Independent Prognostic Marker Research Articles

Impact of stress hyperglycemia ratio, derived from glycated albumin or hemoglobin A1c, on mortality among ST-segment elevation myocardial infarction patients

BackgroundStress hyperglycemia ratio (SHR), associated with adverse outcomes in patients with ST-segment elevation myocardial infarction (STEMI), has several definitions. This study aims to assess the prognostic value of SHR, derived from hemoglobin A1c (HbA1c) or glycated albumin (GA), to mortality.MethodsThe study comprised 1,643 STEMI patients who underwent percutaneous coronary intervention (PCI) in two centers. SHR1 was calculated using fasting blood glucose (FBG)/GA, while SHR2 was calculated using the formula FBG/(1.59*HbA1c-2.59). The primary endpoints were in-hospital death and all-cause mortality, with a median follow-up duration of 1.56 years.ResultsHigher SHR1 and SHR2 values are associated with increased risks of in-hospital death and all-cause mortality. Each standard deviation increase in SHR1 corresponded to a 39% and 22% escalation in in-hospital death and all-cause mortality, respectively. The respective increases for SHR2 were 51% and 26%. Further examinations validated these relationships as linear. Additionally, the areas under the curve (AUC) for in-hospital death were not significantly different between SHR1 and SHR2 (p > 0.05). Incorporating SHR1 or SHR2 into the base model significantly improved the discrimination and risk reclassification for in-hospital and all-cause mortality. A subgroup analysis revealed that the effects of SHR1 and SHR2 were more pronounced in patients with hypercholesteremia.ConclusionSHR1 and SHR2 have emerged as robust and independent prognostic markers for STEMI patients undergoing PCI. The SHR calculation based on either HbA1c or GA can provide additional predictive value for mortality beyond traditional risk factors, helping to identify high-risk STEMI patients.

Abstract C030: Gene expression profiling tumors of patients with clear cell renal carcinoma: May identify race-specific therapeutic targets

Abstract Black/African Americans are second only to Native Americans in having the highest incidence of renal cancer. Despite this statistic, very few studies have examined whether patients’ tumors exhibit racial/ethnic differences in both gene and/or protein expression. Using KIRC (clear cell renal carcinoma) TCGA dataset (n=542), we identified several genes that were significantly overexpressed in ccRCC derived from Blacks. Genes which showed significantly higher expression in tumors from Blacks (n=56) relative to Whites (n=463) included ADCK5 (adj p<0.001) ADRM1 (adj p=0.013), ANKRD13D (adj p=0.003), ARFGAP1 (adj p=.003), LZTS2 (adj p<0.001), FAM160B2 (adj p<0.001), and NCKAP5L (adj p=0.029). In addition, these genes were also significantly overexpressed (p<0.05) in advanced stages, higher grades as well as in metastatic tissues. Multivariate cox regression analysis of gene expression, ethnicity, racial demographics and clinicopathological features relative to overall survival time revealed that ARFGAP1 (HRadj=2.40 [95% CI: 1.37 to 4.22]), ANKRD13D (HRadj=2.56 [95% CI:1.43 to 4.59]), FAM160B2 (HRadj= 1.74 [95%CI: 1.01 to 2.99]), and NCKAP5L (HRadj= 1.88 [95% CI:1.04 to 3.41]) were independent prognostic markers for survival. Higher expression of ADCK5 (HRadj= 5.98[CI:1.26-28.35], p=0.012), ADMR1 (HRadj= 1.74 [95%CI: 1.05 to 11.38], p=0.03), ANKRD13D (HRadj= 9.05 [95%CI: 1.13 to 72.66], p=0.014), ARFGAP1 (HRadj=3.79 [95% CI: 0.097-14.54], p=0.041), FAM160B2 (HRadj=5.09 [95%CI: 1.01-25.75], p=0.033), LZTS2 (HRadj=3.32 [95%CI: 1.01-10.99], p=0.037) was associated with lower ccRCC survival in Blacks. ROC analysis indicated that expression of FAM160B2 and LZTS2 are strong predictors of ccRCC survival among Blacks (AUC= 0.737, p<0.001, AUC= 0.731, p<0.001, respectively), whereas ADCK5 and ADRM1, (AUC=0.692, p<0.001, AUC=0.625, p=0.001, respectively) were moderate predictors of ccRCC survival among Blacks. FAM160B2 (family with sequence similarity 160 member B2) is a protein kinase A anchoring protein (AKAP) and participates in PKA (protein kinase A) phosphorylation of HSP70 (heat shock protein 70). FAM160B2 is also involved in hepatocellular carcinoma and promotes tumorigenesis. LZTS2, which encodes leucine zipper tumor suppressor 2, was identified as a tumor susceptibility gene within the 10q24.3 chromosomal region and is approximately 15Mb from the PTEN locus. This region containing both loci is frequently deleted in a variety of human malignancies, including prostate cancer. Taken collectively our data suggest that the aforementioned genes, FAM160B2 and LZTS2 may be useful both as clinical biomarkers and should be explored as potential therapeutic targets for ccRCC, with possible greater impact among Black patients. Citation Format: Romonia Renee Reams, Simone O. Heyliger, Marilyn Saulsbury, Emanuela Taioli, Diana Wilkie, John Carpten. Gene expression profiling tumors of patients with clear cell renal carcinoma: May identify race-specific therapeutic targets [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C030.

The prognostic significance of tumor-immune microenvironment in ascites of patients with high-grade serous carcinoma.

