Premalignant Transformation Research Articles

Germ cells positive for PLAP and c-Kit in 11-16 year old normal boys with ongoing spermatogenesis.

Positive staining of testicular germ cells for PLAP and c-Kit beyond infancy may be associated with the presence of GCNIS (Germ Cell Neoplasia In Situ). We recently reported our findings of positive staining of normal, infantile germ cells for PLAP, and c-Kit up to 2years of age, contrary to previous studies. The present study aims to elucidate whether otherwise normal testes of boys undergoing puberty express PLAP, c-Kit, Oct3/4, or D2-40. Biopsies were taken from 31 boys (11.5-16.5years of age, mean and median of 13.5years), who underwent surgery either for torsion of the testis (15) or a history suspicious of intermittent torsion of the testis (16). 21 were biopsied on both sides, making a total of 52 biopsies. Four testes were necrotic. The biopsies were fixed in Stieve's medium, cut into 2μm sections, and mounted on coated slides. One slide was processed for H-E, and the others incubated with primary antibody for PLAP, c-Kit, D2-40, and Oct3/4. 87% of the boys stained positive for both PLAP and c-Kit. None were positive for either D2-40 or Oct3/4. None had any histological features characteristic of GCNIS. Only two boys showed no signs of having initiated spermatogenesis. Those positive for PLAP were likewise for c-Kit, and vice versa, except 2; one boy, 13years, was positive for PLAP, but negative for c-KIT, another, 16years, was negative for PLAP and positive for c-Kit. Three boys stained positive for PLAP and c-Kit on the right side, and negative on the left. One boy was negative for c-Kit on the right side, positive on the left, and positive for PLAP bilaterally. Positive staining of testicular germ cells for PLAP and c-Kit seems to be a normal finding in boys not having completed puberty. Rather than indicating pre-malignant transformation, the positivity is indicative of an ongoing maturational process of the germ cells.

Clinical analysis of 30 patients with atypical glandular hyperplasia or malignant transformation in foci of adenomyosis

Objective: To explore the clinical characteristics of patients with pre-malignant or malignant transformation in ectopic endometrium of adenomyosis. Methods: A retrospective study was performed on 4 818 patients received surgical treatment in Peking Union Medical College Hospital and pathologically confirmed as adenomyosis from January 2010 to October 2016. The clinical features and prognosis of 30 patients among these with pre-malignant or malignant transformation in ectopic endometrium were reviewed. Results: A total of 4 818 patients pathologically diagnosed with adenomyosis. Among these,30 (0.62%,30/4 818) patients were diagnosed with pre-malignant or malignant transformation in foci of adenomyosis, including 16 (0.33%, 16/4 818) of atypical endometrial hyperplasia and 14 (0.29%, 14/4 818) of endometrioid carcinoma. The average age of 30 patients was (50.8±3.0) years (range: 27-65 years). Patients with atypical hyperplasia in foci of adenomyosis coexist with 13 cases of endometrioid carcinoma in situ, 1 case of borderline serous papilloma in eutopic endometrium, 1 case of atypical hyperplasia in eutopic endometrium, and 1 case of normal eutopic endometrium. Fourteen patients with endometrioid carcinoma in foci of adenomyosis coexist with endometrioid carcinoma in situ. Among the 27 patients with pre-malignant or malignant transformation in adenomyosis and coexisting with endometrioid carcinoma in situ, 23 of them were in International Federation of Gynecology and Obstetrics (FIGO) stage Ⅰa (85%, 23/27), 2 in stage Ⅰb (7%, 2/27), and 2 in stage Ⅲa (7%, 2/27). Seven percent (2/27) of patients had deep myometrial infiltration, and 7% (2/27) of patients had lymphovascular space invasion. The median follow-up time was 60 months (range: 32-104 months), and there were no recurrence or death. Conclusions: The incidence of atypical glandular hyperplasia or malignant transformation in foci of adenomyosis is rare, and it may be closely related to endometrial disease in situ. Pre-malignant or malignant change in foci of adenomyosis seems not to increase the risk of deep myometrial invasion, lymphovascular space invasion, and recurrence or mortality in patients of adenomyosis with endometrial adenocarcinoma.

