Next-Generation Digital Histopathology of the Tumor Microenvironment

Progress in cancer research is substantially dependent on innovative technologies that permit a concerted analysis of the tumor microenvironment and the cellular phenotypes resulting from somatic mutations and post-translational modifications. In view of a large number of genes, multiplied by differential splicing as well as post-translational protein modifications, the ability to identify and quantify the actual phenotypes of individual cell populations in situ, i.e., in their tissue environment, has become a prerequisite for understanding tumorigenesis and cancer progression. The need for quantitative analyses has led to a renaissance of optical instruments and imaging techniques. With the emergence of precision medicine, automated analysis of a constantly increasing number of cellular markers and their measurement in spatial context have become increasingly necessary to understand the molecular mechanisms that lead to different pathways of disease progression in individual patients. In this review, we summarize the joint effort that academia and industry have undertaken to establish methods and protocols for molecular profiling and immunophenotyping of cancer tissues for next-generation digital histopathology—which is characterized by the use of whole-slide imaging (brightfield, widefield fluorescence, confocal, multispectral, and/or multiplexing technologies) combined with state-of-the-art image cytometry and advanced methods for machine and deep learning.

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The Nutritional Cytokine Leptin Promotes NSCLC by Activating the PI3K/AKT and MAPK/ERK Pathways in NSCLC Cells in a Paracrine Manner

BioMed Research International ◽

10.1155/2019/2585743 ◽

2019 ◽

Vol 2019 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Fengzhou Li ◽

Shilei Zhao ◽

Tao Guo ◽

Jinxiu Li ◽

Chundong Gu

Keyword(s):

Lung Cancer ◽

Tumor Microenvironment ◽

Western Blot ◽

Signaling Pathways ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Lung Fibroblasts ◽

Normal Lung ◽

Serum Leptin ◽

Coculture System

Purpose.Leptin is a nutritional cytokine encoded by the obesity gene whose concentration in the tumor microenvironment is closely related to the occurrence and progression of cancer. However, previous evidence has suggested that there is no clear relationship between serum leptin concentrations and lung cancer progression. Cancer-associated fibroblasts (CAFs), the most abundant component of the tumor microenvironment in a variety of solid tumors, were recently reported to produce leptin. Therefore, it was inferred that leptin is most likely to affect non-small-cell lung cancer (NSCLC) through an autocrine and paracrine mechanism. In the current study, we investigated the paracrine effect and mechanism of leptin produced by CAFs on NSCLC by establishing a novel in vitro cell coculture system.Methods.A noncontact coculture device was designed and made by 3D printing. CAFs and paired normal lung fibroblasts (NLFs) from 5 patients were successfully isolated and cocultured with two NSCLC cell lines in a coculture system. The background expression of leptin was detected by western blot. The in situ expression of leptin and its receptor (Ob-R) in NSCLC tissues and paired normal lung tissues was analyzed by immunohistochemistry. Furthermore, we downregulated the expression of leptin in CAFs and assessed changes in its promotion on NSCLC cells in the coculture system. Finally, changes in the phosphorylation of ERK1/2 and AKT were examined to investigate the molecular mechanisms responsible for the paracrine promotion of NSCLC cells by leptin.Results.Leptin was overexpressed in nearly all five primary CAF lines compared with its expression in paired NLFs. IHC staining showed that the expression of leptin was high in NSCLC cells, slightly lower in CAF, and negative in normal lung tissue. Ob-R was strongly expressed in NSCLC cells. The ability of A549 and H1299 cells to proliferate and migrate was enhanced by high leptin levels in both the cocultured fibroblasts and the culture medium. Furthermore, western blot assays suggested that the MAPK/ERK1/2 and PI3K/AKT signaling pathways were activated by leptin produced by CAFs, which demonstrated that the functions of paracrine leptin in NSCLC are as those of the serum leptin to other cancers.Conclusion.Leptin produced by CAF promotes proliferation and migration of NSCLC cells probably via PI3K/AKT and MAPK/ERK1/2 signaling pathways in a paracrine manner.

