Late Tumor Research Articles

Late relapsing germ cell tumors with elevated tumor markers

PurposeLate relapsing germ cell tumors (LR-GCT) are considered a rare distinct biologic entity as their clinical presentation and response to treatment is different to early recurrences. While serum tumor markers (AFP and ß-HCG) play an important role at the time of first diagnosis to correctly classify prognosis and treatment of germ cell tumors, they may not have the same significance in a late relapse situation.Patients and methodsThirty-seven patients with LR-GCT with elevated serum tumor markers were identified in our database. Twenty-six patients underwent primary surgical resection of the late relapsing tumor. Eleven patients received salvage chemotherapy and a post-chemotherapy residual tumor resection. Serum tumor markers, histological findings and oncological outcome were analyzed.ResultsIn the histopathological specimen, viable cancer was found in 20 cases (54%) and teratoma was found in 16 cases (43%). In nine cases (24%), a somatic-type malignant transformation was present. In 19 of 37 patients (51.4%), the late relapse specimen presented a histological type of GCT, which was not present in the primary histology. Twenty-two patients (59.5%) were included in follow-up analysis. Mean and median follow-up time was 62.2 and 53 months, respectively. Seventeen patients (77.3%) suffered a relapse or had progressive disease after LR therapy. Five patients (22.7%) have been relapse-free after LR therapy (mean FU 61.6 months). Ten patients died of disease during follow-up (45.5%) and had a mean time from LR to death of 66.4 months. Eleven patients were alive at last follow-up (mean FU 62.2 months). Relapse and survival rate were similar between patients who received primary resection of LR tumor and patients who received salvage chemotherapy followed by surgery.ConclusionPatients with a late relapsing germ cell tumor and elevated markers have a poor prognosis and a high risk for another relapse independent on primary treatment. The histological type and aggressiveness of a late relapsing tumor cannot be predicted with serum tumor marker levels at the time of diagnosis of LR. In up to 54% of cases, primary histology did not coincide with LR histology. Therefore, we propose primary surgical resection of a late relapsing tumor if a complete resection is feasible in order to gain exact histology and tailor further treatment.

Development and validation of a novel survival model for acute myeloid leukemia based on autophagy-related genes.

BackgroundAcute myeloid leukemia (AML) is one of the most common blood cancers, and is characterized by impaired hematopoietic function and bone marrow (BM) failure. Under normal circumstances, autophagy may suppress tumorigenesis, however under the stressful conditions of late stage tumor growth autophagy actually protects tumor cells, so inhibiting autophagy in these cases also inhibits tumor growth and promotes tumor cell death.MethodsAML gene expression profile data and corresponding clinical data were obtained from the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases, from which prognostic-related genes were screened to construct a risk score model through LASSO and univariate and multivariate Cox analyses. Then the model was verified in the TCGA cohort and GEO cohorts. In addition, we also analyzed the relationship between autophagy genes and immune infiltrating cells and therapeutic drugs.ResultsWe built a model containing 10 autophagy-related genes to predict the survival of AML patients by dividing them into high- or low-risk subgroups. The high-risk subgroup was prone to a poorer prognosis in both the training TCGA-LAML cohort and the validation GSE37642 cohort. Univariate and multivariate Cox analysis revealed that the risk score of the autophagy model can be used as an independent prognostic factor. The high-risk subgroup had not only higher fractions of CD4 naïve T cell, NK cell activated, and resting mast cells but also higher expression of immune checkpoint genes CTLA4 and CD274. Last, we screened drug sensitivity between high- and low-risk subgroups.ConclusionThe risk score model based on 10 autophagy-related genes can serve as an effective prognostic predictor for AML patients and may guide for patient stratification for immunotherapies and drugs.

Correlation between preoperative inflammatory biomarkers and postoperative pneumonia or long-term prognosis in patients with esophageal cancer after neoadjuvant therapy