High-grade serous carcinoma (HGSC) is often associated with ascites at presentation. Our objective was to quantify immune cells (ICs) in ascites prior to any treatment was given and evaluate their impact on progression-free survival (PFS) and overall survival (OS). Forty-seven patients with primary HGSC and ascites were included. Flow-cytometric analysis was performed to detect percentages of CD3+ T cells (CD4+, CD8+, Tregs, and NKT cells), B cells, NK cells (CD56brightCD16- and CD56dimCD16+ subsets), macrophages and dendritic cells (DCs). Furthermore, CD103 expression was analyzed on T cells and their subsets, while PD-1 and PD-L1 expression on all ICs. Cut-off of low and high percentages of ICs was determined by the median of variables, and correlation with PFS and OS was calculated. CD3+ cells were the predominant ICs (median 51%), while the presence of other ICs was much lower (median ≤10%). CD103+ expression was mostly present on CD8+, and not CD4+ cells. PD-1 was mainly expressed on CD3+ T cells (median 20%), lower expression was observed on other ICs (median ≤10%). PD-L1 expression was not detected. High percentages of CD103+CD3+ T cells, PD-1+ Tregs, CD56brightCD16- NK cells, and DCs correlated with prolonged PFS and OS, while high percentages of CD8+ cells, macrophages, and PD-1+CD56brightCD16- NK cells, along with low percentages of CD4+ cells, correlated with better OS only. DCs were the only independent prognostic marker among all ICs. Our results highlight the potential of ascites tumor-immune microenvironment to provide additional prognostic information for HGSC patients. However, a larger patient cohort and longer follow-up are needed to confirm our findings.

Untargeted metabolomics identifies metabolic dysregulation of sphingolipids associated with aggressive chronic lymphocytic leukaemia and poor survival.

Metabolic dependencies of chronic lymphocytic leukaemia (CLL) cells may represent new personalized treatment approaches in patients harbouring unfavourable features. Here, we used untargeted metabolomics and lipidomics analyses to isolate metabolomic features associated with aggressive CLL and poor survival outcomes. We initially focused on profiles associated with overexpression of the adverse metabolic marker glycosyltransferase (UGT2B17) associated with poor survival and drug resistance. Leukaemic B-cell metabolomes indicated a significant perturbation in lipids, predominantly bio-active sphingolipids. Expression of numerous enzyme-encoding genes of sphingolipid biosynthesis pathways was significantly associated with shorter patient survival. Targeted metabolomics further exposed higher circulating levels of glucosylceramides (C16:0 GluCer) in CLL patients relative to healthy donors and an aggressive cancer biology. In multivariate analyses, C16:0 GluCer and sphinganine were independent prognostic markers and were inversely linked to treatment-free survival. These two sphingolipid species function as antagonistic mediators, with sphinganine being pro-apoptotic and GluCer being pro-proliferative, tested in leukemic B-CLL cell models. Blocking GluCer synthesis using ceramide glucosyltransferase inhibitors induced cell death and reduced the proliferative phenotype, which further sensitized a leukaemic B-cell model to the anti-leukaemics fludarabine and ibrutinib in vitro. Specific sphingolipids may serve as prognostic markers in CLL, and inhibiting enzymatic pathways involved in their biosynthesis has potential as a therapaeutic approach.

Prognostic Significance of Alpha-Fetoprotein in Staging of Chronic Hepatitis B Infection

Background Ascertaining the stage of chronic hepatitis-B infection (CHBI) remains one of the major predicaments to effective therapeutic decision. There is pressing need to forestall dearth of such reliable biomarker(s). Despite the promising tendency of alpha-fetoprotein (AFP), it has not been assessed in staging CHBI.ObjectiveThis study was to determine the prognosis of serum AFP as a biomarker for staging CHBI. MethodsParticipants were grouped into three based on their hepatitis-B envelope antigen (HBeAg) status and alanine aminotransferase (ALT) level. By denoting HbeAg(+) as (EP), HbeAg(-) as (EN), elevated ALT as (H) and normal ALT as (I), the stages were EPH, ENH and ENI. AFP was assayed, One-way ANOVA, Multivariate linear regression and area under curve were adopted for the analysis. ResultsAFP was significantly elevated, (P < 0.05) in EPH, which equally has the highest prevalence of elevated AFP (64.7%). After adjusting for confounding factors, odds ratio was 1.438 (95% CI, 0.62–1.948), while area under the curve for predicting EPH was (0.828, 95% CI, 0.778 –0.895).ConclusionsThe finding of elevated AFP in CHBI is an independent prognostic marker of EPH. It is often associated with necroinflammation; thus, it is a reliable indicator for treatment initiation.Rwanda J Med Health Sci 2023;6(3):355-366

Myosteatosis as a Prognostic Marker for Postoperative Mortality in Adult Patients Undergoing Surgery in General—A Systematic Review

