Prognostic Indicator Of Poor Survival Research Articles

A novel approach ofdifferentiation of adenoma and carcinoma in lung cancer based on biogenic in situ synthesis of gold nanostructures on various oligonucleotide motifs.

Aunique approach is introducedfor constructing gold nanocrystals (AuNCs) with RNA motif-directed morphologies in a sequence-independent manner and itsapplications in theclinical areaare described. By using this method, a label-free LSPR-based detection method for the SOX2OT transcript, long non-coding RNAs (lncRNAs), which is a prognostic indicator of poor survival in lung cancer patientsis presented. For the first time, we examined how the structural changes of RNA after the heteroduplex formation with a specific DNA probe can change the morphology and LSPR band of AuNCs. Using this method, is was possible to differentiate lung squamous cell carcinoma from adenocarcinoma samples without a need for a prior amplification of the target lncRNA. The approach of using specific DNA probe enables the in situ synthesis of nanocrystals in a different way and expands this method for future translational medicine, particularly detection of specific RNA.

SNAI1: a key modulator of survival in lung squamous cell carcinoma and its association with metastasis

BackgroundSnail family zinc finger 1 (SNAI1) has been implicated in cancer progression and prognosis across various malignancies. This study aims to elucidate the prognostic significance of SNAI1 expression in Lung Squamous Cell Carcinoma (LUSC) using data from The Cancer Genome Atlas (TCGA) database.MethodsSNAI1 expression levels in LUSC patients were stratified using X-tile software to establish optimal cut-off values. Kaplan-Meier survival analysis was performed to assess the impact of SNAI1 expression on overall survival (OS). Univariate and multivariate Cox regression analyses were conducted to evaluate the prognostic value of SNAI1, considering clinical parameters such as age, clinical stage, and TNM classification. Additionally, we explored the interaction between SNAI1 expression and metastatic status, and performed Gene Set Enrichment Analysis (GSEA) to investigate associated cellular pathways. Correlations between SNAI1 and immune checkpoint molecules were also examined.ResultsKaplan-Meier analysis revealed significant differences in OS among high, medium, and low SNAI1 expression groups (p < 0.001), with median survival times of 1.6, 3.0, and 5.8 years, respectively. Dichotomizing patients into high and low SNAI1 expression groups confirmed that high SNAI1 expression was associated with significantly poorer OS (p < 0.001). SNAI1 remained an independent prognostic factor in multivariate analysis. High SNAI1 expression correlated with poorer survival outcomes regardless of metastatic status, and the combination of high SNAI1 expression and metastasis resulted in the poorest survival. GSEA identified significant associations between SNAI1 and inflammatory, immune response pathways. Positive correlations were observed between SNAI1 and key immune checkpoint molecules, suggesting an interplay with immune checkpoint mechanisms.ConclusionsHigh SNAI1 expression is a robust prognostic indicator of poor survival in LUSC, independent of other clinical factors. Its association with immune checkpoint molecules highlights its potential as a therapeutic target. These findings underscore the prognostic and therapeutic relevance of SNAI1 in LUSC and possibly other cancers. Further research is warranted to explore targeted therapies against SNAI1.

Comprehensive Analysis of Metastatic Prostate Cancer: Real-World Data From the Middle East.

Metastatic prostate cancer (Pca) is a complex disease with diverse clinical characteristics and outcomes across the geographical distribution. Herein, we present a series of patients from the Middle East, aiming at identifying disease outcomes and prognostic factors specific to this regional context. This is a retrospective study of patients with metastatic Pca, diagnosed at King Hussein Cancer Center, Jordan, between 2006 and 2018. Survival was estimated using the Kaplan-Meier method and compared using the log-rank test. Factors that significantly affected overall survival (OS) in the univariable analysis were examined in a multivariable Cox regression analysis. A total of 188 patients with metastatic Pca were included in this analysis, of whom 168 (89%) had de novo metastatic disease. The median age at diagnosis was 68 years, 144 (77%) had bone metastasis, 32 (17%) had visceral metastasis, and 126 (67%) had high-volume disease. At a median follow-up of 67 months, the median OS was 44.3 months. The following factors predicted inferior OS in univariable analysis: smoking, normal BMI, high-volume disease, high alkaline phosphatase (ALP), previous local therapy for prostate, and orchiectomy versus medical androgen deprivation therapy (ADT). On multivariable analysis, high-volume disease (hazard ratio [HR], 1.92 [95% CI, 1.17 to 3.13]; P = .0094), high ALP (HR, 2.136 [95% CI, 1.38 to 3.31]; P < .001), and orchiectomy (HR, 2.40 [95% CI, 1.51 to 3.82]; P < .001) emerged as independent factors for inferior OS. Metastatic Pca outcomes in our population closely align with the global benchmark. High volume status, elevated ALP, and performance of surgical as opposed to medical ADT emerge as prognostic indicators of poor survival.