Objective: To examine the correlation between neutrophil-lymphocyte ratio (NLR), lymphocyte-monocyte ratio (LMR) and neutrophil-monocyte ratio (NMR) for postoperative pneumonia or long-term overall survival in patients with esophageal cancer after neoadjuvant therapy. Methods: The clinical data of 137 patients, including 111 males and 26 females, with the age of (M(QR))61(10) years (range: 45 to 75 years), undergoing radical resection of esophageal cancer after neoadjuvant therapy admitted at Department of Thoracic Surgery, West China Hospital from January 2016 to May 2019 were analyzed retrospectively. The blood routine one or two days before surgery and the occurrence of pneumonia after surgery were collected via hospital information system. The absolute count of neutrophils, lymphocytes and monocytes was recorded, to calculate NLR, LMR and NMR. The survival of patients was recorded systematically via follow-up. In the first part, the influencing factors of postoperative inflammation were analyzed, to group the patients into two groups according to the occurrence of postoperative pneumonia. χ2 test, t-test or rank-sum test were conducted for inter-group comparison. In the second part, cutoff values of inflammatory biomarkers were obtained with the receiver operating characteristic (ROC) curve and grouped, with postoperative pneumonia as endpoint criteria. Independent factors correlated with postoperative pneumonia were determined through univariate and multivariate Logistic regression analysis. In the third part, the analysis on prognosis factors was carried on, with the survival as endpoint criteria. Cutoff values of inflammatory biomarkers were obtained with X-Tile software and grouped. The survival analysis was carried on with univariate and multivariate Cox proportional hazards regression model, and the Kaplan-Meier curve was drawn finally. The results of survival analysis were verified by Log-rank test. Results: Median follow-up time was 614 (299) days (range: 382 to 1 612 days). Cutoff values of NLR, LMR, and NMR obtained via the ROC curve were 3.0, 3.9, and 6.2, respectively. According to the multivariate Logistic regression analysis, NLR>3.0 (OR=2.740, 95% CI: 1.221 to 6.152, P=0.015) and LMR>3.9 (OR=0.140, 95% CI: 0.022 to 0.890, P=0.037) were independent prognosis factors for postoperative pneumonia in patients with esophageal cancer after neoadjuvant therapy. Cutoff values of NLR, LMR, and NMR obtained with X-Tile software were 3.3, 4.2, and 7.2, respectively. Through multivariate Cox proportional risk regression analysis, late tumor ypTNM staging (8th AJCC) (HR=2.087, 95% CI:1.079 to 4.038, P=0.029), poor pathologic response (HR=2.251, 95% CI: 1.117 to 4.538, P=0.023), and LMR>4.2 (HR=0.347, 95% CI: 0.127 to 0.946, P=0.039) could be independent prognosis factors for overall survival. Kaplan-Meier survival analysis indicated that the overall survival of patients with LMR ≤4.2 was worse (P=0.002), with the 1-year overall survival rate of 82.9%, and the 1-year overall survival rate of patients with LMR>4.2 was 94.6%. Conclusion: Preoperative LMR ≤3.9 and NLR>3.0 can be considered as independent prognosis factors for postoperative pneumonia, while LMR≤4.2 as one of independent prognosis factors for overall survival.

Evaluation of the Prognostic Value of Solute Carrier Family 34 Member 2 "SLC34A2" in Papillary Thyroid Carcinoma: An Immunohistochemical Study.

Background Papillary thyroid carcinoma (PTC) usually has an indolent clinical course, yet a subset of patients might show an aggressive course. Thus, better stratification of at-risk patients is mandatory for proper management. Solute carrier family 34 member 2 (SLC34A2) is an independent prognostic indicator in several cancers. However, only a few studies have been conducted to evaluate the prognostic value of SLC34A2 in PTC, with none of them assessing its immunohistochemical (IHC) expression in a large cohort of patients with PTC or exploring its possible relationship with tumor progression. Aim of the Study. We aimed to evaluate the IHC expression of SLC34A2 in a large series of PTC patients, correlate its expression with established clinicopathological factors, and find any possible relationship between this marker and patient prognosis. Material and Methods. A total of 476 samples (including 238 samples of PTC and 238 samples of normal thyroid tissue) collected between 2002 and 2005 were extracted from the archives of the Pathology Lab, Ain Shams University Hospitals. IHC analysis was performed using an anti-SLC34A2 antibody. Follow-up data were obtained. Results High SLC34A2 expression significantly correlated with important adverse clinicopathological parameters of PTC—i.e., late tumor stage, positive extrathyroid extension, tumor size > 4 cm, and age ≥ 55 years (p ≤ 0.001 for each). Kaplan–Meier analysis revealed that high SLC34A2 expression significantly correlated with shorter disease-free survival (DFS; p = 0.005), but not with overall survival (p = 0.111). Multivariate analysis showed SLC34A2 to be an independent prognostic factor affecting DFS. Conclusions High SLC34A2 IHC expression correlated with adverse clinicopathological prognostic parameters. Furthermore, SLC34A2 was identified as an independent factor for DFS that could serve to improve risk stratification of PTC patients for better management.

Abstract 2311: Identification of therapeutic targets in HNSCC

Abstract HNSCC is 6th most common malignancy in the world. Despite advances in diagnosis and treatment, the survival rates remain low due in large part to metastatic disease. The underlying biology associated with metastatic disease and poor outcome in HNSCC remains unclear. Importantly, metastatic cells acquire new properties that permit them to invade surrounding tissues and seed metastasis at distant sites. While these acquired properties contribute to aggressiveness of metastatic cancer, they can be exploited to target metastatic cells selectively, sparing toxicity in normal tissues. We used functional genomic technologies to identify new therapeutic targets for advanced disease in HNSCC. These targets were identified by conducting whole genome shRNA screens in matched sets of cell lines derived from primary HNSCC tumors and their respective metastatic sites/recurrences. Since hypoxia is an important attribute of aggressive and therapy resistant subpopulations of HNSCC tumor cells, we also aimed to identify genes that became essential when cells are exposed to hypoxia. To complement the data from the functional screens and to further characterize our cell line collection, we utilized several high through put approaches including mutational analysis, proteomics and gene expression profiling. Moreover, we performed chemical screens using libraries of over 4000 FDA approved drugs with the aim to combine our functional genomic data with the results from the drug screens to discover drug/gene “hit” combinations that would provide a basis for development of novel anti-cancer therapies.While HNSCC derived cell lines represent a valuable tool to study this disease, their biology might not always accurately represent the biology of the tumors they were derived from. Therefore, we looked at the expression of the components of several key pathways that we discovered using the in vitro characterization in a patient material from over HNSCC surgical samples constructed into a TMA. Five of the hits belonged to the Notch signalling pathway and 4 others came from the proteomic analysis as they showed higher levels of surface expression in metastatic cells and/or under hypoxic conditions. Interestingly, higher proportion of patients with late stages of HNSCC belonged to the high Jag2 and Hey1 populations as compared to patients with low stages of HNSCC that mostly expressed low or intermediate levels of these proteins. This data is in line with our observations that Jag2 and Hey1 expression is elevated in cells derived from metastatic sites and is low in cells derived from primary tumors. Proportion of cells with high CD66 and TRAIL proteins was also higher among the late stage tumors suggesting that these biomarkers are of interest for further investigation. We discovered several molecular pathways that are important in metastatic/hypoxic HNSCC. We are investigating our ability to target these pathways using FDA approved drugs in order to achieve durable cures. Citation Format: Maria Kondratyev, Aleksandra Pesic, Anna Dvorkin-Sheva, Troy Ketela, Natalie Stickle, Laurie Aiiles, Reidar Grenman, Marianne Koritzinsky, Brad Wouters. Identification of therapeutic targets in HNSCC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2311.

