Process Of Malignant Transformation Research Articles

TMT-based quantitative proteomic analysis reveals the underlying mechanisms of glycidyl methacrylate-induced 16HBE cell malignant transformation

Glycidyl methacrylate (GMA) has been widely used as tackifying/crosslinking copolymer monomer in the industrial section. Occupational and environmental exposure to GMA is inevitable. GMA is classified as a Group 2 A carcinogen. However, it still lacks a sufficient understanding of its carcinogenicity at the protein level. The major pathways and players during the malignant transformation process remain unknown. In this study, we first established and characterized a malignant transformation model using human bronchial epithelial (16HBE) cells exposed to 8 μg/mL GMA. Then the proteomics approach, western-blot analysis as well as quantitative PCR (qPCR) analysis were employed to investigate its underlying mechanisms of carcinogenicity. Our results showed that the 16HBE cells exposed to GMA and passaged to the 40th generation had undergone a malignant transformation. Proteomic analysis revealed that 123 proteins were significantly up-regulated while 160 proteins were down-regulated during the process of malignant transformation. Importantly, further pathway analysis identified the extracellular matrix-receptor (ECM-receptor) interaction pathway to be one of the major players mediating the process and most of the differentially expressed proteins (DEPs) were up-regulated, including two vital proteins, CD44 and MMP14, as well as members from integrin family. These results provide direct proteomic evidence that DEPs related to the ECM-receptor interaction pathway play an active role in reinforcing the carcinogenicity of GMA. The findings of this study might deepen our understanding of the underlying mechanisms of GMA carcinogenicity and thus facilitate the risk assessment of GMA.

Role of Tumor Suppressor P53 Family in Glucose Metabolism in Association with Diabetes

Subsequent manuscripts have shown that p53, a 391 amino acid protein, was associated with tumour suppression by inducing cell cycle arrest, senescence, or programmed cell death, and mainly focused on oncology. But recent studies are proclaiming that the p53 family of genes is also associated with regulating the balance of the glucose uptake via the many glucose transporters by directly repressing the expression of the GLUT1 & GLUT4, use of glycolysis and oxidative phosphorylation. The p53 regulates phenotypes of diabetes as it is interjecting in many points in regulating the pancreatic function and apoptosis, dysregulation of the beta cells, and developing insulin resistance. In the context of pancreatic beta cells, which are functioning for insulin secretion, it is regulated by the p53 by multiple signalling pathways. Endoplasmic stress results in the accumulation of the p53 that interacts with PARK2 (parkin), leading to insulin deficiency. Additionally, several proteins (N-terminal truncated isoforms of p53, factor TCFL2, the microRNA miR 200) regulate the p53 pathway, which can also regulate its ability to mediate pancreatic beta cell death. While in glucose haemostasis, wild-type p53 negatively and mutant p53 positively regulate glucose transporters and glucose importers. P21/CDKNIA plays an important role in developing insulin resistance as the p53-mediated senescence of adipose tissue marks it. Also, excessive O-GLcynated p53 inhibits the insulin signalling pathway from acting on the liver, generating insulin resistance. Here, we present a literature review on the role of p53 in metabolism, diabetes, pancreatic function, glucose homeostasis, and insulin resistance. Interestingly, mutations in the p53 gene were shown to occur at different phases of the multistep process of malignant transformation.

Single-Cell Multi-Omic and Spatial Analysis of Nodal B Cell Non-Hodgkin Lymphomas Reveals Plasticity in B Cell Maturation As a Driver of Intratumor Heterogeneity

