BACKGROUND & AIMS: :The reactive oxygen species (ROS) producing enzyme, NADPH oxidase 4 (Nox4), is upregulated in response to TGFβ in lens epithelial cells in vitro, and its selective inhibition was shown to block aspects of TGFβ-induced epithelial-mesenchymal transition (EMT). In the present in situ study we validate the role(s) of Nox4 in TGFβ-induced lens EMT leading to anterior subcapsular cataract (ASC) formation. Mice overexpressing TGFβ in the lens, that develop ASC, were crossed to Nox4-deficient mice. When comparing mice overexpressing TGFβ in lens, to mice that were also deficient for Nox4, we see the delayed onset of cataract, along with a delay in EMT protein markers normally associated with TGFβ-induced fibrotic cataracts. In the absence of Nox4, we also see elevated levels of ERK1/2 activity that was shown to be required for TGFβ/Smad2/3-signaling. qRT-PCR revealed upregulation of Nox2 and its regulatory subunit in TGFβ-overexpressing lens epithelial cells devoid of Nox4. Taken together, these findings provide an improved platform to delineate putative Nox4 (and ROS) interactions with Smad2/3 and/or ERK1/2, in particular in the development of fibrotic diseases, such as specific forms of cataract.

背景与目标： : 活性氧 (ROS) 产生酶NADPH氧化酶4 (Nox4) 在体外晶状体上皮细胞中响应tgf β 而上调，其选择性抑制被证明可阻断tgf β 诱导的上皮间质转化 (EMT)。在目前的原位研究中，我们验证了Nox4在tgf β 诱导的晶状体EMT中导致前囊下白内障 (ASC) 形成的作用。将在晶状体中过表达tgf β 的小鼠杂交至Nox4-deficient小鼠，以形成ASC。当将晶状体中过表达tgf β 的小鼠与也缺乏Nox4的小鼠进行比较时，我们看到白内障的延迟发作，以及通常与tgf β 诱导的纤维化性白内障相关的EMT蛋白标记的延迟。在没有Nox4的情况下，我们还看到ERK1/2活性水平升高，这被证明是tgf β/Smad2/3信号传导所必需的。qRT-PCR揭示了在没有nox4的tgf β 过表达的晶状体上皮细胞中Nox2及其调节亚基的上调。总之，这些发现提供了一个改进的平台来描述假定的Nox4 (和ROS) 与Smad2/3和/或ERK1/2的相互作用，特别是在纤维化疾病 (例如特定形式的白内障) 的发展中。

BACKGROUND & AIMS: :Brain damage due to stroke or traumatic brain injury (TBI), both leading causes of serious long-term disability, often leads to the development of epilepsy. Patients who develop post-injury epilepsy tend to have poor functional outcomes. Emerging evidence highlights a potential role for blood-brain barrier (BBB) dysfunction in the development of post-injury epilepsy. However, common mechanisms underlying the pathological hyperexcitability are largely unknown. Here, we show that comparative transcriptome analyses predict remodeling of extracellular matrix (ECM) as a common response to different types of injuries. ECM-related transcriptional changes were induced by the serum protein albumin via TGFβ signaling in primary astrocytes. In accordance with transcriptional responses, we found persistent degradation of protective ECM structures called perineuronal nets (PNNs) around fast-spiking inhibitory interneurons, in a rat model of TBI as well as in brains of human epileptic patients. Exposure of a naïve brain to albumin was sufficient to induce the transcriptional and translational upregulation of molecules related to ECM remodeling and the persistent breakdown of PNNs around fast-spiking inhibitory interneurons, which was contingent on TGFβ signaling activation. Our findings provide insights on how albumin extravasation that occurs upon BBB dysfunction in various brain injuries can predispose neural circuitry to the development of chronic inhibition deficits.

背景与目标： : 由于中风或创伤性脑损伤 (TBI) 引起的脑损伤，都是导致严重长期残疾的主要原因，通常会导致癫痫的发展。发生伤后癫痫的患者往往功能不良。新出现的证据强调了血脑屏障 (BBB) 功能障碍在损伤后癫痫发展中的潜在作用。然而，病理性过度兴奋的常见机制在很大程度上是未知的。在这里，我们显示比较转录组分析预测细胞外基质 (ECM) 的重塑是对不同类型损伤的常见反应。血清蛋白白蛋白通过tgf β 信号传导在原发性星形胶质细胞中诱导ECM相关的转录变化。根据转录反应，我们发现在TBI的大鼠模型以及人类癫痫患者的大脑中，在快速刺激的抑制性中间神经元周围，保护性ECM结构 (称为神经周网 (pnn)) 持续降解。幼稚的大脑暴露于白蛋白足以诱导与ECM重塑有关的分子的转录和翻译上调，以及快速刺激的抑制性中间神经元周围pnn的持续分解，这取决于tgf β 信号激活。我们的发现提供了有关各种脑损伤中BBB功能障碍时发生的白蛋白外渗如何使神经回路容易发展为慢性抑制缺陷的见解。

