Orthogonally oriented scaffolds with aligned fibers for engineering intestinal smooth muscle

Controlling cellular alignment is critical in engineering intestines with desired structure and function. Although previous studies have examined the directional alignment of cells on the surface (x–y plane) of parallel fibers, quantitative analysis of the cellular alignment inside implanted scaffolds with oriented fibers has not been reported. This study examined the cellular alignment in the x–z and y–z planes of scaffolds made with two layers of orthogonally oriented fibers. The cellular orientation inside implanted scaffolds was evaluated with immunofluorescence. Quantitative analysis of coherency between cell orientation and fiber direction confirmed that cells aligned along the fibers not only on the surface (x–y plane) but also inside the scaffolds (x–z & y–z planes). Our study demonstrated that two layers of orthogonally aligned scaffolds can generate the histological organization of cells similar to that of intestinal circular and longitudinal smooth muscle.     

Imaging chemical exchange saturation transfer (CEST) effects following tumor‐selective acidification using lonidamine

Increased lactate production through glycolysis in aerobic conditions is a hallmark of cancer. Some anticancer drugs have been designed to exploit elevated glycolysis in cancer cells. For example, lonidamine (LND) inhibits lactate transport, leading to intracellular acidification in cancer cells. Chemical exchange saturation transfer (CEST) is a novel MRI contrast mechanism that is dependent on intracellular pH. Amine and amide concentration‐independent detection (AACID) and apparent amide proton transfer (APT*) represent two recently developed CEST contrast parameters that are sensitive to pH. The goal of this study was to compare the sensitivity of AACID and APT* for the detection of tumor‐selective acidification after LND injection. Using a 9.4‐T MRI scanner, CEST data were acquired in mice approximately 14 days after the implantation of 10⁵ U87 human glioblastoma multiforme (GBM) cells in the brain, before and after the administration of LND (dose, 50 or 100 mg/kg). Significant dose‐dependent LND‐induced changes in the measured CEST parameters were detected in brain regions spatially correlated with implanted tumors. Importantly, no changes were observed in T₁‐ and T₂‐weighted images acquired before and after LND treatment. The AACID and APT* contrast measured before and after LND injection exhibited similar pH sensitivity. Interestingly, LND‐induced contrast maps showed increased heterogeneity compared with pre‐injection CEST maps. These results demonstrate that CEST contrast changes after the administration of LND could help to localize brain cancer and monitor tumor response to chemotherapy within 1 h of treatment. The LND CEST experiment uses an anticancer drug to induce a metabolic change detectable by endogenous MRI contrast, and therefore represents a unique cancer detection paradigm which differs from other current molecular imaging techniques that require the injection of an imaging contrast agent or tracer. Copyright © 2015 John Wiley & Sons, Ltd.     

免疫细胞在急性髓系白血病骨髓微环境中的浸润模式及其预后价值

目的:探究免疫细胞在急性髓系白血病骨髓微环境中的浸润模式，以及分析其与临床特征和预后间的关系。方法:从GEO数据库中下载422例急性髓系白血病患者的基因表达谱数据和相关临床数据，采用CIBERSORT算法计算急性髓系白血病患者骨髓微环境中22种免疫细胞的构成比例，然后分别分析浸润性免疫细胞与年龄、FAB分型等临床特征间的相关性，同时采用Kaplan-Meier生存曲线分析浸润性免疫细胞和总体生存时间之间的关系。结果:急性髓系白血病骨髓微环境中单核细胞，CD8+T细胞和嗜酸性粒细胞浸润丰富，其中调节性T细胞在老年急性髓系白血病患者的浸润程度较高（P=0.021）,而M2巨噬细胞的分布水平较低（P=0.014）。另外在不同病理类型的患者间有11种免疫细胞的浸润水平有显著差异（P＜0.01）。生存分析结果显示未活化的肥大细胞和M2巨噬细胞低表达以及活化的树突状细胞和调节性T细胞高表达可以预测AML患者的不良预后。结论:免疫细胞浸润模式与急性髓系白血病的年龄和病理类型间存在显著相关性，同时可以预测急性髓系白血病患者的预后。     

Upregulation of LAGE3 correlates with prognosis and immune infiltrates in colorectal cancer: A bioinformatic analysis

BackgroundColorectal cancer (CRC) is the primary cause of cancer-related deaths worldwide. Identification of new CRC biomarkers is imperative to improve the prognosis and development of therapies against the disease. LAGE3 (L Antigen Family Member 3) functions as a tRNA modifier, although its potential role in CRC has not been fully elucidated.MethodsRNA-seq matrix and corresponding clinical information were downloaded from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases, then subjected to survival, enrichment, and tumor microenvironment analyses using packages implemented in R.ResultsWe found that LAGE3 was upregulated and significantly correlating with poor prognosis in multiple CRC cohorts. Additionally, multivariate Cox regression analysis revealed that LAGE3 was an independent prognostic factor in patients with CRC, whereas functional enrichment analysis indicated that it could regulate protein targeting, tRNA processing, and the PD-1/PD-L1 checkpoint pathway. Furthermore, CIBERSORT analysis indicated a negative relationship between LAGE3 and levels of infiltration for multiple immune cells, especially CD8 + T cells in CRC. Particularly, LAGE3 expression was inversely correlated with the expression of immune checkpoints as well as that of various immune cell types of signature genes.ConclusionCollectively, our results indicate that high LAGE3 expression correlates with adverse prognosis and poor immune infiltration in CRC patients.     