Introduction Intratumor heterogeneity (ITH) is increasingly recognized as an important factor in the variability of B cell non-Hodgkin lymphoma (B-NHL) patient outcomes. It is well established that distinct nodal B cell non-Hodgkin lymphoma (NBNHL) entities originate from different stages of the humoral affinity maturation process, but drivers of ITH are not well understood. In this study, we aimed to investigate whether the maturation process is a driver of ITH. Methods 51 diffuse large B cell lymphoma (DLBCL), follicular lymphoma (FL), mantle cell lymphoma (MCL), marginal zone lymphoma (MZL), and non-malignant reactive lymph node (rLN) samples from distinct patients were sequenced with CITE-Seq (coupling single-cell RNA-seq with surface epitope profiling), highly multiplexed immunohistochemistry (IHC) with 57 proteins, and single-cell B cell receptor sequencing. Transcription factor activites were inferred from gene expression using the SCENIC package. (Fig 1a)Results Using the rLN samples (n = 8), a single-cell reference map of nodal B cell maturation states was constructed based on known B cell maturation states and their markers. This reference map was used to classify maturation states in malignant samples, which were verified and refined against their maturation marker profiles. Comparable to the non-malignant context in rLN samples, variability in maturation states was also observed for malignant cells in most NBNHL tumors. Although commonalities in composition existed between samples within entities, such as a dominant pre-GC mantle cell population in MCL and abundance of memory B cells in MZL, we observed high variability in maturation state proportions between these samples (Fig 1b). Several GCB and ABC DLBCL samples contained subpopulations of both GC and post-GC phenotypes. BCR-sequencing confirmed that different intra-tumor maturation states observed were monoclonal, and therefore arose from the same cell of origin. Common epigenetic drivers of the maturation processes, such as the transcription factors EZH2 and EGR1, were upregulated in subpopulations advanced in maturation. Known pathways common driving malignant transformation in NBNHL pathogenesis, BCL6 and BCL2, were upregulated in centroblasts and post-GC subpopulations respectively in tumors of the same entities. Conclusions Our findings reveal maturation is not halted during malignancy, but rather remains a plastic and divergent process. Ongoing maturation and malignant transformation processes are intertwined as the driving forces of intratumor heterogeneity, blurring the known boundaries of NBNHL entities. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract A069: Hi-C analysis in patient-derived organoids reveals the impact of three-dimensional chromatin structures on pancreatic cancer transcriptional subtypes

Abstract Three-dimensional (3D) chromatin structures connect genes to regulatory elements, thereby contributing to cell-type-specific gene regulation. However, its role in pancreatic cancer remains unclear. Inspired by our previous finding that open chromatin profiles analyzed by ATAC-seq well characterize the biological features and transcriptional status of pancreatic neoplasms, we hereby demonstrate the previously overlooked impact of 3D epigenetic regulation in pancreatic cancers. Through integrative analysis of Hi-C, ChIP-seq, and RNA-seq using patient-derived organoids, 3D genome modules, including compartments, topologically associating domains, and enhancer-promoter loops were found to play important roles in characterizing pancreatic tumors. Interestingly, intraductal papillary mucinous neoplasm (IPMN) lineages tended to have unique 3D chromatin structure profiles. We further found that transcriptional subtypes in pancreatic cancer, either classical/progenitor-like or basal/squamous-like, were largely underpinned by 3D structures. Compartment was responsible for supporting many subtype-specific genes, by synchronously activating genes that were in the vicinity, as represented by CYP2C family genes. Meanwhile, enhancer-promoter loop fine-tuned some key subtype genes, such as GATA6. By exploring the driving forces behind these 3D structures, we found that HNF1B, a transcription factor implicated in the progenitor subtype, was sufficient to induce subtype switch, together with their underlying 3D genome structure alterations. We demonstrate that HNF1B overexpression in squamous-type pancreatic cancer organoids induced the upregulation and downregulation of genes associated with the progenitor and squamous subtype, respectively. Mechanistically, these dynamic transcriptional and 3D structural modulation were accompanied by HNF1B-induced long-range genomic interactions and H3K27ac modification at HNF1B bound sites, implying HNF1B as a driving force. We also provide evidence suggesting that this HNF1B-induced subtype switch was mediated through the phase-separating property of HNF1B, which was enabled by its intrinsically disordered region (IDR). Induction of mutant HNF1B, whose IDR was either deleted (HNF1B-dIDR) or substituted by transcriptional phase-separation forming IDR from another peptide AKAP95 (HNF1B-AKAP95), showed that subtype switching effects of HNF1B were diminished by HNF1B-dIDR or recapitulated by HNF1B-AKAP95. Thus, HNF1B contributes to the transcriptional subtype formation in pancreatic cancers through its phase-separating structural ability. Collectively, the mapping of 3D structural changes induced by transcription factors, such as HNF1B, is useful for further understanding the features of pancreatic cancers. Our study may imply the potential of 3D structures to effectively suppress or unwind the malignant transformation process and to discover subtype-specific therapeutic targets. Citation Format: Hiroyuki Kato, Dosuke Iwadate, Keisuke Yamamoto, Hiroaki Fujiwara, Hideaki Ijichi, Mitsuhiro Fujishiro, Keisuke Tateishi. Hi-C analysis in patient-derived organoids reveals the impact of three-dimensional chromatin structures on pancreatic cancer transcriptional subtypes [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr A069.