BACKGROUND & AIMS: :Diabetes mellitus is a serious metabolic condition associated with a multitude of cardiovascular complications. Moreover, the prevalence of diabetes in heart failure populations is higher than that in control populations. However, the role of cardiomyocyte alterations in type 2 diabetes mellitus (T2DM) has not been well characterized and the underlying mechanisms remain elusive. In this study, two patients who were diagnosed as T2DM were recruited and patient-specific induced pluripotent stem cells (iPSCs) were generated from urine epithelial cells using nonintegrated Sendai virus. The iPSC lines derived from five healthy subjects were used as controls. All iPSCs were differentiated into cardiomyocytes (iPSC-CMs) using the monolayer-based differentiation protocol. T2DM iPSC-CMs exhibited various disease phenotypes, including cellular hypertrophy and lipid accumulation. Moreover, T2DM iPSC-CMs exhibited higher susceptibility to high-glucose/high-lipid challenge than control iPSC-CMs, manifesting an increase in apoptosis. RNA-Sequencing analysis revealed a differential transcriptome profile and abnormal activation of TGFβ signaling pathway in T2DM iPSC-CMs. We went on to show that inhibition of TGFβ significantly rescued the hypertrophic phenotype in T2DM iPSC-CMs. In conclusion, we demonstrate that the iPSC-CM model is able to recapitulate cellular phenotype of T2DM. Our results indicate that iPSC-CMs can therefore serve as a suitable model for investigating molecular mechanisms underlying diabetic cardiomyopathies and for screening therapeutic drugs.

背景与目标： 糖尿病是一种与多种心血管并发症相关的严重代谢性疾病。此外，心力衰竭人群中糖尿病的患病率高于对照组。然而，心肌细胞改变在2型糖尿病 (T2DM) 中的作用尚未得到很好的表征，其潜在机制仍然难以捉摸。在这项研究中，招募了两名被诊断为T2DM的患者，并使用非整合仙台病毒从尿液上皮细胞中产生了患者特异性诱导多能干细胞 (iPSCs)。来自五名健康受试者的iPSC品系被用作对照。使用基于单层的分化方案将所有iPSCs分化为心肌细胞 (ipsc-cms)。T2DM ipsc-cms表现出各种疾病表型，包括细胞肥大和脂质积累。此外，与对照ipsc-cms相比，T2DM ipsc-cms对高糖/高脂攻击的敏感性更高，表现出细胞凋亡的增加。RNA测序分析揭示了T2DM ipsc-cms中存在差异的转录组谱和tgf β 信号通路的异常激活。我们继续证明，抑制tgf β 可显着挽救T2DM ipsc-cms中的肥厚表型。总之，我们证明了ipsc-二1212模型能够概括T2DM的细胞表型。我们的结果表明，ipsc-cms可以作为研究糖尿病心肌病潜在分子机制和筛选治疗药物的合适模型。

BACKGROUND & AIMS: :Transforming growth factor-β (TGFβ) signaling regulates multiple cellular processes, including extracellular matrix production, cell growth, apoptosis and differentiation. Dysfunction of TGFβ signaling has been implicated in various human disorders ranging from vascular diseases to cancer. TGFβ signaling is negatively regulated by the transcriptional repressor TGFβ-induced factor 1 (TGIF1). The tumor suppressor Fbxw7 is the substrate-recognition factor of a ubiquitin ligase that targets multiple proteins for degradation, including c-Myc, cyclin E, c-Jun and Notch. Here, we describe that TGIF1 is targeted for degradation by Fbxw7 in a phosphorylation-dependent manner. Inactivation of Fbxw7 results in the accumulation of phosphorylated TGIF1 molecules and repression of TGFβ-dependent transcription. Cancer cell lines with inactivating mutations in Fbxw7 show enhanced levels of TGIF1 and attenuated TGFβ-dependent signaling. Importantly, inactivation of Fbxw7 attenuates TGFβ-dependent regulation of cell growth and migration. Taken together, our results suggest that Fbxw7 is a novel regulator of TGFβ signaling.