Abstract A12: The establishment, maintenance, and maladaptive role of epigenetic memory in mediating pancreatic tumorigenesis

Abstract Cellular decision-making is highly dependent on cues present in the environment. As such, biologic systems have evolved efficient ways to respond to salient and often transient stimuli by encoding and transducing relevant extracellular information to chromatin. Additionally, accumulating evidence suggests that cells are capable of remembering previously experienced stimuli, allowing them to adjust their behaviors appropriately when confronted in the future with a similar or different stimulus. However, the ability of cells to respond to a transient stimulus and to encode a persistent record of that experience following its resolution has yet to be explored comprehensively in a preneoplastic context. Here we determine the lasting effect in the pancreas of an acinar-to-ductal metaplasia (ADM) event, to elucidate the impact of prior inflammation on tissue function and subsequent tumorigenesis. In a lineage-traced model of acinar cell dedifferentiation, we demonstrate that following ADM, histologically normal acinar cells exhibit broad gains in chromatin accessibility that persist well after exposure to the initial ADM-inducing stimulus. We show that the effect on chromatin is not reflected in the transcriptional state of acinar cells. Moreover, we employ extensive flow-cytometry based analysis of immune cell and niche acinar populations to show there is no long-term cell-extrinsic population, nor a significant expansion of either the Dclk1+ or Nk1r+ acinar populations. Finally, we employ single-cell RNA-seq analyses to define the alterations to epithelial cells and the associated microenvironment long after regeneration following an episode of pancreatitis. These data suggest that inflammation-exposed acinar cells are capable of establishing an altered, persistently accessible chromatin state that lasts long after the initial lineage destabilizing event. Currently we are defining if, following resolution of pancreatitis, activation of mutant Kras will be sufficient to induce premalignant transformation within the acinar compartment that manifests as pancreatic intraepithelial neoplasia (PanIN)—the most common precursor lesion to pancreatic ductal adenocarcinoma (PDAC). Understanding how this postinflammatory chromatin state synergizes with oncogenic stress will further explain why inflammation is a hallmark risk factor for cancer development. Together, our studies will characterize the establishment, maintenance, and maladaptive role of epigenetic memory in the development of pancreatic cancer. Citation Format: David J. Falvo, Rohit Chandwani. The establishment, maintenance, and maladaptive role of epigenetic memory in mediating pancreatic tumorigenesis [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr A12.

Identification of FASN-Dependent Onco-Metabolic Regulation of the Pentose Phosphate Pathway (PPP) and Nucleotide Metabolism in Non-Hodgkin Lymphoma (NHL)

Introduction: FASN catalyzes de novo fatty acid (FA) biosynthesis, which is an oncogenic function observed in many cancers, including NHL. HIF1a inducible FASN activity is responsible for overcoming negative hypoxic influence and upregulation of glucose metabolism as observed during premalignant transformation. FASN catalytic activity is also dependent on precursors derived from glucose metabolism. However, implications of FASN targeted therapies on these interdependent metabolic interactions and the overall impact on cancer cell proliferation remains unknown. Methods: FASN small molecule inhibitors cerulenin, orlistat, TVB3657 and TVB3166 were evaluated using a diverse panel of B cell NHL lines and primary NHL cells for the impact on FASN inhibition signaling & induction of cell death (MTT, caspases & AnnexinV/PI). Global transcriptomics were done with Affymetrix Human 2.0 ST Genechip with Gene Set Enrichment and Ingenuity Pathway Analysis (GSEA & IPA). Metabolomic profiling was performed using mass spectrometry. TXNRD1, GSR, NQO1 expression and activity were evaluated by western blot and using enzyme assay kits. Global DNA & RNA synthesis were evaluated using ClickIt Edu and EU assay kits. RNAseq data available from 775 NHL patients were utilized for metabolic transcriptome mapping. Results: Treatment with all FASN inhibitors resulted in dose- and time-dependent reduction (>90%) in cell viability and cell death in all NHL cell lines. GSEA and IPA identified cell cycle & RNA metabolism as downregulated biological processes with carbohydrate and oxidative stress as upregulated biological processes observed. "Key gene" analysis predicted TNF signaling as a prominent response to FASN inhibition, validated as increased TNF secretion by ELISA with cell fractionation and western blot analysis revealing activation of TNF-PI3K signaling. Activation of PI3K with FASN inhibition also corresponded with increased expression of PI3K-dependent metabolic genes associated with glucose and lipid metabolism. Furthermore, metabolomic profiling in SUDHL10 cells revealed accumulation of the FASN precursor acetyl-CoA with FASN inhibition that was accompanied by increased ketogenic glycolytic and citric acid cycle activity. In addition, NADPH accumulation (FASN substrate) occurred with FASN inhibition, which was accompanied with reduction in ribose-phosphate and nucleotide pools as these processes are relevant to NADPH-generating PPP function. G6PD, TXNRD1, GSR & NQO1 were identified as "key genes" responsive to FASN inhibition. Interestingly, this cluster of "key genes" represented the entire enzymatic activity related to the first rate-limiting step in PPP. Subsequently, we determined that increased NADP(H) pools were responsible for impaired NADPH regenerating functions of TXNRD1 and GSR antioxidant enzymes, with PI3K-dependent activation of NQO1 and uptake of glucose facilitating de novo glutathione synthesis, which substituted for the loss of antioxidant functions in SUDHL10, SUDHL4 and Raji cells. All responses were sensitive to inhibition by PI3K inhibition (e.g., BKM120) with co-targeting via FASN & PI3K inhibition resulting in markedly increased oxidative stress, loss of mitochondrial membrane potential & synergistic cell death in NHL cell lines and primary NHL cells. Finally, FASN inhibition was associated with reduction in ribo/deoxy-ribonucleotide pools that decreased global transcriptional activity (de novo RNA synthesis, by EU labeling) and replication (by EdU incorporation in DNA) (Fig 1). Analysis of RNAseq data and mapping of metabolic transcriptome from 772 NHL patients showed consistently elevated expression of genes related to glycolysis, citric acid cycle, fatty acid and nucleotide metabolism in patients with mutations in p53, MYC, BCL2, mTOR, MYD88, PIM2 & CREBP genes, suggesting that these onco-metabolic interactions may be important for lymphomagenesis. Conclusions: Taken together, FASN oncogenic activity appears to extend beyond de novo fatty acid biosynthesis, serving as a central onco-metabolic regulator of malignant cell proliferation vis-à-vis integrating glucose, nucleotides, and antioxidant metabolic functions in NHL. In addition, co-targeting FASN and PI3K induced synergistic cell death. Altogether, blocking FASN and the dependent onco-metabolic functions represent highly novel targets for therapeutic strategies in NHL. Disclosures Chen: Oncomics, LLC: Consultancy, Patents & Royalties. Dave:Data Driven Bioscience: Equity Ownership. Evens:Seattle Genetics: Consultancy, Honoraria, Research Funding; Pharmacyclics: Consultancy, Honoraria; Epizyme: Consultancy, Honoraria; Tesaro: Research Funding; Verastem: Consultancy, Honoraria.