Abstract 463: Features consistent with a small-cell neuroendocrine like (SC/NE-L) transformation can be readily detected and quantified in circulating tumor cells (CTCs)

Abstract Background: The increasing availability and earlier use of life prolonging drugs targeting the androgen receptor signaling axis (ARSI) has resulted in an increase in the frequency of late state tumors with “small cell/neuroendocrine like (SC/NE-L) phenotypes” similar to small-cell lung cancer (SCLC). These tumors are typically heterogenous wherein the small-cell component represents only a portion of CTC's present. Definitive pathologic criteria to diagnose small-cell neuroendocrine transformation are lacking, and the eligibility criteria across trials in this space inconsistent, limiting the ability to relate outcomes between studies. We hypothesize that an analytically validated assay for a rigorously defined “small-cell CTC” phenotype would clarify the identification and diagnosis of SC/NE-L tumors. Methods: Using the WHO guidelines for small-cell diagnosis in tissue as reference, we defined an equivalent set of single-cell criteria for defining a CTC with small cell or neuroendocrine like (SC/NE) histology that include: a circulating CD45- cell with a high nuclear to cytoplasm area ratio, small size, high circularity, lack of detectable nucleoli, salt and pepper chromatin, SC/NE or tumor-related protein expression (DLL3, CD56, CK etc.), and other features associated with lineage plasticity (i.e., CK speckling). Clinical feasibility of feature extraction was explored on CTCs detected using the Epic Sciences platform in the blood of patients with small-cell lung and late state prostate cancers. Results: Using the Epic Sciences platform, morphologic features associated with small-cell or neuroendocrine transformation can be readily identified and quantified automatically in CTCs in a liquid biopsy from patients with small-cell lung and prostate cancers. Conclusions: Identifying small-cell to neuroendocrine like (SC/NE-L) CTC subtypes may aid in establishing that lineage transformation to a SC/NE-L phenotype has occurred where invasive single site tissue biopsies may not be informative or feasible. Citation Format: Howard I. Scher, Adam Jendrisak, Cole Gilbertson, Audrey Gill, Ethan Barnett, Anuradha Gopalan, Niamh Keegan, Samir Zaidi, Hannah Benoliel, Carbone Emily, Eva Wang, James Lu, Alisa Tubbs, Amanda Anderson, Joshua Jones, Joseph D. Schonhoft, Rick Wenstrup. Features consistent with a small-cell neuroendocrine like (SC/NE-L) transformation can be readily detected and quantified in circulating tumor cells (CTCs) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 463.

Abstract 1718: Tumor eradicated by an effective in situ vaccine based on a TLR agonist and an agonistic anti-OX40 antibody

Abstract As cancers are antigenic, numerous efforts have been devoted to harnessing the exquisite specificity of the immune system to destruct tumors over the past two decades. In this study, we have employed a practical approach called in situ vaccination without the need to previously identify and isolate antigen targets. Intratumoral injection of a TLR7 ligand R837 and an agonistic anti-OX40 antibody showed impressive synergistic anti-tumor effects. R837 upregulated the expression of OX40 on CD4+ T cells in the treated tumor. Then, anti-OX40 antibody triggered systemic tumor-specific T cell responses, with one-fold increase of effector memory T cells properties and stronger toxicity of lymphocytes in spleen. Remarkably, both the injected tumor and the noninjected tumor in the distant lesion of the mice were eliminated after vaccination. IL-1α is a pro-inflammatory cytokine, however IL-10 is an important anti-inflammatory cytokine that plays a central role as a negative immune regulator. In the tumor microenvironment, we found that the level of IL-1α increased and IL-10 decreased after vaccination. From RNA-seq analysis, many immune-related pathways in the tumor microenvironment were activated after vaccination, such as TNF signaling pathway, NOD-like receptor signaling pathway, C-type lectin receptor signaling pathway, Toll-like receptor signaling pathway and so on. This in situ vaccine based on two low doses of immunoomdulators caused little harm to normal organs and provided long-term protection against the same syngeneic tumor rechallenge. It could be applied to various types of late stage solid tumors and worthy of further clinical research. Citation Format: Yanhong Chu, Qin Liu, Baorui Liu. Tumor eradicated by an effective in situ vaccine based on a TLR agonist and an agonistic anti-OX40 antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1718.

