MiR-92b-3p ameliorates inflammation and autophagy by targeting TRAF3 and suppressing MKK3-p38 pathway in caerulein-induced AR42J cells

Acute pancreatitis (AP) is an inflammatory disease with high morbidity and mortality. Dysregulation of microRNAs (miRNAs) was involved in human diseases, including AP. However, the effects of miR-92b-3p on AP process and its mechanism remain not been fully clarified. The expression levels of miR-92b-3p and tumor necrosis factor receptor-associated factor-3 (TRAF3) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of TRAF3, tumor necrosis factor α (TNF-α) TNF-α, interleukin-6 (IL-6), phosphorylated mitogen-activated protein kinase kinase 3 (p-MKK3), MKK3, p38 and phosphorylated p38 (p-p38) were detected by western blot. The concentration of TNF-α and IL-6 in the medium was measured using ELISA kits. The possible binding sites of miR-92b-3p and TRAF3 were predicted by TargetScan and verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The expression level of miR-92b-3p was decreased and TRAF3 expression was increased in AR42J cells stimulated with caerulein. Moreover, the protein levels of pro-inflammatory cytokines (TNF-α and IL-6) were markedly elevated, and the expression levels of autophagy-related markers Beclin1 as well as the ratio of LC3-II/I were obviously increased in AR42J cells treated with caerulein. In addition, overexpression of miR-92b-3p or knockdown of TRAF3 significantly suppressed the release of pro-inflammatory cytokines and autophagy in caerulein-induced AR42J cells. Furthermore, TRAF3 was a direct target of miR-92b-3p and its upregulation reversed the effects of miR-92b-3p overexpression on inflammatory response and autophagy. Besides, overexpression of miR-92b-3p inhibited the activation of the MKK3-p38 pathway by affecting TRAF3 expression. In conclusion, miR-92b-3p attenuated inflammatory response and autophagy by downregulating TRAF3 and suppressing MKK3-p38 pathway in caerulein-induced AR42J cells, providing a novel avenue for treatment of AP.     

靶向PSCA的CAR-NK-92细胞对宫颈癌的抗瘤作用

目的：构建并验证靶向前列腺干细胞抗原（prostate stem cell antigen，PSCA）的嵌合抗原受体（chimeric antigen receptor，CAR）修饰的NK-92细胞对宫颈癌的抗瘤活性。方法：构建PSCA靶向的CAR慢病毒表达载体，利用慢病毒感染法获得PSCA CAR-NK-92细胞。用流式细胞术及Western blotting实验检测宫颈癌细胞中PSCA的表达水平。通过体外效靶细胞共孵育实验以及体内裸鼠移植瘤模型中的治疗情况，验证PSCA CAR-NK-92细胞对宫颈癌Hela及MS751细胞杀伤能力及其裸鼠移植瘤生长的抑制能力。结果：成功构建 PSCA CAR-NK-92 细胞，PSCA 在宫颈癌细胞中高表达（均 P<0.01）。体外共孵育结果显示，PSCA CAR-NK-92细胞可以以剂量依赖的方式裂解PSCA+的宫颈癌细胞；体内抗肿瘤数据表明，PSCA CAR-NK-92细胞较转染空载体的NK-92细胞显著抑制宫颈癌移植瘤的生长（P<0.01），并且可以有效地浸润肿瘤组织并高水平分泌TNF-α和IFN-（γ 均P<0.01）。结论：靶向PSCA的CAR-NK-92细胞在体内外都显示出了良好的对PSCA+肿瘤细胞的杀伤能力，有潜力成为一种针对宫颈癌的潜在治疗策略。     

肿瘤化疗所致恶心呕吐的发生机制和药物治疗的研究进展

化疗所致恶心、呕吐(CINV)是肿瘤患者最常见的不良反应。如果没有镇吐治疗,70%～80%接受化疗的患者会出现恶心、呕吐症状。其程度受化疗药物致吐强弱等多因素的影响。在20世纪90年代5-HT3受体阻滞剂和地塞米松的联合应用使得70%的急性CINV得到了有效的控制。近年来开发了新一代半衰期更长、亲和力更高的5-HT3受体阻滞剂palonosetron(帕洛诺司琼)。此外,随着对P物质和NK-1受体研究的深入以及NK-1受体阻滞剂aprepitant的问世,急性和迟发性CINV的完全缓解率有了进一步提高。根据近年的新进展和NCCN止吐治疗指南新版本的修订,本文综述了CINV的机制和药物治疗的研究进展。     

The expression of Mirc1/Mir17–92 cluster in sputum samples correlates with pulmonary exacerbations in cystic fibrosis patients

Introduction

Cystic fibrosis (CF) is a multi-organ disorder characterized by chronic sino-pulmonary infections and inflammation. Many patients with CF suffer from repeated pulmonary exacerbations that are predictors of worsened long-term morbidity and mortality. There are no reliable markers that associate with the onset or progression of an exacerbation or pulmonary deterioration. Previously, we found that the Mirc1/Mir17–92a cluster which is comprised of 6 microRNAs (Mirs) is highly expressed in CF mice and negatively regulates autophagy which in turn improves CF transmembrane conductance regulator (CFTR) function. Therefore, here we sought to examine the expression of individual Mirs within the Mirc1/Mir17–92 cluster in human cells and biological fluids and determine their role as biomarkers of pulmonary exacerbations and response to treatment.