背景与目标： 转化生长因子-β (tgf β) 信号调节多种细胞过程，包括细胞外基质的产生，细胞生长，凋亡和分化。Tgf β 信号传导功能障碍与从血管疾病到癌症的各种人类疾病有关。Tgf β 信号传导受转录阻遏因子tgf β 诱导因子1 (TGIF1) 的负调节。肿瘤抑制因子Fbxw7是泛素连接酶的底物识别因子，该酶靶向多种降解蛋白，包括c-Myc，细胞周期蛋白E，c-6月和Notch。在这里，我们描述了TGIF1以磷酸化依赖性方式被Fbxw7降解。Fbxw7的失活导致磷酸化的TGIF1分子的积累和tgf β 依赖性转录的抑制。Fbxw7中具有失活突变的癌细胞系显示出TGIF1水平增强和tgf β 依赖性信号减弱。重要的是，Fbxw7的失活减弱了tgf β 依赖性细胞生长和迁移的调节。总之，我们的结果表明Fbxw7是tgf β 信号传导的新型调节剂。

BACKGROUND & AIMS: :Signaling pathway(s) responsible for transforming growth factor β (TGFβ)-induced epithelial mesenchymal transition (EMT), invasion and migration of H460 cells (non-small cell lung cancer/NSCLC) was identified in the study. The results showed that TGFβ-induced p(38)/β-catenin/PPARγ signaling pathway played a critical role in the promotion of EMT, invasion and migration of H460 cells. All these pathological outcomes attributed to PPARγ-increased expression of p-EGFR, p-c-MET and Vimentin and the decrease of E-cadherin. Transforming growth factor β and p(38)-induced β-catenin not only stimulated the expression of PPARγ but also physically interacted with it. Blocking the ligand binding domain of PPARγ (with GW9662) could significantly interfere the binding between PPARγ and β-catenin, and interrupt the nuclear infiltration of both factors. These findings suggested that β-catenin was an upstream regulator and a ligand of PPARγ, and the binding between these two molecules was critical for their nuclear infiltration. Transforming growth factor β-induced tumor invasion and migration was also seen in U373 cells (brain glioma, with high inducible PPARγ) in a PPARγ-dependent manner, but not in CH27 cells (squamous NSCLC, with low PPARγ). PPARγ shRNA, GW9662, JW67 and 2,4-diaminoquinazoline were all revealed to have important values in the control of the intrinsic and TGFβ-induced EMT, tumor invasion and migration of H460 cells. The results further suggested that PPARγ and β-catenin may be the potential markers for the early diagnosis and/or treatment of metastatic tumors.

背景与目标： : 研究确定了负责转化生长因子 β (tgf β) 诱导的上皮间质转化 (EMT)，H460细胞 (非小细胞肺癌/NSCLC) 侵袭和迁移的信号通路。结果表明，tgf β 诱导的p(38)/β-catenin/ppar γ 信号通路在促进H460细胞的EMT，侵袭和迁移中起关键作用。所有这些病理结果均归因于ppar γ-p-EGFR，p-c-MET和波形蛋白的表达增加以及E-钙粘蛋白的减少。转化生长因子 β 和p(38) 诱导的 β-连环蛋白不仅刺激ppar γ 的表达，而且与ppar γ 的物理相互作用。阻断ppar γ 的配体结合结构域 (带有GW9662) 可以显着干扰ppar γ 与 β-catenin之间的结合，并中断两种因子的核渗透。这些发现表明 β-catenin是ppar γ 的上游调节剂和配体，这两个分子之间的结合对于它们的核渗透至关重要。转化生长因子 β 诱导的肿瘤侵袭和迁移也以ppar γ 依赖性方式在U373细胞 (脑胶质瘤，具有高诱导型ppar γ) 中观察到，但在CH27细胞 (鳞状NSCLC，具有低ppar γ) 中未观察到。Ppar γ shRNA，GW9662，JW67和2,4-二氨基喹唑啉均被发现在控制内源性和tgf β 诱导的EMT，肿瘤侵袭和H460细胞迁移方面具有重要价值。结果进一步表明，ppar γ 和 β-catenin可能是早期诊断和/或治疗转移性肿瘤的潜在标志物。

BACKGROUND & AIMS: :Renal fibrosis, a common pathological manifestation of virtually all types of chronic kidney diseases (CKD), often results in diffuse kidney scarring and predisposes to end-stage renal disease. Currently, there is no effective therapy against renal fibrosis. Recently, our laboratory identified an ER-resident protein, thioredoxin domain containing 5 (TXNDC5), as a critical mediator of cardiac fibrosis. Transcriptome analyses of renal biopsy specimens from CKD patients revealed marked TXNDC5 upregulation in fibrotic kidneys, suggesting a potential role of TXNDC5 in renal fibrosis. Employing multiple fluorescent reporter mouse lines, we showed that TXNDC5 was specifically upregulated in collagen-secreting fibroblasts in fibrotic mouse kidneys. In addition, we showed that TXNDC5 was required for TGFβ1-induced fibrogenic responses in human kidney fibroblasts (HKF), whereas TXNDC5 over-expression was sufficient to promote HKF activation, proliferation and collagen production. Mechanistically, we showed that TXNDC5, transcriptionally controlled by ATF6-dependent ER stress pathway, mediates its pro-fibrogenic effects by enforcing TGFβ signaling activity through post-translational stabilization and upregulation of type I TGFβ receptor in kidney fibroblasts. Using a tamoxifen-inducible, fibroblast-specific Txndc5 knockout mouse line, we demonstrated that deletion of Txndc5 in kidney fibroblasts mitigated the progression of established kidney fibrosis, suggesting the therapeutic potential of TXNDC5 targeting for renal fibrosis and CKD. .