Abstract 2300: Overcoming TKI-resistance in NSCLC using SphK2 inhibitors

Abstract In 2020, 228,820 lung cancer cases and 135,720 deaths have been anticipated. NSCLC is currently treated with Tyrosine Kinase Inhibitors (TKIs) such as Erolitinib (ER; second generation) and Osimertinib (OR; third generation). However, patients often develop resistance to these drugs overtime. This acquired resistance to TKIs is due to a secondary T790M mutation in the kinase domain which could be responsible for inducing Epithelial Mesenchymal Transition (EMT). EMT is a vital process in development of metastasis and occurs, by downregulation of E-cadherin, and upregulation of proteins like N-cadherin and vimentin. Therefore, it is imperative to find other molecularly targeted therapies to combat TKI resistance. Sphingosine kinase (SphK) is a type of lipid kinase that phosphorylates sphingosine to sphingosine-phosphates. There are two functional SphK isoforms (SphK1 and SphK2) that have been identified and characterized in mammalian cells. As a member of the SphK family, SphK2 activity can be stimulated by a series of cytokines such as EGF and VEGF, and acts as an oncogenic enzyme in tumor cells. Overexpression of SphK2 has been demonstrated in many cancer types and is closely related with cancer progression and the poor survival of patients. Currently minimal studies have been done on lung cancer and SphK2. This study investigated the roles of SphK2 in OR-resistant wild-type EGFR H2170 and H358, EGFR-mutated and TKI-sensitive H3255 cells (L858R mutation), and EGFR mutated TKI-resistant H1975 cells (L858R and T790M mutation). The modulation of EMT biomarkers was determined by qPCR. Key EMT regulators such as vimentin and N-cadherin were upregulated by 2.5 and 1.5-fold, respectively, while E-cadherin was downregulated by 1.5-fold. Immunofluorescence studies with Vimentin showed that H1975OR cells were elongated and have a mesenchymal phenotype compared to the H3255OR and their respective parental counterparts. MTT studies done using inhibitors of SphK2 in NSCLC OR-resistant cells showed a 3.0-fold decrease in cell viability in comparison to a 2.0-fold decrease in their parental counterparts. In wound closure assays at 24 hours the wound closure was 1.5-fold less in cells treated with SphK2 inhibitor as compared to parental cells treated with SphK2 inhibitors. Currently studies are being done to determine expression of SphK2 in late and early stage NSCLC tumors. In conclusion SphK2 was found to be a crucial positive regulator of different modes of cell migration and a key biomarker in combating TKI-resistance in NSCLC. Citation Format: Nabiha Haleema Khan, Neelu Puri. Overcoming TKI-resistance in NSCLC using SphK2 inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2300.

Abstract 3169: The therapeutically inflamed tumor microenvironment drives melanoma progression

Abstract Even the most therapy responsive melanomas eventually evade immune checkpoint inhibition. Interestingly, initial responses seem to benefit from localized inflammation, but this can become prohibitive when treating advanced disease. Subsequently, there is an urgent need to elucidate how inflammation impacts the tumor microenvironment (TME). Here, inhibition of EGFR and Ras/Raf/MEK is shown to drive the release of a pro-inflammatory secretome that activates the TME to promote progression. This secretome was enriched with immune recruiting cytokines and members of the transforming growth factor beta (TGFβ) superfamily. Both healthy and cancer associated fibroblasts (CAFs) were hyperactivated in response to the inflamed secretome with induction of the inflammatory marker, alpha smooth muscle actin (αSMA). Pharmacologic inhibition of TGFβ receptor type I (TGFβRI) using SB431542 or clinically relevant TEW-7197, diminished CAF αSMA phenotype, disrupted TGFβ and phospo-SMAD2/3 signaling, and blunted CAF expression of tumor promoting extracellular matrix and adhesion molecules. Invasion of melanoma spheroids was enhanced ~2.5-fold in CAF coculture transwells. However, treatment with MEK inhibitor AZD6244 and TEW-7197 only limited invasion of mixed melanoma and CAF multi-cell spheroids in vitro. Prompted by this, the extracellular matrix of patient-derived and melanoma xenograft tumors was analyzed for inflammatory protein expression. Micrometastatic foci were flanked by peri-tumor αSMA+ CAFs and enriched for inflammatory Tenascin-C (TNC) at the invasive front. The analysis of human melanoma tissue microarrays corroborated the experimentally observed expression of αSMA CAFs and TNC which statistically increase with malignant tumor grade. Conversely, the anti-invasive Decorin (DCN), decreased in malignant melanoma while collagen-1 increased during late stage tumor fibrosis. These findings prompted the use of ex vivo human skin organ cultures (SOCs) to define how cell matrix impacted melanoma. Addition of DCN to the SOC eliminated melanoma vertical invasion whereas pathological TNC promoted progression. Thus, a BRAFV600E resistant patient-derived metastatic cell line was used to establish intrasplenic xenografts that were treated after one week using AZD6244 and TEW-7197. Both monotherapy and combination therapy were well tolerated and resulted in reduced splenic tumor burden, αSMA staining, and halted metastasis to the liver. Citation Format: Andrew M. Bradshaw, Erica Kuo, Jelena Grahovac, Kyle Sylakowski, Cindy Sander, Howard Edington, John M. Kirkwood, Alan Wells. The therapeutically inflamed tumor microenvironment drives melanoma progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3169.