Abstract 7649: Inflammatory biomarkers by sarcopenia status in diverse individuals with lung cancer

Abstract Background: Sarcopenia and inflammation are independent prognostic indicators of poor survival in lung cancer. Systemic inflammation contributes to sarcopenia or low muscle mass through increased catabolism and decreased synthesis of muscle protein. In turn, sarcopenia leads to inflammation in the muscle and exacerbation of systemic inflammation. Specific Aim: The aim of this study was to determine the association between sarcopenia and systemic biomarkers of inflammation including neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), and albumin in diverse individuals with lung cancer and determine their use as prognostic indicators of sarcopenia. Methods: This retrospective cross-sectional study included 218 non-Hispanic Black (n=111) and White (n=107) individuals with lung cancer. Computed tomography scans were analyzed to calculate skeletal mass index (SMI) at the third lumbar vertebra. Sarcopenia was defined using published sex-specific criteria for SMI in healthy individuals then categorized into the binary variable of sarcopenia/no sarcopenia. NLR, PLR, and albumin were analyzed as both continuous and categorical variables, with NLR&amp;gt; 3, PLR &amp;gt;150, and albumin &amp;lt;3.5 defining moderate to high inflammation. T-tests, Mann Whitney U, Chi square, and adjusted Logistic Regression were used as appropriate. Results: Subjects presented with a mean age of 68.5 + 9.8 years, BMI of 26.3 + 5.9 kg/m2, 50.9% non-Hispanic Black, and 49.5% female with significantly higher prevalence of late-stage lung cancer (61.9% versus 38.1% early-stage, p&amp;lt;0.001) and no differences in age and BMI by race/ethnicity. Mean NLR (6.2 + 7.2), PLR (226.9 + 157.9), and albumin (3.2 + 0.6) indicated an inflammatory state in diverse individuals with lung cancer, with significantly higher NLR (8.5 + 9.7 versus 4.7 + 4.2, p&amp;lt;0.001) and PLR (268.9 + 204.5 versus 198.7 + 108.9, p&amp;lt;0.01) and significantly lower albumin (3.1 + 0.7 versus 3.4 + 0.5, p&amp;lt;0.01) in those with sarcopenia compared to those without sarcopenia. The odds of sarcopenia were 2.2 higher with NLR &amp;gt; 3 compared to NLR &amp;lt;3 (95% CI: 1.1, 4.4) and 2.5 higher with albumin &amp;lt;3.5 compared to albumin &amp;gt;3.5 (95% CI: 1.2, 5.3) after controlling for race/ethnicity, stage of cancer, age, and BMI. PLR &amp;gt; 150 was not a significant predictor of sarcopenia. Conclusion: Diverse individuals with sarcopenia of lung cancer present with higher inflammation compared to individuals with lung cancer but without sarcopenia as indicated by NLR, PLR, and albumin levels. Moderate to high levels of inflammation as indicated by high NLR and low albumin may help predict sarcopenia status in diverse individuals with lung cancer. Citation Format: Cecily A. Byrne, Giamila Fantuzzi, Vanessa M. Oddo, Sage Kim, Timothy J. Koh, Sandra L. Gomez. Inflammatory biomarkers by sarcopenia status in diverse individuals with lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7649.

SOX13 is a novel prognostic biomarker and associates with immune infiltration in breast cancer.

The transcription factor, SOX13 is part of the SOX family. SOX proteins are crucial in the progression of many cancers, and some correlate with carcinogenesis. Nonetheless, the biological and clinical implications of SOX13 in human breast cancer (BC) remain rarely known. We evaluated the survival and expression data of SOX13 in BC patients via the UNLCAL, GEPIA, TIMER, and Kaplan-Meier plotter databases. Immunohistochemistry (IHC) was used to verify clinical specimens. The gene alteration rates of SOX13 were acquired on the online web cBioportal. With the aid of the TCGA data, the association between SOX13 mRNA expression and copy number alterations (CNA) and methylation was determined. LinkedOmics was used to identify the genes that co-expressed with SOX13 and the regulators. Immune infiltration and tumor microenvironment evaluations were assessed by ImmuCellAI and TIMER2.0 databases. SOX13 correlated drug resistance analysis was performed using the GDSC2 database. Higher SOX13 expression was discovered in BC tissues in comparison to normal tissues. Moreover, increased gene mutation and amplification of SOX13 were found in BC. Patients with increased SOX13 expression levels showed worse overall survival (OS). Cox analysis showed that SOX13 independently served as a prognostic indicator for poor survival in BC. Further, the expression of SOX13 was also confirmed to be correlated with tumor microenvironment and diverse infiltration of immune cells. In terms of drug sensitivity analysis, we found higher expression level of SOX13 predicts a high IC50 value for most of 198 drugs which predicts drug resistance. The present findings demonstrated that high expression of SOX13 negatively relates to prognosis and SOX13 plays an important role in cancer immunity. Therefore, SOX13 may potentially be adopted as a biomarker for predicting BC prognosis and infiltration of immune cells.

