析。
结果：与出生后暴露或未暴露的婴儿相比，孕期暴露于饥荒的年轻成年人体重不足。相比之下，与未妊娠或未妊娠相比，出生后遭受饥荒的年轻人更多。体重过轻的成年人在子宫内妊娠时接受饥荒后，进行了葡萄糖耐量试验，血糖升高，与未暴露的人相比，出生后暴露的人的空腹血糖升高。确定了在七个亚稳态的等位基因（VTRNA2-1，PAX8，PRDM-9，ZFP57附近，BOLA附近，EXD3）的DNA甲基化存在显着差异，这些差异已知随妊娠饥荒的暴露而变化。
结论：成年期孟加拉国后代在成年后患糖尿病和肥胖的过程中经历了饥荒暴露，但是妊娠和出生后的暴露时间对表型有不同的影响。 DNA甲基化差异在先前确定的对概念性饥荒暴露敏感的亚稳态表位等位基因处复制。

BACKGROUND & AIMS: :Breast Cancer 1 (BRCA1) gene is a well-characterized tumor suppressor gene, mutations of which are primarily found in women with breast and ovarian cancers. BRCA1-associated RING domain 1 (BARD1) gene has also been identified as an important tumor suppressor gene in breast, ovarian, and uterine cancers. Underscoring the functional significance of the BRCA1 and BARD1 interactions, prevalent mutations in the BRCA1 gene are found in its RING domain, through which it binds the RING domain of BARD1. BARD1-BRCA1 heterodimer plays a crucial role in a variety of DNA damage response (DDR) pathways, including DNA damage checkpoint and homologous recombination (HR). However, many mutations in both BARD1 and BRCA1 also exist in other domains that significantly affect their biological functions. Intriguingly, recent genome-wide studies have identified various single nucleotide polymorphisms (SNPs), genetic alterations, and epigenetic modifications in or near the BARD1 gene that manifested profound effects on tumorigenesis in a variety of non-breast and non-gynecological cancers. In this review, we will briefly discuss the molecular functions of BARD1, including its BRCA1-dependent as well as BRCA1-independent functions. We will then focus on evaluating the common BARD1 related SNPs as well as genetic and epigenetic changes that occur in the non-BRCA1-dominant cancers, including neuroblastoma, lung, and gastrointestinal cancers. Furthermore, the pro- and anti-tumorigenic functions of different SNPs and BARD1 variants will also be discussed.

背景与目标： ：乳腺癌1（BRCA1）基因是特征明确的抑癌基因，其突变主要在患有乳腺癌和卵巢癌的女性中发现。 BRCA1相关的RING域1（BARD1）基因也已被确定为乳腺癌，卵巢癌和子宫癌的重要抑癌基因。突出BRCA1和BARD1相互作用的功能重要性，发现BRCA1基因在其RING域中普遍存在突变，通过该突变与BARD1的RING域结合。 BARD1-BRCA1异二聚体在包括DNA损伤检查点和同源重组（HR）在内的各种DNA损伤反应（DDR）途径中起着至关重要的作用。但是，BARD1和BRCA1中的许多突变也存在于其他域中，这些突变会显着影响其生物学功能。有趣的是，最近的全基因组研究已经确定了BARD1基因内或附近的各种单核苷酸多态性（SNP），遗传变异和表观遗传修饰，这些变异对各种非乳腺癌和非妇科癌症的肿瘤发生均产生了深远的影响。在这篇综述中，我们将简要讨论BARD1的分子功能，包括其依赖于BRCA1的功能以及不依赖于BRCA1的功能。然后，我们将专注于评估常见的BARD1相关SNP以及在非BRCA1占主导地位的癌症（包括神经母细胞瘤，肺癌和胃肠道癌症）中发生的遗传和表观遗传学变化。此外，还将讨论不同SNP和BARD1变体的促肿瘤和抗肿瘤功能。

BACKGROUND & AIMS: :Genistein (4',5,7-trihydroxyisoflavone) is the most abundant isoflavone found in the soybean. The effects of genistein on various cancer cell lines have been extensively studied but the precise molecular mechanisms are not known. We report here the epigenetic mechanism of the action of genistein on androgen-sensitive (LNCaP) and androgen-insensitive (DuPro) human prostate cancer cell lines. Genistein induced the expression of tumor suppressor genes p21 (WAF1/CIP1/KIP1) and p16 (INK4a) with a concomitant decrease in cyclins. There was a G(0)-G(1) cell cycle arrest in LNCaP cells and a G(2)-M arrest in DuPro cells after genistein treatment. Genistein also induced apoptosis in DuPro cells. DNA methylation analysis revealed the absence of p21 promoter methylation in both cell lines. The effect of genistein on chromatin remodeling has not been previously reported. We found that genistein increased acetylated histones 3, 4, and H3/K4 at the p21 and p16 transcription start sites. Furthermore, we found that genistein treatment also increased the expression of histone acetyl transferases that function in transcriptional activation. This is the first report on epigenetic regulation of various genes by genistein through chromatin remodeling in prostate cancer. Altogether, our data provide new insights into the epigenetic mechanism of the action of genistein that may contribute to the chemopreventive activity of this dietary isoflavone and have important implications for epigenetic therapy.