背景与目标： : 肾脏纤维化是几乎所有类型的慢性肾脏疾病 (CKD) 的常见病理表现，通常会导致弥漫性肾脏瘢痕形成并易患终末期肾脏疾病。目前，尚无针对肾脏纤维化的有效疗法。最近，我们的实验室确定了ER驻留蛋白，含有5 (TXNDC5) 的硫氧还蛋白结构域，是心脏纤维化的关键介质。CKD患者肾活检标本的转录组分析显示，纤维化肾脏中TXNDC5明显上调，提示TXNDC5在肾脏纤维化中的潜在作用。使用多个荧光报告小鼠品系，我们显示TXNDC5在纤维化小鼠肾脏中分泌胶原蛋白的成纤维细胞中特异性上调。此外，我们发现TXNDC5是tgfβ1诱导的人肾脏成纤维细胞 (HKF) 纤维化反应所必需的，而TXNDC5的过表达足以促进HKF的活化，增殖和胶原蛋白的产生。从机制上讲，我们表明，受ATF6-dependent ER应激途径转录控制的TXNDC5通过翻译后稳定和肾脏成纤维细胞中I型tgf β 受体的上调来增强tgf β 信号活性，从而介导其促纤维化作用。使用他莫昔芬诱导的成纤维细胞特异性Txndc5敲除小鼠系，我们证明了肾脏成纤维细胞中Txndc5的缺失减轻了已建立的肾脏纤维化的进展，提示TXNDC5靶向肾脏纤维化和CKD的治疗潜力。

BACKGROUND & AIMS: :Secreted cytokines of the TGFβ family are found in all multicellular organisms and implicated in regulating fundamental cell behaviors such as proliferation, differentiation, migration and survival. Signal transduction involves complexes of specific type I and II receptor kinases that induce the nuclear translocation of Smad transcription factors to regulate target genes. Ligands of the BMP and Nodal subgroups act at a distance to specify distinct cell fates in a concentration-dependent manner. These signaling gradients are shaped by multiple factors, including proteases of the proprotein convertase (PC) family that hydrolyze one or several peptide bonds between an N-terminal prodomain and the C-terminal domain that forms the mature ligand. This review summarizes information on the proteolytic processing of TGFβ and related precursors, and its spatiotemporal regulation by PCs during development and various diseases, including cancer. Available evidence suggests that the unmasking of receptor binding epitopes of TGFβ is only one (and in some cases a non-essential) function of precursor processing. Future studies should consider the impact of proteolytic maturation on protein localization, trafficking and turnover in cells and in the extracellular space.

背景与目标： : tgf β 家族的分泌细胞因子存在于所有多细胞生物中，并与调节基本细胞行为 (例如增殖，分化，迁移和存活) 有关。信号转导涉及特定的I型和II型受体激酶的复合物，这些复合物诱导Smad转录因子的核易位以调节靶基因。BMP和节点亚组的配体在一定距离上起作用，以浓度依赖性方式指定不同的细胞命运。这些信号传导梯度由多种因素形成，包括前蛋白转化酶 (PC) 家族的蛋白酶，该蛋白酶水解N末端前结构域与形成成熟配体的C末端结构域之间的一个或几个肽键。这篇综述总结了有关tgf β 和相关前体的蛋白水解加工以及PCs在发育和各种疾病 (包括癌症) 中的时空调控的信息。现有证据表明，tgf β 受体结合表位的揭开只是前体加工的一种 (在某些情况下是非必需的) 功能。未来的研究应考虑蛋白水解成熟对细胞和细胞外空间中蛋白质定位，运输和周转的影响。

