Mapping of Human Polyomavirus in Renal Cell Carcinoma Tissues.

Worldwide, the incidence of renal cell carcinoma (RCC) is rising, accounting for approximately 2% of all cancer diagnoses and deaths. The etiology of RCC is still obscure. Here, we assessed the presence of HPyVs in paraffin-embedded tissue (FFPE) resected tissue from patients with RCC by using different molecular techniques. Fifty-five FFPE tissues from 11 RCC patients were included in this study. Consensus and HPyV-specific primers were used to screen for HPyVs. Both PCR approaches revealed that HPyV is frequently detected in the tissues of RCC kidney resections. A total of 78% (43/55) of the tissues tested were positive for at least one HPyV (i.e., MCPyV, HPyV6, HPyV7, BKPyV, JCPyV, or WUyV). Additionally, 25 tissues (45%) were positive for only one HPyV, 14 (25%) for two HPyVs, 3 (5%) for three HPyVs, and 1 one (1%) tissue specimen was positive for four HPyVs. Eleven (20%) RCC specimens were completely devoid of HPyV sequences. MCPyV was found in 24/55 RCC tissues, HPyV7 in 19, and HPyV6 in 8. The presence of MCPyV and HPyV6 was confirmed by specific FISH or RNA-ISH. In addition, we aimed to confirm HPyV gene expression by IHC. Our results strongly indicate that these HPyVs infect RCC and nontumor tissues, possibly indicating that kidney tissues serve as a reservoir for HPyV latency. Whether HPyVs possibly contribute to the etiopathogenesis of RCC remains to be elucidated.

Single-cell transcriptomic landscape and the microenvironment of normal adjacent tissues in hypopharyngeal carcinoma.

BACKGROUND: The cellular origin of hypopharyngeal diseases is crucial for further diagnosis and treatment, and the microenvironment in tissues may also be associated with specific cell types at the same time. Normal adjacent tissues (NATs) of hypopharyngeal carcinoma differ from non-tumor-bearing tissues, and can influenced by the tumor. However, the heterogeneity in kinds of disease samples remains little known, and the transcriptomic profile about biological information associated with disease occurrence and clinical outcome contained in it has yet to be fully evaluated. For these reasons, we should quickly investigate the taxonomic and transcriptomic information of NATs in human hypopharynx. RESULTS: Single-cell suspensions of normal adjacent tissues (NATs) of hypopharyngeal carcinoma were obtained and single-cell RNA sequencing (scRNA-seq) was performed. We present scRNA-seq data from 39,315 high-quality cells in the hypopharyngeal from five human donors, nine clusters of normal adjacent human hypopharyngeal cells were presented, including epithelial cells, endothelial cells (ECs), mononuclear phagocyte system cells (MPs), fibroblasts, T cells, plasma cells, B cells, mural cells and mast cells. Nonimmune components in the microenvironment, including epithelial cells, endothelial cells, fibroblasts and the subpopulations of them were performed. CONCLUSIONS: Our data provide a solid basis for the study of single-cell landscape in human normal adjacent hypopharyngeal tissues biology and related diseases.

Endometrial Cancer-Adjacent Tissues Express Higher Levels of Cancer-Promoting Genes than the Matched Tumors.

Molecular alterations in tumor-adjacent tissues have recently been recognized in some types of cancer. This phenomenon has not been studied in endometrial cancer. We aimed to analyze the expression of genes associated with cancer progression and metabolism in primary endometrial cancer samples and the matched tumor-adjacent tissues and in the samples of endometria from cancer-free patients with uterine leiomyomas. Paired samples of tumor-adjacent tissues and primary tumors from 49 patients with endometrial cancer (EC), samples of endometrium from 25 patients with leiomyomas of the uterus, and 4 endometrial cancer cell lines were examined by the RT-qPCR, for MYC, NR5A2, CXCR2, HMGA2, LIN28A, OCT4A, OCT4B, OCT4B1, TWIST1, STK11, SNAI1, and miR-205-5p expression. The expression levels of MYC, NR5A2, SNAI1, TWIST1, and STK11 were significantly higher in tumor-adjacent tissues than in the matched EC samples, and this difference was not influenced by the content of cancer cells in cancer-adjacent tissues. The expression of MYC, NR5A2, and SNAI1 was also higher in EC-adjacent tissues than in samples from cancer-free patients. In addition, the expression of MYC and CXCR2 in the tumor related to non-endometrioid adenocarcinoma and reduced the risk of recurrence, respectively, and higher NR5A2 expression in tumor-adjacent tissue increased the risk of death. In conclusion, tissues proximal to EC present higher levels of some cancer-promoting genes than the matched tumors. Malignant tumor-adjacent tissues carry a diagnostic potential and emerge as new promising target of anticancer therapy.