背景与目标： ：Genistein（4'，5,7-三羟基异黄酮）是大豆中含量最高的异黄酮。金雀异黄素对各种癌细胞系的作用已被广泛研究，但确切的分子机制尚不清楚。我们在这里报告染料木黄酮对雄激素敏感（LNCaP）和雄激素不敏感（DuPro）人前列腺癌细胞系的作用的表观遗传机制。金雀异黄素诱导了抑癌基因p21（WAF1 / CIP1 / KIP1）和p16（INK4a）的表达，并伴随着细胞周期蛋白的降低。金雀异黄素处理后，LNCaP细胞中有G（0）-G（1）细胞周期停滞，而DuPro细胞中有G（2）-M停滞。金雀异黄素还诱导DuPro细胞凋亡。 DNA甲基化分析显示两种细胞系均不存在p21启动子甲基化。染料木黄酮对染色质重塑的影响以前尚未见报道。我们发现染料木黄酮增加了p21和p16转录起始位点处的乙酰化组蛋白3、4和H3 / K4。此外，我们发现金雀异黄素处理还增加了在转录激活中起作用的组蛋白乙酰基转移酶的表达。这是关于染料木黄酮通过染色质重塑在前列腺癌中对各种基因进行表观遗传调控的首次报道。总之，我们的数据为染料木黄酮作用的表观遗传机制提供了新的见解，这可能有助于这种饮食异黄酮的化学预防活性，并且对表观遗传疗法具有重要意义。

BACKGROUND & AIMS: BACKGROUND:Ipilimumab (Ipi) improves the survival of advanced melanoma patients with an incremental long-term benefit in 10-15 % of patients. A tumor signature that correlates with this survival benefit could help optimizing individualized treatment strategies. METHODS:Freshly frozen melanoma metastases were collected from patients treated with either Ipi alone (n: 7) or Ipi combined with a dendritic cell vaccine (TriMixDC-MEL) (n: 11). Samples were profiled by immunohistochemistry (IHC), whole transcriptome (RNA-seq) and methyl-DNA sequencing (MBD-seq). RESULTS:Patients were divided in two groups according to clinical evolution: durable benefit (DB; 5 patients) and no clinical benefit (NB; 13 patients). 20 metastases were profiled by IHC and 12 were profiled by RNA- and MBD-seq. 325 genes were identified as differentially expressed between DB and NB. Many of these genes reflected a humoral and cellular immune response. MBD-seq revealed differences between DB and NB patients in the methylation of genes linked to nervous system development and neuron differentiation. DB tumors were more infiltrated by CD8(+) and PD-L1(+) cells than NB tumors. B cells (CD20(+)) and macrophages (CD163(+)) co-localized with T cells. Focal loss of HLA class I and TAP-1 expression was observed in several NB samples. CONCLUSION:Combined analyses of melanoma metastases with IHC, gene expression and methylation profiling can potentially identify durable responders to Ipi-based immunotherapy.

背景与目标： 背景：伊立木单抗（Ipi）可改善晚期黑色素瘤患者的生存率，并在10-15％的患者中增加长期获益。与此生存获益相关的肿瘤特征可能有助于优化个体化治疗策略。
方法：从单独用Ipi（n：7）或Ipi联合树突状细胞疫苗（TriMixDC-MEL）（n：11）治疗的患者中收集新鲜冷冻的黑色素瘤转移灶。通过免疫组织化学（IHC），全转录组（RNA-seq）和甲基-DNA测序（MBD-seq）对样品进行分析。
结果：根据临床进展将患者分为两组：持久获益（DB； 5例）和无临床获益（NB； 13例）。通过IHC分析了20个转移灶，通过RNA-和MBD-seq分析了12个转移灶。鉴定出325个基因在DB和NB之间差异表达。这些基因中的许多反映了体液和细胞的免疫反应。 MBD-seq揭示了DB和NB患者之间在与神经系统发育和神经元分化有关的基因甲基化方面的差异。与NB肿瘤相比，DB肿瘤更易被CD8（）和PD-L1（）细胞浸润。 B细胞（CD20（））和巨噬细胞（CD163（））与T细胞共定位。在几个NB样品中观察到了HLA I类和TAP-1表达的局灶性丧失。
结论：将黑色素瘤转移与IHC，基因表达和甲基化分析相结合，可以潜在地确定基于Ipi的免疫治疗的持久应答者。