Abstract A008: Mechanisms of immune escape in NF1-associated peripheral nerve sheath tumors

Abstract Background: Neurofibromatosis type 1 (NF1) is a neurogenetic condition characterized by neurocognitive symptoms, cutaneous findings, and a predisposition for benign and malignant tumors including peripheral nerve sheath tumors (PNST). About half of patients with NF1 develop plexiform neurofibromas (pNF), non-malignant tumors that often grow rapidly during childhood and can cause significant deformity, disruption of function, and pain. Some lesions, denoted atypical neurofibromatous neoplasms of uncertain biologic potential (ANNUBP), exhibit atypia, loss of neurofibroma architecture, high cellularity and mitotic activity, as an immediate precursor to malignant transformation. For people with NF1 there is a 10-15% overall lifetime risk of developing the aggressive soft tissue sarcoma malignant peripheral nerve sheath tumors (MPNST). Despite many clinical trials of chemotherapy and targeted agents, there has been little advancement in treatment outcomes and overall patient survival remains poor; therefore, new therapeutic approaches are needed. PNST are made up of transformed Schwann cell precursors, which do not grow and survive in isolation but rather interact with infiltrating immune cells. A deeper understanding of the relationship between the pre-existing immunity and the tumor microenvironment (TME) will help unveil potential for new combinations and adjuvant therapies for patients with PNST. Methods: We have developed a unique Johns Hopkins biospecimen repository of human NF1-associated PNST specimens. We use quantitative and spatial resolution of the geography and nature of tumor infiltrating immune cells in human PNST and have determined the interactions of T cells, myeloid cells, and immunoregulatory molecules, using a combination of multiplex chromogenic, high-dimensional flow cytometry and gene expression profiling studies. We have also analyzed existing transcriptomic datasets from 21 pNF and 34 MPNST cases. Results: RNA sequence analysis revealed an accumulation of immunosuppressive Th2 cells and tumor infiltrating myeloid cells (TIM) in the TME of PNST, which we postulate generates an anti-inflammatory response against tumors. Immunophenotyping of 17 pNF, 8 ANNUBP, and 15 MPNST human specimens confirmed the higher presence of infiltrating myeloid compared to lymphoid cells, with a predominance of CD163+ myeloid cells (TIM) during progression to malignancy. We also detected a significant increase in regulatory T cells and cytotoxic CD8+ T cells in MPNST vs ANNUBP vs pNF and near absence of CD19+ B cells in all tumor types. Multiparameter flow cytometry of single cells suspensions are being studied to further investigate the association of myeloid inflammation leading to the immune evasion of PNST. Conclusions: An immunosuppressive microenvironment characterizes PNST during the process of malignant transformation, generating an immune-excluded phenotype. Leveraging the immune contexture and the mechanisms of immune modulation in PNST will inform interventions to stimulate anti-tumor immunity in this dire disease. Citation Format: Lindy Zhang, Kai Pollard, Ana Calizo, Alexandre Maalouf, Aditya Suru, Jiawan Wang, Jineeta Banerjee, Christine A. Pratilas, Nicolas J. Llosa. Mechanisms of immune escape in NF1-associated peripheral nerve sheath tumors [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A008.

LABIAL PHOTOCARCINOGENESIS

<h3>Objectives</h3> Determine the damage caused by photocarcinogenesis in the lower lip region. <h3>Study Design</h3> We conducted a retrospective study and literature review, based on published studies on photocarcinogenesis in the labial region, to determine the damage caused by photocarcinogenesis in the lower lip region. <h3>Results</h3> The lower lip region is affected by changes caused by chronic exposure to solar radiation. Photocarcinogenesis can be considered as a process of malignant transformation caused by ultraviolet (UV) radiation. The etiology of actinic cheilitis and lip squamous cell carcinoma is attributed to mutations in DNA and damage to proteins in epithelial cells caused by the cumulative action of radiation that culminates in malignant transformation. UV radiation has both mutagenic and immunosuppressive effects, which may be able to initiate and maintain neoplastic processes, and is therefore considered a complete carcinogen. The dose, exposure time, and wavelength are closely linked to this process. <h3>Conclusions</h3> UV radiation is the most relevant etiologic factor for the occurrence of skin and lip neoplasms. Understanding the process of photocarcinogenesis in the lip region is of paramount importance for the dental surgeon to be part of the front line of early diagnosis and prevention.

Upregulation of Circ_0035266 Contributes to the Malignant Progression of Inflammation-Associated Malignant Transformed Cells Induced by Tobacco-Specific Carcinogen NNK.