Recent Advances in Pathology: the 2022 Annual Review Issue of The Journal of Pathology.

The 2022 Annual Review Issue of The Journal of Pathology, Recent Advances in Pathology, contains 15 invited reviews on research areas of growing importance in pathology. This year, the articles include those that focus on digital pathology, employing modern imaging techniques and software to enable improved diagnostic and research applications to study human diseases. This subject area includes the ability to identify specific genetic alterations through the morphological changes they induce, as well as integrating digital and computational pathology with 'omics technologies. Other reviews in this issue include an updated evaluation of mutational patterns (mutation signatures) in cancer, the applications of lineage tracing in human tissues, and single cell sequencing technologies to uncover tumour evolution and tumour heterogeneity. The tissue microenvironment is covered in reviews specifically dealing with proteolytic control of epidermal differentiation, cancer-associated fibroblasts, field cancerisation, and host factors that determine tumour immunity. All of the reviews contained in this issue are the work of invited experts selected to discuss the considerable recent progress in their respective fields and are freely available online (https://onlinelibrary.wiley.com/journal/10969896). © 2022 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Field cancerization in breast cancer.

Breast cancer affects one in seven women worldwide during their lifetime. Widespread mammographic screening programs and education campaigns allow for early detection of the disease, often during its asymptomatic phase. Current practice in treatment and recurrence monitoring is based primarily on pathological evaluations but can also encompass genomic evaluations, both of which focus on the primary tumor. Although breast cancer is one of the most studied cancers, patients still recur at a rate of up to 15% within the first 10 years post-surgery. Local recurrence was originally attributed to tumor cells contaminating histologically normal (HN) tissues beyond the surgical margin, but advances in technology have allowed for the identification of distinct aberrations that exist in the peri-tumoral tissues themselves. One leading theory to explain this phenomenon is the field cancerization theory. Under this hypothesis, tumors arise from a field of molecularly altered cells that create a permissive environment for malignant evolution, which can occur with or without morphological changes. The traditional histopathology paradigm dictates that molecular alterations are reflected in the tissue phenotype. However, the spectrum of inter-patient variability of normal breast tissue may obfuscate recognition of a cancerized field during routine diagnostics. In this review, we explore the concept of field cancerization focusing on HN peri-tumoral tissues: we present the pathological and molecular features of field cancerization within these tissues and discuss how the use of peri-tumoral tissues can affect research. Our observations suggest that pathological and molecular evaluations could be used synergistically to assess risk and guide the therapeutic management of patients. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Meta-analysis of 16S rRNA Microbial Data Identified Distinctive and Predictive Microbiota Dysbiosis in Colorectal Carcinoma Adjacent Tissue.