RNA helicase DDX5 modulates sorafenib sensitivity in hepatocellular carcinoma via the Wnt/β-catenin–ferroptosis axis

Reduced expression of the RNA helicase DDX5 associated with increased hepatocellular carcinoma (HCC) tumor grade and poor patient survival following treatment with sorafenib. While immunotherapy is the first-line treatment for HCC, sorafenib and other multi-tyrosine kinase inhibitors (mTKIs) are widely used when immunotherapy is contra-indicated or fails. Herein, we elucidate the role of DDX5 in sensitizing HCC to sorafenib, offering new therapeutic strategies. Treatment of various human HCC cell lines with sorafenib/mTKIs downregulated DDX5 in vitro and in preclinical HCC models. Conversely, DDX5 overexpression reduced the viability of sorafenib-treated cells via ferroptosis, suggesting a role for DDX5 in sorafenib sensitivity. RNAseq of wild-type vs. DDX5-knockdown cells treated with or without sorafenib identified a set of common genes repressed by DDX5 and upregulated by sorafenib. This set significantly overlaps with Wnt signaling genes, including Disheveled-1 (DVL1), an indispensable Wnt activator and prognostic indicator of poor survival for sorafenib-treated patients. DDX5-knockout (DDX5KO) HCC cells exhibited DVL1 induction, Wnt/β-catenin pathway activation, and ferroptosis upon inhibition of canonical Wnt signaling. Consistently, xenograft HCC tumors exhibited reduced growth by inhibition of Wnt/β-catenin signaling via induction of ferroptosis. Significantly, overexpression of DDX5 in HCC xenografts repressed DVL1 expression and increased ferroptosis, resulting in reduced tumor growth by sorafenib. We conclude that DDX5 downregulation by sorafenib mediates adaptive resistance by activating Wnt/β-catenin signaling, leading to ferroptosis escape. Conversely, overexpression of DDX5 in vivo enhances the anti-tumor efficacy of sorafenib by suppressing Wnt/β-catenin activation and induction of ferroptosis. Thus, DDX5 overexpression in combination with mTKIs is a promising therapeutic strategy for HCC.

Prognostic and Clinicopathological Significance of Telomerase Reverse Transcriptase Upregulation in Oral Cancer: A Systematic Review and Meta-Analysis.

Simple SummaryHanahan and Weinberg proposed to the scientific community a series of distinctive characteristics that all malignant neoplastic cells should possess, which had an enormous impact on the scientific community. One of the most relevant hallmarks of cancer involves the ability of tumor cells to acquire replicative immortality, achieved essentially through the maintenance of telomere length, dependent essentially on telomerase activity. Telomerase lengthens telomeres through the reverse transcription of a 6-bp telomere repeat sequence at the 3′ end of the telomere, which requires the enzymatic component of telomerase, i.e., the telomerase reverse transcriptase (TERT). This mechanism is essential for tumor cells to survive for a long time, and, as has been pointed out, it is frequently activated in malignant cells. This is the first systematic review and meta-analysis, based on 21 studies and 1698 oral squamous cell carcinoma (OSCC) patients, demonstrating that TERT protein overexpression behaves as a prognostic biomarker, significantly associated with poor survival in oral cancer.The aim of this systematic review and meta-analysis was to evaluate the current evidence on the prognostic and clinicopathological significance value of telomerase reverse transcriptase (TERT) upregulation in patients with oral squamous cell carcinoma (OSCC). PubMed, Embase, Web of Science, and Scopus were searched for studies published before April 2022, not restricted by date or publication language. The methodological quality of primary-level studies was critically assessed using the Quality in Prognosis Studies (QUIPS) tool. We carried out meta-analyses, explored heterogeneity and its sources, and performed subgroup, meta-regression, sensitivity, and small-study effects analyses. Twenty-one studies (1698 patients) met inclusion criteria. TERT protein overexpression was significantly associated with worse overall survival (hazard ratio [HR] = 3.01, 95% CI = 1.70–5.35, p < 0.001), disease-free survival (HR = 4.03, 95% CI = 1.80–9.05, p = 0.001), and higher histological grade OSCC (odds ratio [OR] = 3.20, 95% CI = 1.83–5.62, p < 0.001). These large effect sizes were consistently obtained by homogeneous subgroups (p > 0.10, I2 = 0.0, respectively), which reflects a high quality of evidence. On the other hand, TERT gene mutations obtained constantly nonsignificant null effect sizes for all outcomes investigated, evidencing no prognostic or clinicopathological value. In conclusion, our findings indicate that TERT upregulation is a prognostic indicator of poor survival in oral cancer. Our findings support the immunohistochemical assessment of TERT overexpression, which could probably be incorporated into the prognostic evaluation of OSCC.

Combining Pre- and Postoperative Lymphocyte-C-Reactive Protein Ratios Can Better Predict Hepatocellular Carcinoma Prognosis After Partial Hepatectomy.