BACKGROUND & AIMS: :Phosphorylation of histone H3 at serine 10 (p-H3S10) is a marker of active gene transcription. Using cognitive models of neural plasticity, p-H3S10 was shown to be downstream of extracellular signal-regulated kinase (ERK) signalling in the hippocampus. In this study, we show that nociceptive signalling after peripheral formalin injection increased p-H3S10 expression in the ipsilateral dorsal horn. This increase was maximal 30 minutes after formalin injection and occurred mainly within p-ERK-positive neurons. Spinal p-H3S10-enhanced expression was also observed in neurokinin 1 receptor (NK1R), c-Fos, and Zif268 positive neurons and was inhibited by ablation of serotonergic descending controls. The mitogen and stress-activated protein kinase 1 (MSK1) is downstream of ERK and can induce p-H3S10. We found that, after formalin injection, most phospho-MSK1 (p-MSK1)-positive cells (87% ± 3%) expressed p-ERK and the majority of p-H3S10-positive cells (85% ± 5%) expressed p-MSK1. Inhibition of ERK activity with the MEK inhibitor SL327 reduced formalin-induced p-ERK, p-MSK1, and p-H3S10, demonstrating that spinal p-MSK1 and p-H3S10 were at least partly downstream of ERK signalling. Crucially, pharmacological blockade of spinal MSK1 activity with the novel MSK1 inhibitor SB727651A inhibited formalin-induced spinal p-H3S10 and nocifensive behaviour. These findings are the first to establish the involvement of p-H3S10 and its main kinase, MSK1, in ERK regulation of nociception. Given the general importance of ERK signalling in pain processing, our results suggest that p-H3S10 could play a role in the response to injury.

背景与目标： ：丝氨酸10（p-H3S10）处的组蛋白H3磷酸化是活性基因转录的标志。使用神经可塑性的认知模型，p-H3S10显示为海马中细胞外信号调节激酶（ERK）信号的下游。在这项研究中，我们显示注射福尔马林外围药后的伤害性信号传导会增加同侧背角中p-H3S10的表达。注射福尔马林后30分钟，这种增加最大，主要发生在p-ERK阳性神经元内。在神经激肽1受体（NK1R），c-Fos和Zif268阳性神经元中也观察到了脊髓p-H3S10增强的表达，并被消融的血清素能下降对照所抑制。有丝分裂原和应激激活蛋白激酶1（MSK1）在ERK的下游，可以诱导p-H3S10。我们发现，注射福尔马林后，大多数磷酸-MSK1（p-MSK1）阳性细胞（87％±3％）表达p-ERK，而大多数p-H3S10阳性细胞（85％±5％）表达p-ERK。 -MSK1。用MEK抑制剂SL327抑制ERK活性可降低福尔马林诱导的p-ERK，p-MSK1和p-H3S10，表明脊髓p-MSK1和p-H3S10至少部分位于ERK信号传导的下游。至关重要的是，用新型MSK1抑制剂SB727651A阻断脊柱MSK1活性的药理作用抑制了福尔马林诱导的脊柱p-H3S10和伤害行为。这些发现是第一个确定p-H3S10及其主要激酶MSK1参与ERK伤害感受调节的研究。考虑到ERK信号在疼痛处理中的重要性，我们的研究结果表明p-H3S10可能在伤害反应中发挥作用。

BACKGROUND & AIMS: :While pralatrexate (PDX) has been successfully developed for the treatment of T-cell lymphoma, the mechanistic basis for its T-cell selectivity and acquired resistance remains elusive. In an effort to potentially identify synergistic combinations that might circumnavigate or delay acquired PDX resistance, we generated resistant cells lines over a broad concentration range. PDX-resistant cell lines H9-12 and H9-200 were developed, each exhibiting an IC50 of 35 and over 1000 nM, respectively. These lines were established in vitro from parental H9 cells. Expression analysis of the proteins known to be important determinants of antifolate pharmacology revealed increase expression of dihydrofolate reductase (DHFR) due to gene amplification, and reduced folate carrier1 downregulation, as the putative mechanisms of resistance in H9-12 and H9-200 cells. Cross resistance was only seen with methotrexate but not with romidepsin, azacitidine (AZA), decitabine, gemcitabine, doxorubicin, or bortezomib. Resistance to PDX was reversed by pretreatment with hypomethylating agents in a concentration-dependent fashion. Comparison of gene expression profiles of parental and resistant cell lines confirmed markedly different patterns of gene expression, and identified the dual specificity phosphatase four (DUSP4) as one of the molecular target of PDX activity. Reduced STAT5 phosphorylation following exposure to PDX was observed in the H9 but not in the H9-12 and H9-200 cells. These data suggest that combination with hypomethylating agents could be potent, and that DUSP4 and STAT5 could represent putative biomarkers of PDX activity.