Background: Assessing frailty is important in treating surgical patients to predict peri- and postoperative events like complications or mortality. The current standard is not optimal; therefore, new prognostic markers are being evaluated to enrich the current frailty assessment. One of these new markers is fat degeneration of the psoas muscle (myosteatosis). This can be assessed by measuring the psoas muscle density (PMD) with computed tomography (CT). The aim of this review is to investigate PMD, and, thus, myosteatosis, as a prognostic marker for postoperative mortality in adult patients undergoing general surgery. Methods: An electronic search was performed in PubMed to identify relevant studies associating PMD with postoperative mortality. The looked-upon period for mortality to occur did not matter for this review. The looked-upon outcome measure for this review was the hazard ratio. Results: From 659 potential articles from PubMed, 12 were included, for a total of 4834 participants. Articles were excluded when not focused on PMD, if the type of intervention was not specified, and when imaging other than with CT on the level of the third vertebra was performed. The included articles were assessed for bias with the Newcastle–Ottawa Scale (NOS). PMD was, after multivariable analyses, identified as an independent significant prognostic marker for several surgical cardiovascular interventions when we looked at the 5-year mortality rate and for fenestrated branched endovascular aortic repair (F-BEVAR) a slight significant protective correlation between postoperative mortality and PMD (when divided by psoas muscle area (PMI)) when we looked at the 30-day and 3-year mortality. Also, PMD was identified as an independent significant prognostic marker for a variety of surgical gastrointestinal interventions when we looked at 30-day/90-day/1-year/3-year/5-year mortality. PMD was not identified as a significant prognostic marker in urologic surgery. Conclusion: Myosteatosis has the potential to be a valuable contribution to the current frailty assessment for patients undergoing cardiovascular, gastrointestinal, or urologic surgery. However, more research must be conducted to further strengthen the prognostic value of myosteatosis, with special attention to, e.g., gender- or age-specific interpretations of the results.

Polypharmacy Independently Predicts Overall Survival in Patients with Lymphoid Cancer

Introduction Comorbidity is common among patients with lymphoid cancer (LC) and may limit treatment options. Often, comorbidity is accompanied by medication leading to polypharmacy in multimorbid patients. Whereas comorbidity is difficult to eliminate, discontinuing inappropriate medication may offer an attractive modifiable risk. Aim To investigate the effect of polypharmacy and comorbidity on overall survival (OS) in patients with LC. Methods We retrieved data on all Danish patients registered with chronic lymphocytic leukemia (CLL) from the Danish CLL Register since 2008 and with mantle cell lymphoma (MCL), diffuse large B-cell lymphoma (DLBCL), and follicular lymphoma (FL) from the Danish Lymphoma Register since 2005. All patients were diagnosed before June 2019; those with more than one LC diagnosis were excluded. Baseline data were used to calculate international prognostic indices (IPI). International Classification of Diseases (ICD10) diagnosis codes were retrieved from the Danish Patient Register, and Charlson comorbidity index (CCI) scores were calculated (Quan et al., Am J Epidemiol, 2011) at time of LC diagnosis. All patients were given a hematological CCI score of 2. Information on medication prescribed within the year up until LC diagnosis was retrieved from the Danish Prescription Register, and the number of drug classes (using the third ATC level, e.g. C03) was used to define polypharmacy as at least 5 drug classes. OS was calculated from time of diagnosis until death or end of follow-up. Multivariable analyses were equally adjusted for CCI, polypharmacy, and appropriate IPI. Results We identified 15,314 patients with LC including 5155 with CLL, 953 with MCL, 6182 with DLBCL, and 3024 with FL. The median age was 69 years (IQR 60;76). CLL-IPI and R-IPI are provided in the Figure. MIPI was low, intermediate, and high in 137 (15%), 297 (32%), and 493 (53%) patients, respectively, and FLIPI2 was low, intermediate, and high in 391 (16%), 1469 (62%), and 518 (22%), respectively. The median CCI score was 2 (including the hematological CCI score of 2, IQR 2;2). Most common comorbidities included chronic pulmonary disease (5%), congestive heart failure (CHF; 3%), rheumatic disease (RD; 3%), and metastatic cancer (MC; 2%). MC (3% vs 1-2%), RD (4% vs 2%), and AIDS (0.3% vs 0.0%) were significantly more prevalent in DLBCL as compared with other LCs ( P≤0.00054). Patients had most frequently been prescribed antibacterial (J01; 50%), analgesic (N02; 40%), renin-angiotensin system (C09; 33%), antithrombotic (B01; 32%), lipid modifying (C10; 30%), anti-inflammatory/rheumatic (M01; 29%), dyspepsia related (A02; 27%), and diuretic (C03; 27%) drugs. Antibacterials (54% vs 46-51%), analgesics (46% vs 33-36%), dyspepsia related (32% vs 22-28%), anti-inflammatory/antirheumatic (33% vs 24-28%), and systemic corticosteroid (H02; 13% vs 6-8%) drugs were being prescribed significantly more often in patients with DLBCL as compared with those with other LCs ( P<0.0001). In multivariable analyses adjusted for disease specific IPIs and CCI (2 vs >2), polypharmacy (<5 vs ≥5) was an independent prognostic marker of OS in CLL (hazard ratio [HR] 1.7, 95% confidence interval [CI]; 1.5;2.0), MCL (HR 1.4, 95% CI 1.2;1.7), DLBCL (HR 1.6, 95% CI 1.5;1.8), and FL (HR 1.8, 95% CI 1.6;2.1; P<0.0001). Using polypharmacy as a continuous variable (0-2 vs 3-4 vs 5-6 vs 7-8 vs 9-10 vs >10), we further demonstrated a dose-response effect on OS in CLL, DLBCL (Figure), MCL, and FL (data not shown). Similar to CLL and DLBCL, FL with >2 drug classes were significantly associated with shorter OS, whereas in MCL, polypharmacy groups were small ( n≤262) and only significantly associated with shorter OS for groups with ≥5 drug classes. Conclusion Polypharmacy is an important prognostic marker of OS independent of the CCI and IPIs in CLL, MCL, DLBCL, and FL. Polypharmacy is most likely a result of an underlying comorbidity, still informing the prognosis further than register based CCI scores. Further studies will seek to clarify the extent of medication that may be considered for discontinuation in newly diagnosed patients. Moreover, these results indicate that consistent collection of data on both comorbidity and polypharmacy at baseline in clinical trials may improve subgroup analyses and clinical translation of results. This is particularly important when extrapolating clinical trial results to ineligible populations.