BackgroundVarious preoperative inflammatory indicators have been identified as potential predictors of poor prognosis in patients with hepatocellular carcinoma (HCC), but the role of postoperative inflammatory indicators remains unclear. This study aimed to explore the prognostic value of the postoperative lymphocyte–C-reactive protein ratio (PostLCR) on its own and combined with preoperative LCR (PreLCR).MethodsA total of 290 patients with primary HCC were retrospectively enrolled in the study. Univariate analysis was used to identify factors significantly associated with poor disease-free survival (DFS) and overall survival (OS), then multivariate analysis was performed to identify independent prognostic indicators of poor survival. Prognostic models based on preoperative, postoperative, and both types of indicators were then constructed, and their predictive performance were evaluated using time-dependent receiver operating characteristic curves and the concordance index (C-index).ResultsPreLCR and PostLCR levels correlated with DFS and OS more strongly than other pre- and postoperative inflammatory indicators, respectively. Decreased PreLCR and PostLCR were independent prognostic factors for both DFS and OS, while HCC patients with decreased PreLCR and PostLCR had worse prognosis than patients with increased PreLCR and PostLCR. Patients into three groups based on their cut-off values of PreLCR and PostLCR, Kaplan–Meier survival analysis indicated that HCC patients with low PreLCR and PostLCR had the worst DFS and OS. The combined model showed better predictive performance at 1 and 3 years post-surgery than individual pre- and postoperative models, the American Joint Committee on Cancer/Tumor-Node-Metastasis (8th edition) staging system and the Barcelona Clinic Liver Cancer system. The combine model demonstrated a markedly superior C-index compared with the other models in DFS and OS.ConclusionOur study showed PreLCR and PostLCR are independent predictors of DFS and OS in HCC patients after partial hepatectomy. Models that include both PreLCR and PostLCR can predict prognosis better than well-established clinical staging systems.

C1QTNF6 is a Prognostic Biomarker and Related to Immune Infiltration and Drug Sensitivity: A Pan-Cancer Analysis.

Background: The discovery of reliable cancer biomarkers could tune a diagnosis and improve the way patients are treated. However, many cancers lack robust biomarkers. C1QTNF6 has been preliminarily elucidated for its role in some tumors. However, no pan-cancer analysis has been performed to comprehensively explore the value of C1QTNF6. Methods: Data from the TCGA database, GTEx database stored in the USUC Xena were used for analyzing the profiles of C1QTNF6 expression in normal and tumor tissues in pan-cancer. Subsequently, the gene alteration rates of C1QTNF6 were acquired on the online web cBioportal. With the aid of the TCGA data, the association between C1QTNF6 mRNA expression and copy number alterations (CNA) and methylation was determined. Survival analyses of C1QTNF6 were carried out. Moreover, the tumor biological and immunological characteristics of C1QTNF6 were clarified in the forms of the correlation between C1QTNF6 expression and hallmark Pathway scores in MsigDB database, immune cell infiltration, immune-related genes. We conducted a GSEA of C1QTNF6 to illustrate its potential biological functions. In addition, GDSC2 data with 198 drugs were adopted to explore drug sensitivity with the change of C1QTNF6 expression. Result: C1QTNF6 was overexpressed in many types of cancer, Survival analysis showed that C1QTNF6 independently served as a prognostic indicator for poor survival in many tumors. Besides, we also identified a positive correlation between C1QTNF6 and cancer hallmark pathway score, tumor microenvironment related pathways score (TMEp score), and immune characteristic. In terms of drug sensitivity analysis, we found higher expression level of C1QTNF6 predicts a high IC50 value for most of 198 drugs which predicts drug resistance. Conclusions: Our study provides a new biological marker for pan-cancer, which is beneficial to the diagnosis and treatment of cancer, which bring a new therapeutic target for tumors.

Metastatic Tumor Cell-Specific FABP7 Promotes NSCLC Metastasis via Inhibiting β-Catenin Degradation.

Metastasis accounts for 90% of cancer-related deaths and represents a prominent malignant feature in non-small cell lung cancer (NSCLC), while tumor cell-specific mechanisms and molecules pivotal for the metastatic capacity remain unclear. By analyzing single-cell RNA sequencing data, we found that fatty acid binding protein 7 (FABP7) was specifically up-regulated in tumor cells of metastatic NSCLC patients and might be a prognostic indicator for poor survival. Experimental studies based on NSCLC cell lines showed that FABP7 promoted the metastatic competencies of NSCLC cells in vitro and in vivo. Mechanistically, we demonstrated that FABP7 was important to canonical Wnt signaling activation and competitively inhibited the interaction between β-catenin and components of its cytoplasmic degradation complex, thereby repressing the phosphorylation-dependent ubiquitination and degradation of β-catenin. Our present study identifies FABP7 as a metastatic tumor cell-specific pro-metastatic gene and uncovers a previously unknown regulatory mechanism underlying Wnt hyperactivation via FABP7-impaired cytoplasmic β-catenin degradation, implicating a novel molecule in regulating NSCLC metastasis.

Single-Cell Multi-Omic Analysis Uncovers Comprised Immune Function and Primary Resistance Mechanism in Acute Myeloid Leukemia