Cigarette smoking-induced chronic inflammation has been considered a vital driver of lung tumorigenesis. The compounds 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco-specific carcinogen, and lipopolysaccharide (LPS), an inflammatory inducer, are important components of tobacco smoke which have been implicated in inflammation-driven carcinogenesis. However, the biological effects and underlying mechanisms of LPS-mediated inflammation on NNK-induced tumorigenesis are still unclear. In this study, BEAS-2B human bronchial epithelial cells were exposed to NNK, LPS or both, for short- or long-term periods. We found that acute LPS exposure promoted the secretion of granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-6 in NNK-treated BEAS-2B cells. In addition, chronic LPS exposure facilitated the NNK-induced malignant transformation process by promoting cell proliferation, cell cycle alteration, migration, and clonal formation. Previously, we determined that circular RNA circ_0035266 enhanced cellular inflammation in response to NNK + LPS by sponging miR-181d-5p and regulating expression of its downstream target DEAD-Box Helicase 3 X-Linked (DDX3X). Here, we found that knockdown of circ_0035266 or DDX3X led to a remarkable inhibition of the proliferation, cell cycle progression, and migration of NNK + LPS-transformed BEAS-2B cells, whereas overexpression of these genes produced the opposite effects, indicating the oncogenic roles of circ_0035266 and DDX3X in the malignant progression of chronic inflammation-driven malignant transformed cells. Moreover, the regulatory relationships among circ_0035266, miR-181d-5p, and DDX3X were further confirmed using a group of lung cancer tissues. Conclusively, our findings provide novel insights into our understanding of inflammation-driven tumorigenesis using a cellular malignant transformation model, and indicate a novel tumor-promoting role for circ_0035266 in chemical carcinogenesis.

Abstract 139: Dock7 regulates the AKT/mTOR pathway to promote survival and sustain the transformative properties of cancer cells

Abstract Akt is a well-known target of mitogenic signaling, which is activated by phosphoinositide 3-kinase, PDK1 and mTORC2, and is commonly regarded as a master regulator of cell survival. However, there is still a good deal to learn regarding how Akt can promote survival when cancer cells are faced by the multitude of challenges they need to overcome during the process of malignant transformation. While investigating the role of Dock7, a Cdc42 and Rac guanine nucleotide exchange factor of the Dock180 family, in tumorigenesis, we discovered an unexpected connection between Dock7, Akt and cell survival. Specifically, we found that Dock7 is necessary for cancer cells to survive under the stressful conditions that arise due to the loss of a substratum or serum deprivation, through its ability to stabilize a low, basal level of Akt kinase activity. Dock7 binds to Akt, as well as associates with the known Akt substrate, TSC2, promoting its phosphorylation. Since the phosphorylation of TSC2 eliminates its ability to work with its partner TSC1 to deactivate the Rheb GTPase, a direct activator of mTORC1, we probed whether mTORC1 function is also necessary for Dock7-dependent cancer cell survival. Indeed, we determined that Dock7, as well as its signaling partner Cdc42, is required to maintain a functional level of S6 kinase activity, a downstream target of mTORC1, which was prevented by the mTORC1 inhibitor rapamycin. We further show an association between mTOR, Rheb and Dock7, but not Raptor, the defining component of mTORC1. Additionally, Raptor is dispensable for the Dock7-dependent S6 kinase activity, despite its rapamycin sensitivity. Together, these findings elucidate a novel Dock7-Akt-mTOR signaling node which promotes cancer cell survival in the absence of classical mTORC1 function to allow cells to overcome stresses faced during the development of cancer. Citation Format: Oriana Y. Teran, Miao-chong J. Lin, Matthew R. Zanotelli, Kristin F. Wilson, Richard A. Cerione. Dock7 regulates the AKT/mTOR pathway to promote survival and sustain the transformative properties of cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 139.

Abstract 3626: Genomic evolutionary patterns in matched pre-invasive and invasive lung disease in TRACERx