背景与目标： ：尽管已成功开发出pralatrexate（PDX）用于治疗T细胞淋巴瘤，但其T细胞选择性和获得性耐药的机制基础仍然难以捉摸。为了潜在地确定可能绕过或延迟获得的PDX耐药性的协同组合，我们在较宽的浓度范围内产生了耐药细胞系。已开发出具有PDX抵抗力的细胞系H9-12和H9-200，它们的IC50分别为35和1000 overnM。这些系是从亲本H9细胞体外建立的。已知蛋白是抗叶酸药理学的重要决定因素的蛋白质的表达分析表明，由于基因扩增，二氢叶酸还原酶（DHFR）的表达增加，并且减少了叶酸携带者1的下调，这是H9-12和H9-200细胞耐药的推定机制。仅在氨甲蝶呤中观察到交叉耐药，而在罗米地辛，阿扎胞苷（AZA），地西他滨，吉西他滨，阿霉素或硼替佐米中则未见到交叉耐药。通过用次甲基化剂预处理以浓度依赖性方式逆转了对PDX的抗性。亲本和抗性细胞系基因表达谱的比较证实了基因表达的显着不同，并鉴定了双重特异性磷酸酶四（DUSP4）作为PDX活性的分子靶标之一。在H9中观察到暴露于PDX后STAT5磷酸化降低，但在H9-12和H9-200细胞中未观察到。这些数据表明，与次甲基化剂联合使用可能是有效的，而DUSP4和STAT5可能代表了PDX活性的推测生物标记。

BACKGROUND & AIMS: :Transforming growth factor-β (TGF-β) promotes tumor invasion and metastasis by inducing epithelial-mesenchymal transition (EMT). EMT is often related with acquisition of stemness characteristics. The objective of this study was to determine whether EMT and stemness characteristics induced by TGF-β might be associated with epigenetic regulation in lung cancer. A human normal lung epithelial cell line and four lung cancer cell lines were treated with TGF-β. Transcriptome analysis of BEAS-2B and A549 cells incubated with TGF-β were analyzed through next-generation sequencing (NGS). Western blotting was carried out to investigate expression levels of epithelial and mesenchymal markers. Wound healing and Matrigel invasion assay, sphere formation assay, and in vivo mice tumor model were performed to evaluate functional characteristics of EMT and stemness acquisition. To investigate whether activation of EMT and stem cell markers might be involved in epigenetic regulation of lung cancer, experiment using a DNA methyltransferase inhibitor (5-azacytidine, AZA), methylation-specific PCR (MSP) and bisulfite sequencing were performed. NGS revealed changes in expression levels of EMT markers (E-cadherin, N-cadherin, fibronectin, vimentin, slug and snail) and stem cell markers (CD44 and CD87) in both BEAS-2B and A549 cells. Functional analysis revealed increased migration, invasion, sphere formation, and tumor development in mice after TGF-β treatment. Expression of slug and CD87 genes was activated following treatment with AZA and TGF-β. MSP and bisulfite sequencing indicated DNA demethylation of slug and CD87 genes. These results suggest that TGF-β induced EMT and cancer stemness acquisition could be associated with activation of slug and CD87 gene by their promoter demethylation.

背景与目标： ：转化生长因子-β（TGF-β）通过诱导上皮-间质转化（EMT）促进肿瘤侵袭和转移。 EMT通常与词干特征的获取有关。这项研究的目的是确定TGF-β诱导的EMT和干性特征是否可能与肺癌的表观遗传调控有关。用TGF-β处理人正常肺上皮细胞系和四种肺癌细胞系。通过下一代测序（NGS）分析了用TGF-β孵育的BEAS-2B和A549细胞的转录组分析。进行了蛋白质印迹以研究上皮和间充质标志物的表达水平。进行伤口愈合和基质胶侵袭测定，球形成测定和体内小鼠肿瘤模型以评估EMT和干性获得的功能特征。为了研究EMT和干细胞标志物的激活是否可能参与肺癌的表观遗传调控，使用DNA甲基转移酶抑制剂（5-氮杂胞苷，AZA），甲基化特异性PCR（MSP）和亚硫酸氢盐测序进行了实验。 NGS揭示了BEAS-2B和A549细胞中EMT标记（E-钙黏着蛋白，N-钙黏着蛋白，纤连蛋白，波形蛋白、,和蜗牛）和干细胞标记（CD44和CD87）表达水平的变化。功能分析显示，在TGF-β处理后，小鼠的迁移，侵袭，球形成和肿瘤发展增加。用AZA和TGF-β处理后，Slug和CD87基因的表达被激活。 MSP和亚硫酸氢盐测序表明，Slug和CD87基因的DNA脱甲基。这些结果表明，TGF-β诱导的EMT和癌症干性获得可能与它们的启动子去甲基化与Slug和CD87基因的激活有关。