Crocetin and Crocin from Saffron in Cancer Chemotherapy and Chemoprevention.

Cancer is a disorder which has a powerful impact on the quality life and life expectancy despite the increase in drugs and treatments available for cancer patients. Moreover, many new therapeutic options are known to have adverse reactions without any improvement in outcome than before. Nowadays, natural products or plant derivatives are used as chemoprevention drugs and chemotherapy is the new approach that uses specific cell premalignant transformation in the malignant form. Natural substances derived from plants, such as polyphenols, flavonoids, carotenoids, alkaloids and others, can be biologically active and have a wide spectrum of effects. The protective effects of Saffron carotenoids (crocin and crocetin) have been extensively studied mainly for their antioxidant properties, however, they have various other biological activities including tumor growth inhibition with the induction of cell death. The relevant information on Saffron and its carotenoids was collected from scientific databases (such as PubMed, Web of Science, Science Direct). To identify all published articles in relation to saffron, crocin and crocetin, in different types of cancer, no language restriction has been used. To date, crossing the words saffron and cancer, approximately 150 articles can be found. If crossing is made between crocin and cancer, approximately 60 articles can be found. With the crossing between crocetin and cancer, the number is approximately 55, while between carotenoids and cancer, the number exceeds 16.000 reports. In all the papers published to date, there are evidences that saffron and its carotenoids exert chemopreventive activity through anti-oxidant activity, cancer cells apoptosis, inhibition of cell proliferation, enhancement of cell differentiation, modulation of cell cycle progression and cell growth, modulation of tumor metabolism, stimulation of cell-to-cell communication and immune modulation. Here, we have tried to offer an up-to-date overview of pre-clinical experimental investigations on the potential use of the main carotenoids of saffron in tumor models and focus the attention on the molecular mechanisms involved.

The Molecular Chaperone Heat Shock Protein 70 Controls Liver Cancer Initiation and Progression by Regulating Adaptive DNA Damage and Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase Signaling Pathways.

Delineating the mechanisms that drive hepatic injury and hepatocellular carcinoma (HCC) progression is critical for development of novel treatments for recurrent and advanced HCC but also for the development of diagnostic and preventive strategies. Heat shock protein 70 (HSP70) acts in concert with several cochaperones and nucleotide exchange factors and plays an essential role in protein quality control that increases survival by protecting cells against environmental stressors. Specifically, the HSP70-mediated response has been implicated in the pathogenesis of cancer, but the specific mechanisms by which HSP70 may support malignant cell transformation remains to be fully elucidated. Here, we show that genetic ablation of HSP70 markedly impairs HCC initiation and progression by distinct but overlapping pathways. This includes the potentiation of the carcinogen-induced DNA damage response, at the tumor initiation stage, to increase the p53-dependent surveillance response leading to the cell cycle exit or death of genomically damaged differentiated pericentral hepatocytes, and this may also prevent their conversion into more proliferating HCC progenitor cells. Subsequently, activation of a mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) negative feedback pathway diminishes oncogenic signals, thereby attenuating premalignant cell transformation and tumor progression. Modulation of HSP70 function may be a strategy for interfering with oncogenic signals driving liver cell transformation and tumor progression, thus providing an opportunity for human cancer control.