BACKGROUND & AIMS: :The mesenchymal, clear cell, and dedifferentiated chondrosarcoma subtypes are extremely rare, together constituting 10% to 15% of all chondrosarcomas. Their poor prognosis and lack of efficacious treatment emphasizes the need to elucidate the pathways playing a pivotal role in these tumors. We constructed tissue microarrays containing 42 dedifferentiated, 23 clear cell, and 23 mesenchymal chondrosarcomas and performed immunohistochemistry to study the expression of growth plate-signaling molecules and molecules shown to be involved in conventional chondrosarcoma. We observed high expression of SOX-9 and FGFR-3, as well as aberrant cellular localization of heparan sulfate proteoglycans, in all subtypes. TGFβ signaling through p-SMAD2 and PAI-1 was highly active in all chondrosarcoma subtypes, which suggests that TGFβ inhibitors as a possible therapeutic strategy in rare chondrosarcoma subtypes. As in conventional chondrosarcoma, antiapoptotic proteins (Bcl-2, and/or Bcl-xl) were highly expressed in all subtypes. Inhibition with the BH-3 mimetic ABT-737 rendered dedifferentiated chondrosarcoma cell lines sensitive to doxorubicin or cisplatin. Our data indicate that antiapoptotic proteins may play an important role in chemoresistance, suggesting a promising role for targeting Bcl-2 family members in chondrosarcoma treatment, irrespective of the subtype.

背景与目标： : 间充质，透明细胞和去分化的软骨肉瘤亚型极为罕见，共同构成所有软骨肉瘤的10% 15%。他们的不良预后和缺乏有效的治疗，强调需要阐明在这些肿瘤中起关键作用的途径。我们构建了包含42个去分化，23个透明细胞和23个间充质软骨肉瘤的组织微阵列，并进行了免疫组织化学研究，以研究生长板信号分子和显示与常规软骨肉瘤有关的分子的表达。我们在所有亚型中观察到SOX-9和FGFR-3的高表达以及硫酸乙酰肝素蛋白聚糖的异常细胞定位。Tgf β 通过p-SMAD2和PAI-1的信号传导在所有软骨肉瘤亚型中均具有很高的活性，这表明tgf β 抑制剂可能是罕见软骨肉瘤亚型的治疗策略。与常规软骨肉瘤一样，抗凋亡蛋白 (Bcl-2和/或Bcl-xl) 在所有亚型中均高表达。用BH-3模拟ABT-737抑制可使去分化的软骨肉瘤细胞系对阿霉素或顺铂敏感。我们的数据表明，抗凋亡蛋白可能在化学耐药性中起重要作用，这表明在软骨肉瘤治疗中靶向Bcl-2家族成员的有希望的作用，而与亚型无关。

BACKGROUND & AIMS: :The molecular mechanisms that regulate and coordinate signaling between the extracellular matrix (ECM) and cells contributing to the developing vasculature are complex and poorly understood. Myocardin-like protein 2 (MKL2) is a transcriptional co-activator that in response to RhoA and cytoskeletal actin signals physically associates with serum response factor (SRF), activating a subset of SRF-regulated genes. We now report the discovery of a previously undescribed MKL2/TGFβ signaling pathway in embryonic stem (ES) cells that is required for maturation and stabilization of the embryonic vasculature. Mkl2(-/-) null embryos exhibit profound derangements in the tunica media of select arteries and arterial beds, which leads to aneurysmal dilation, dissection and hemorrhage. Remarkably, TGFβ expression, TGFβ signaling and TGFβ-regulated genes encoding ECM are downregulated in Mkl2(-/-) ES cells and the vasculature of Mkl2(-/-) embryos. The gene encoding TGFβ2, the predominant TGFβ isoform expressed in vascular smooth muscle cells and embryonic vasculature, is activated directly via binding of an MKL2/SRF protein complex to a conserved CArG box in the TGFβ2 promoter. Moreover, Mkl2(-/-) ES cells exhibit derangements in cytoskeletal organization, cell adhesion and expression of ECM that are rescued by forced expression of TGFβ2. Taken together, these data demonstrate that MKL2 regulates a conserved TGF-β signaling pathway that is required for angiogenesis and ultimately embryonic survival.

背景与目标： : 调节和协调细胞外基质 (ECM) 与有助于发展的脉管系统的细胞之间信号传导的分子机制很复杂，而且知之甚少。Myocardin样蛋白2 (MKL2) 是一种转录共激活剂，可响应RhoA和细胞骨架肌动蛋白信号与血清反应因子 (SRF) 物理相关，从而激活SRF调控基因的一部分。我们现在报告在胚胎干 (ES) 细胞中发现了先前未描述的MKL2/tgf β 信号通路，这是胚胎脉管系统成熟和稳定所必需的。Mkl2(-/-) 无效胚胎在选定的动脉和动脉床的中膜中表现出严重的紊乱，从而导致动脉瘤扩张，解剖和出血。值得注意的是，在Mkl2(-/-) ES细胞和Mkl2(-/-) 胚胎的脉管系统中，tgf β 表达，tgf β 信号传导和编码ECM的tgf β 调节基因被下调。编码TGFβ2 (在血管平滑肌细胞和胚胎脉管系统中表达的主要TGFβ 亚型) 的基因通过将MKL2/SRF蛋白复合物与TGFβ2启动子中的保守的CArG盒结合而直接激活。此外，Mkl2(-/-) ES细胞在细胞骨架组织，细胞粘附和ECM表达方面表现出紊乱，这些紊乱被tgfβ2的强制表达所挽救。总之，这些数据表明，MKL2调节保守的TGF-β 信号通路，这是血管生成和最终胚胎存活所必需的。