Abstract 2523: Novel genomic characterization in late stage colorectal cancer

Abstract Introduction: Colorectal cancer patients with high-level microsatellite instability benefit more from immune checkpoint inhibitor therapy, but they only account for around 15% of all colorectal cancer patients. The remaining patients are still lack of effective therapeutic biomarkers. For patients in late stage without optimal treatment, it is essential to find novel biomarkers which can be used to select appropriate therapies and therefore improve patients' overall survival. Methods: We used whole-exome sequencing to analyze 114 colorectal cancer samples. The samples included 86 primary late stage tumors and 28 metastatic tumors from liver, lung and other organs. The primary samples were further divided into different groups based on their tumor location (sideness) and microsatellite status (MS) since these indications are of great significance in clinical presentation and treatment. We then assessed somatic genomic lesions, copy number variations, germline gene alterations and mutational signatures in the above three pairs of comparisons respectively (primary vs. metastatic, right- vs. left-sided, MS instable vs. stable). Results: There were significant differences between the analyzed pairs. For the primary and metastatic samples, the most prominent difference was the copy number variation which happened in 25 genes in the metastatic samples and only one gene in the primary samples (gain of IL12) (p = 0.036). For the right- and left-sided comparison, SNP/Indel mutations were the most prominent. They happened more frequently in the right-sided than the left-sided tumors and those are rare genes in colorectal cancer, including BCL11A (p = 0.018), DICER1 (p = 0.038), NR4A3 (p = 0.004), NIN (p = 0.008) and FBXW7 (p = 0.019). For the MSI-MSS comparison, tumor mutation burden (TMB) stands out high in the MSI group compared to the MSS group (p < 0.0001). In addition, dozens of significantly mutated genes were observed in the MSI samples (p < 0.0001), such as KMT2B, ACVR2A, KMT2D, ASXL1, TCF7L2 and etc. However, TP53 occurred mainly in the MSS patients (p = 0.0007). Conclusion: Our study enables a deeper understanding of the genomic characterization of colorectal cancer from an integrated view. Copy number variations, gene mutations, and TMB were found to be the most significant differences between the comparison groups of primary vs. metastatic samples, left vs. right sided samples, and MSI vs. MSS samples respectively. Genomic differences between metastatic and primary samples may be potential therapeutic targets for metastatic colorectal cancer. Citation Format: Jianxia Li, Qianchao Wu, Jianwei Zhang, Huabin Hu, Yue Cai, Jiayu Lin, Jun Liu, Yanhong Deng. Novel genomic characterization in late stage colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2523.

Abstract 2155: BRCAness in HGSC cancer initiation

Abstract Introduction: The luteal phase of BRCA mutation carrier tissues has distinct profiles of differentially expressed genes compared to wild-type fimbria. High Grade Serous Ovarian Carcinoma (HGSOC) is one of the most lethal gynecological diseases and presents at late tumor stage. Studies focusing on the early events of the disease have implicated the fimbriated end of the fallopian tube as the high-risk zone for cancer development. In this study, we aim to identify BRCA transcriptomic signatures in the fimbria of patients with a BRCA mutation and compare the luteal and follicular phases to capture the effect of the extrinsic milieu in response to stressors. Experimental Procedures: Laser capture microdissection (LCM) was performed on 68 formalin fixed paraffin embedded (FFPE) fallopian tube samples based on BRCA mutation status and ovulatory cycle: Normal-Follicular, Normal-Luteal, BRCA-Follicular, BRCA-Luteal. Nine HGSC samples were derived from patients with a mutation in BRCA1. Six-Twelve, 10um FFPE sections were cut and stained with hematoxylin prior to LCM. RNA was extracted using the Roche High Pure FFPE Micro Kit and samples and were processed using Illumina Tru-Seq Stranded Total RNA Kit with RiboGold ready and sequenced on the Illumina Hi-seq 2000 V3. Bioinformatic analysis performed on multiple comparisons highlighted differentially expressed genes. Gene Set Enrichment Analysis (GSEA) performed on subgroups highlighted pathways of interest for each comparison. Genes highly expressed or strongly suppressed were shortlisted for further analysis. Western blot analysis performed on wildtype patients (WT), BRCA1 mutation and BRCA2 mutation showed protein expression of selected genes. Results: The fimbriae compared to ampullae have more stem-like features, have up-regulated antioxidant/xenobiotic genes, and have more inflammatory cells intercalated within the epithelia. The fimbria of FTE-BRCA have distinctly up-regulated genes involved in DDR (DNA damage response) which are exacerbated in the luteal phase of the ovulatory cycle. BRCA1/2 luteal versus follicular phases are involved in endoplasmic reticulum stress, migration, MAPKK. Whereas decreased differentially expressed genes are involved with G2/M checkpoint, DNA replication and ER-associated degradation pathway. Post-ovulatory phase of the ovarian cycle drives inflammation, differentiation and DNA damage-repair in FTE cells. Furthermore, GSEA analysis of the luteal and follicular phases showed positive enrichment of the inflammatory response pathway and negative enrichment of oxidative phosphorylation in the luteal phase. Conclusions: Our results highlight distinct pathways related to repair and inflammation for the BRCA1 mutation carriers and luteal phase of the fimbria. These results are consistent with previous examination of the fallopian tube and provide insight into targetable genes associated with the development of HGSC. Citation Format: Ramlogan Sowamber, Iru Paudel, Alex Sanchez, Alexandra Diaz, Patricia Shaw, Matthew Schlumbrecht, Sophia George. BRCAness in HGSC cancer initiation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2155.