BACKGROUND & AIMS: :The aim of this study was to identify novel antimelanogenic drugs from an epigenetic screening library containing various modulators targeting DNA methyltransferases, histone deacetylases, and other related enzymes/proteins. Of 141 drugs tested, K8 (4-((hydroxyamino)carbonyl)-N-(2-hydroxyethyl)-N-phenyl-benzeneacetamide; HPOB) was found to effectively inhibit the α-melanocyte-stimulating hormone (α-MSH)-induced melanin synthesis in B16-F10 murine melanoma cells without accompanying cytotoxicity. Additional experiments showed that K8 did not significantly reduce the mRNA and protein level of tyrosinase (TYR) or microphthalmia-associated transcription factor (MITF) in cells, but it potently inhibited the catalytic activity TYR in vitro (IC50, 1.1-1.5 µM) as compared to β-arbutin (IC50, 500-700 µM) or kojic acid (IC50, 63 µM). K8 showed copper chelating activity similar to kojic acid. Therefore, these data suggest that K8 inhibits cellular melanin synthesis not by downregulation of TYR protein expression through an epigenetic mechanism, but by direct inhibition of TYR catalytic activity through copper chelation. Metal chelating activity of K8 is not surprising because it is known to inhibit histone deacetylase (HDAC) 6 through zinc chelation. This study identified K8 as a potent inhibitor of cellular melanin synthesis, which may be useful for the treatment of hyperpigmentation disorders.

背景与目标： ：这项研究的目的是从表观遗传学筛选文库中鉴定出新型的产炭黑药物，其中包含针对DNA甲基转移酶，组蛋白脱乙酰基酶和其他相关酶/蛋白质的各种调节剂。在测试的141种药物中，发现K8（4-（（（羟基氨基）羰基）-N-（2-羟乙基）-N-苯基苯乙酰胺; HPOB）有效抑制α-黑素细胞刺激激素（α-MSH）-诱导B16-F10鼠黑色素瘤细胞中黑色素的合成而没有细胞毒性。其他实验表明，K8不会显着降低细胞中酪氨酸酶（TYR）或小眼症相关转录因子（MITF）的mRNA和蛋白水平，但它可以有效抑制TYR的体外催化活性（IC50，1.1-1.5 µM）。与β-熊果苷（IC50，500-700 µM）或曲酸（IC50，63 µM）相比。 K8表现出与曲酸相似的铜螯合活性。因此，这些数据表明，K8不是通过表观遗传机制抑制TYR蛋白表达，而是通过铜螯合直接抑制TYR催化活性来抑制细胞黑色素合成。 K8的金属螯合活性不足为奇，因为已知它可以通过锌螯合抑制组蛋白脱乙酰基酶（HDAC）6。这项研究确定K8是细胞黑色素合成的有效抑制剂，可能对治疗色素沉着过度有用。

BACKGROUND & AIMS: :To investigate the detailed functions and underlying mechanisms of miR-518a-5p/CCR6 in diffuse large B cell lymphoma (DLBCL) is needed. In this study, CCR6 expression levels were tested both in DLBCL cell lines and specimens. Through bioinformatics analysis and quantitative real-time PCR (qRT-PCR) validation, CCR6's targeted miRNA was obtained. Dual luciferase assay was used to verify their targeted relationship. Futhermore, using qRT-PCR, western blot, CCK8, Transwell assays, flow cytometry, pyrosequencing, chromatin immunoprecipitation, and azacitidine/C646 treatment, the detailed functions and underlying mechanisms of CCR6 and its targeted miRNA in DLBCL were detected. We found that negative correlation existed between CCR6 and miR-518a-5p in DLBCL. Both up-regulated miR-518a-5p and down-regulated CCR6 inhibited cell proliferation and invasion in vitro. Experiment then verified the regulatory relationship between miR-518a-5p and CCR6. JAK2 and STAT6 levels were reduced in DLBCL cells transfected with miR-518a-5p mimic or CCR6 small interfering RNA. Interestingly, we showed for the first time that a hyper-methylated condition existed at the promoter region of miR-518a-5p and azacitidine changed levels of miR-518a-5p in a time- and concentration-dependent manner. Finally, we found an enriched histone H3 on lysine 27 acetylation existed in the promoter of CCR6, whose expression could also be changed via C646 in a time- and concentration-dependent manner. The above results suggest that miR-518a-5p-CCR6 feedback loop plays a critical role in DLBCL development. The overexpression of CCR6 is mainly mediated by epigenetic modification through transcriptional and post-transcriptional activation, which provides new directions for DLBCL treatment.