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Iron as a Central Player and Promising Target in Cancer Progression

International Journal of Molecular Sciences ◽

10.3390/ijms20020273 ◽

2019 ◽

Vol 20 (2) ◽

pp. 273 ◽

Cited By ~ 41

Author(s):

Michaela Jung ◽

Christina Mertens ◽

Elisa Tomat ◽

Bernhard Brüne

Keyword(s):

Tumor Microenvironment ◽

Tumor Cell ◽

Cancer Progression ◽

Crucial Role ◽

Iron Homeostasis ◽

Molecular Mechanisms ◽

Tumor Therapy ◽

Essential Element ◽

Major Function ◽

Iron Pool

Iron is an essential element for virtually all organisms. On the one hand, it facilitates cell proliferation and growth. On the other hand, iron may be detrimental due to its redox abilities, thereby contributing to free radical formation, which in turn may provoke oxidative stress and DNA damage. Iron also plays a crucial role in tumor progression and metastasis due to its major function in tumor cell survival and reprogramming of the tumor microenvironment. Therefore, pathways of iron acquisition, export, and storage are often perturbed in cancers, suggesting that targeting iron metabolic pathways might represent opportunities towards innovative approaches in cancer treatment. Recent evidence points to a crucial role of tumor-associated macrophages (TAMs) as a source of iron within the tumor microenvironment, implying that specifically targeting the TAM iron pool might add to the efficacy of tumor therapy. Here, we provide a brief summary of tumor cell iron metabolism and updated molecular mechanisms that regulate cellular and systemic iron homeostasis with regard to the development of cancer. Since iron adds to shaping major hallmarks of cancer, we emphasize innovative therapeutic strategies to address the iron pool of tumor cells or cells of the tumor microenvironment for the treatment of cancer.

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High TRAF6 Expression Is Associated With Esophageal Carcinoma Recurrence and Prompts Cancer Cell Invasion

Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics ◽

10.3727/096504016x14749340314441 ◽

2017 ◽

Vol 25 (4) ◽

pp. 485-493 ◽

Cited By ~ 12

Author(s):

Xinyang Liu ◽

Zhichao Wang ◽

Guoliang Zhang ◽

Qikun Zhu ◽

Hui Zeng ◽

...

Keyword(s):

Esophageal Cancer ◽

Cancer Progression ◽

Poor Prognosis ◽

Molecular Mechanisms ◽

Tumor Necrosis Factor Receptor ◽

Migration And Invasion ◽

Loss Of Function ◽

Underlying Mechanisms ◽

Patient Prognosis ◽

Cancer Tissues

Esophageal cancer is one of the most common types of cancer, and it has a poor prognosis. The molecular mechanisms of esophageal cancer progression remain largely unknown. In this study, we aimed to investigate the clinical significance and biological function of tumor necrosis factor receptor-associated factor 6 (TRAF6) in esophageal cancer. Expression of TRAF6 in esophageal cancer was examined, and its correlation with clinicopathological factors and patient prognosis was analyzed. A series of functional and mechanism assays were performed to further investigate the function and underlying mechanisms in esophageal cancer. Expression of TRAF6 was highly elevated in esophageal cancer tissues, and patients with high TRAF6 expression have a significantly shorter survival time than those with low TRAF6 expression. Furthermore, loss-of-function experiments showed that knockdown of TRAF6 significantly reduced the migration and invasion abilities of esophageal cancer cells. Moreover, the pro-oncogenic effects of TRAF6 in esophageal cancer were mediated by the upregulation of AEP and MMP2. Altogether, our data suggest that high expression of TRAF6 is significant for esophageal cancer progression, and TRAF6 indicates poor prognosis in esophageal cancer patients, which might be a novel prognostic biomarker or potential therapeutic target in esophageal cancer.

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Extracellular HSP90 Machineries Build Tumor Microenvironment and Boost Cancer Progression

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.735529 ◽

2021 ◽

Vol 9 ◽

Author(s):

Pietro Poggio ◽

Matteo Sorge ◽

Laura Seclì ◽

Mara Brancaccio

Keyword(s):

Tumor Microenvironment ◽

Cell Surface ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Diagnostic Tools ◽

Surface Receptors ◽

And Behavior ◽

Cancer Cell Signaling ◽

The Impact ◽

Stimulation Of

HSP90 is released by cancer cells in the tumor microenvironment where it associates with different co-chaperones generating complexes with specific functions, ranging from folding and activation of extracellular clients to the stimulation of cell surface receptors. Emerging data indicate that these functions are essential for tumor growth and progression. The understanding of the exact composition of extracellular HSP90 complexes and the molecular mechanisms at the basis of their functions in the tumor microenvironment may represent the first step to design innovative diagnostic tools and new effective therapies. Here we review the impact of extracellular HSP90 complexes on cancer cell signaling and behavior.