Interrogating the microenvironmental landscape of tumors with computational image analysis approaches

The tumor microenvironment is an interacting heterogeneous collection of cancer cells, resident as well as infiltrating host cells, secreted factors, and extracellular matrix proteins. With the growing importance of immunotherapies, it has become crucial to be able to characterize the composition and the functional orientation of the microenvironment. The development of novel computational image analysis methodologies may enable the robust quantification and localization of immune and related biomarker-expressing cells within the microenvironment. The aim of the review is to concisely highlight a selection of current and significant contributions pertinent to methodological advances coupled with biomedical or translational applications. A further aim is to concisely present computational advances that, to our knowledge, have currently very limited use for the assessment of the microenvironment but have the potential to enhance image analysis pipelines; on this basis, an example is shown for the detection and segmentation of cells of the microenvironment using a published pipeline and a public dataset. Finally, a general proposal is presented on the conceptual design of automation-optimized computational image analysis workflows in the biomedical and clinical domain.     

皮肤T细胞淋巴瘤免疫微环境研究进展

【摘要】 皮肤T细胞淋巴瘤（CTCL）是一组具有不同临床表现、病理学改变、免疫表型和分子生物学特征的异质性肿瘤，其发病机制不完全清楚。越来越多的证据表明，肿瘤免疫微环境的组成和功能对于CTCL的发生发展起着重要作用，深入了解其微环境的组成有助于寻找更有效的抗肿瘤免疫治疗方法，同时帮助临床医生更加准确地判断疾病预后。本文就近年来关于CTCL免疫微环境的基础和临床研究进展作一综述。     

Astrocytes enhance glioblastoma growth

Glioblastoma (GBM) is a deadly disease with a need for deeper understanding and new therapeutic approaches. The microenvironment of glioblastoma has previously been shown to guide glioblastoma progression. In this study, astrocytes were investigated with regard to their effect on glioblastoma proliferation through correlative analyses of clinical samples and experimental in vitro and in vivo studies. Co-culture techniques were used to investigate the GBM growth enhancing potential of astrocytes. Cell sorting and RNA sequencing were used to generate a GBM-associated astrocyte signature and to investigate astrocyte-induced GBM genes. A NOD scid GBM mouse model was used for in vivo studies. A gene signature reflecting GBM-activated astrocytes was associated with poor prognosis in the TCGA GBM dataset. Two genes, periostin and serglycin, induced in GBM cells upon exposure to astrocytes were expressed at higher levels in cases with high “astrocyte signature score”. Astrocytes were shown to enhance glioblastoma cell growth in cell lines and in a patient-derived culture, in a manner dependent on cell–cell contact and involving increased cell proliferation. Furthermore, co-injection of astrocytes with glioblastoma cells reduced survival in an orthotopic GBM model in NOD scid mice. In conclusion, this study suggests that astrocytes contribute to glioblastoma growth and implies this crosstalk as a candidate target for novel therapies.     

结直肠癌微环境中细胞因子的表达及其与CD16a mRNA表达的相关性研究

目的探讨结直肠癌肿瘤微环境中细胞因子的表达及其与CD16a mRNA表达的关系。方法分别采用实时荧光定量PCR法和流式细胞术微球阵列法(CBA法)检测42例结直肠癌组织及其癌旁组织中CD16a mRNA和8种细胞因子[包括白细胞介素(IL)-2、IL-4、IL-6、IL-10、IL-12、肿瘤坏死因子-α(TNF-α)、γ-干扰素(IFN-γ)和血管内皮生长因子(VEGF)]的表达水平,分析两者之间的相关性。结果结直肠癌组织中IL-6、TNF-α和VEGF的表达水平高于癌旁组织(P<0.05),而IL-2、IL-4、IL-10、IL-12和IFN-γ在2种组织中的表达水平比较差异均无统计学意义(P>0.05)。术前CEA正常组的IL-6和VEGF的表达水平高于术前CEA升高组(P<0.05)。相关性分析结果显示:结直肠癌组中IL-6的相对表达水平与CD16a mRNA的表达水平呈负相关(P<0.05)。结论结直肠癌组织中IL-6、TNF-α和VEGF的表达水平较癌旁组织明显升高,提示促血管生成作用及免疫抑制增强。此外,结直肠癌肿瘤微环境中CD16a mRNA的表达与IL-6的表达呈负相关。