背景与目标： ：研究miR-518a-5p / CCR6在弥漫性大B细胞淋巴瘤（DLBCL）中的详细功能和潜在机制。在这项研究中，在DLBCL细胞系和标本中均测试了CCR6表达水平。通过生物信息学分析和定量实时PCR（qRT-PCR）验证，获得了CCR6的靶向miRNA。使用双重荧光素酶测定法来验证它们的靶向关系。此外，使用qRT-PCR，western blot，CCK8，Transwell分析，流式细胞术，焦磷酸测序，染色质免疫沉淀和阿扎胞苷/ C646处理，检测了CCR6及其靶向miRNA在DLBCL中的详细功能和潜在机制。我们发现DLBCL中CCR6和miR-518a-5p之间存在负相关。上调的miR-518a-5p和下调的CCR6均在体外抑制细胞增殖和侵袭。然后，实验验证了miR-518a-5p与CCR6之间的调控关系。在用miR-518a-5p模拟物或CCR6小干扰RNA转染的DLBCL细胞中，JAK2和STAT6水平降低。有趣的是，我们首次证明了miR-518a-5p启动子区域存在高甲基化条件，而阿扎胞苷以时间和浓度依赖性方式改变了miR-518a-5p的水平。最后，我们发现CCR6启动子中存在赖氨酸27乙酰化富集的组蛋白H3，其表达也可通过C646以时间和浓度依赖性的方式改变。以上结果表明，miR-518a-5p-CCR6反馈回路在DLBCL的发展中起着至关重要的作用。 CCR6的过表达主要是通过转录和转录后激活的表观遗传修饰介导的，这为DLBCL治疗提供了新的方向。

BACKGROUND & AIMS: :Previous studies suggest epigenetic alterations may contribute to the association between maternal prenatal depression and adverse offspring outcomes. Developmental researchers have recently begun to examine these associations in relation to epigenetic age acceleration/deceleration, a biomarker of developmental risk that reflects the deviation between epigenetic age and chronological age. In the perinatal period, preliminary studies indicate that maternal prenatal depression may lead to epigenetic age deceleration in newborns, which may predict adverse developmental outcomes. The present study examined the relationship between maternal prenatal exposures (i.e., depression, stress, and SSRI use) and offspring epigenetic age deceleration in 303 mother-offspring dyads. Women were recruited in the first trimester of pregnancy and followed longitudinally until delivery. Maternal depression, perceived stress, and SSRI use were assessed at each prenatal visit. Newborn epigenetic age was determined via cord blood samples. Results indicated maternal prenatal stress was not associated with newborn epigenetic age deceleration (ΔR2 = 0.002; p = 0.37). Maternal prenatal depression was associated with decelerated epigenetic age (ΔR2 = 0.01, p = 0.04), but this relationship did not hold when accounting for maternal use of SSRIs (ΔR2 = 0.002, p = 0.43). Conversely, maternal SSRI use significantly predicted newborn epigenetic age deceleration over and above the influence of maternal depression (ΔR2 = 0.03, p = 0.001). These findings suggest maternal prenatal SSRI use may significantly contribute to the previously documented association between maternal prenatal depression and epigenetic age deceleration. Further studies are needed to examine how these epigenetic differences at birth may contribute to adverse outcomes in later development.