As research focusing on the colorectal cancer fecal microbiome using shotgun sequencing continues, increasing evidence has supported correlations between colorectal carcinomas (CRCs) and fecal microbiome dysbiosis. However, large-scale on-site and off-site (surrounding adjacent) tissue microbiome characterization of CRC was underrepresented. Here, considering each taxon as a feature, we demonstrate a machine learning-based method to investigate tissue microbial differences among CRC, colorectal adenoma (CRA), and healthy control groups using 16S rRNA data sets retrieved from 15 studies. A total of 2,099 samples were included and analyzed in case-control comparisons. Multiple methods, including differential abundance analysis, random forest classification, cooccurrence network analysis, and Dirichlet multinomial mixture analysis, were conducted to investigate the microbial signatures. We showed that the dysbiosis of the off-site tissue of colonic cancer was distinctive and predictive. The AUCs (areas under the curve) were 80.7%, 96.0%, and 95.8% for CRC versus healthy control random forest models using stool, tissue, and adjacent tissue samples and 69.9%, 91.5%, and 89.5% for the corresponding CRA models, respectively. We also found that the microbiota ecologies of the surrounding adjacent tissues of CRC and CRA were similar to their on-site counterparts according to network analysis. Furthermore, based on the enterotyping of tissue samples, the cohort-specific microbial signature might be the crux in addressing classification generalization problems. Despite cohort heterogeneity, the dysbiosis of lesion-adjacent tissues might provide us with further perspectives in demonstrating the role of the microbiota in colorectal cancer tumorigenesis.IMPORTANCE Turbulent fecal and tissue microbiome dysbiosis of colorectal carcinoma and adenoma has been identified, and some taxa have been proven to be carcinogenic. However, the microbiomes of surrounding adjacent tissues of colonic cancerous tissues were seldom investigated uniformly on a large scale. Here, we characterize the microbiome signatures and dysbiosis of various colonic cancer sample groups. We found a high correlation between colorectal carcinoma adjacent tissue microbiomes and their on-site counterparts. We also discovered that the microbiome dysbiosis in adjacent tissues could discriminate colorectal carcinomas from healthy controls effectively. These results extend our knowledge on the microbial profile of colorectal cancer tissues and highlight microbiota dysbiosis in the surrounding tissues. They also suggest that microbial feature variations of cancerous lesion-adjacent tissues might help to reveal the microbial etiology of colonic cancer and could ultimately be applied for diagnostic and screening purposes.

In situ analysis of Her2 DNA and RNA in retinoblastoma and adjacent retina.

Retinoblastoma (RB) is an ocular tumor of early childhood. Current treatments attempt to preserve visual function, but may spare chemoresistant tumor cells. One potential therapeutic target for RB is HER2, (ERBB2), expressed in RB in truncated form. In this study, we tested the hypothesis that Her2 DNA and RNA are expressed in RB tumors and adjacent retina. We examined 24 human RB tumors as well as normal-appearing adjacent retinal tissues for Her2 DNA and RNA expression by in situ hybridization. We also examined 28 RB tumors for HER2 protein immunoreactivity. 21/22 RB tumors expressed Her2 DNA and 14/19 tumors expressed Her2 RNA. In 17 paired cases, there were three cases in which Her2 DNA was detected, but not RNA. We also saw Her2 RNA signal in six instances of "normal" adjacent retinal tissue. Heterogeneous HER2 protein expression in specific tumor regions also was confirmed by quantitative HER2 immunohistochemistry. In summary, Her2 DNA and RNA are expressed in many RB tumors, and in some adjacent ocular tissues, with hetereogenous protein expression throughout. These results may provide important insights regarding RB tumor progression, and drug targeting approaches designed to spare the eye, preserve vision and improve quality of life for RB patients.

Novel reference genes in colorectal cancer identify a distinct subset of high stage tumors and their associated histologically normal colonic tissues.

BACKGROUND: Reference genes are often interchangeably called housekeeping genes due to 1) the essential cellular functions their proteins provide and 2) their constitutive expression across a range of normal and pathophysiological conditions. However, given the proliferative drive of malignant cells, many reference genes such as beta-actin (ACTB) and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) which play critical roles in cell membrane organization and glycolysis, may be dysregulated in tumors versus their corresponding normal controls METHODS: Because Next Generation Sequencing (NGS) technology has several advantages over hybridization-based technologies, such as independent detection and quantitation of transcription levels, greater sensitivity, and increased dynamic range, we evaluated colorectal cancers (CRC) and their histologically normal tissue counterparts by NGS to evaluate the expression of 21 "classical" reference genes used as normalization standards for PCR based methods. Seventy-nine paired tissue samples of CRC and their patient matched healthy colonic tissues were subjected to NGS analysis of their mRNAs. RESULTS: We affirmed that 17 out of 21 classical reference genes had upregulated expression in tumors compared to normal colonic epithelial tissue and dramatically so in some cases. Indeed, tumors were distinguished from normal controls in both unsupervised hierarchical clustering analyses (HCA) and principal component analyses (PCA). We then identified 42 novel potential reference genes with minimal coefficients of variation (CV) across 79 CRC tumor pairs. Though largely consistently expressed across tumors and normal control tissues, a subset of high stage tumors (HSTs) as well as some normal tissue samples (HSNs) located adjacent to these HSTs demonstrated dysregulated expression, thus identifying a subset of tumors with a potentially distinct and aggressive biological profile. CONCLUSION: While classical CRC reference genes were found to be differentially expressed between tumors and normal controls, novel reference genes, identified via NGS, were more consistently expressed across malignant and normal colonic tissues. Nonetheless, a subset of HST had profound dysregulation of such genes as did many of the histologically normal tissues adjacent to such HSTs, indicating that the HSTs so distinguished may have unique biological properties and that their histologically normal tissues likely harbor a small population of microscopically undetected but metabolically active tumors.