Role of ten-eleven translocation proteins and 5-hydroxymethylcytosine in hepatocellular carcinoma.

In mammals, methylation of the 5th position of cytosine (5mC) seems to be a major epigenetic modification of DNA. This process can be reversed (resulting in cytosine) with high efficiency by dioxygenases of the ten‐eleven translocation (TET) family, which perform oxidation of 5mC to 5‐hydroxymethylcytosine (5hmC), 5‐formylcytosine and 5‐carboxylcytosine. It has been demonstrated that these 5mC oxidation derivatives are in a dynamic state and have pivotal regulatory functions. Here, we comprehensively summarized the recent research progress in the understanding of the physiological functions of the TET proteins and their mechanisms of regulation of DNA methylation and transcription. Among the three TET genes, TET1 and TET2 expression levels have frequently been shown to be low in hepatocellular carcinoma (HCC) tissues and received most attention. The modulation of TET1 also correlates with microRNAs in a post‐transcriptional regulatory process. Additionally, recent studies revealed that global genomic 5hmC levels are down‐regulated in HCC tissues and cell lines. Combined with the reported results, identification of 5hmC signatures in HCC tissues and in circulating cell‐free DNA will certainly contribute to early detection and should help to design therapeutic strategies against HCC. 5hmC might also be a novel prognostic biomarker of HCC. Thus, a detailed understanding of the molecular mechanisms resulting in the premalignant and aggressive transformation of TET proteins and cells with 5hmC disruption might help to develop novel epigenetic therapies for HCC.

DNA-PKc deficiency drives pre-malignant transformation by reducing DNA repair capacity in concert with reprogramming the epigenome in human bronchial epithelial cells

The expression of DNA-dependent protein kinase catalytic subunit (DNA-PKc) is highly variable in smokers and reduced enzyme activity has been associated with risk for lung cancer. An in vitro model of lung pre-malignancy was used to evaluate the role of double-strand break DNA repair capacity in transformation of hTERT/CDK4 immortalized human bronchial epithelial cells (HBECs) and reprograming of the epigenome. Here we show that knockdown of DNA-PKc to levels simulating haploinsufficiency dramatically reduced DNA repair capacity following challenge with bleomycin and significantly increased transformation efficiency of HBEC lines exposed weekly for 12 weeks to this radiomimetic. Transformed HBEC lines with wild type or knockdown of DNA-PKc showed altered expression of more than 1,000 genes linked to major cell regulatory pathways involved in lung cancer. While lung cancer driver mutations were not detected in transformed clones, more than 300 genes that showed reduced expression associated with promoter methylation in transformed clones or predictive for methylation in malignant tumors were identified. These studies support reduced DNA repair capacity as a key factor in the initiation and clonal expansion of pre-neoplastic cells and double-strand break DNA damage as causal for epigenetic mediated silencing of many lung cancer-associated genes. The fact that DNA damage, repair, and epigenetic silencing of genes are causal for many other cancers that include colon and prostate extends the generalizability and impact of these findings.

Myelodysplastic Syndrome related alterations of MAPK signaling in the bone marrow of experimental mice including stem/progenitor compartment

Myelodysplastic syndrome is considered globally as heterogenous group of neoplasm which often proclaims leukemic progression. The heterogeneity is reflected not only in clinical manifestations of the disease but also in salient causes of disease development. In spite of multiple therapeutic modalities, shortfall towards treatment of this disorder still persists. The focal point of tussle suggested toward defects, which are not confined to any unifying cellular signalling. The pathobiology of the disease often experiences an intriguing paradox involving ‘hyperproliferative bone marrow with pancytopenic peripheral blood’. In our present study we have reported about MAPK signaling in the hematopoietic stem progenitor compartmental (HSPC) dysregulation during the course of alkylator(ENU) induced myelodysplasia. The phospho-protein status of RTK’s(FLT3, PDGFR, EGFR) were markedly increased that activated MAPK signaling proteins which finally executed their tasks by transcription of c-Myc and Rb leading to uncontrolled cellular proliferation, simultaneously the activated c-Jun revealed stress related apoptosis. Altogether, the role of activated MAPK signaling in the HSPC’s may have led to hyperproliferation and concurrent enhanced apoptosis of abnormal cells which gradually headed towards premalignant transformations during the course of disease. The phenotypic expression of the HSPC markers CD 150 and CD 90 also established a mechanistic correlation with MAPK signalling alterations and overall scenario.