Abstract Introduction: The genomic landscape of pre-invasive lung disease, including AAH/AIS/MIA (atypical adenomatous hyperplasia, adenocarcinoma in situ, minimally invasive carcinoma) and squamous CIS (carcinoma in situ) which are considered to be precursors to lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC), has been profiled to a limited degree, due to the lack of matched samples within the same patient. Using TRACERx study we investigated the phylogenetic evolutionary patterns between matched pre-invasive and invasive lesions. Methods: Post-operative pathology reports for 746 primary invasive tumors collected within TRACERx were screened to identify pre-invasive lesions that had been resected at the time of primary surgery. 50 pre-invasive lesions (21 AAH, 7 AIS, 6 MIA, 16 CIS) from 36 pts were subjected to whole exome sequencing with 119 matched tumor regions from 22 LUAD, 13 LUSC and 1 pleomorphic carcinoma. Phylogenetic relationships between pre-invasive and invasive lesions, timing of clonal divergence, mutational processes were investigated. Results: Two major types of phylogenetic relationships were identified based on whether pre-invasive and invasive lesions shared a somatic common ancestor (SCA group; 7/22 LUAD, 11/13 LUSC) or not (non-SCA group; 15/22 LUAD, 2/13 LUSC, 1/1 pleomorphic carcinoma). Within SCA group, two evolutionary patterns were identified based on whether only clonal mutations or clonal and subclonal mutations were shared between pre-invasive and invasive lesions. 3/16 SCA pre-invasive lesions from ex-smokers were found to diverge from the invasive tumor years prior to diagnosis. Compared with non-SCA, SCA pre-invasive lesions harbored elevated mutational burden, APOBEC mutagenesis, chromosomal instability featured by increased loss of heterozygosity and somatic copy number alterations (SCNAs). Whole genome doubling was only detected in SCA pre-invasive lesions. In SCA LUAD, 3/7 exhibited clonal TP53 mutations, 3/7 exhibited clonal co-mutations of KEAP1 and STK11. In 5/15 non-SCA LUADs harboring a KRAS mutation, convergent KRAS mutation was detected in 4 matched pre-invasive and invasive lesions. Clonal focal gains on 3q21-29 (containing PIK3CA, ATR, SOX2) were detected in 10/11 SCA LUSC, suggesting that 3q gains may represent early events in tumor initiation. In the non-SCA group, convergent SCNAs encompassing oncogenes (PIK3CA, SOX2, FGFR3) or tumor suppressor genes (STK11, KEAP1, MGA) were detected. Conclusions: We demonstrate that matched pre-invasive and invasive lesions fall in two distinct phylogenetic evolutionary groups: those that have a shared somatic common ancestor and those that do not. SCA pre-invasive lesions exhibit elevated APOBEC mutagenesis and chromosomal instability, shedding some light on the process of malignant transformation. Citation Format: Haoran Zhai, Clare Puttick, David A. Moore, Michelle Dietzen, Sophia Ward, Maise Al Bakir, Thomas Watkins, Andrew Rowan, Selvaraju Veeriah, Robert Hynds, Oriol Pich, Nnennaya Kanu, TRACERx Consortium, Nicholas McGranahan, Charles Swanton, Mariam Jamal-Hanjani. Genomic evolutionary patterns in matched pre-invasive and invasive lung disease in TRACERx [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3626.

Encouraging specific biomarkers-based therapeutic strategies for hepatocellular carcinoma.

The prevention, early discovery and effective treatment of patients with hepatocellular carcinoma (HCC) remain a global medical challenge. At present, HCC is still mainly treated by surgery, supplemented by vascular embolization, radio frequency, radiotherapy, chemotherapy and biotherapy. The application of multikinase inhibitor sorafenib, chimeric antigen receptor T cells, or PD-1/PD-L1 inhibitors can prolong the median survival of HCC patients. However, the treatment efficacy is still unsatisfactory due to HCC metastasis and postoperative recurrence. During the process of hepatocyte malignant transformation, HCC tissues can express and secrete many types of specific biomarkers, or oncogenic antigen molecules into blood, for example, alpha-fetoprotein, glypican-3, Wnt3a (one of the key signaling molecules in the Wnt/β-catenin pathway), insulin-like growth factor (IGF)-II or IGF-I receptor, vascular endothelial growth factor, secretory clusterin and so on. In addition, combining immunotherapy with non-coding RNAs might improve anti-cancer efficacy. These biomarkers not only contribute to HCC diagnosis or prognosis, but may also become molecular targets for HCC therapy under developing or clinical trials. This article reviews the progress in emerging biomarkers in basic research or clinical trials for HCC immunotherapy.

O6-methylguanine DNA methyltransferase is upregulated in malignant transformation of gastric epithelial cells via its gene promoter DNA hypomethylation.