Background: Approximately 20% of AML patients do not respond to induction chemotherapy (primary resistance) and 40-60% of patients develop secondary resistance, eventually leading to relapse followed by refractory disease (RR-AML). Diversified molecular mechanisms have been proposed for drug resistance and RR phenotype. However, we still cannot predict when relapse will occur, nor which patients will become resistant to therapy.Single-cell multi-omic (ScMo) profiling may provide new insights into our understanding of hematopoietic stem cell (HSC) differentiation trajectories, tumor heterogeneity and clonal evolution. Here we applied ScMo to profile bone marrow (BM) from AML patients and healthy controls.Methods: AML samples were collected at diagnosis with institutional IRB approval. Cells were stained with a panel of 62 DNA barcoded antibodies and 10x Genomics Single Cell 3' Library Kit v3 was used to generate ScMo data. After normalization, clusters were identified using Uniform Manifold Approximation and Projection (UMAP) and annotated using MapCell (Koh and Hoon, 2019). We analyzed 23,933 cells from 4 adult AML BM samples, and 39,522 cells from 2 healthy adults and 3 sorted CD34+ normal BM samples. Gene set enrichment analysis (GSEA) and Enrichr program were used to examine underlying pathways among differentially expressed genes between healthy and AML samples.Results: We identified 16 cell types between the AML and normal samples (Fig 1a) amongst 45 clusters in the UMAP projection (Fig 1b). Comparative analysis of the T cell clusters in AML samples with healthy BM cells identified an “AML T-cell signature” with over-expression of genes such as granzymes, NK/T cell markers, chemokine and cytokine, proteinase and proteinase inhibitor (Fig 2a). Among them, IL32 is known to be involved in activation-induced cell death in T cells and has immunosuppressive role, while CD8+ GZMB+ and CD8+ GZMK+ cells are considered as dysfunctional or pre-dysfunctional T cells. Indeed, Enrichr analysis showed the top rank of phenotype term - “decreased cytotoxic T cell cytolysis”.We next examined whether NK cells, are similarly dysfunctional in the AML ecosystem. The “AML NK cell signature” includes Fc Fragment family, IFN-stimulated genes (ISGs), the effector protein-encoding genes and other genes when compared to normal NK cells (Fig 2b). GSEA analysis revealed “PD-1 signalling” among the top 5 ranked pathways in AML-NK cells, though no increase in PD-1 protein nor PDCD1 gene were identified in these cells. Inhibitory receptor CD160 was expressed higher in AML samples along with exhaustion (dysfunction) associated genes TIGIT, PRF1 and GZMB (Fig 2c). Enrichr analysis uncovered enrichment of “abnormal NK cell physiology and ”impaired natural killer cell mediated cytotoxicity“. Similarly, the ”AML monocyte signature“ was significantly enriched with genes in ”Tumor Infiltrating Macrophages in Cancer Progression and Immune Escape“ and ”Myeloid Derived Suppressor Cells in Cancer Immune Escape“.We also analyzed HSPC component in one pair of cytogenetically matched, untreated complete remission (CR) /RR AML pair (Fig 2d). Notably, half of the 10 genes overexpressed in RR-AML, CXCR4, LGALS1, S100A8, S100A9, SRGN (Serglycin), regulate cell-matrix interaction and play pivotal roles in leukemic cells homing bone marrow niche. The first 4 of these genes have been demonstrated as prognostic indicators of poor survival and associated with chemo-resistance and anti-apoptotic function. Furthermore, single-cell trajectory analysis of this CR/RR pair illustrated a change in differentiation pattern of HSPCs in CR-AML to monocytes in RR-AML. We are currently analyzing more AML samples to validate these findings.Conclusions: Our ScMo analysis demonstrates that the immune cells are systematically reprogrammed and functionally comprised in the AML ecosystem. Upregulation of BM niche factors could be the underlying mechanism for RR-AML. Thus, reversing the inhibited immune system is an important strategy for AML therapy and targeting leukemic cell-BM niche interaction should be considered for cases with high expression of these molecules on AML HSPCs.Note: J.Z. and J.A.S. share co-first authorship. [Display omitted] DisclosuresScolnick: Proteona Pte Ltd: Current holder of individual stocks in a privately-held company. Xu: Proteona Pte Ltd: Current Employment. Ooi: Jansen: Honoraria; Teva Pharmaceuticals: Honoraria; GSK: Honoraria; Abbvie: Honoraria; Amgen: Honoraria. Lovci: Proteona Pte Ltd: Current Employment. Chng: Aslan: Research Funding; Takeda: Honoraria; Johnson & Johnson: Honoraria, Research Funding; BMS/Celgene: Honoraria, Research Funding; Amgen: Honoraria; Novartis: Honoraria, Research Funding; Antengene: Honoraria; Pfizer: Honoraria; Sanofi: Honoraria; AbbVie: Honoraria.

High levels of Tim-3+Foxp3+Treg cells in the tumor microenvironment is a prognostic indicator of poor survival of diffuse large B cell lymphoma patients

Foxp3+Treg cells display phenotypic and functional heterogeneity, which express high levels of T cell immunoglobulin and mucin-domain containing-3 (Tim-3) in the tumor microenvironment (TME) of colorectal and lung cancer. High abundance of Tim-3+Foxp3+Treg (TFT) cells are associated with poor prognosis in these patients. However, the expression patterns and roles of TFT cells in TME of diffuse large B cell lymphoma (DLBCL) remain to be established. Double immunofluorescence and flow cytometry analyses were employed to investigate TFT cell enrichment in paraffin-embedded fresh tumor tissues from patients with DLBCL. Spearman's or Pearson's correlation and Kaplan-Meier survival analyses were further applied to decide the prognostic value of TFT cell levels in DLBCL. The IL-10-secreting function of TFT cells in vitro was examined via flow cytometry and ELISA. Our results showed for the first time that TFT cells are highly enriched in TME of DLBCL patients and associated with predictions of poor prognoses. TFT cell-induced secretion of IL-10 in the TME was suppressed by an anti-Tim-3 antibody in vitro. In conclusion, high abundance of TFT cells in the TME is predictive of poor outcomes of DLBCL. TFT cells promote DLBCL development partly by secreting IL-10 in the TME. Anti-Tim-3 antibodies (that block IL-10 secretion) may present an effective therapeutic agent for DLBCL.