Alteration of the PD-1/PD-L1 axis in cervical intraepithelial neoplasia-preliminary study

Introduction: Programmed cell death 1 (PD-1) and its ligand PD-L1 have an important function in the maintenance of the immune homeostasis. PD-1 as the inhibitory checkpoint controls damage of the healthy tissues during infection and promotes self-tolerance. High PD-1 expression on the surface of T cells changes its ability to eliminate of cancer and infectious disease. Cervical intraepithelial neoplasia is a premalignant transformation associated with infection with oncogenic types of HPV human papillomavirus, which is a direct precursor to cervical cancer. The relationship of the PD-1/PD-L1 signaling pathway to the development of CIN has not yet been established.

Expression of Psoriasin in Human Papillomavirus-Induced Cervical High-Grade Squamous Intraepithelial Lesions.

Persistent infection with human papillomavirus causes cervical high-grade squamous intraepithelial lesions (HSILs). The role of antimicrobial peptides (AMPs) in premalignant and malignant transformation is not fully understood. In this study, we examined the expression of human β-defensin 1 (HBD-1), HBD-2, HBD-3, LL37, psoriasin, and interleukin 8 (IL-8) in women with HSIL before and 6 months after surgery. Biopsies and secretion samples from the cervical canal were collected from 19 patients with HSIL and 14 healthy controls. The mRNA expression of HBD-1, HBD-2, HBD-3, LL37, psoriasin, and IL-8 was analyzed before and 6 months after surgery excision using reverse transcriptase real time polymerase chain reaction. For protein analyses, ELISA and immunohistochemistry were used for psoriasin and ELISA for IL-8. The mRNA expression of psoriasin was lower in patients before treatment compared with healthy controls (p = .05). After surgery, when the infection was cleared, psoriasin increased on mRNA (p = .04) and protein (p = .03) levels compared with before treatment. Immunostaining for psoriasin after treatment was prominent and localized in the cytoplasm of the epithelial cells. After treatment, IL-8 mRNA was reduced compared with before treatment (p = .05), but not on the protein level. No changes in mRNA expression of the other AMPs analyzed were observed in pretreatment and posttreatment samples. In this study of AMP expression in human papillomavirus-induced HSIL, we observed lower psoriasin levels before surgery compared with after treatment, when both mRNA and protein levels were similar to healthy controls. Interleukin 8, on the other hand, was increased before treatment, indicating an inflammatory response.

Determination of Thymidine Kinase 1 (TK1) Level as a Risk Warning Biomarker to Improve Early Detection of Breast Cancer

Early detection of breast cancer (BC) is a global target to reduce mortality and morbidity also to improve therapeutic and survival outcomes. Currently, mammography is the gold standard in BC diagnosis followed by biopsy when warranted. Thymidine Kinase 1 (TK1) is a proliferative biomarker that succeeded in discovering premalignant transformations of breast cancer before the appearance of any symptoms. This study aimed to provide a non-invasive method to early detect BC by measuring TK1 in sera of women with breast lesions alongside mammography. The study included 271 women divided into five BIRADS categories. Methods and Material: only one blood sample was collected from each woman to detect TK1 concentration, before undergoing mammography and Fine Needle Aspiration Cytology (FNAC) or true cut. Results: TK1 levels were significantly different between BI-RADS categories. It was correlated with clinical stage, histological grade, lymph node metastasis, and vascular invasion. TK1 levels could distinguish between healthy individuals and patients who had breast lesions with a sensitivity and a specificity as follows 91.3 and 87.5%, respectively. Furthermore, this test could discriminate between benign and malignant breast lesions with a sensitivity of 92.5% and a specificity of 91.2%. Conclusion: These findings suggest the determination of TK1 levels as a risk warning biomarker to improve early detection of BC.