Clinical Outcomes Associated with Del(17p) in Newly Diagnosed Multiple Myeloma Treated with Triplet and Daratumumab-Based Quadruplet Induction Regimens

Background: del(17p) is an independent prognostic marker associated with poor clinical outcome identified in a number of studies and has been incorporated into risk stratification models for multiple myeloma (Palumbo A et al J Clin Oncol 2015). Recent studies have reported that the percentage of myeloma cells with del(17p) by fluorescent in situ hybridization (FISH) can have a significant impact on outcomes (Corre J et al. Blood 2021; Thakurta A et al. Blood 2019; Thanendrarajan S et al. Haematologica 2017). Different collaborative groups have recommended different cut-points for the presence of del(17p) with clinical significance ranging from ≥20% or >55% of affected cells. Herein, we examined clinical outcomes of newly diagnosed multiple myeloma (NDMM) patients with del(17p) treated with novel combination triplet and quadruplet induction regimens, including bortezomib, lenalidomide, dexamethasone (VRd), carfilzomib, lenalidomide, dexamethasone (KRd), +/- daratumumab (DVRd and DKRd). Methods: We conducted a chart review study of 578 NDMM patients treated with VRd, KRd, DVRd and DKRd at Memorial Sloan Kettering (MSK) from 1/1/2015 to 5/1/23. Cutoff date for analysis was 7/19/23. Only patients with available cytogenetic results (FISH and/or SNP-array) were included. Patients who received ≤1 prior cycle of another regimen for MM were included. We analyzed the percentage of cells with del(17p) by inter-phase FISH analyses and/or SNP-array with CD138 enrichment when FISH was not available. A cut-point of 20% for del(17p) was used (del(17p) <20% vs del(17p) ≥20%) and compared with patients with standard-risk disease. Discrete patient characteristics were summarized by frequency (percentage) and continuous characteristics were summarized by median (Interquartile Range, IQR). PFS and OS were evaluated by Kaplan-Meier method. Results: There were 391 patients treated with triplet combination, of which 46 patients had del(17p) on baseline bone marrow biopsy. Among these patients, the presence of del(17p) in <20% of cells was detected in 13 patients, and the presence of del(17p) in ≥20% of affected cells was detected in 30 patients. Three patients did not have measurements. There were 187 patients treated with quadruplet combination, of which 20 had del(17p). Among these patients, 9 had del(17p) <20% of cells, 10 had del(17p) in ≥20% of cells, and one patient did not have measurements. There were 212 standard-risk NDMM patients treated with triplet induction and 99 patients treated with quadruplet regimens. The median age was 65 (IQR 54-73) for patients with del(17p) <20%, 61 (IQR 56-68) for del(17p) ≥20%, and 64 (IQR 56-69) for standard-risk patients (P=0.53). Other patient's baseline characteristics are summarized in Table 1. Best ORR among the three groups after triplet or quadruplet induction regimens were high: 95% for patients with del(17p) <20%, 93% for patients with del(17p) ≥20%, and 90% for standard-risk patients (P=0.53). After a median follow-up of 56.9 months, 2-year estimated PFS was 76% (95%CI, 59%-97%), 69% (95%CI, 55%-86%), and 85% (95%CI, 81%-90%) for del(17p) <20%, del(17p) ≥20%, and standard-risk groups, respectively. Median PFS was 48.9 months (95%CI, 26.9-not reached [NR]) for patients with del(17p) <20%, 34.3 months (29.8-66.7) for del(17p) ≥20%, and not reached (72.7-NR) for patients with standard-risk NDMM (P<0.0001). Median PFS was not significantly different between patients with del(17p) <20% and del(17p) ≥20% treated with triplet and quadruplet induction regimens (P=0.33). 2-year OS was 89% (95%CI 76%-100%), 85% (74%-98%), and 95% (93%-98%) for patients with del(17p) <20%, del(17p) ≥20%, and standard-risk (P=0.043). Conclusion: In the MSK experience, the presence of del(17p) was associated with poorer outcomes compared to standard-risk disease in patients with NDMM treated with both triplet and dara-based quadruplet induction regimens. We did not detect a significant difference in PFS and OS amongst del(17p) patients in our cohort using a cut-point of ≥20% or <20% of cells. These findings suggest that the presence of del(17p) alone portend poor prognosis, regardless of FISH cutoff threshold utilized. Data on TP53 mutation status and longer follow-up outcomes will be presented at the meeting.