Mapping the current flow in sacral nerve stimulation using computational modelling.

Sacral nerve stimulation (SNS) is an established treatment for faecal incontinence involving the implantation of a quadripolar electrode into a sacral foramen, through which an electrical stimulus is applied. Little is known about the induced spread of electric current around the SNS electrode and its effect on adjacent tissues, which limits optimisation of this treatment. The authors constructed a 3-dimensional imaging based finite element model in order to calculate and visualise the stimulation induced current and coupled this to biophysical models of nerve fibres. They investigated the impact of tissue inhomogeneity, electrode model choice and contact configuration and found a number of effects. (i) The presence of anatomical detail changes the estimate of stimulation effects in size and shape. (ii) The difference between the two models of electrodes is minimal for electrode contacts of the same length. (iii) Surprisingly, in this arrangement of electrode and neural fibre, monopolar and bipolar stimulation induce a similar effect. (iv) Interestingly when the active contact is larger, the volume of tissue activated reduces. This work establishes a protocol to better understand both therapeutic and adverse stimulation effects and in the future will enable patient-specific adjustments of stimulation parameters.

Expression and association of IL-21, FBXL20 and tumour suppressor gene PTEN in laryngeal cancer.

OBJECTIVE: To study the expression of three genes IL-21, FBXL20 and tumour suppressor gene PTEN in laryngeal cancer; analyse the differences in their expression in laryngeal cancer and adjacent tissues; by using pEGFP-N1-IL21 and pGPU/GFP/Neo-FBXL20 expression vectors, to analyse the characteristics in their expression in laryngeal cancer cells outside the body as well as the associations among them. METHODS: The expression of the three genes in laryngeal cancer and adjacent tissues from 30 cases and in normal laryngeal tissues from 20 healthy persons was detected with the RT-PCR; laryngeal cancer cell line (HEp-2 cells) transfection was also performed with the pEGFP-N1-IL21 and pGPU/GFP/Neo-FBXL20 expression vectors we constructed, to detect the mRNA expression of the three genes. Cell proliferation, apoptosis and cell cycle were measured by the MTT assay. RESULTS: The results of RT-PCR showed that the expression of IL-21 and FBXL20 was up-regulated in laryngeal cancer, while the expression of tumour suppressor gene PTEN was significantly decreased (p < 0.01). In HEp-2 cells transfected with pGPU/GFP/Neo-IL-21 and pGPU/GFP/Neo-FBXL20 expression vectors, the mRNA expression of PTEN was restored to some extent (p < 0.05); in addition, the ability of HEp-2 cells in proliferation and invasion was also reduced. CONCLUSIONS: IL-21 and FBXL20 genes are important in the occurrence and development of laryngeal cancer; the expression of PTEN gene can suppress laryngeal cancer, and there's a certain association among IL-21, FBXL20 and PTEN.

Association Between Shear Wave Elastography of Virtual Touch Tissue Imaging Quantification Parameters and the Ki-67 Proliferation Status in Luminal-Type Breast Cancer.