DeepSMILE: Contrastive self-supervised pre-training benefits MSI and HRD classification directly from H&E whole-slide images in colorectal and breast cancer

We propose a Deep learning-based weak label learning method for analyzing whole slide images (WSIs) of Hematoxylin and Eosin (H&E) stained tumor tissue not requiring pixel-level or tile-level annotations using Self-supervised pre-training and heterogeneity-aware deep Multiple Instance LEarning (DeepSMILE). We apply DeepSMILE to the task of Homologous recombination deficiency (HRD) and microsatellite instability (MSI) prediction. We utilize contrastive self-supervised learning to pre-train a feature extractor on histopathology tiles of cancer tissue. Additionally, we use variability-aware deep multiple instance learning to learn the tile feature aggregation function while modeling tumor heterogeneity. For MSI prediction in a tumor-annotated and color normalized subset of TCGA-CRC (n=360 patients), contrastive self-supervised learning improves the tile supervision baseline from 0.77 to 0.87 AUROC, on par with our proposed DeepSMILE method. On TCGA-BC (n=1041 patients) without any manual annotations, DeepSMILE improves HRD classification performance from 0.77 to 0.81 AUROC compared to tile supervision with either a self-supervised or ImageNet pre-trained feature extractor. Our proposed methods reach the baseline performance using only 40% of the labeled data on both datasets. These improvements suggest we can use standard self-supervised learning techniques combined with multiple instance learning in the histopathology domain to improve genomic label classification performance with fewer labeled data.     

结直肠癌患者血清miR-92a-3p表达水平及其临床意义

目的分析miR-92a-3p在体检健康者、结直肠腺瘤息肉患者以及结直肠癌(CRC)患者手术前后血清中的相对表达水平,探讨其在CRC临床诊断中的应用价值。方法用Trizol法分别对30例体检健康者、25例结直肠腺瘤息肉患者和40例CRC患者术前及术后的血清进行总RNA提取,用实时荧光定量PCR检测miR-92a-3p的相对表达水平,分析其在各组中的表达差异及其与CRC临床病理参数的关系,以受试者工作曲线确定血清miR-92a-3p的CRC诊断效能。结果 CRC术前组血清miR-92a-3p的表达水平高于结直肠腺瘤息肉组(Z=4.75,P0.01)、CRC术后组(Z=4.28,P0.01)和健康人对照组(Z=8.00,P0.01);健康人对照组血清miR-92a-3p的表达水平低于CRC术后组(Z=4.08,P0.01)和结直肠腺瘤息肉组(Z=2.70,P=0.01);结直肠腺瘤息肉组和CRC术后组的血清miR-92a-3p的表达水平差异无统计学意义(Z=0.99,P=0.32);血清miR-92a-3p鉴别CRC的ROC曲线下面积(AUCROC)为0.899(95%CI:0.840~0.959),当cut off值为2.75时,敏感性和特异性均为80%;血清miR-92a-3p的高低与CRC的浸润深度(Z=2.14,P=0.03)、淋巴结转移(Z=2.86,P=0.04)、TNM分期(Z=2.93,P0.01)等临床病理参数关系密切。结论血清miR-92a-3p可能为临床上CRC的筛查、病情监测等方面提供实验室依据。     

Exosomal microRNA in serum is a novel biomarker of recurrence in human colorectal cancer

Background:

Functional microRNAs (miRNAs) in exosomes have been recognised as potential stable biomarkers in cancers. The aim of this study is to identify specific miRNAs in exosome as serum biomarkers for the early detection of recurrence in human colorectal cancer (CRC).

Methods:

Serum samples were sequentially obtained from six patients with and without recurrent CRC. The miRNAs were purified from exosomes, and miRNA microarray analysis was performed. The miRNA expression profiles and copy number aberrations were explored using microarray and array CGH analyses in 124 CRC tissues. Then, we validated exosomal miRNAs in 2 serum sample sets (90 and 209 CRC patients) by quantitative real-time RT–PCR.

Results:

Exosomal miR-17-92a cluster expression level in serum was correlated with the recurrence of CRC. Exosomal miR-19a expression levels in serum were significantly increased in patients with CRC as compared with healthy individuals with gene amplification. The CRC patients with high exosomal miR-19a expression showed poorer prognoses than the low expression group (P<0.001).

Conclusions:

Abundant expression of exosomal miR-19a in serum was identified as a prognostic biomarker for recurrence in CRC patients.