BACKGROUNDO6-methylguanine-DNA methyltransferase (MGMT) is a suicide enzyme that repairs the mispairing base O6-methyl-guanine induced by environmental and experimental carcinogens. It can transfer the alkyl group to a cysteine residue in its active site and became inactive. The chemical carcinogen N-nitroso compounds (NOCs) can directly bind to the DNA and induce the O6-methylguanine adducts, which is an important cause of gene mutation and tumorigenesis. However, the underlying regulatory mechanism of MGMT involved in NOCs-induced tumorigenesis, especially in the initiation phase, remains largely unclear.AIMTo investigate the molecular regulatory mechanism of MGMT in NOCs-induced gastric cell malignant transformation and tumorigenesis.METHODSWe established a gastric epithelial cell malignant transformation model induced by N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) or N-methyl-N-nitroso-urea (MNU) treatment. Cell proliferation, colony formation, soft agar, cell migration, and xenograft assays were used to verify the malignant phenotype. By using quantitative real-time polymerase chain reaction (qPCR) and Western blot analysis, we detected the MGMT expression in malignant transformed cells. We also confirmed the MGMT expression in early stage gastric tumor tissues by qPCR and immunohistochemistry. MGMT gene promoter DNA methylation level was analyzed by methylation-specific PCR and bisulfite sequencing PCR. The role of MGMT in cell malignant transformation was analyzed by colony formation and soft agar assays. RESULTSWe observed a constant increase in MGMT mRNA and protein expression in gastric epithelial cell malignant transformation induced by MNNG or MNU treatment. Moreover, we found a reduction of MGMT gene promoter methylation level by methylation-specific PCR and bisulfite sequencing PCR in MNNG/MNU-treated cells. Inhibition of the MGMT expression by O6-benzylguanine promoted the MNNG/MNU-induced malignant phenotypes. Overexpression of MGMT partially reversed the cell malignant transformation process induced by MNNG/MNU. Clinical gastric tissue analysis showed that MGMT was upregulated in the precancerous lesions and metaplasia tissues, but downregulated in the gastric cancer tissues.CONCLUSIONOur finding indicated that MGMT upregulation is induced via its DNA promoter hypomethylation. The highly expressed MGMT prevents the NOCs-induced cell malignant transformation and tumorigenesis, which suggests a potential novel approach for chemical carcinogenesis intervention by regulating aberrant epigenetic mechanisms.

Decoding Insulin-Like Growth Factor Signaling Pathway From a Non-coding RNAs Perspective: A Step Towards Precision Oncology in Breast Cancer.

Breast cancer (BC) is a highly complex and heterogenous disease. Several oncogenic signaling pathways drive BC oncogenic activity, thus hindering scientists to unravel the exact molecular pathogenesis of such multifaceted disease. This highlights the urgent need to find a key regulator that tunes up such intertwined oncogenic drivers to trim the malignant transformation process within the breast tissue. The Insulin-like growth factor (IGF) signaling pathway is a tenacious axis that is heavily intertwined with BC where it modulates the amplitude and activity of vital downstream oncogenic signaling pathways. Yet, the complexity of the pathway and the interactions driven by its different members seem to aggravate its oncogenicity and hinder its target-ability. In this review, the authors shed the light on the stubbornness of the IGF signaling pathway and its potential regulation by non-coding RNAs in different BC subtypes. Nonetheless, this review also spots light on the possible transport systems available for efficient delivery of non-coding RNAs to their respective targets to reach a personalized treatment code for BC patients.

Telomerase in Cancer: Function, Regulation, and Clinical Translation.

Simple SummaryCells undergoing malignant transformation must circumvent replicative senescence and eventual cell death associated with progressive telomere shortening that occurs through successive cell division. To do so, malignant cells reactivate telomerase to extend their telomeres and achieve cellular immortality, which is a “Hallmark of Cancer”. Here we review the telomere-dependent and -independent functions of telomerase in cancer, as well as its potential as a biomarker and therapeutic target to diagnose and treat cancer patients.During the process of malignant transformation, cells undergo a series of genetic, epigenetic, and phenotypic alterations, including the acquisition and propagation of genomic aberrations that impart survival and proliferative advantages. These changes are mediated in part by the induction of replicative immortality that is accompanied by active telomere elongation. Indeed, telomeres undergo dynamic changes to their lengths and higher-order structures throughout tumor formation and progression, processes overseen in most cancers by telomerase. Telomerase is a multimeric enzyme whose function is exquisitely regulated through diverse transcriptional, post-transcriptional, and post-translational mechanisms to facilitate telomere extension. In turn, telomerase function depends not only on its core components, but also on a suite of binding partners, transcription factors, and intra- and extracellular signaling effectors. Additionally, telomerase exhibits telomere-independent regulation of cancer cell growth by participating directly in cellular metabolism, signal transduction, and the regulation of gene expression in ways that are critical for tumorigenesis. In this review, we summarize the complex mechanisms underlying telomere maintenance, with a particular focus on both the telomeric and extratelomeric functions of telomerase. We also explore the clinical utility of telomeres and telomerase in the diagnosis, prognosis, and development of targeted therapies for primary, metastatic, and recurrent cancers.

Hereditary Ovarian Carcinoma: Cancer Pathogenesis Looking beyond BRCA1 and BRCA2.

Besides BRCA1 and BRCA2, several other inheritable mutations have been identified that increase ovarian cancer risk. Surgical excision of the fallopian tubes and ovaries reduces ovarian cancer risk, but for some non-BRCA hereditary ovarian cancer mutations the benefit of this intervention is unclear. The fallopian tubes of women with hereditary ovarian cancer mutations provide many insights into the early events of carcinogenesis and process of malignant transformation. Here we review cancer pathogenesis in hereditary cases of ovarian cancer, the occurrence of pre-invasive lesions and occult carcinoma in mutation carriers and their clinical management.