Abstract PS6-42: Prognostic value of neutrophil to lymphocyte ratio in patients with breast cancer in a Mexican population

Abstract Background: Inflammatory serum markers (IBM) have emerged as prognostic factors in solid tumors. In breast cancer (BC) in particular, an elevated neutrophil to lymphocyte ratio (NLR) has been linked to lower overall survival (OS) and disease-free survival (DFS), although a standard cutoff value has not been established. In Mexico there is no information regarding NLR as a prognostic marker in BC. The aim of the present study was to assess NLR as prognostic factor in BC patients in a Mexican population. Methods: This single-center retrospective and descriptive study included patients, &amp;gt;18 years old with histological diagnosis of BC who were treated at Medica Sur oncological center in Mexico City between January 2008 and December 2019. The patients were divided into two groups according to their NLR. NLR was calculated using the following formula: absolute neutrophil count / absolute lymphocyte count (mm3). NLR was considered as elevated (&amp;gt;2), and as low (≤2) based on data from previous studies. The primary endpoint was OS. Statistical analysis was performed with SPSS v25. The associations between PNI and clinicopathologic characteristics were analyzed using Pearson's χ2 test. Kaplan-Meier and log-rank test methods were used for survival analysis. The prognostic value of the pre-treatment NLR was assessed by univariate and multivariate analysis. P&amp;lt;0.05 was considered to indicate a statistically significant difference. This study was approved by our scientific and bioethical committee. Results: A total of 110 patients were included in the final analysis. All patients had infiltrating ductal carcinoma; 18.2% were triple negative, 23.6% over-expressed HER 2 and around 51% had hormone receptor expression. 15.5% had metastatic disease. Median follow-up was 65 months. Mean NLR at diagnosis was 2.82 (SD 2.59). Mean NLR in patients with triple negative BC (TNBC) was significantly higher than in patients with non TNBC (p= 0.055), no other statistically significant associations were found between mean NLR and clinical characteristics. Median OS was not reached in two groups, but the 100 months OS was higher in NLR ≤2 group vs NLR &amp;gt; 2 group (90 vs 78%) (p= 0.079). In univariate and multivariate analysis, triple negative histological subtype and elevated NLR were independent prognostic indicators of poor survival. Conclusion: An elevated pretreatment NLR was an independent prognostic factor, associated with lower OS in patients with BC with local and advanced disease, although statistically significant association was not found. NLR is an accessible and minimally invasive marker with clinical value for evaluation of the prognosis of BC patients, especially in resource-limited settings. Citation Format: Andrés Mauricio Arroyave-Ramírez, Edgar Varela-Santoyo, Pamela Denisse Soberanis-Piña, Hector Hugo Buerba-Vieregge, Daniel Motola-Kuba. Prognostic value of neutrophil to lymphocyte ratio in patients with breast cancer in a Mexican population [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS6-42.

Long Noncoding RNA AFAP1-AS1 Is a Critical Regulator of Nasopharyngeal Carcinoma Tumorigenicity.

BackgroundThe long noncoding RNA actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) is a critical player in various cancers. However, the clinical value and functional mechanisms of AFAP1-AS1 during the tumorigenicity of nasopharyngeal carcinoma (NPC) remain unclear. Here, we investigated the clinical application and potential molecular mechanisms of AFAP1-AS1 in NPC tumorigenesis and progression.MethodsThe expression level of AFAP1-AS1 was determined by qRT-PCR in 10 paired fresh human NPC tissues and adjacent normal tissues. RNAscope was performed on 100 paired paraffin-embedded NPC and adjacent nontumor specimens. The biological functions of AFAP1-AS1 were assessed by in vitro and in vivo functional experiments. RNA-protein pull-down assays were performed to detect and identify the AFAP1-AS1-interacting protein KAT2B. Protein-RNA immunoprecipitation (RIP) assays were conducted to examine the interaction of AFAP1-AS1 and KAT2B. Chromatin immunoprecipitation (ChIP) and luciferase analyses were utilized to identify the binding site of transcription intermediary factor 1 alpha (TIF1α) and H3K14ac on the RBM3 promoter.ResultsAFAP1-AS1 is upregulated in NPC and is a poor prognostic indicator for survival in NPC patients. AFAP1-AS1 was required for NPC proliferation in vitro and tumorigenicity in vivo. Mechanistic investigations suggested that AFAP1-AS1 binds to KAT2B and promotes acetyltransferase activation at two residues (E570/D610). KAT2B further promotes H3K14 acetylation and protein binding to the bromo domain of TIF1α. Consequently, TIF1α acts as a nuclear transcriptional coactivator of RBM3 transcription, leading to YAP mRNA stabilization and enhanced NPC tumorigenicity.ConclusionsOur findings suggest that AFAP1-AS1 functions as an oncogenic biomarker and promotes NPC tumorigenicity through enhanced KAT2B acetyltransferase activation and YAP mRNA stabilization.

The Proportion of Tumour-Stroma in Metastatic Lymph Nodes is An Accurately Prognostic Indicator of Poor Survival for Advanced-Stage Colon Cancers.