Post-CAR-T Minimal Residual Disease (MRD) Monitoring in Mantle Cell Lymphoma Enables Early Relapse Detection

INTRODUCTION: Over 40% of patients with mantle cell lymphoma (MCL) will progress within 1 year after treatment with brexucabtagene autoleucel (Brexu-cel)(Michael Wang et al. ZUMA-2 NEJM 2020). Monitoring minimal residual disease (MRD) of circulating tumor cells (CTC) via next-generation sequencing of the immunoglobulin sequences (IgHTS) with a sensitivity of 1 in million (ClonoSeq, Adaptive Biotechnologies) has previously been shown to identify patients at high risk of MCL relapse (Simone Ferrero et al. Blood 2022). However, there is a limited understanding of MRD monitoring after Brexu-cel. A sub-analysis of 27 patients post-Brexu-cel in ZUMA-2 showed predictive performance of MRD measured at months 1, 3, and 6 in estimating relapse potential. METHODS: Clonotypes were identified from paraffin-embedded tissues and underwent PCR amplification of the IgH, IgH-DJ, and Ig Kappa/lambda regions. CTC MRD assessments were monitored at 1, 3, 6, and 12 months (if applicable) following standard-of-care brexu-cel infusion in all patients with MCL at our center who had at least 3 months of follow-up. PFS and overall survival (OS) were determined by Kaplan-Meier analysis and significance by log-rank test. The comparison between the two groups was conducted using a two-tailed Mann-Whitney test. Significances were calculated using a Fisher exact test between two categorical variables or as otherwise specified. Statistical analysis and plots were generated using Prism 8.4.1 (GraphPad) RESULTS: Between January 2020 and December 2022, all treated patients (n=34) were analyzed. 3 patients were excluded as they were unable to reach day 28 (D28) assessments due to death, progressive disease, or refusal of CAR-T infusion. 5 patients were excluded due to a follow-up of <3 months. The median follow-up time for the 26 included patients was 18.9 months. The median age was 68, 73% were male and 27% were female, 4 median lines of therapy prior, including AutoHSCT (34.6%) or BTKi (92%). The best overall response rate at D28 was 100% with a CR rate of 73% which was similar to ZUMA-2. The median OS was 18.9 months and the median PFS was 15.3 months. We chose to analyze the Day 28 MRD assessment and found 17 of 23 patients (74%) had undetectable MRD and 6 patients had detectable MRD (26%). Due to the relatively small sample size, no significant differences were found between Age, Gender, Pre-treatment LDH, MIPI score, Ki-67, TP53 mutational status, and probability of Day 28 undetectable MRD. Notably, however, 3 of the 6 patients with TP53 mutations had detectable Day 28 MRD. Those patients with detectable MRD compared with those with undetectable MRD had a lower median PFS of 10.74 months vs 17.69 months (p-value= 0.21, Figure 1); although the median OS was similar of 17.69 and 17.84 months, (p-value= 0.75) respectively. 88% of patients (15 out of 17) with undetectable MRD on Day 28 not only maintained this status at 6 months but also showed no radiological disease progression in 93% (14 out of 15) of cases at the time of longest follow-up. A total of 7 patients relapsed during follow-up. MRD predicted 6 impending relapses earlier than PET CT with a median of 294 days (range 15-332 days) prior to progression (Figure 2). CONCLUSION: Evidence from the ZUMA-2 study and our study highlights the significance of early post-CAR-T MRD as an independent prognostic marker for predicting disease recurrence and durable remissions in MCL. Combining MRD measurements with PET-CT at months 1, 3, and 6 after CAR-T therapy can effectively identify high-risk patients for disease progression, offering promising potential to improve outcomes in MCL.

The impact of the association between Val16Ala-SOD2 SNP and SOD2 immunohistochemistry expression in the prognosis of patients with esophageal cancer

IntroductionEsophageal cancer is an extensive public health issue worldwide, warranting the search for biomarkers related to its risk and progression. Previous studies have indicated an association between Val16AlaSOD2 single nucleotide polymorphism in the gene encoding the enzyme superoxide dismutase 2 and esophageal cancer. However, further investigations are needed to clarify its role in disease risk and progression. ObjectiveTo investigate the role of Val16AlaSOD2-SNP in esophageal cancer progression and in the survival of patients MethodsTumor samples were utilized for Val16Ala-SNP genotyping, while SOD2 expression levels in tissue were assessed using immunohistochemistry. A SOD2 Val16Ala-SNP database was used to obtain information on the genotype of healthy individuals. Risk and overall survival analyzes were performed. ResultsThe Val16Ala SNP was associated with an increased risk of esophageal cancer (RR 2.18, 95%CI 1.23–3.86), regardless of age and gender, but did not have a significant effect on patient survival. In contrast, weak SOD2 expression demonstrated a significantly associated with poor overall survival after treatment, independent of other clinicopathological variables (HR, 0.41; 95% CI, 0.22–0.79 P = 0.007). ConclusionsVal16Ala SNP was positively associated with esophageal cancer, and the expression of SOD2 was an independent prognostic marker.

Baseline metastatic growth rate is an independent prognostic marker in patients with advanced BRAF V600 mutated melanoma receiving targeted therapy

BackgroundTargeted therapy (TT) of BRAF V600 mutated unresectable melanoma with inhibitors of the MAPK pathway achieves response rates of up to 76%, but most patients develop secondary resistance. Albeit TT is strikingly efficacious during the first days of treatment, even in advanced cases, long-term survival is highly unlikely, especially in patients with unfavorable baseline characteristics like elevated lactate dehydrogenase (LDH). In patients treated with anti-PD-1 immune checkpoint inhibitors, elevated baseline metastatic growth rate (MGR) was the most important prognostic factor. Here, we aimed at investigating the prognostic impact of MGR in patients with unresectable melanoma receiving TT. MethodsClinical records of 242 patients with at least one measurable target lesion (TL) receiving TT at seven skin cancer centers were reviewed. Baseline MGR was determined measuring the largest TL at baseline and at one earlier timepoint. ResultsOverall survival (OS) and progression-free survival (PFS) were significantly impaired in patients with an MGR > 3.9 mm/month (median OS: 11.4 vs. 35.5 months, P < 0.0001; median PFS: 4.8 vs. 9.2 months, P < 0.0001). Multivariable analysis of OS and PFS revealed that the prognostic impact of elevated MGR was independent of LDH, presence of brain and liver metastases, tumor burden, and line of treatment. The prognostic significance of elevated MGR was highest in patients with normal LDH. ConclusionsBaseline MGR is an important independent prognostic marker for OS and PFS in melanoma patients treated with TT. Its implementation in clinical routine is easy and could facilitate the prognostic stratification.