Audit of Lymph Node Sampling in Colorectal Carcinoma Resections-implication for Prognosis and Management

Aims: To review lymph node yield in colorectal carcinoma (CRC) resections and its associated factors in a Nigerian Teaching Hospital practice.
 Study design: This was a retrospective cross-sectional study.
 Place and duration of study: Department of Pathology, University College Hospital Ibadan Nigeria and colectomies from January 2014 to December 2018 were reviewed.
 Methodology: Surgical Pathology reports of CRC resections at the University College Hospital Ibadan over 5 years (2014-2018) were reviewed. Colectomy lengths, tumour location (colon/rectum), tumour size, comment on presence of lymph node (yes/no), lymph node count, presence of attached mesentery (yes/no), histological subtype, tumour grade, presence of tumour-positive node and count, and pT stage were documented. Fisher’s Exact test was employed to test the effects of these variables on presence of lymph node and tumour-positive node at histology using SPSS 20. Significance level was set at P < .05.
 Results: Of 66 histology reports retrieved, 62 (93.9%) had comments on search for lymph nodes and attached mesentery was documented in 25 (37.9%). The median colectomy length and tumour size were 25cm (6cm-152cm) and 6.75cm (3-30cm) respectively. Lymph nodes were present in 52 (78.8%) specimens; 28 (53.8%) of these had tumour-positive lymph nodes. Adenocarcinoma NOS was the commonest histological subtype 53 (80.3%), mucinous carcinoma 12 (18.2%) and signet ring carcinoma 1 (1.5%). Eighteen, 9 and 1of adenocarcinoma NOS, mucinous carcinoma, and signet ring carcinoma respectively had tumour-positive lymph nodes. Finding of lymph node was significantly associated with comment on search for lymph node (p < .01) while finding tumour-positive nodes was associated with histological subtype, presence of mesentery, late tumour stage and lymph node count ≥ 12 (p < .05).
 Conclusion: If lymph nodes were present, more than likely there will be metastatic involvement. To increase Lymph node yield in CRC resections, submission of mesentery and search for lymph nodes is indicated. When nodes are absent, a mention is required for practice audit. It is imperative to include both clinical and grossing notes for lymph nodes to certify and guide precise staging of the cancer.

High Levels of Serum IgG for Opisthorchis viverrini and CD44 Expression Predict Worse Prognosis for Cholangiocarcinoma Patients after Curative Resection.

BackgroundOpisthorchis viverrini (OV)-associated cholangiocarcinoma (CCA) has a high immune response with chronic inflammation and oxidative stress. CD44 and Nestin, two cancer stem cell (CSC) markers, play major roles in cancer cell survival. Effects of immune response and expression CSC markers on survival of patients with CCA remain unclear.ObjectiveTo investigate the effects of level of OV IgG together with CSC marker expression and also the combination of these markers on survival of CCA patients after curative resection.MethodsAll serum specimens from CCA patients who underwent curative surgery from 2005 to 2015 were examined for IgG for OV antigen by ELISA. Tissue specimens were studied for CD44 and Nestin expression. Survival analysis by Cox proportional hazard model was used for estimating hazard ratio (HR) with a 95% confidence interval (CI).ResultsIn this study, 122 (69.3%) of 176 were positive for OV IgG, and 35 (19.9%) were considered to have high-positive OV IgG. CD44s positive expression was found in 54 (40%), CD44v6 high expression in 96 (69.6%), CD44v8-10 high expression in 87 (63.5%) and Nestin high expression in 21 (16.1%). Multivariate survival analysis found that high-positive OV IgG and late stage tumor were independent prognostic factors with the adjusted HR of 2.24 (95% CI 1.27–3.93) and 2.78 (95% CI 1.46–5.29), respectively. Subgroup analysis in early and late stage CCA showed that a combined positive OV IgG and CD44s expression with the high expression of CD44v8-10 had the significantly poorest prognosis with HR of 3.75 (95% CI 1.61–8.72) and HR of 1.76 (95% CI 1.02–3.03), respectively.ConclusionA high level of OV IgG as well as a high level of CSC markers resulted in an aggressive CCA. OV IgG level together with CSC markers can be used as the prognostic markers for CCA patients’ survival. The study of the CD44 pathway is promising for adjuvant treatment.

Congenital tumor of the oral cavity – epulis neonatorum. Case report

A newborn baby with congenital epulis can be a startling sight for both parents and medics involved in the care of a neonate. These benign neonatal tumours can be exceptionally big, occupying a significant portion of the oral cavity and posing a risk of airway obstruction and significantly hindering feeding. Prenatal diagnosis remains difficult due to late tumour development and non-specific findings (ultrasound/MRI). The treatment of choice remains surgical excision of the tumour. In our report, we present a description of two cases of neonates operated at the Department of Pediatric Otorhinolaryngology of the Medical University of Warsaw, at the turn of 2020/2021. In both cases the diagnosis was made at birth and histopathological examination confirmed the diagnosis of congenital granular cell tumour (CGCT). The growths were excised under general anaesthesia with a good postoperative outcome, which allowed oral feeding on the second postoperative day.