Long-isoform NFE2L1 silencing inhibits acquisition of malignant phenotypes induced by arsenite in human bronchial epithelial cells

Chronic arsenic exposure is associated with the increased risk of several types of cancer, among which, lung cancer is the most deadly one. Nuclear factor erythroid 2 like 1 (NFE2L1), a transcription factor belonging to CNC-bZIP family, regulates multiple important cellular functions in response to acute arsenite exposure. However, the role of NFE2L1 in lung cancer induced by chronic arsenite exposure is unknown. In this study, we firstly showed that chronic arsenite exposure (36 weeks) led to epithelial-mesenchymal transition (EMT) and malignant transformation in human bronchial epithelial cells (BEAS-2B). During the process of malignant transformation, the expression of long isoforms of NFE2L1 (NFE2L1-L) was elevated. Thereafter, BEAS-2B cells with NFE2L1-L stable knockdown (NFE2L1-L-KD) was chronically exposed to arsenite. As expected, silencing of NFE2L1-L gene strikingly inhibited the arsenite-induced EMT and the subsequent malignant transformation. Additionally, NFE2L1-L silencing suppressed the transcription of EMT-inducer SNAIL1 and increased the expression of E-cadherin. Conversely, NFE2L1-L overexpression increased SNAIL1 transcription but decreased E-cadherin expression. Collectively, our data suggest that NFE2L1-L promotes EMT by positively regulating SNAIL1 transcription, and is involved in malignant transformation induced by arsenite.

The dysregulation of unsaturated fatty acid-based metabolomics in the MNNG-induced malignant transformation of Het-1A cells.

Studies have shown that environmental carcinogens exerted an important function in the high incidence of esophageal cancer (EC). Nitrosamines have been identified as important environmental carcinogens for EC. This study aimed to investigate the metabolic disturbances and new key toxicological markers in the malignant transformation process of normal esophageal epithelial cells (Het-1A) induced by MNNG (N-methyl-N'-nitro-N-nitrosoguanidine). Untargeted metabolomic and lipidomic profiling analysis by using ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC-MS) were applied to explore the metabolic network alterations of Het-1A cells. The metabolomic results showed that significant alterations were observed in metabolic signatures between different generations (P5, P15, P25, P35) and the control cell group (P0). A total of 48 differential endogenous metabolites were screened and identified, mainly containing fatty acids, amino acids, and nucleotides. The differential metabolites were predominantly linked to the pathway of biosynthesis of unsaturated fatty acids metabolism. The cell lipidomic profiling revealed that the most differential lipids contained fatty acids (FAs), phosphatidylcholines (PC), phosphatidylethanolamines (PE), and phosphatidylserines (PS). The enrichment of the lipidomic pathway also confirmed that the lipid metabolism of biosynthesis of unsaturated fatty acids was the significant variation during the cell malignant transformation. Furthermore, we detected the expression of the upstream regulatory enzymes related to the unsaturated fatty acids to explore the regulation mechanism. The expression of stearoyl-CoA desaturase (SCD), ELOVL fatty acid elongase 1 (ELOVL1) promoted, and fatty acid desaturase 1 (FADS1) inhibited the key fatty acids of unsaturated fatty acids metabolism compared to the control cell group. Overall, our results revealed that lipid fatty acid metabolism was involved in the malignant transformation of Het-1A cells induced by MNNG and deepened the awareness of the carcinogenic mechanism of environmental exposure pollutants.

Interleukin-17 and vascular endothelial growth factor: new biomarkers for the diagnosis of papillary thyroid carcinoma in patients with Hashimoto's thyroiditis.

ObjectiveThe incidences of papillary thyroid carcinoma (PTC) and Hashimoto’s thyroiditis (HT) have shown increasing trends. Numerous studies have shown a close relationship between the two diseases, but the exact mechanism linking PTC with HT is still unclear. Interleukin-17 (IL-17) plays an important role in the development of malignant tumors. However, information on the association between IL-17 and thyroid disease is lacking.MethodsTissue samples were collected from patients with thyroid diseases admitted to the thyroid surgery department of our hospital between May 2015 and December 2017. The characteristics of the thyroid were observed by ultrasonography, hematoxylin-eosin staining, enzyme-linked immunosorbent assays, and immunohistochemistry.ResultsWe found that HT with carcinoma (HTC) showed unique characteristics in two-dimensional ultrasound images. Moreover, IL-17 and vascular endothelial growth factor (VEGF) levels showed gradually increasing trends during the process of HT malignant transformation, with a significant positive correlation between the two cytokines. Serum IL-17 and VEGF levels could distinguish between HTC and HT with benign adenoma.ConclusionOur data suggest that serum IL-17 and VEGF levels may represent novel biomarkers for the diagnosis of HT malignant nodules.