NEIL3 promotes cell proliferation of ccRCC via the cyclin D1-Rb-E2F1 feedback loop regulation

Nei endonuclease VIII-like 3 (NEIL3), a novel tumor-related gene, is differentially expressed and involved in pathophysiological processes in multiple tumors. However, the potential biological functions and molecular mechanisms of NEIL3 in human clear cell renal cell carcinoma (ccRCC) have not been identified. In this research, we demonstrated that NEIL3, transcriptionally activated by E2F1, served as an oncogene to facilitate cell proliferation and cell cycle progression and contribute to tumorigenesis via the cyclin D1-Rb-E2F1 feedback loop in ccRCC. First, we found that NEIL3 expression was upregulated in ccRCC tissues and cell lines compared with matched adjacent nontumor tissues and renal tubular epithelial cells and was also positively correlated with adverse clinicopathological characteristics, such as advanced cancer stages and higher tumor grades, and acted as an independent prognostic marker in ccRCC. Mechanistically, we demonstrated that NEIL3 promoted cell proliferation, DNA replication and cell cycle progression in vitro and tumor growth in vivo. Furthermore, we found that NEIL3 overexpression activated the cyclin D1-Rb-E2F1 pathway, and the E2F1 upregulation transcriptionally activated NEIL3 expression, thus forming a feedback loop. In addition, there was a positive correlation between NEIL3 and E2F1 expression in clinical specimens of ccRCC. Taken together, our results suggest that NEIL3 serves as a proto-oncogene in ccRCC and presents as a novel candidate for ccRCC diagnosis and treatment.

Discovery of a new marker to identify myeloid cells associated with metastatic breast tumours

BackgroundMyeloid cells play an essential role in cancer metastasis. The phenotypic diversity of these cells during cancer development has attracted great interest; however, their functional heterogeneity and plasticity have limited their role as prognostic markers and therapeutic targets.MethodsTo identify markers associated with myeloid cells in metastatic tumours, we compared transcriptomic data from immune cells sorted from metastatic and non-metastatic mammary tumours grown in BALB/cJ mice. To assess the translational relevance of our in vivo findings, we assessed human breast cancer biopsies and evaluated the association between arginase 1 protein expression in breast cancer tissues with tumour characteristics and patient outcomes.ResultsAmong the differentially expressed genes, arginase 1 (ARG1) showed a unique expression pattern in tumour-infiltrating myeloid cells that correlated with the metastatic capacity of the tumour. Even though ARG1-positive cells were found almost exclusively inside the metastatic tumour, ARG1 protein was also present in the plasma. In human breast cancer biopsies, the presence of ARG1-positive cells was strongly correlated with high-grade proliferating tumours, poor prognosis, and low survival.ConclusionOur findings highlight the potential use of ARG1-positive myeloid cells as an independent prognostic marker to evaluate the risk of metastasis in breast cancer patients.Graphical

Epigenetic alterations in urothelial bladder cancer associated with disease outcomes.

Bladder cancer (BLCA) is a molecular heterogeneous disease with known genetic distinctive signatures. However, DNA methylation is highly prevalent across a wide range of tumors, suggesting its potential in oncogenesis. Here, we aimed to interrogate the role of nine epigenetic alterations as diagnostic and prognostic markers in BLCA. DNA methylation, gene expression, and clinicopathological information were retrieved from The Cancer Genome Atlas data portal. Methylation values and gene expression were assessed to determine their association with normal and malignant tissue. Additionally, we studied the association between methylation values and clinicopathological variables. For the prognostic model, Kaplan-Meier Survival curves were generated. Lastly, univariate and multivariate analysis were performed to evaluate the simultaneous impact of methylation and clinicopathological variables on the risk of tumor progression and survival. Nine CpG sites' methylation -values involved in our study demonstrated different methylation signatures between normal and malignant urothelium. Hypermethylated CpGs were overrepresented in tumor tissue (p < 0.0001). Opposingly, 4 CpG sites showed lower methylation values in tumor samples (p < 0.0001). Cg12743248high and cg17192862low are risk factors for progression-free survival, whereas cg12374721high (HR:3.003 (1.283-7.030)) also demonstrated to be the most valuable independent risk factor for disease progression and a risk factor for overall survival. We have identified that methylated cg12374721 shows promise as a diagnostic and independent prognostic marker in BLCA progression.

Overexpression of VEGFA mediated by HIF-1 is associated with higher rate of spread through air spaces in resected lung adenocarcinomas.