BACKGROUND & AIMS: INTRODUCTION:Systemic sclerosis (SSc)-related pulmonary arterial hypertension (PAH) has emerged as a major mortality prognostic factor. Mutations of transforming growth factor beta (TGFβ) receptor genes strongly contribute to idiopathic and familial PAH. OBJECTIVE:To explore the genetic bases of SSc-PAH, we combined direct sequencing and genotyping of candidate genes encoding TGFβ receptor family members. MATERIALS AND METHODS:TGFβ receptor genes, BMPR2, ALK1, TGFR2 and ENG, were sequenced in 10 SSc-PAH patients, nine SSc and seven controls. In addition, 22 single-nucleotide polymorphisms (SNP) of these four candidate genes were tested for association in a first set of 824 French Caucasian SSc patients (including 54 SSc-PAH) and 939 controls. The replication set consisted of 1516 European SSc (including 219 SSc-PAH) and 3129 controls from the European League Against Rheumatism Scleroderma Trials and Research group network. RESULTS:No mutation was identified by direct sequencing. However, two repertoried SNP, ENG rs35400405 and ALK1 rs2277382, were found in SSc-PAH patients only. The genotyping of 22 SNP including the latter showed that only rs2277382 was associated with SSc-PAH (p=0.0066, OR 2.13, 95% CI 1.24 to 3.65). Nevertheless, this was not replicated with the following result in combined analysis: p=0.123, OR 0.79, 95% CI 0.59 to 1.07. CONCLUSIONS:This study demonstrates the lack of association between these TGFβ receptor gene polymorphisms and SSc-PAH using both sequencing and genotyping methods.

背景与目标：

BACKGROUND & AIMS: STUDY QUESTION:Is there an effect of the TGFβ inhibitor SB431542 (SB) on the epiblast compartment of human blastocysts, and does it affect subsequent human embryonic stem cell (hESC) derivation? SUMMARY ANSWER:SB increases the mean number of NANOG-positive cells in the inner cell mass (ICM), and allows for subsequent hESC derivation. WHAT IS KNOWN ALREADY:It is known that inhibition of TGFβ by SB has a positive effect on mouse ESC self-renewal, while active TGFβ signalling is needed for self-renewal of primed ESC. STUDY DESIGN, SIZE, DURATION:From December 2011 until March 2012, 263 donated spare embryos were used from patients who had undergone IVF/ICSI in our centre. PARTICIPANTS/MATERIALS, SETTING, METHODS:Donated human embryos were cultured in the presence of SB or Activin A, and immunocytochemistry was performed on Day 6 blastocysts for NANOG and GATA6. Moreover, blastocysts were used for the derivation of hESC, with or without exposure to SB. MAIN RESULTS AND THE ROLE OF CHANCE:Immunocytochemistry revealed a significantly higher number of NANOG-positive ICM cells in the SB group compared with the control (12.0 ± 5.9 versus 6.1 ± 4.7), while no difference was observed in the Activin A group compared with other groups (6.7 ± 3.7). The number of GATA6-positive ICM cells did not differ between the SB, Activin A and control group (8.8 ± 4.3, 8.0 ± 4.6 and 7.2 ± 4.0, respectively). Blocking TGFβ signalling did not prevent subsequent hESC line derivation. LIMITATIONS, REASONS FOR CAUTION:The number of human blastocysts available for this study was too low to reveal if the observed increase in NANOG-positive epiblast cells after exposure to SB affected the efficiency of hESC derivation (12.5% compared with 16.7%). WIDER IMPLICATIONS OF THE FINDINGS:This work can contribute to the derivation of naive hESC lines in the future. STUDY FUNDING/COMPETING INTEREST(S):M.V.d.J. is holder of a Ph.D. grant of the Agency for Innovation by Science and Technology (IWT, grant number SB093128), Belgium. G.D. and this research are supported by the Research Foundation Flanders (FWO), grant number FWO-3G062910) and a Concerted Research Actions funding from BOF (Bijzonder Onderzoeksfonds University Ghent, grant number BOF GOA 01G01112). S.M.C.d. S.L. is supported by the Netherlands Organization of Scientific Research (NWO) (ASPASIA 015.007.037) and the Interuniversity Attraction Poles (PAI) (no. P7/07). P.D.S. is holder of a fundamental clinical research mandate by the FWO. We would like to thank Ferring Company (Aalst, Belgium) for financial support of this study. The authors do not have any competing interests to declare. TRIAL REGISTRATION NUMBER:Not applicable.