Histone Modifications and Their Targeting in Lymphoid Malignancies.

In a wide range of lymphoid neoplasms, the process of malignant transformation is associated with somatic mutations in B cells that affect the epigenetic machinery. Consequential alterations in histone modifications contribute to disease-specific changes in the transcriptional program. Affected genes commonly play important roles in cell cycle regulation, apoptosis-inducing signal transduction, and DNA damage response, thus facilitating the emergence of malignant traits that impair immune surveillance and favor the emergence of different B-cell lymphoma subtypes. In the last two decades, the field has made a major effort to develop therapies that target these epigenetic alterations. In this review, we discuss which epigenetic alterations occur in B-cell non-Hodgkin lymphoma. Furthermore, we aim to present in a close to comprehensive manner the current state-of-the-art in the preclinical and clinical development of epigenetic drugs. We focus on therapeutic strategies interfering with histone methylation and acetylation as these are most advanced in being deployed from the bench-to-bedside and have the greatest potential to improve the prognosis of lymphoma patients.

Pleomorphic adenoma: detection of PLAG1 rearrangement–positive tumor components using whole-slide fluorescence in situ hybridization

Pleomorphic adenoma (PA) consists of heterogeneous histological architecture mixed with epithelioid and mesenchymal forms. Various types of epithelial or myoepithelial malignancies arise from PA, but sarcomas are extremely rare. A human androgen receptor gene (HUMARA) clonality assay has suggested that PA is clonal in nature. However, clonality of various tumor components of PA would be difficult to determine with this assay. In addition, the results obtained should be carefully interpreted. PLAG1 rearrangements are considered a good molecular marker for neoplasticity in PA. We aimed to clarify the neoplasticity of the various tumor components present in PA using whole-slide fluorescence in situ hybridization (FISH). Five PA cases positive for PLAG1 rearrangements were examined. Using an immunohistochemistry panel, cell components in PA were classified into eight cell types. To precisely localize PLAG1 rearrangement-positive cell components at the cellular level, sequential retrieval of whole-slide imaging (WSI) data of HE histology and FISH for PLAG1 rearrangement was carried out. PLAG1 rearrangements were detected in ductal cells, myoepithelial spindle cells, myoepithelial oncocytic cells, myoepithelial plasmacytoid cells, and mesenchymal chondroid cells, but not in mesenchymal lipid cells, mesenchymal fibrous cells, or vascular endothelial cells. Immunohistochemical PLAG1 expression was restricted to cell components harboring PLAG1 rearrangements.The results of the present study indicate that ductal and myoepithelial, chondroid cells are neoplastic but lipid, fibrous, and endothelial cells are not. PLAG1 immunohistochemistry is useful in discriminating neoplastic from non-neoplastic cell components. These findings may be important for elucidating tumorigenesis and the process of malignant transformation in PA.

Genetic alterations associated with malignant transformation of sporadic vestibular schwannoma

IntroductionMalignant peripheral nerve sheath tumor of the vestibulocochlear nerve (VN-MPNST) is exceedingly rare and carries a poor prognosis. Little is known about its underlying genetics and in particular the process of malignant transformation. There is an ongoing debate on whether the transformation is initiated by ionizing radiation. We present here the analysis and comparison of two post-radiation VN-MPNST and one undergoing spontaneous transformation.MethodsFour tumors from three patients (radiation-naïve vestibular schwannoma before (VS) and after (VN-MPNST) malignant transformation in addition to two post-radiation VN-MPNST) were subjected to DNA whole-genome microarray and whole-exome sequencing and tumor-specific mutations were called. Mutational signatures were characterized using MuSiCa.ResultsThe tumor genomes were characterized predominantly by copy-number aberrations with 36–81% of the genome affected. Even the VS genome was grossly aberrated. The spontaneous malignant transformation was characterized by a near-total whole-genome doubling, disappearance of NF2 mutation and new mutations in three cancer-related genes (GNAQ, FOXO4 and PDGFRB). All tumors had homozygous loss of the tumor suppressor CDKN2A. Neither mutational signature nor copy number profile was associated with ionizing radiation.ConclusionThe VN-MPNST genome in our cases is characterized by large copy-number aberrations and homozygous deletion of CDKN2A. Our study demonstrates a VS with genetic alterations similar to its malignant counterpart, suggesting the existence of premalignant VS. No consistent mutational signature was associated with ionizing radiation.