Spread through air spaces (STAS), a newly identified pattern of invasion in lung adenocarcinomas (LACs), is an unfavorable prognostic factor for patients with LAC, but the molecular characteristics and mechanisms underlying STAS have not been adequately explored. In total, 650 pathologically confirmed invasive LAC patients who underwent curative resection between December 2019 and April 2020 were reviewed. Disease-free survival (DFS) and overall survival (OS) were analyzed using the log-rank test and the Cox proportional hazards model. A comparative deep sequencing analysis was conducted to explore the molecular characteristics underlying STAS. Vascular endothelial growth factor A (VEGFA) expression was evaluated by immunoblotting and immunohistochemical analysis using fresh tumor tissue and tissue microarray. STAS was more prevalent in patients with a smoking history (p < 0.001), high pathological TNM stage (p < 0.001), lymphovascular invasion (p < 0.001), visceral pleural invasion (p < 0.001) and micropapillary/solid histological subtypes (p < 0.001). STAS-negative patients had better DFS (p < 0.001) and OS (p = 0.003) compared to STAS-positive patients with invasive LACs, especially in the lymph node-negative population (p < 0.001). After RNA-sequencing analysis, hypoxia-inducible factor-1 (HIF-1) signaling was enriched and appeared to be strongly correlated with STAS, and more STAS-positive individuals were detected in the higher VEGFA-expressing group (p = 0.042). We demonstrated that STAS was an independent prognostic marker of poor clinical outcome, especially in lymph node-negative patients, and that higher VEGFA expression mediated by HIF-1 signaling was associated with an increased STAS rate.

Could mesangial C3 deposition be an independent prognostic marker in immunoglobulin A nephropathy?

Immunoglobulin A nephropathy (IgAN) is a common primary glomerulonephropathy. There is evidence that mesangial C3 deposition plays a role in the development of the disease. The aim of this study was to examine the effect of C3 deposition on the prognosis of IgAN patients. The study included 1135 patients with biopsy-confirmed IgAN from the database of the Turkish Nephrology Association Glomerular Diseases Working Group (TSN-GOLD). Patients were excluded from the study if they were aged < 18 or > 75years or if C3 staining had not been performed in the immunofluorescent analysis. C3 deposition was defined as an immunofluorescence intensity of C3 ≥ 2 + within the mesangium. The primary endpoints were the development of end-stage renal disease, a 30% decrease in glomerular filtration rate compared to the basal value or an elevation in proteinuria to a nephrotic level (3.5 gr/day). Mesangial C3 deposition was observed in 603 (53.1%) patients. No statistically significant difference was foundatbaselinebetween the groups with and without mesangial C3deposition, as for age, sex, BMI, proteinuria level, or the presence of hypertension. In the follow-up period with a meanduration of 78months, no significant difference was found between the two groups regarding the primary endpoints (p = 0.43). A significant correlation between C3 deposition and segmental glomerulosclerosis (S1) according to the Oxford MEST-C classification was found (p = 0.001). Although a correlation was observed between mesangial C3 deposition and the S1 MEST-C classification, mesangial C3 deposition was not aprognostic factor in IgAN.

BIOM-54. VALIDATING MCM2 IMMUNOHISTOCHEMISTRY AS A SURROGATE MARKER FOR AGGRESSIVE MENINGIOMA

Abstract Increasing efforts have focused to improve meningioma classification through various multi-omic analyses. One such recent study (Nassiri, et al, Nature, 2021) proposed an integrative molecular classification for meningiomas that included 4 molecular subtypes (MG1-4), with possible immunohistochemical surrogates for each subtype including S100B, ACADL, SCGN, and MCM2, respectively. While these immunohistochemical surrogates have been proposed for meningioma risk-stratification, their clinical utility has not been thoroughly tested. In this study, we sought to evaluate the utility of these immunohistochemical markers for risk-stratification in an independent cohort of meningioma. Tissue microarrays containing meningioma tissues were constructed, immunohistochemically stained, and blindly scored. Our cohort included n = 142 tumors (CNS WHO: 1=93, 2=45, 3=4). Median age was 49 years (range 14-80 years) with 89 female and 53 male patients. We found that MCM2 immunopositivity was a univariate marker of poor prognosis (median progression-free survival 1.2 years) when compared to negative tumors (median progression-free survival 13.0 years). The other three markers (S100B, ACADL, SCGN) were not associated with survival. MCM2 staining held up as a significantly independent (p = 0.03) poor prognostic marker on multivariate Cox proportional hazard modeling including other histopathological features: CNS WHO grade, brain invasion, mitotic index, and tumor size. Although MCM2 staining was independent of CNS WHO grade, all 4 of the CNS WHO 3 meningiomas were positive for MCM2. MCM2-positive tumors had a higher average mitotic count (6.1 mitoses/10hpf) compared to MCM2-negative tumors (1.8 mitoses/10hpf) (p = 0.0002, two-tailed t test). There was no difference between MCM2-positive and MCM2-negative meningiomas in relation to maximum tumor size (4.9cm and 4.2cm, respectively) and mean age (51.71 years and 50.34 years, respectively). In summary, in our cohort we found MCM2 immunopositive staining (surrogate marker for high-risk molecular subtype M4) to be an independently poor prognostic biomarker in meningioma when accounting for other well-established histopathological characteristics.

NIMG-65. METHYLTHIOADENOSINE PHOSPHORYLASE STATUS CORRELATES WITH CLINICAL OUTCOME AND11C-METHIONINE UPTAKE IN ASTROCYTOMA, IDH-MUTANT