背景与目标：

BACKGROUND & AIMS: :Treatment of muscle-invasive bladder cancer remains a major clinical challenge. Aberrant HGF/c-MET upregulation and activation is frequently observed in bladder cancer correlating with cancer progression and invasion. However, the mechanisms underlying HGF/c-MET-mediated invasion in bladder cancer remains unknown. As part of a negative feedback loop SMAD7 binds to SMURF2 targeting the TGFβ receptor for degradation. Under these conditions, SMAD7 acts as a SMURF2 agonist by disrupting the intramolecular interactions within SMURF2. We demonstrate that HGF stimulates TGFβ signalling through c-SRC-mediated phosphorylation of SMURF2 resulting in loss of SMAD7 binding and enhanced SMURF2 C2-HECT interaction, inhibiting SMURF2 and enhancing TGFβ receptor stabilisation. This upregulation of the TGFβ pathway by HGF leads to TGFβ-mediated EMT and invasion. In vivo we show that TGFβ receptor inhibition prevents bladder cancer invasion. Furthermore, we make a rationale for the use of combinatorial TGFβ and MEK inhibitors for treatment of high-grade non-muscle-invasive bladder cancers.

背景与目标： : 肌肉浸润性膀胱癌的治疗仍然是一个主要的临床挑战。在膀胱癌中经常观察到异常的HGF/c-MET上调和激活与癌症的进展和侵袭相关。然而，HGF/c-MET介导的膀胱癌侵袭的潜在机制仍然未知。作为负反馈回路的一部分，SMAD7与靶向tgf β 受体的SMURF2结合以降解。在这些条件下，SMAD7通过破坏SMURF2内的分子内相互作用而充当SMURF2激动剂。我们证明HGF通过c-SRC介导的SMURF2磷酸化刺激tgf β 信号传导，导致SMAD7结合的丧失和SMURF2 C2-HECT相互作用的增强，抑制SMURF2并增强tgf β 受体的稳定。HGF对tgf β 途径的这种上调导致tgf β 介导的EMT和侵袭。在体内，我们显示tgf β 受体抑制可防止膀胱癌侵袭。此外，我们为使用组合tgf β 和MEK抑制剂治疗高级别非肌肉浸润性膀胱癌提供了理论依据。

BACKGROUND & AIMS: :Esophageal cancer (EC) is known as a type of common malignant tumor, with the incidence ranking eighth worldwide. Because of the high metastasis of advanced EC, the total survival rate has been quite low. Esophageal squamous cell carcinoma (ESCC) is a main type of EC. Targeted therapy for ESCC has become a new direction; however, newly therapeutic targets are also badly needed. Shc SH2 domain-binding protein (SHCBP1) is located on 16q11.2, which is a downstream protein of the Shc adaptor. SHCBP1 participates in the regulation of several physiological and pathologic processes, such as cytokinesis. Recent studies have found that SHCBP1 was abnormally upregulated in multiple types of tumors, such as breast cancer and liver cancer, and that it affects the proliferation and motility of cancer cells in vitro. However, it remains unclear whether SHCBP1 is related to the progression of EC. Herein, we found the upregulation of SHCBP1 in human EC tissues. Our findings further demonstrated that SHCBP1 expression was related to the clinical features of ESCC patients. We found that SHCBP1 depletion inhibited the proliferation and motility of ESCC cells via the transforming growth factor β pathway and that it suppressed the growth of tumors in mice. We, therefore, concluded that SHCBP1 could serve as a promising EC molecular target.

背景与目标： 食管癌 (EC) 是一种常见的恶性肿瘤，发病率在全球排名第八。由于晚期EC的高转移，总生存率一直很低。食管鳞状细胞癌 (ESCC) 是EC的主要类型。靶向治疗ESCC已成为一个新的方向; 然而，也迫切需要新的治疗靶点。Shc SH2结构域结合蛋白 (SHCBP1) 位于16q11.2上，是Shc衔接子的下游蛋白。SHCBP1参与多种生理和病理过程的调节，例如胞质分裂。最近的研究发现，SHCBP1在多种类型的肿瘤 (如乳腺癌和肝癌) 中异常上调，并且在体外影响癌细胞的增殖和运动。然而，目前尚不清楚SHCBP1是否与EC的进展有关。在此，我们发现SHCBP1在人EC组织中的上调。我们的发现进一步证明SHCBP1的表达与ESCC患者的临床特征有关。我们发现SHCBP1耗竭通过转化生长因子 β 途径抑制了ESCC细胞的增殖和运动，并抑制了小鼠肿瘤的生长。因此，我们得出结论，SHCBP1可以作为一个有前途的EC分子靶标。