背景与目标： ：先前的研究表明，表观遗传学改变可能会导致产前产妇抑郁与不良后代结局之间的关联。发育研究人员最近开始研究与表观遗传年龄加速/减速有关的这些关联，表观遗传年龄的加速/减速是反映表观遗传年龄与年代年龄之间差异的发育风险生物标志。在围产期，初步研究表明，母亲的产前抑郁症可能导致新生儿的表观遗传年龄下降，这可能预示不良的发育结果。本研究检查了303个母子二胎母亲的产前暴露量（即抑郁，压力和SSRI的使用）与后代表观遗传年龄减慢之间的关系。在妊娠的头三个月招募妇女，然后纵向随访直至分娩。每次产前检查时都要评估产妇的抑郁，感觉到的压力和使用SSRI的情况。新生儿表观遗传年龄是通过脐带血样本确定的。结果表明，产前产前压力与新生儿表观遗传年龄减慢无关（ΔR2= 0.002; p = 0.37）。孕妇产前抑郁症与表观遗传年龄降低有关（ΔR2= 0.01，p = 0.04），但是当考虑到母亲使用SSRI时这种关系不成立（ΔR2= 0.002，p = 0.43）。相反，母亲SSRI的使用显着预测了新生儿后生年龄的减速，超过了母亲抑郁的影响（ΔR2= 0.03，p = 0.001）。这些发现表明，孕妇产前使用SSRI可能大大促进了孕妇产前抑郁与表观遗传年龄减慢之间的关联。需要进行进一步的研究，以检查出生时的这些表观遗传差异如何对以后的发育产生不利影响。

BACKGROUND & AIMS: :In mammals, profound changes in the population and functions of adult stem cells occur with age and these changes are thought to underlie functional decline and pathophysiology at the tissue and organismal levels associated with aging. SIRT1, a member of the conserved sirtuin family, functions as an anti-aging regulator for adult stem cells. Mediated through its regulatory roles in AMPK and mTORC1 pathways as well as gene expression, SIRT1 modulate the activities of genes maintaining stem cell functions and delays cellular senescence. Further investigation of the cross-talk between SIRT1 and other longevity target genes under different physiological conditions of stem cells may help us better design intervention strategies to antagonize stem cells aging.

背景与目标： ：在哺乳动物中，成年干细胞的数量和功能随着年龄的增长而发生深刻变化，这些变化被认为是与衰老相关的组织和机体水平的功能下降和病理生理的基础。 SIRT1是sirtuin保守家族的成员，可作为成人干细胞的抗衰老调节剂。通过其在AMPK和mTORC1途径以及基因表达中的调控作用介导，SIRT1调节维持干细胞功能并延缓细胞衰老的基因活性。进一步研究SIRT1与其他寿命目标基因在干细胞不同生理条件下的串扰可能有助于我们更好地设计干预策略，以对抗干细胞衰老。

BACKGROUND & AIMS: :Purpose: Abnormal activation of the NF-κB pathway induces a more aggressive phenotype of cutaneous melanoma. Understanding the mechanisms involved in melanoma NF-κB activation may identify novel targets for this pathway. KPC1, an E3 ubiquitin ligase, is a regulator of the NF-κB pathway. The objective of this study was to investigate the mechanisms regulating KPC1 expression and its clinical impact in melanoma.Experimental Design: The clinical impact of KPC1 expression and its epigenetic regulation were assessed in large cohorts of clinically well-annotated melanoma tissues (tissue microarrays; n = 137, JWCI cohort; n = 40) and The Cancer Genome Atlas database (TCGA cohort, n = 370). Using melanoma cell lines, we investigated the functional interactions between KPC1 and NF-κB, and the epigenetic regulations of KPC1, including DNA methylation and miRNA expression.Results: We verified that KPC1 suppresses melanoma proliferation by processing NF-κB1 p105 into p50, thereby modulating NF-κB target gene expression. Concordantly, KPC1 expression was downregulated in American Joint Committee on Cancer stage IV melanoma compared with early stages (stage I/II P = 0.013, stage III P = 0.004), and low KPC1 expression was significantly associated with poor overall survival in stage IV melanoma (n = 137; HR 1.810; P = 0.006). Furthermore, our data showed that high miR-155-5p expression, which is controlled by DNA methylation at its promoter region (TCGA; Pearson's r -0.455; P < 0.001), is significantly associated with KPC1 downregulation (JWCI; P = 0.028, TCGA; P = 0.003).Conclusions: This study revealed novel epigenetic regulation of KPC1 associated with NF-κB pathway activation, promoting metastatic melanoma progression. These findings suggest the potential utility of KPC1 and its epigenetic regulation as theranostic targets. Clin Cancer Res; 23(16); 4831-42. ©2017 AACR.