Abstract PURPOSE Methylthioadenosine phosphorylase (MTAP) immuno histochemistry staining has been proposed as a surrogate of Cyclin-dependent kinase inhibitor 2A homozygous deletion (CDKN2A HD) in various types of tumors. This study was designed to determine whether MTAP status correlates with clinical outcome and uptake of 11C-methionine in astrocytoma, IDH-mutant. METHODS We conducted a MTAP immunohistochemistry staining of 30 astrocytoma, IDH-mutant patients who underwent 11C-methionine positron emission tomography scans prior to surgical resection from 2000 to 2020. The tumor-to-normal (T/N) ratio of 11C-methionine uptake was calculated by dividing the mean standardized uptake value (SUV) for the tumor by the mean SUV of the normal brain. RESULTS Eight (26.7%) of the thirty astrocytoma, IDH-mutant patients exhibited loss of cytoplasmic MTAP expression, which is accompanied with a poor prognosis. The median progression free survival (PFS) in astrocytoma, IDH-mutant patients with loss of MTAP was 1.88 years and significantly shorter than that of those with MTAP retention (6.80 years, p = 0.003). The median Overall survival in astrocytoma, IDH-mutant patients with loss of MTAP was 5.23 years, again significantly shorter than that of those with MTAP retention (10.69 years, p = 0.0191). Multivariate analysis revealed that MTAP status was an independent prognostic marker for PFS in astrocytoma, IDH-mutant patients. The median T/N ratio in tumors with loss of cytoplasmic MTAP expression was 2.12 (IQR 1.92-2.50), i.e., significantly higher than that of tumors with MTAP retention (1.65, IQR 1.23-1.94, p = 0.0116, U test). Thirteen (59.1%) of twenty-two patients with MTAP retention recurred and nine patients underwent a second surgery after radiation treatment with or without chemotherapy. Three (33.3%) of the nine recurrent patients showed loss of MTAP immunoreactivity, suggesting a role for this loss in tumor progression. CONCLUSION Our study showed MTAP status to correlate with clinical outcome and T/N ratio of 11C-methionine in astrocytoma, IDH-mutant patients.

IMMU-08. EOSINOPHILS ARE IMPORTANT MODULATORS OF AN IMMUNE-RESPONSIVE TUMOR-MICROENVIRONMENT IN GLIOBLASTOMA

Abstract Eosinophils, bone marrow-derived granulocytes involved in allergic asthma and autoimmunity, have attracted increasing attention due to their assumed role in effective responses to immune checkpoint blockade in some extracranial tumors. However, the role of this rare type of immune cells in immunologically cold tumors such as glioblastoma (GBM) has not been studied so far. When quantifying immune infiltrates and levels of 30 immune-relevant cytokines in 60 matched pairs of human primary and recurrent GBM, strongest correlations (r= 0.71-0.93) were observed for the eosinophil-associated cytokines Eotaxin and Il-5 as well as for TNFa, IFNg and Il2 which are important cytokines for cytotoxic T cell responses. A combined score of these cytokines (Eo score) turned out to be an independent prognostic marker for improved overall survival of GBM patients. Furthermore, multicolor immunofluorescent staining revealed that Eo score high GBM are significantly higher infiltrated by effector T cells and show a significantly lower infiltration with anti-inflammatory M2-like tumor-associated macrophages / microglia cells (M2-TAM) in recurrent GBM. Flowcytometry analysis of GBM-derived single cell suspensions confirmed a positive correlation of higher eosinophil numbers with higher numbers of tumor-infiltrating T cells, while an inverse relation between eosinophils and monocytes could be demonstrated in the TCGA data set. Spatial transcriptomic data of human GBM confirmed that areas with a higher eosinophil infiltration showed higher numbers of T cells and pro-inflammatory M1-TAM, lower numbers of M2-TAM and a remarkable vessel normalization. Finally, in a newly developed patient-derived GBM organoid model we could corroborate the functional impact of eosinophils on the tumor-microenvironment. An increase of eosinophil numbers in these patient avatars resulted in a significantly decreased number of M2-TAM. Altogether, tumor-infiltrating eosinophils seem to be a novel type of immune-modulator in GBM and may enhance immune responses by shaping the tumor niche from an immunosuppressive to an immunepermissive microenvironment.

The Clinical Relevance of Selected Cytokines in Newly Diagnosed Multiple Myeloma Patients.

Multiple myeloma (MM) is the second most common hematological neoplasm. Cytokines, chemokines, and their receptors, induced by the microenvironment of MM, participate in tumor growth, the attraction of leukocytes, cell homing, and bone destruction. This study aimed to assess the correlation between the pretreatment serum concentrations of interleukin-6 (IL-6), interleukin-8 (IL-8), angiogenic chemokine monocyte chemoattractant protein-1 (MCP-1), and vascular endothelial growth factor (VEGF) and the clinical outcomes and survival of patients newly diagnosed with MM. The study group consisted of 82 individuals. The IL-8 concentration was significantly positively correlated with the age of onset (p = 0.007), the International Staging System (ISS) stage (p = 0.03), the Eastern Cooperative Oncology Group (ECOG) performance status (p < 0.001), the degree of anemia before treatment (p < 0.0001), the degree of kidney disease (p < 0.001), and VEGF (p = 0.0364). Chemotherapy responders had significantly lower concentrations of IL-8 (p < 0.001), IL-6 (p < 0.001), and VEGF (p = 0.04) compared with non-responders. Patients with treatment-induced polyneuropathy had significantly higher levels of IL-8 (p = 0.033). Patients with a high level of IL-6 had a 2-fold higher risk of progression-free survival (PFS) reduction (17 vs. 35 months; HR = 1.89; p = 0.0078), and a more than 2.5-fold higher risk of overall survival (OS) reduction (28 vs. 78 months; HR = 2.62; p < 0.001). High levels of IL-6, IL-8, and VEGF demonstrated significant predictive values for some clinical conditions or outcomes of newly diagnosed MM patients. Patients with an early response to chemotherapy had a significantly lower concentration of these cytokines. A high pretreatment IL-6 concentration was an independent negative prognostic marker for newly diagnosed MM patients.