The importance of tumour microenvironment in tumour behaviour has now become clearer. This study aimed to determine the prognostic effect of the proportion of tumour-stroma (PTS) in metastatic lymph nodes of advanced-stage colon cancers (CCs). We investigated PTS in positive lymph nodes of stage III-IV CC patients who underwent surgical treatment between 2004 and 2014. We used a standard approach in methodology. PTS was significantly associated with prognostic factors in the metastatic lymph nodes (perineural invasion [p = 0.031], lymphatic invasion [p = 0.032], invasive margin [p = 0.043], advanced pT [p = 0.020], and margin involvement [p = 0.034]). In addition, the correlations between PTS estimates (R = 0.704 to 0.617, p < 0.001), the reproducibility of the research (Κappa = 0.72-0.68) and the usefulness of the cut-off value (ROC: 50.33%; AUC = 0.752 [0.667-0.857]) were successful. In univariate analysis, 5-year survival was poor for RFS (p < 0.001), OS (p = 0.001) and LR (p = 0.013) in high PTS patients. Multivariate analysis confirmed that high PTS was an independent worse parameter for RFS (HR = 1.32, 95% CI: 1.17-2.55, p = 0.001) and OS (HR = 1.37, 95% CI: 1.25-1 - 2.56, p = 0.009). In this study, we showed that high PTS in metastatic lymph nodes was a successful prognostic marker for advanced-stage CCs. Also, the standard approach we used for the methodology was successful.

Cyclophilin B overexpression predicts a poor prognosis and activates metastatic pathways in colon cancer.

BackgroundCyclophilin B (CypB) has been found overexpressed in various malignant tumors. To date, there are few studies on CypB in colon cancer. In this study, we aimed to analyze the CypB expression pattern and to further evaluate its clinical significance, especially its prognostic value for colon cancer.MethodsCypB expression was investigated in colon cancer tissue microarrays (TMA) by RNAscope in situ hybridization and immunohistochemical (IHC) staining. The correlation between CypB and clinicopathological characteristics was analyzed. The Cancer Genome Atlas (TCGA) RNA-seq dataset of colon adenocarcinoma (COAD) was further analyzed to validate our main findings. Gene Set Enrichment Analysis (GSEA) and Search Tool for the Retrieval of Interacting Genes (STRING) analysis were performed to enrich CypB related biological pathways. In vitro experiments by knockdown of CypB in colon cancer cell HCT116 were performed to verify the bioinformatics results and analyze its role in the metastatic pathways in colon cancer.ResultsWe found that CypB expression was highly upregulated in colon cancer tissues (P<0.05). Importantly, the overall survival (OS) time of patients with high CypB expression was significantly shorter than those with low CypB expression, and overexpressed CypB was identified as an independent prognostic indicator for poor survival (P=0.015). Subgroup analysis indicated that a high level of CypB was associated with a shorter OS time, especially for advanced cancer patients, such as later T stage, lymph node metastasis, larger tumor size (P<0.05). Analysis of TCGA RNA-seq dataset of COAD provided us with a larger clinical sample verification. Bioinformatics analysis and the following in vitro study revealed that CypB was involved in tumor metastatic associated signaling pathways.ConclusionsCypB overexpression predicts a poor prognosis and may activate metastatic pathways in colon cancer.

DJ‑1 promotes cell proliferation and tumor metastasis in esophageal squamous cell carcinoma via the Wnt/β‑catenin signaling pathway.

DJ-1, an oncogene, has been reported to be an independent prognostic indicator of poor survival in patients with esophageal squamous cell carcinoma (ESCC). The aim of the present study was to investigate the role of DJ-1 in tumor cell proliferation and invasion in ESCC and its underlying mechanisms. It was observed that the expression level of DJ-1 was upregulated and positively associated with EMT biomarkers in 84 human ESCC tissue specimens. Overexpression and knockdown experiments demonstrated that DJ-1 was involved in proliferation, migration, invasion and EMT in ECA-109 cells in vitro and extensive peritoneal seeding in a peritoneal dissemination mice model. Furthermore, the present data revealed that DJ-1 could activate the Wnt/β-catenin signaling pathway, which mediates the EMT and metastasis in ESCC. In conclusions, DJ-1 promoted proliferation, invasion, metastasis and the EMT in ESCC via activation of the Wnt/β-catenin signal pathway. The present results suggested DJ-1 could represent a promising therapeutic target for the prevention and treatment of ESCC-related metastasis.