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RalGPS2 Interacts with Akt and PDK1 Promoting Tunneling Nanotubes Formation in Bladder Cancer and Kidney Cells Microenvironment

Cancers ◽

10.3390/cancers13246330 ◽

2021 ◽

Vol 13 (24) ◽

pp. 6330

Author(s):

Alessia D’Aloia ◽

Edoardo Arrigoni ◽

Barbara Costa ◽

Giovanna Berruti ◽

Enzo Martegani ◽

...

Keyword(s):

Bladder Cancer ◽

Tumor Microenvironment ◽

Cell Lines ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Cell Communication ◽

Time Lapse ◽

Nucleotide Exchange ◽

Class Iii ◽

Tunneling Nanotubes

RalGPS2 is a Ras-independent Guanine Nucleotide Exchange Factor for RalA GTPase that is involved in several cellular processes, including cytoskeletal organization. Previously, we demonstrated that RalGPS2 also plays a role in the formation of tunneling nanotubes (TNTs) in bladder cancer 5637 cells. In particular, TNTs are a novel mechanism of cell–cell communication in the tumor microenvironment, playing a central role in cancer progression and metastasis formation. However, the molecular mechanisms involved in TNTs formation still need to be fully elucidated. Here we demonstrate that mid and high-stage bladder cancer cell lines have functional TNTs, which can transfer mitochondria. Moreover, using confocal fluorescence time-lapse microscopy, we show in 5637 cells that TNTs mediate the trafficking of RalA protein and transmembrane MHC class III protein leukocyte-specific transcript 1 (LST1). Furthermore, we show that RalGPS2 is essential for nanotubes generation, and stress conditions boost its expression both in 5637 and HEK293 cell lines. Finally, we prove that RalGPS2 interacts with Akt and PDK1, in addition to LST1 and RalA, leading to the formation of a complex that promotes nanotubes formation. In conclusion, our findings suggest that in the tumor microenvironment, RalGPS2 orchestrates the assembly of multimolecular complexes that drive the formation of TNTs.

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Low Dose Cyclophosphamide Modulates Tumor Microenvironment by TGF-β Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms21030957 ◽

2020 ◽

Vol 21 (3) ◽

pp. 957 ◽

Cited By ~ 2

Author(s):

Hui Zhong ◽

Yifan Lai ◽

Rui Zhang ◽

Abdelkader Daoud ◽

Qingyuan Feng ◽

...

Keyword(s):

T Cells ◽

Tumor Microenvironment ◽

Immune Function ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Low Dose ◽

Histopathological Analysis ◽

Lewis Lung Cancer ◽

Continuous Administration ◽

Nude Mouse Model

The tumor microenvironment has been recently recognized as a critical contributor to cancer progression and anticancer therapy-resistance. Cyclophosphamide (CTX) is a cytotoxic agent commonly used in clinics for the treatment of cancer. Previous reports demonstrated that CTX given at low continuous doses, known as metronomic schedule, mainly targets endothelial cells and circulating Tregs with unknown mechanisms. Here, we investigated the antitumor activity of two different metronomic schedules of CTX along with their corresponding MTD regimen and further explored their effect on immune function and tumor microenvironment. Toxicity evaluation was monitored by overall survival rate, weight loss, and histopathological analysis. A nude mouse model of Lewis lung cancer was established to assess the anti-metastatic effects of CTX in vivo. CD4+, CD8+, and CD4+CD25+FoxP3 T cells were selected by flow cytometry analysis. Low and continuous administration of CTX was able to restore immune function via increase of CD4+/CD8+ T cells and depletion of T regulatory cells, not only in circulatory and splenic compartments, but also at the tumor site. Low-dose CTX also reduced myofibroblasts, accompanied with an increased level of E-cadherin and low N-cadherin, both in the primary tumor and lung through the TGF-β pathway by the downregulated expression of TGF-β receptor 2. Our data may indicate that several other molecular mechanisms of CTX for tumor may be involved in metronomic chemotherapy, besides targeting angiogenesis and regulatory T cells.