Developing Applicable and Cost-Efficient Screens for Early Detection of AML

Introduction The limited improvement of acute myeloid leukemia (AML) patient survival rates over the last few decades reveals that the strategy of targeting AML after diagnosis provides limited success. In many solid cancers, early detection leads to decreased morbidity and improved survival suggesting that a similar approach may benefit AML patients. Recently, we showed that the initial genomic events that drive AML can be detected years before diagnosis and that individuals at increased risk share distinct features that differentiate them from those with benign age related clonal hematopoiesis (ARCH). However, the relative high incidence of ARCH and the low incidence of AML, along with the fact that ARCH is recurrently being driven by mutations that occur in genes associated with AML, impedes complete discrimination. High sequencing costs for the relative large number of mutated genes implicated with the disease further hinders clinical adaptation. Therefore, a better strategic approach that is focussed on the most informative mutations and the development of accurate tools for data mining would better estimate the risk for AML development and create applicable and cost-efficient screens for early detection of AML.Methods We hypothesize that the majority of the power to accurately predict the development of AML can be derived by a minimal number of pre-leukemic hotspot mutations (pLHM) and that improved accuracy of mutation calling algorithms will increase the discrimination between high and low risk cases. We developed a novel approach to differentiate technical errors from true mutations by accounting for local sequence features to derive contextual error signatures. We demonstrate that Error Correction by Signatures Integration (ECSI) detects mutations at a higher sensitivity and specificity when compared to other techniques. 320 blood samples taken years before AML diagnosis and 856 controls were interrogated for the presence of pLHM. This data was used to construct an AML prediction model that was tuned to achieve 100% positive predictive value. To estimate the frequency of individuals at the highest risk for AML development in the general population, we applied the model to a total of 42,838 individuals whose blood was sequenced in four independent ARCH studies.Results ECSI revealed that some pLHM lie within signatures with particularly high error rates. For example, DNMT3A-R882H is defined by the signature G[C>T]G that has the highest sequencing error rate of all. Our analysis suggests that this mutation and others might be over reported when error signatures are not being considered. As compared with other mutation calling techniques, our analysis unbiasedly increased the discrimination between cases and controls by accurately indicating the presence of mutations in pre-AML cases and flagging others that are not significantly above their corresponding signature's error-rate in the controls. We show that a highly specific AML prediction model can be generated by targeting a small number of genomic loci corresponding to changes in only 46 amino-acids previously reported to define AML with poor outcome. Testing our model on the four ARCH datasets pointed to 103 individuals who are at the highest risk for AML development. At least 6 could have been confirmed to developed hematological cancer after sampling.Conclusions The development of a technically simple tool for data mining that is reliable and has a rapid turnaround is a critical step for clinical adaptation of sequencing-based screens for early cancer detection. We present a novel mutation calling method that fulfills such criteria and provides an improved analytical sensitivity and specificity. We found that a minimal group of pLHM carries most of the information needed to accurately predict AML development. Focusing on such a small yet an informative number of mutations provides a ‘two birds, one stone’ strategic approach that enables better discrimination between pre-AML and benign ARCH at reduced costs. Implementing a highly effective and affordable genomic assay is an important step enabling a wider population screen to identify the portion of individuals at the highest risk for pre-malignant cell transformation with minimal false discovery rate. Populations targeted by our prediction model should be prioritized for frequent follow-up, clinical studies and further research to elucidate additional risk-parameters. DisclosuresNo relevant conflicts of interest to declare.

HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk.

Breast cancer (BrCa) is the malignant tumor that most seriously threatens female health; however, the molecular mechanism underlying its progression remains unclear. Here, we found that conditional deletion of hypermethylated in cancer 1 (HIC1) in the mouse mammary gland might contribute to premalignant transformation in the early stage of tumor formation. Moreover, the chemokine (C-X-C motif) ligand 14 (CXCL14) secreted by HIC1-deleted BrCa cells bound to its cognate receptor GPR85 on mammary fibroblasts in the microenvironment and was responsible for activating these fibroblasts via the ERK1/2, Akt, and neddylation pathways, whereas the activated fibroblasts promoted BrCa progression via the induction of epithelial-mesenchymal transition (EMT) by the C-C chemokine ligand 17 (CCL17)/CC chemokine receptor 4 (CCR4) axis. Finally, we confirmed that the HIC1-CXCL14-CCL17 loop was associated with the malignant progression of BrCa. Therefore, the crosstalk between HIC1-deleted BrCa cells and mammary fibroblasts might play a critical role in BrCa development. Exploring the progression of BrCa from the perspective of microenvironment will be beneficial for identifying the potential prognostic markers of breast tumor and providing more effective treatment strategies.

Pre-malignant transformation by senescence evasion is prevented by the PERK and ATF6alpha branches of the Unfolded Protein Response

The incidence of carcinomas highly increases with age. However, the initial steps of the age-related molecular carcinogenic processes remain poorly characterized. We previously showed that normal human epidermal keratinocytes spontaneously and systematically escape from senescence to give rise to preneoplastic emerging cells through a process called post-senescence neoplastic emergence (PSNE). To identify molecular pathways involved in the switch from senescence to pre-transformation, we performed Connectivity Map analyses and DAVID functional annotations followed by hierarchical clustering and multidimensional scaling of the gene expression signature of PSNE cells. We identified endoplasmic reticulum stress related pathways as key regulators of PSNE. Invalidation by RNA interference of the UPR sensors PERK, ATF6α, but not IRE1α, delayed the occurrence of senescence when performed in pre-senescent cells, and increased the PSNE frequency when performed in already senescent cells. Conversely, endoplasmic reticulum stress inducers applied to already senescent cells decreased the frequency of PSNE. In conclusion, these results indicate that the activation of the UPR could protect from the early carcinogenic steps by senescence evasion. This opens new avenues to explore therapeutics that could be useful in decreasing the age-associated tumor incidence.