BACKGROUND & AIMS: :Testis development is dependent on the key sex-determining factors SRY and SOX9, which activate the essential ligand FGF9. Although FGF9 plays a central role in testis development, it is unable to induce testis formation on its own. However, other growth factors, including activins and TGFβs, also present testis during testis formation. In this study, we investigated the potential of FGF9 combined with activin and TGFβ to induce testis development in cultured XX gonads. Our data demonstrated differing individual and combined abilities of FGF9, activin and TGFβ to promote supporting cell proliferation, Sertoli cell development and male germ line differentiation in cultured XX gonads. FGF9 promoted proliferation of supporting cells in XX foetal gonads at rates similar to those observed in vivo during testis cord formation in XY gonads but was insufficient to initiate testis development. However, when FGF9, activin and TGFβ were combined, aspects of testicular development were induced, including the expression of Sox9, morphological reorganisation of the gonad and deposition of laminin around germ cells. Enhancing β-catenin activity diminished the testis-promoting activities of the combined growth factors. The male promoting activity of FGF9 and the combined growth factors directly or indirectly extended to the germ line, in which a mixed phenotype was observed. FGF9 and the combined growth factors promoted male germ line development, including mitotic arrest, but expression of pluripotency genes was maintained, rather than being repressed. Together, our data provide evidence that combined signalling by FGF9, activin and TGFβ can induce testicular characteristics in XX gonads.

背景与目标： : 睾丸发育取决于关键的性别决定因素SRY和SOX9，它们激活了必需配体fgf9。尽管FGF9在睾丸发育中起着核心作用，但它无法自行诱导睾丸形成。然而，其他生长因子，包括激活素和转化生长因子 β，在睾丸形成过程中也存在睾丸。在这项研究中，我们研究了FGF9与激活素和tgf β 结合在培养的XX性腺中诱导睾丸发育的潜力。我们的数据表明，FGF9，激活素和tgf β 在促进培养的XX性腺中支持细胞增殖，支持细胞发育和雄性种系分化方面具有不同的个体和综合能力。FGF9促进XX胎儿性腺中支持细胞的增殖，其速率与XY性腺中睾丸索形成过程中体内观察到的速率相似，但不足以启动睾丸发育。然而，当FGF9，激活素和tgf β 结合时，会诱导睾丸发育的各个方面，包括Sox9的表达，性腺的形态重组和层粘连蛋白在生殖细胞周围的沉积。增强 β-catenin活性会降低联合生长因子的睾丸促进活性。FGF9和组合生长因子的雄性促进活性直接或间接扩展到种系，其中观察到混合表型。FGF9和组合的生长因子促进了雄性种系的发育，包括有丝分裂停滞，但多能性基因的表达得以维持，而不是被抑制。总之，我们的数据提供了证据，表明FGF9，激活素和tgf β 的联合信号可以诱导XX性腺的睾丸特征。

BACKGROUND & AIMS: :The TGFβ superfamily is composed of more than 33 growth and differentiation factors, including TGFβ1, β2, β3, BMPs, GDFs, nodal-related proteins, and activins. These members usually exert pleiotropic actions on several tissues and control multiple cellular processes, such as cell growth, cell survival, cell migration, cell fate specification, and differentiation, both during embryonic development and postnatal life. Although the effects of these factors on immune responses were elucidated long ago, most studies have been focused on the actions of TGFβs on T cells, as major regulators of adaptive immunity. In this review, we discuss new findings about the involvement of TGFβ superfamily members in the control of B cell development and function. Moreover, the potential contribution of TGFβ signaling to control B cell-mediated autoimmune diseases and its utility in the design of new therapies are also discussed.

背景与目标： : tgf β 超家族由超过33个生长和分化因子组成，包括TGFβ1，β2，β3，BMPs，GDFs，节点相关蛋白和激活素。这些成员通常在几个组织上发挥多效性作用，并控制多个细胞过程，例如胚胎发育和出生后的细胞生长，细胞存活，细胞迁移，细胞命运规范和分化。尽管这些因素对免疫反应的影响早已阐明，但大多数研究都集中在tgf β 对T细胞的作用上，作为适应性免疫的主要调节剂。在这篇综述中，我们讨论了有关tgf β 超家族成员参与控制b细胞发育和功能的新发现。此外，还讨论了tgf β 信号传导对控制b细胞介导的自身免疫性疾病的潜在贡献及其在设计新疗法中的效用。