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The circular RNA circDLG1 promotes gastric cancer progression and anti-PD-1 resistance through the regulation of CXCL12 by sponging miR-141-3p

Molecular Cancer ◽

10.1186/s12943-021-01475-8 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Dong-Liang Chen ◽

Hui Sheng ◽

Dong-Sheng Zhang ◽

Ying Jin ◽

Bai-Tian Zhao ◽

...

Keyword(s):

Gastric Cancer ◽

Cancer Progression ◽

Immune Evasion ◽

Molecular Mechanisms ◽

Circular Rnas ◽

Gastric Cancer Cells ◽

Cancer Tissues ◽

Gastric Cancer Progression

Abstract Background Dysregulation of circular RNAs (circRNAs) plays an important role in the development of gastric cancer; thus, revealing the biological and molecular mechanisms of abnormally expressed circRNAs is critical for identifying novel therapeutic targets in gastric cancer. Methods A circRNA microarray was performed to identify differentially expressed circRNAs between primary and distant metastatic tissues and between gastric cancer tissues sensitive or resistant to anti-programmed cell death 1 (PD-1) therapy. The expression of circRNA discs large homolog 1 (DLG1) was determined in a larger cohort of primary and distant metastatic gastric cancer tissues. The role of circDLG1 in gastric cancer progression was evaluated both in vivo and in vitro, and the effect of circDLG1 on the antitumor activity of anti-PD-1 was evaluated in vivo. The interaction between circDLG1 and miR-141-3p was assessed by RNA immunoprecipitation and luciferase assays. Results circDLG1 was significantly upregulated in distant metastatic lesions and gastric cancer tissues resistant to anti-PD-1 therapy and was associated with an aggressive tumor phenotype and adverse prognosis in gastric cancer patients treated with anti-PD-1 therapy. Ectopic circDLG1 expression promoted the proliferation, migration, invasion, and immune evasion of gastric cancer cells. Mechanistically, circDLG1 interacted with miR-141-3p and acted as a miRNA sponge to increase the expression of CXCL12, which promoted gastric cancer progression and resistance to anti-PD-1-based therapy. Conclusions Overall, our findings demonstrate how circDLG1 promotes gastric cancer cell proliferation, migration, invasion and immune evasion and provide a new perspective on the role of circRNAs during gastric cancer progression.

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The molecular mechanisms of LncRNA-correlated PKM2 in cancer metabolism

Bioscience Reports ◽

10.1042/bsr20192453 ◽

2019 ◽

Vol 39 (11) ◽

Author(s):

Ting Tao ◽

Shiyuan Wu ◽

Zheng Sun ◽

Wei Ma ◽

Sichun Zhou ◽

...

Keyword(s):

Cancer Progression ◽

Molecular Mechanisms ◽

Cancer Metabolism ◽

Metabolic Reprogramming ◽

Functional Studies ◽

Genome Wide ◽

Non Coding Rnas ◽

Cancer Tissues ◽

Rate Limiting ◽

Important Marker

Abstract Reprogrammed metabolism is an important hallmark of cancer cells. Pyruvate kinase (PK) is one of the major rate-limiting enzymes in glucose metabolism. The M2 isoform of PK (PKM2), is considered to be an important marker of metabolic reprogramming and one of the key enzymes. Recently, through the continuous development of genome-wide analysis and functional studies, accumulating evidence has demonstrated that long non-coding RNAs (LncRNAs) play vital regulatory roles in cancer progression by acting as either potential oncogenes or tumor suppressors. Furthermore, several studies have shown that up-regulation of PKM2 in cancer tissues is associated with LncRNAs expression and patient survival. Thus, scientists have begun to unveil the mechanism of LncRNA-associated PKM2 in cancer metabolic progression. Based on these novel findings, in this mini-review, we summarize the detailed molecular mechanisms of LncRNA related to PKM2 in cancer metabolism. We expect that this work will promote a better understanding of the molecular mechanisms of PKM2, and provide a profound potential for targeting PKM2 to treat tumors.