Association between VEGF-634G>C Gene Polymorphism with Gastric Premalignant Lesions and Serum VEGF Levels in Helicobacter pylori Gastritis Patients.

AIM:To evaluate the association between VEGF-634G>C gene polymorphism with premalignant gastric lesions as well as the level of VEGF.METHODS:This cross-sectional study included patients with H. pylori gastritis at Haji Adam Malik General Hospital, Permata Bunda General Hospital, and Universitas Sumatera Utara Hospital, Medan, Indonesia. Detection of H. pylori infection was made using positive results of 14C-UBT, rapid urease test, and/or immunohistochemistry. Gastric premalignant lesion diagnosis was made when one or more of the following were present: chronic atrophic gastritis, intestinal metaplasia, or dysplasia. Real-time polymerase chain reaction (RT-PCR) was used to examine VEGF-634G>C gene polymorphism. Additionally, serum samples of patients with H. pylori gastritis were obtained to determine the level of circulating VEGF. Data were analysed using SPSS version 22.RESULTS:A total number of 87 patients with H. pylori gastritis were included in this study. Of all participants, 26 patients (29.9%) showed gastric premalignancy. There was a significant association between GG+GC genotype of VEGF-634G>C and gastric premalignant lesions (P = 0.003; OR (CI 95%) = 6.07 (1.88-41.71)). VEGF-634 G>C polymorphism also showed an association with VEGF serum levels (P = 0.005). Patients with the GG+GC genotype would be at risk of 3.16 times to have high VEGF levels compared to CC genotypes.CONCLUSION:VEGF-634G>C polymorphism, in particular, GG+GC genotype was associated with an increased risk of gastric premalignant transformation as well as having high VEGF levels in patients with H.pylori gastritis.

Altered microRNA profiles during early colon adenoma progression in a porcine model of familial adenomatous polyposis.

MicroRNAs are dysregulated in various cancers including colorectal cancer, and are potential useful biomarkers of disease development. We used next generation sequencing to investigate miRNA expression profiles in low- and high-grade intraepithelial dysplastic polyps from pigs carrying a mutation in the adenomatous polyposis coli tumour suppressor (APC1311, orthologous to human APC1309) that model an inherited predisposition to colorectal cancer, familial adenomatous polyposis. We identified several miRNAs and their isomiRs significantly (P < 0.05) differentially expressed between low and high-grade intraepithelial dysplastic polyps. Of these, ssc-let-7e, ssc-miR-98, ssc-miR-146a-5p, ssc-miR-146b, ssc-miR-183 and ssc-miR-196a were expressed at higher level and ssc-miR-126-3p at lower level in high-grade intraepithelial dysplastic polyps. Functional miRNA target analysis revealed significant (P < 0.001) over-representation of cancer-related pathways, including ‘microRNAs in cancer’, ‘proteoglycans in cancer’, ’pathways in cancer’ and ‘colorectal cancer’. This is the first study to reveal miRNAs associated with premalignant transformation of colon polyps.

The Relationship Between COPD Phenotypes and Severity and Lung Cancer Development: Where We Have to Focused?

SESSION TITLE: COPD 1 SESSION TYPE: Original Investigation Poster PRESENTED ON: Wednesday, November 1, 2017 at 01:30 PM - 02:30 PM PURPOSE: Cross-sectional studies show that the prevalence of chronic obstructive pulmonary disease(COPD) is around 50% of those diagnosed with lung cancer(LC),although the prevalence might change depending on the patient′s age sex,and smoking exposure.The aim of our study was to analyze the clinical and histological characteristics of LC in COPD patients. METHODS: From February 2011 to March 2016 we have evaluated 416 patients with COPD and in all patients with COPD the diagnosis was confirmed by recommendations of GOLD guidelines.All patients with COPD were treated by GOLD guidelines and heve received bronchdilators.The patients were older than 35 years(range:35-85 years) and the diagnosis of LC among COPD patients was confirmed by histological or cytological specimens in all cases.All patients were smokers(ex-or current). RESULTS: Among 416 COPD patients in 89(21%) we have found LC,and 70(79%) were not treated with surgery because of the disseminated disease(stages 3B and IV).In COPD patients poor lung function and chronic cough with sputum significantly impact to the development of LC.In 61(69%) patients the LC was associated with advanced stages of the COPD(GOLD groups C and D) and only in 28(31%) patients the LC was associated with early stages of the disease(GOLD groups A and B; P<0.001). In LC-COPD patients chronic cough with sputum and airway disease predominant phenotype of COPD was common finding compared with emphysema predominant phenotype of the disease(P<0.001).Our this finding might be explain with airway tickening and peribronchial fibrosis in such patients which leads to airway obstruction and remodeling in COPD with excess metalloproteinase and growth factors that may potentially cause a premalignant transformation ,epithelial mesenchymal transition.Advanced stages of the COPD most common were associated with squamous cell lung cancer compared with early stages of the disease (58[65%] vs 9[10%];P<0.001)which most common were associated with adenocarcinoma .Small cell lung cancer was more common finding in COPD patients with groups Cand D(P<0.05). CONCLUSIONS: Our study shown that poor lung function and severity of the COPD were strongly risk factors for development of lung cancer.Airway disease predominant phenotype of the COPD and chronic cough with sputum although increased the risk of the lung cancer in COPD patients.The predominance of airway tickening and peribronchial fibrosis in COPD patients increased risk of lung cancer development.Poor lung function and more severe disease most common was associated with squamous cell lung cancer. CLINICAL IMPLICATIONS: Our findings might be useful for respiratory therapist, thoracic oncologists,pulmonary medicine specialists,nurse specialists. DISCLOSURE: The following authors have nothing to disclose: Alizamin Sadigov I ready discuss any information about our investigation