OBJECTIVES: To evaluate the association between shear wave elastography parameters using virtual touch tissue imaging quantification (VTIQ) and the Ki-67 index in luminal-type breast cancer. METHODS: Eighty-one patients with 82 lesions of pathologic confirmed luminal-type breast cancer underwent virtual touch tissue imaging quantification examination before surgery between December 2015 and June 2016. Patients were divided into 2 groups according to the Ki-67 index (≥14% versus < 14%), which is used to define luminal type B and luminal type A, respectively. The mean shear wave velocity (SWVmean ) and lesion-to-adjacent tissues ratio (SWV ratio) were calculated for each lesion. RESULTS: The SWVmean , SWV ratio, histologic grade, axillary lymph node involvement, and lymphovascular invasion showed a significant positive association with a high Ki-67 index (all P < .05). Receiver operating characteristic curve analysis for the differential diagnosis between high (≥14%) and low (<14%) Ki-67 groups displayed that the optimal cutoff value for SWVmean and SWV ratio were 3.99 meters per second and 2.861, with sensitivity 94% and 72%, specificity 40.6% and 56.2%, and area under the receiver operating characteristic curve of 0.689 and 0.651, respectively. Univariate analysis showed that SWVmean (P = .005), SWV ratio (P = .029), histologic grade (P = .011), presence of axillary node involvement (P = .004), and lymphovascular invasion (P = .008) were significantly associated with high Ki-67 status. Multivariable analysis displayed that SWVmean (hazard ratio [HR], 1.459, 95% confidence interval [CI], 1.028-2.072; P = .035), histologic grade (HR, 4.105; 95% CI, 1.142-14.763; P = .031), and presence of axillary node involvement (HR, 3.75; 95% CI, 1.228-11.453; P = .020) maintained significance for predicting high Ki-67 status. CONCLUSIONS: The SWVmean using the virtual touch tissue imaging quantification method showed significant correlation with the Ki-67 index, suggesting the potential to assess tumor proliferation status in luminal-type breast cancer with a noninvasive manner.

Prostate cancer induces C/EBPß expression in surrounding epithelial cells which relates to tumor aggressiveness and patient outcome.

BACKGROUND: Implantation of rat prostate cancer cells into the normal rat prostate results in tumor-stimulating adaptations in the tumor-bearing organ. Similar changes are seen in prostate cancer patients and they are related to outcome. One gene previously found to be upregulated in the non-malignant part of tumor-bearing prostate lobe in rats was the transcription factor CCAAT/enhancer-binding protein-ß (C/EBPß). METHODS: To explore this further, we examined C/EBPß expression by quantitative RT-PCR, immunohistochemistry, and Western blot in normal rat prostate tissue surrounding slow-growing non-metastatic Dunning G, rapidly growing poorly metastatic (AT-1), and rapidly growing highly metastatic (MatLyLu) rat prostate tumors-and also by immunohistochemistry in a tissue microarray (TMA) from prostate cancer patients managed by watchful waiting. RESULTS: In rats, C/EBPß mRNA expression was upregulated in the surrounding tumor-bearing prostate lobe. In tumors and in the surrounding non-malignant prostate tissue, C/EBPß was detected by immunohistochemistry in some epithelial cells and in infiltrating macrophages. The magnitude of glandular epithelial C/EBPß expression in the tumor-bearing prostates was associated with tumor size, distance to the tumor, and metastatic capacity. In prostate cancer patients, high expression of C/EBPß in glandular epithelial cells in the surrounding tumor-bearing tissue was associated with accumulation of M1 macrophages (iNOS+) and favorable outcome. High expression of C/EBPß in tumor epithelial cells was associated with high Gleason score, high tumor cell proliferation, metastases, and poor outcome. CONCLUSIONS: This study suggest that the expression of C/EBP-beta, a transcription factor mediating multiple biological effects, is differentially expressed both in the benign parts of the tumor-bearing prostate and in prostate tumors, and that alterations in this may be related to patient outcome.

Expression of galectin-9 mRNA in hepatocellular carcinoma.

Liver cancer, with a very high prevalence, is one of the most common causes of death worldwide. Galectin-9, a semi-galactoside-binding protein, was demonstrated to be involved in the formation and metastasis processes of tumors such as breast cancer, and has significant impact on the development and prognosis of tumor. In this study, 90 cases of liver cancer patients who had liver cancer resection surgery treatment, were selected. Samples of liver cancer tissues and cancer-adjacent tissues from the surgery resection of liver cancer patients, which also confirmed by pathology after operation as specimens, were obtained to detect the expression level of Galectin-9 mRNA. The comparing results showed that there were significant differences between the expression of Galectin-9 mRNA in cancer-adjacent tissues and that in cancer tissues (P < 0.05), in terms of pathology differentiation, TNM, and recurrence transfer aspects. However, there were no obvious correlations with gender, age, the size of tumor, and HBsAg. The expression of Galectin-9 mRNA has a close relationship with pathological differentiation, TNM, and recurrence metastasis. Our data presented here provide theoretical basis for new target of liver cancer diagnosis as well as potential prognosis.