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The Role of miR-4256/HOXC8 Signaling Axis in the Gastric Cancer Progression: Evidence From lncRNA-miRNA-mRNA Network Analysis

Frontiers in Oncology ◽

10.3389/fonc.2021.793678 ◽

2022 ◽

Vol 11 ◽

Author(s):

Haijuan Gu ◽

Yuejiao Zhong ◽

Jibin Liu ◽

Qian Shen ◽

Rong Wei ◽

...

Keyword(s):

Gastric Cancer ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Differentially Expressed ◽

Gastric Cancer Cells ◽

Functional Studies ◽

Cancer Tissues ◽

Gastric Cancer Progression

Gastric cancer is a deadly human malignancy and the molecular mechanisms underlying gastric cancer pathophysiology are very complicated. Thus, further investigations are warranted to decipher the underlying molecular mechanisms. With the development of high-throughput screening and bioinformatics, gene expression profiles with large scale have been performed in gastric cancer. In the present study, we mined The Cancer Genome Atlas (TCGA) database and analyzed the gene expression profiles between gastric cancer tissues and normal gastric tissues. A series of differentially expressed lncRNAs, miRNAs and mRNAs between gastric cancer tissues and normal gastric tissues were identified. Based on the differentially expressed genes, we constructed miRNA-mRNA network, lncRNA-mRNA network and transcriptional factors-mRNA-miRNA-lncRNA network. Furthermore, the Kaplan survival analysis showed that high expression levels of EVX1, GBX2, GCM1, HOXC8, HOXC9, HOXC10, HOXC11, HOXC12 and HOXC13 were all significantly correlated with shorter overall survival of the patients with gastric cancer. On the other hand, low expression level of HOXA13 was associated with shorter overall survival of patients with gastric cancer. Among these hub genes, we performed the in vitro functional studies of HOXC8 in the gastric cancer cells. Knockdown of HOXC8 and overexpression of miR-4256 both significantly repressed the gastric cancer cell proliferation and migration, and miR-4256 repressed the expression of HOXC8 via targeting its 3’ untranslated region in gastric cancer cells. Collectively, our results revealed that a complex interaction networks of differentially expressed genes in gastric cancer, and further functional studies indicated that miR-4256/HOXC8 may be an important axis in regulating gastric cancer progression.

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LncRNA AC245100.4 affects the tumorigenesis of prostate cancer via regulating the STAT3/NR4A3 axis

10.21203/rs.3.rs-131948/v1 ◽

2020 ◽

Author(s):

Chi Liu ◽

Ping Lin ◽

Jiabin Zhao ◽

Hui Xie ◽

Tianhu Zheng ◽

...

Keyword(s):

Prostate Cancer ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Rna Seq ◽

Rna Immunoprecipitation ◽

Cancer Tissues ◽

Promising Avenue

Abstract Background Prostate cancer (PCa) is the second most common cancer and the fifth leading cause of cancer deaths among men globally. However, the molecular mechanisms leading to the progression have not been fully elucidated. Methods The expression and location of AC245100.4 were examined by RT-qPCR and nuclear-cytoplasmic separation assay. RNA-seq analysis was performed to identify the downstream of AC245100.4. RNA immunoprecipitation was performed to identify the proteins those bind to AC245100.4. Western blotting was performed to quantify the expression of proteins. Finally, a series of gain- or loss-functional assays were done to prove the precise role of AC245100.4 and NR4A3 in PCa. Results We identify a critical lncRNA AC245100.4, which is significantly up-regulated in prostate cancer tissues and cells. Knockdown of AC21500.4 can significantly inhibit prostate cancer progression in vitro and in vivo. Further RNA-seq analysis shows that NR4A3 may be the potential target of AC245100.4. Mechanistically, AC245100.4 de-regulates NR4A3 transcriptionally via increasing p-STAT3, which is a transcriptional repressor of NR4A3. Conclusion Our results demonstrated that AC245100.4 was a critically oncogenic lncRNA in PCa via inhibiting NR4A3 and paved a promising avenue to combat PCa progression by targeting AC245100.4 or NR4A3.

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