Xanthogranulomatous Cholecystitis: The Needle in the Haystack

Introduction: The rise of laparoscopic cholecystectomy (LC) as the gold standard for management of gallstone disease has exponentially increased the number of sacrificed gallbladders. In this gargantuan pool of specimens, several rare variants of inflammation, infection, premalignant and malignant transformation have become manifest. One of these is the xanthogranulomatous variety of cholecystitis (XGC), an aggressive inflammatory condition known for throwing the surgeon off course from a simple LC to aggressive liver resection by virtue of its malignant mimicry. Methods: We retrospectively analysed 1000 consecutive LCs performed in a tertiary care hospital from January 2010 to December 2016. In the cohort found to have XGC on histopathology, we analysed the gender predilection, age preponderance, associated comorbidities, clinical presentation, radiological features, intraoperative findings, complications and postoperative course. We then compared this with the rest of the source population. Results: Of the 1000 patients that constituted the study population, 35 patients were found to have XGC. We found a male predilection of 5:4, with age ranging from 32 to 76 years, in opposition to the study population that varied from 2 to 85 years. The cohort had a 20% incidence of diabetes mellitus, comparable to the source population. No difference was found in either duration or severity of symptoms in the two groups. Preoperative radiological diagnosis of XGC was not done in any of the cases. Intraoperatively, 14.3% of the patients were found to have Mirizzi's syndrome, which formed the majority of the Mirizzi's in the source population. Empyema of the gall bladder was found in 5 of the patients, with incidence similar to the source population. Operative time in the XGC cohort was significantly higher (80±23 vs 43±7 minutes) which also formed a sizable fraction of the subtotal excisions performed (26.7%). While the incidence of complications was similar in both groups, postoperative stay was significantly longer in the XGC cohort (2.7±1.3 days vs 1.1±0.3 days). Only 3 cases of gall bladder malignancy were found in the entire study population, none of whom exhibited features of XGC. Conclusion: XGC continues to be the bane of the surgeon, confounding findings and causing even seasoned professionals to second guess their clinical decisions. Albeit rare, a working knowledge of the condition is of paramount importance in order to provide the patient the best and most appropriate care.

Potential ultrastructure predicting factors for hepatocellular carcinoma in HCV infected patients

ABSTRACTHepatitis C virus represents one of the rising causes of hepatocellular carcinoma (HCC). Although the early diagnosis of HCC is vital for successful curative treatment, the majority of lesions are diagnosed in an irredeemable phase. This work deals with a comparative ultrastructural study of experimentally gradually induced HCC, surgically resected HCC, and potential premalignant lesions from HCV-infected patients, with the prospect to detect cellular criteria denoting premalignant transformation. Among the main detected pathological changes which are postulated to precede frank HCC: failure of normal hepatocyte regeneration with star shape clonal fragmentation, frequent elucidation of hepatic progenitor cells and Hering canals, hepatocytes of different electron density loaded with small sized rounded monotonous mitochondria, increase junctional complexes bordering bile canaliculi and in between hepatocyte membranes, abundant cellular proteinaceous material with hypertrophied or vesiculated rough endoplasmic reticulum (RER), sequestrated nucleus with proteinaceous granular material or hypertrophied RER, formation of lipolysosomes, large autophagosomes, and micro-vesicular fat deposition.In conclusion, the present work has visualized new hepatocytic division or regenerative process that mimic splitting or clonal fragmentation that occurs in primitive creature. Also, new observations that may be of value or assist in predicting HCC and identifying the appropriate patient for surveillance have been reported. Moreover, it has pointed to the possible malignant potentiality of liver stem/progenitor cells.For reliability, the results can be subjected to cohort longitudinal study.