Cartilage oligomeric matrix protein (COMP)-mediated cell differentiation to proteolysis mechanism networks from human normal adjacent tissues to lung adenocarcinoma.

BACKGROUND: To understand cartilage oligomeric matrix protein (COMP) mechanism network from human normal adjacent tissues to lung adenocarcinoma. METHODS: COMP complete different activated (all no positive correlation, Pearson CC < 0.25) and uncomplete (partly no positive correlation except COMP, Pearson CC < 0.25) network were identified in higher lung adenocarcinoma compared with lower human normal adjacent tissues from the corresponding COMP-stimulated (≥0.25) or inhibited (Pearson CC ≤ -0.25) overlapping molecules of Pearson correlation coefficient (CC) and GRNInfer, respectively. COMP complete different activated and inhibited (all no positive correlation, Pearson CC < 0.25) mechanisms networks of higher lung adenocarcinoma and lower human normal adjacent tissues were constructed by integration of Pearson CC, GRNInfer and GO. As visualized by integration of GO, KEGG, GenMAPP, BioCarta and Disease, we deduced COMP complete different activated and inhibited network in higher lung adenocarcinoma and lower human normal adjacent tissues. RESULTS: As visualized by GO, KEGG, GenMAPP, BioCarta and disease database integration, we proposed mainly that the mechanism and function of COMP complete different activated network in higher lung adenocarcinoma was involved in COMP activation with matrix-localized insulin-like factor coupling carboxypeptidase to metallopeptidase-induced proteolysis, whereas the corresponding inhibited network in lower human normal adjacent tissues participated in COMP inhibition with nucleus-localized vasculogenesis, B and T cell differentiation and neural endocrine factors coupling pyrophosphatase-mediated proteolysis. However, COMP complete different inhibited network in higher lung adenocarcinoma included COMP inhibition with nucleus-localized chromatin maintenance, licensing and assembly factors coupling phosphatase-inhibitor to cytokinesis regulators-mediated cell differentiation, whereas the corresponding activated network in lower human normal adjacent tissues contained COMP activation with cytolplasm-localized translation elongation factor coupling fucosyltransferase to ubiquitin-protein ligase-induced cell differentiation. CONCLUSION: COMP different networks were verified not only by complete and uncomplete COMP activated or inhibited networks within human normal adjacent tissues or lung adenocarcinoma, but also by COMP activated and inhibited network between human normal adjacent tissues and lung adenocarcinoma.

Renal cell carcinoma-adjacent tissues enhance mobilization and recruitment of endothelial progenitor cells to promote the invasion of the neoplasm.

AIMS: The aim of this study was to detect the levels of CD34(+)/Flk-1(+) endothelial progenitor cells (EPCs) in renal cell carcinoma (RCC)-adjacent tissues and explore the correlation of RCC-adjacent tissues EPCs and tumor invasion. METHODS: An orthotopic renal tumor model was successfully established. At days 7, 12, 17 and 21, eight mice were put to death respectively. Tumor diameters were measured and RCC-adjacent tissues were collected. The percentage of EPCs within the kidney mononuclear cell population was detected. The expression levels of Stromal cell-derived factor-1 (SDF-1), vascular endothelial growth factor (VEGF) and their receptors CXCR4, Flk-1 mRNA and protein were probed respectively. And then, mean microvascular density (MVD) was examined. RESULTS: EPC numbers in RCC-adjacent tissues were significantly higher than those in control groups. The ratios of EPCs were increased gradually, and so were tumor diameters. The levels of SDF-1 and VEGF were also increased gradually, but significantly reduced compared with control group at each time point. In addition, CXCR4 and Flk-1 expression were decreased gradually. CONCLUSIONS: Our investigation suggested that EPCs in RCC-adjacent tissues play an important role in early stage RCC invasion, involving the promotion on angiogenesis through releasing several angiogenic factors.