Comparison of long-term survivals following hepatectomy for hepatocellular carcinoma according to the time-point of recurrence and treatment modalities for recurrent disease

Abstract Objective Disease recurrence following curatively intended hepatectomy for hepatocellular carcinoma (HCC) limits oncologic outcome. Based on the extent, location and time-point of tumor recurrence, different therapeutic modalities are available to treat recurrent HCC. Therefore, our aim was to investigate the role of these treatments and the time-point of recurrence on long-term survival. Methods Clinicopathological data of patients, who underwent hepatectomy for HCC at a major hepatobiliary center in Switzerland between 2012 and 2019, were assessed. Patients suffering tumor recurrence were stratified according to the treatment modalities for recurrent HCC including surgical treatment (repeat hepatectomy or liver transplantation), interventional treatment, and conservative treatment (chemotherapy or best supportive care). Groups were compared regarding to overall survival (OS). Additionally, long-term outcomes were compared between patients with early (≤ 12 months) and late (> 12 months) tumor recurrence. Results During the study period, 159 patients underwent hepatectomy for HCC. Median follow-up time was 53 months. After a median time of seven (1-64) months, 74 patients were diagnosed with tumor recurrence (47 %). The majority of patients developed early recurrence (n = 49) and 58 patients had intrahepatic recurrence only. Treatment options were re-resection, liver transplantation, interventional methods, and palliative therapy in 5, 15, 23, and 31 patients, respectively. Surgical treatment was significantly associated with improved OS compared to interventional and conservative treatment (5-year OS: 84% vs. 39% vs. 30%, p < 0.0001). OS was significantly better among patients with late recurrence compared to patients with early recurrence, irrespective of the treatment modality used for the recurrent disease (5-year OS: 70% vs. 38%, p = 0.008). Conclusion Repeat hepatectomy or liver transplantation for recurrent HCC following hepatectomy is associated with better long-term survival compared to interventional or conservative therapies, especially for patients with late tumor recurrence. Patients with intrahepatic HCC recurrence should be evaluated according to the extent of tumor burden, liver function, and functional status to identify the best candidates for a surgical treatment.

The process of urinary bladder cancer recurrence in 33 patients with 113 tumors, analyzed by changes in IHC features of 80 pairs of subsequent tumors.

e16507 Background: The aim was to define IHC changes between the two subsequent urinary bladder cancers (UBC). Methods: IHC data on EGFR, HER2, HER3, Ki-67, MLH1, MSH2, MSH6 and PMS2 in 113 UHC from 24 male and 9 female patients (1 to six recurrences) were used. Except for the Ki-67 value, other markers were stratified: “0” for no positive cells; “1” < = 10% positive cells; “2” 1%-30% positive cells; and “3” 31%-100% positive cells. Data of consecutive tumors were paired in 80 processes of recurrence (PoR). Changes between the latter and the former tumor were calculated: in +/- % for Ki-67 values, and as integer sums of absolute changes in expression for HER markers and for Lynch markers. EM clustering was applied for recognition of relevant IHC changes. Results: Three aspects were tested: the speed of recurrence, was it the first, or later recurrence and whether PoRs depended on the total number of tumors in that patient. Early and late PoRs clustered along the Lynch score, while the intermediate clustered along the delta Ki-67 value. The first PoRs clustered along the HER score and all subsequent PoRs depended on the Lynch score. Conclusions: Even in this limited group of patients Lynch and HER markers showed complex differences between early and late recurrent tumors. The speed of the recurrence and changes of IHC features depended mainly on the rate of change in Lynch markers, suggesting that they should be tested as predictors of UBC recurrence.[Table: see text]

The Impact of IDH1 Mutation and MGMT Promoter Methylation on Recurrence-Free Interval in Glioblastoma Patients Treated With Radiotherapy and Chemotherapeutic Agents.

The aim of this study is to investigate the relationship between isocitrate dehydrogenase-1 (IDH1) mutation and O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation with recurrence-free interval in glioblastoma patients treated with chemoradiotherapies. Clinical data were collected from 82 patients with totally resected glioblastoma and treated with adjuvant therapies from 2014 to 2019. IDH1 mutation was assessed by immunohistochemistry and MGMT promoter methylation was assessed by different sequencing methods. IDH1 mutation was present in 32 cases and 50 cases were IDH1 wildtype; 54 and 28 patients had unmethylated and methylated MGMT promoter, respectively, Of the 82 patients, 62 patients received chemoradiotherapy while 20 patients only received radiation. Approximately, 61% of patients had a tumor recurrence after 1 year, and 39% showed a recurrence before 1 year of treatment. There was no significant relationship between IDH1 mutation and MGMT promoter methylation (p-value = 0.972). Patients with IDH1 mutation and their age <50 years showed a significant difference in recurrence-free interval (p-value = 0.014). Difference in recurrence-free interval was also statistically observed in patients with unmethylated MGMT promoter and treated with chemoradiotherapies (p-value = 0.031), by which they showed a late tumor recurrence (p-value = 0.016). This revealed that IDH1 mutation and MGMT methylation are independent prognostic factors in glioblastoma. Although IDH1-mutant glioblastomas showed late tumor recurrence in patients less than 50 years old, the type of treatment modalities may not show additional beneficial outcome. Patients with unmethylated MGMT and IDH1 mutation, treated with different chemoradiotherapies, showed a late tumor recurrence.