Abstract 1967: Nab-paclitaxel chemotherapy of breast tumors upregulates chemokines that increase chemotaxis of myeloid-lymphatic progenitors

Abstract Introduction: Lymph node metastasis is the main prognostic indicator for poor survival of breast cancer patients. Metastasis is promoted by tumor-induced lymphangiogenesis mediated, in part, through recruitment of myeloid-derived lymphatic endothelial cell progenitors (M-LECP). We previously showed that differentiation of M-LECP is induced by activated Toll-like receptor-4 (TLR4). The goal of this study was to identify TLR4-induced chemokines produced by tumors alone or in response to nab-paclitaxel (nab-PXL) chemotherapy that enhance bone marrow (BM) differentiation and tumor recruitment of M-LECP. Methods: Profile of BM composition for lymphatic-like, myeloid and progenitor cells was assessed by flow cytometry in normal and tumor-bearing mice treated either with saline or nab-PXL. We used two TLR4 overexpressing tumor models: MDA-MB-231 and HCC1806-TLR4+. RT-qPCR was used to determine differences in expression of 130 cytokines in cultured tumor cells and tumors from mice treated either with saline or nab-PXL. Targets with &amp;gt;2-fold upregulation were analyzed for protein expression by ELISA. ELISA-validated cytokines were tested for their ability to induce migration of M-LECP. Results: Both MDA-MB-231 or HCC1806-TLR4+ tumors significantly increased the levels of differentiated M-LECP in the BM, and that was further enhanced by treatment with taxane chemotherapy. Expression of progenitor markers CD117, CD33, and Sca-1 was increased up to 7-fold in both tested tumor models, and further upregulated by nab-PXL treatment by additional 3 to 9-fold. Out of 23 cytokines measured in tumor lysates by ELISA, 40% were upregulated by nab-PXL. Migration assay showed that several of these cytokines increased chemotaxis of M-LECP with CXCL1 being the strongest. Neutralization of CXCL1 in the conditioned medium of nab-PXL-treated MDA-MB-231 cells inhibited migration of lymphatic progenitors by 75%. Conclusion: Taxane chemotherapy acting presumably as a ligand for TLR4 significantly expanded M-LECP population in the BM. Our analysis suggests that these lymphatic precursors can mobilize to tumors by chemokines such as CXCL1 that are highly upregulated in TLR4-positive tumor cells, particularly when activated by paclitaxel. These data suggest that CXCL1 plays an important role in recruiting myeloid-lymphatic progenitors to breast tumors, which, in turn, promotes lymphatic vessel formation and metastasis. Blocking this cytokine might have clinical utility for prevention of tumor mobilization of M-LECP, and by extension, breast cancer metastasis to lymph nodes. Citation Format: Lisa D. Volk-Draper, Sophia Ran. Nab-paclitaxel chemotherapy of breast tumors upregulates chemokines that increase chemotaxis of myeloid-lymphatic progenitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1967.

Survival, disease progression and prognostic factors in elderly patients with mycosis fungoides and Sézary syndrome: a retrospective analysis of 174 patients.

Advanced age at diagnosis is considered a poor prognostic factor in mycosis fungoides (MF) and Sézary syndrome (SS). To evaluate the outcomes and prognostic factors in patients diagnosed at an advanced age (≥65years) with MF/SS. Survival, progression rates and various clinical and histopathological variables were studied in a group of 174 elderly patients diagnosed with MF/SS between 1992 and 2015 at a single referral cancer center in the United States. Kaplan-Meier estimates were used to determine survival and progression and Cox proportional hazards regression univariate and multivariate models were used to identify prognostic factors. Of 174 elderly patients, 76.4% were diagnosed with early-stage (clinical stages IA-IIA) and 23.6% with late-stage MF/SS (IIB-IV). Advanced age was associated with poor overall survival, but not with disease-specific survival (DSS) or progression-free survival (PFS). Gender, increasing clinical stage, T and B classifications, elevated lactate dehydrogenase (LDH) levels and development of large cell transformation (LCT) were significant predictors of poor survival or disease progression. Patients with early-stage MF and <10% total skin involvement (T1 classification) or patch-only disease (T1a/T2a) showed better PFS with no observed disease-specific mortality. Folliculotropic MF was associated with poor DSS in patients with early-stage disease. Older age at diagnosis of MF/SS does not predict worse disease-specific outcomes. Elderly patients with early-stage disease, specifically involving less than 10% of the skin surface with patches but without plaques or folliculotropism, have an excellent prognosis. However, the development of LCT is a strong prognostic indicator of poor survival in elderly patients with MF/SS.

A high neutrophil to lymphocyte ratio prior to BRAF inhibitor treatment is a predictor of poor progression-free survival in patients with metastatic melanoma.

Some studies have shown that a high neutrophil/lymphocyte ratio (NLR) ≥4 before initiating ipilimumab treatment is an independent prognostic indicator of poor survival in patients with metastatic melanoma (MM). To determine whether the NLR before starting BRAF inhibitor (BRAFi) treatment in patients with (MM) is associated with progression-free survival (PFS). This retrospective study included 49 patients consecutively receiving BRAFi for MM between July 2012 and December 2014. Cox proportional hazards regression was used to analyse the relationship between NLR and other factors, such as lactate dehydrogenase (LDH), performance status, BRAFi as first- or second-line therapy, and corticosteroid intake with PFS. The NLR before starting BRAFi was significantly associated with PFS based on univariate analysis and multivariate analysis adjusted for potential confounding factors, such as LDH activity, ulceration, performance status, first-line therapy, and corticosteroid intake. A high NLR (continuous variable) was associated with short PFS (HR: 1.35; 95%CI: 1.07-1.70; p=0.01), and NLR ≥4 was associated with shorter PFS (HR: 3.24; 95%CI: 1.30-8.12; p=0.01). Corticosteroid intake was not associated with short PFS based on multivariate analysis. An NLR >4, before starting BRAFi treatment, is an independent prognostic indicator of poor progression-free survival.