背景与目标： 目的：NF-κB途径的异常激活引起皮肤黑色素瘤更具侵略性的表型。了解黑色素瘤NF-κB激活所涉及的机制可能会确定该途径的新靶标。 KPC1是E3泛素连接酶，是NF-κB途径的调节剂。本研究的目的是研究调节KPC1表达的机制及其在黑色素瘤中的临床影响。实验设计：在大量临床上注明正确的黑素瘤组织（组织芯片； n）中评估了KPC1表达的临床影响及其表观遗传学调控。 = 137，JWCI队列； n = 40）和《癌症基因组图集》数据库（TCGA队列，n = 370）。我们使用黑色素瘤细胞系研究了KPC1和NF-κB之间的功能相互作用，以及KPC1的表观遗传调控，包括DNA甲基化和miRNA表达。调节NF-κB靶基因表达。一致地，与早期阶段相比，美国癌症四期黑色素瘤联合委员会中的KPC1表达下调了（I / II P = 0.013，III P = 0.004），而低的KPC1表达与IV期黑素瘤的总体生存率低显着相关。 （n = 137； HR 1.810； P = 0.006）。此外，我们的数据显示，miR-155-5p高表达受KPC1下调（JWCI； P = 0.028， TCGA； P = 0.003）。结论：这项研究揭示了KPC1的新表观遗传调控与NF-κB途径激活有关，促进了转移性黑色素瘤的进展。这些发现表明，KPC1及其表观遗传调控可能成为治疗诊断的靶标。临床癌症研究； 23（16）; 4831-42。 ©2017 AACR。

BACKGROUND & AIMS: :The causal mechanism for cancer predisposition in Lynch-like syndrome (LLS) remains unknown. Our aim was to elucidate the constitutional basis of mismatch repair (MMR) deficiency in LLS patients throughout a comprehensive (epi)genetic analysis. One hundred and fifteen LLS patients harboring MMR-deficient tumors and no germline MMR mutations were included. Mutational analysis of 26 colorectal cancer (CRC)-associated genes was performed. Pathogenicity of MMR variants was assessed by splicing and multifactorial likelihood analyses. Genome-wide methylome analysis was performed by the Infinium Human Methylation 450K Bead Chip. The multigene panel analysis revealed the presence of two MMR gene truncating mutations not previously found. Of a total of 15 additional MMR variants identified, five -present in 6 unrelated individuals- were reclassified as pathogenic. In addition, 13 predicted deleterious variants in other CRC-predisposing genes were found in 12 probands. Methylome analysis detected one constitutional MLH1 epimutation, but no additional differentially methylated regions were identified in LLS compared to LS patients or cancer-free individuals. In conclusion, the use of an ad-hoc designed gene panel combined with pathogenicity assessment of variants allowed the identification of deleterious MMR mutations as well as new LLS candidate causal genes. Constitutional epimutations in non-LS-associated genes are not responsible for LLS.

背景与目标： ：林奇样综合征（LLS）的癌症易感性的致病机制仍然未知。我们的目的是通过全面的（表观）遗传分析来阐明LLS患者错配修复（MMR）缺陷的构成基础。 115名LLS患者携带MMR缺陷型肿瘤且没有种系MMR突变。进行了26个大肠癌（CRC）相关基因的突变分析。通过剪接和多因素似然分析评估了MMR变异的致病性。全基因组甲基化分析是通过Infinium Human Methylation 450K Bead Chip进行的。多基因组分析揭示了以前未发现的两个MMR基因截短突变的存在。在总共鉴定出的15种其他MMR变体中，有5种（存在于6个无关的个体中）被重新分类为致病性。此外，在其他12位先证者中发现了其他CRC易感基因中的13种预测有害变体。甲基化组分析检测到一个结构性MLH1突变，但与LS患者或无癌症的个体相比，在LLS中未发现其他甲基化差异区域。总之，通过使用临时设计的基因组结合变异的致病性评估，可以鉴定有害的MMR突变以及新的LLS候选致病基因。非LS相关基因的结构性表位突变与LLS不相关。

BACKGROUND & AIMS: :The ability of an environmental exposure (i.e. endocrine disruptor) during sex determination to reprogramme the male germline and promote an epigenetic transgenerational disease phenotype suggests that environmental factors and compounds may permanently alter the germline epigenome. This epigenetic transgenerational phenomenon will be discussed with respect to adult-onset germline disease (e.g. testicular cancer). A thorough literature review is not provided, rather a perspective is provided on how this epigenetic transgenerational toxicology should be considered in germ cell disease and testicular cancer.

背景与目标： 性别确定过程中环境暴露（即内分泌干扰物）重新编程雄性种系并促进表观遗传性跨世代疾病表型的能力表明，环境因素和化合物可能会永久改变种系表观基因组。将针对成年发病的种系疾病（例如睾丸癌）讨论这种表观遗传的世代相传现象。没有提供详尽的文献综述，而是提供了在生殖细胞疾病和睾丸癌中应如何考虑这种表观遗传学的跨代毒理学的观点。