High Transaldolase 1 expression predicts poor survival of patients with upper tract urothelial carcinoma

Upper tract urothelial carcinoma (UTUC) is a rare tumor with an incidence that varies greatly between Eastern and Western countries. Transaldolase 1 (TALDO1) is a rate-limiting enzyme of the pentose phosphate pathway. In humans, aberrant TALDO1 activity has been implicated in various autoimmune diseases and malignancies; however, the function of TALDO1 in UTUC has not been previously investigated. Here we evaluated the clinical significance of TALDO1 expression in 115 paraffin-embedded tumor samples from patients with UTUC using immunohistochemistry. Our results demonstrated that there was an association between high TALDO1 expression and advanced stage (P = 0.011), tumor size (P = 0.005), tumor location (P = 0.047), distant metastases (P = 0.023), local recurrence (P = 0.002), and cancer death (P = 0.003). Using univariate and multivariate analyses, we found that chemotherapy was an independent factor for bladder recurrence-free survival. Late stage (III/IV) and high TALDO1 expression were independent prognostic factors for progression-free and cancer-specific survival. In summary, increased TALDO1 expression in UTUC was significantly correlated with late stage, tumor size, tumor location, distant metastases, local recurrence, and cancer death. Therefore, high TALDO1 expression could be a predictor of poor survival in patients with UTUC. Further studies are necessary to investigate the role of TALDO1 in UTUC development.

Plasma biomarkers for prediction of early tumor recurrence after resection of pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is a disease with a very unfavorable prognosis. Surgical resection represents the only potentially curative treatment option, but recurrence after complete resection is almost certain. In an exploratory attempt we here aimed at identifying preoperative plasma protein biomarkers with the potential to predict early recurrence after resection of PDAC. Peripheral blood samples from 14 PDAC patients divided into three groups according to their time to tumor recurrence after curatively intended resection (early: < 6 months, medium: 6–12 months, late: > 12 months) underwent targeted proteome analysis. Proteins most strongly discriminating early and late recurrence were then examined in a number of established PDAC cell lines and their culture supernatants. Finally, PDAC organoid lines from primary tumors of patients with early and late recurrence were analyzed for confirmation and validation of results. In total, 23 proteins showed differential abundance in perioperative plasma from PDAC patients with early recurrence when compared to patients with late recurrence. Following confirmation of expression on a transcriptional and translational level in PDAC cell lines we further focused on three upregulated (MAEA, NT5E, AZU1) and two downregulated proteins (ATP6AP2, MICA). Increased expression of NT5E was confirmed in a subset of PDAC organoid cultures from tumors with early recurrence. MICA expression was heterogeneous and ATP6AP2 levels were very similar in both organoids from early and late recurrent tumors. Most strikingly, we observed high MAEA expression in all tested PDAC (n = 7) compared to a non-cancer ductal organoid line. MAEA also demonstrated potential to discriminate early recurrence from late recurrence PDAC organoids. Our study suggests that identification of plasma protein biomarkers released by tumor cells may be feasible and of value to predict the clinical course of patients. Prediction of recurrence dynamics would help to stratify up-front resectable PDAC patients for neoadjuvant chemotherapy approaches in an individualized fashion. Here, MAEA and NT5E were the most promising candidates for further evaluation.

Effects of Laparoscopic Radical Gastrectomy on Peritoneal Micrometastases of Gastric Cancer.

Some reports asserted that the stimulation of ultrasonic scalpel and the persistent state of carbon dioxide (CO2) pneumoperitoneum in laparoscopic surgery may affect the adhesion and invasion of gastric cancer (GC) cells. This study aimed to reveal the effects of laparoscopic radical gastrectomy on peritoneal micrometastases (PM) of GC. Fifty-three patients who underwent laparoscopic radical gastrectomy for GC were enrolled in the study. The expressions of carcinoembryonic antigen (CEA) mRNA and dopa decarboxylase (DDC) mRNA in peritoneal lavage fluid were detected by reverse transcription-polymerase chain reaction. The positive rates of CEA mRNA and DDC mRNA in preoperative peritoneal lavage fluid (pre-CEA, pre-DDC) were compared with those in postoperative lavage fluid (post-CEA, post-DDC). The correlation between the expressions of pre-CEA and pre-DDC and clinicopathologic factors and disease-free survival was analyzed. There was no significant difference in the positive rates of pre-CEA and pre-DDC compared with those of post-CEA and post-DDC (all P>0.05). The positive rates of pre-CEA and pre-DDC increased with the increase of TNM stage, deepening of invasion, lymph node metastasis, and serosal invasion (all P<0.05), but had no correlation with tumor location, size, degree of differentiation, nerve invasion, and vascular invasion (all P>0.05). The disease-free survival in the combined positive patients was lower than that in the negative patients. Laparoscopic radical gastrectomy for GC is safe and feasible, without increasing the risk of PM. The PM of GC may be associated with late tumor stage, deep infiltration, lymph node metastasis, and serosal invasion.