Negative Prognostic Biomarker Research Articles

Broad Superiority of Zanubrutinib (Zanu) Over Bendamustine + Rituximab (BR) Across Multiple High-Risk Factors: Biomarker Subgroup Analysis in the Phase 3 SEQUOIA Study in Patients With Treatment-Naive (TN) Chronic Lymphocytic Leukemia (CLL)/Small Lymphocytic Lymphoma (SLL) without del(17p)

Introduction Zanu is a Bruton tyrosine kinase (BTK) inhibitor with high potency, selectivity, and efficacy and a favorable toxicity profile. In the phase 3 SEQUOIA study (NCT03336333; Tam et al. Lancet Oncol. 2022), zanu treatment demonstrated superior progression-free survival (PFS) in TN patients (pts) with CLL/SLL and without del(17p) (cohort 1) compared with BR treatment (hazard ratio [HR], 0.42; 95% CI, 0.28-0.63; 2-sided P<.0001). We evaluated PFS in both treatment arms in pt subgroups based on various biomarkers, including several known negative prognostic factors for CLL. Methods Details of the SEQUOIA study were described previously (Tam et al. Lancet Oncol. 2022). A total of 479 pts in cohort 1 were randomized to either zanu (n=241) or BR (n=238). Blood (CLL) or bone marrow (SLL) samples collected at screening were used for fluorescence in situ hybridization for chromosome abnormalities, cytogenetic analysis for complex karyotype (CK), next-generation sequencing (NGS) per the European Research Initiative on CLL (ERIC) for immunoglobulin heavy chain variable (IGHV) gene mutations and expressed clones, and ultrasensitive targeted NGS for mutation analysis of 106 genes. For NGS, all pathogenic mutations with variant allele frequency ≥1% were analyzed. The association between biomarkers and PFS was quantified using the log-rank test and HR and summarized by the Kaplan-Meier method. Data cutoff was October 31, 2022. Results Overall, the CLL features analyzed were similarly distributed between both treatment arms (Table). In pts with cytogenetic abnormalities, zanu demonstrated significantly better PFS than BR in those with del(11q) ( P<.001), del(13q) ( P<.001), trisomy 12 ( P<.01), or CKT ≥3 ( P<.01) (Table). Furthermore, zanu treatment conferred a similar PFS benefit to pts with the negative prognostic factor, del(11q) ( P=.05) or intermediate prognostic factor, trisomy 12 ( P=.40), compared to those without these abnormalities (Table). Notably, in the zanu arm, CKT ≥3 was not associated with worse PFS ( P=.18; Table). Pts with either mutated IGHV (mIGHV) or unmutated IGHV (uIGHV) in the zanu arm had significantly better PFS than pts in the BR arm (uIGHV: P<.0001; mIGHV: P<.01; Table); analysis was confirmed by an ERIC-certified provider. Furthermore, PFS was unaffected by IGHV mutational status in pts treated with zanu ( P=.12). In pts with uIGHV, IGHV1-69 was the most prevalent expressed clone in both arms (zanu: 24.6% [31/126]; BR: 30.7% [39/127]). In the present study, pts with uIGHV harboring IGHV1-69 clones showed significantly better PFS with zanu vs BR ( P<.0001). Across all pts, the most frequently mutated genes were ATM (zanu: 11%; BR: 16%), SF3B1 (zanu: 17%; BR: 21%), NOTCH1 (zanu: 20%; BR: 20%), and BRAF (zanu: 11%; BR: 7%; Table). Pts with mutations associated with poor prognosis in CLL had significantly better PFS with zanu than with BR (eg, ATM [ P=.02], BRAF [ P=.01], NOTCH1 [ P<.001], SF3B1 [ P<.001]; Table). Furthermore, zanu-treated pts with or without mutations in ATM ( P=.58), NOTCH1 ( P=.38), and SF3B1 ( P=.39) had similar PFS. Conclusions TN del(17p)-negative pts with CLL/SLL were evenly distributed between zanu and BR arms in SEQUOIA cohort 1 based on the biomarkers analyzed. PFS in zanu-treated pts was superior to that of BR-treated pts in all biomarker subgroups analyzed, including some known negative prognostic markers in CLL such as del11q, CKT≥3, and uIGHV. Importantly, within the zanu-treatment arm, pts with negative prognostic biomarkers showed comparable PFS benefit to pts without those markers. Like with other BTK inhibitors, ibrutinib and acalabrutinib, IGHV mutational status did not affect PFS outcome (Bartosz et al. Hematology in Clinical Practice 2022). However, contrary to what has been reported for TN or relapsed/refractory pts treated with ibrutinib-based regimens, CKT ≥3 was not associated with worse PFS in zanubrutinib-treated patients (Rigolin et al. Blood. 2021; Thompson et al. Cancer. 2015). This study provides further evidence that zanu is a valuable first-line treatment option for pts with CLL/SLL.

Stroma AReactive Invasion Front Areas (SARIFA) proves prognostic relevance in gastric carcinoma and is based on a tumor–adipocyte interaction indicating an altered immune response

BackgroundRecently, we presented Stroma AReactive Invasion Front Areas (SARIFA) as a new histomorphologic negative prognostic biomarker in gastric cancer. It is defined as direct contact between tumor cells and fat cells. The aim of this study was to further elucidate the underlying genomic, transcriptional, and immunological mechanisms of the SARIFA phenomenon.MethodsTo address these questions, SARIFA was classified on H&E-stained tissue sections of three cohorts: an external cohort (n = 489, prognostic validation), the TCGA-STAD cohort (n = 194, genomic and transcriptomic analysis), and a local cohort (n = 60, digital spatial profiling (whole transcriptome) and double RNA in situ hybridization/immunostaining of cytokines).ResultsSARIFA status proved to be an independent negative prognostic factor for overall survival in an external cohort of gastric carcinomas. In TCGA-STAD cohort, SARIFA is not driven by distinct genomic alterations, whereas the gene expression analyses showed an upregulation of FABP4 in SARIFA-positive tumors. In addition, the transcriptional regulations of white adipocyte differentiation, triglyceride metabolism, and catabolism were upregulated in pathway analyses. In the DSP analysis of SARIFA-positive tumors, FABP4 and the transcriptional regulation of white adipocyte differentiation were upregulated in macrophages. Additionally, a significantly lower expression of the cytokines IL6 and TNFα was observed at the invasion front.ConclusionsSARIFA proves to be a strong negative prognostic biomarker in advanced gastric cancer, implicating an interaction of tumor cells with tumor-promoting adipocytes with crucial changes in tumor cell metabolism. SARIFA is not driven by tumor genetics but is very likely driven by an altered immune response as a causative mechanism.

Prognostic Significance of Serum Lipids in Patients with Non-Small Cell Lung Cancer Treated with Radiotherapy: A Multicenter Prospective Study

Although lipids have been assessed for their possible roles in cancer survival prediction, studies on the association between serum lipids levels and the prognosis of non-small cell lung cancer (NSCLC) patients are limited. This study aimed to evaluate whether serum lipids are associated with outcomes in patients with NSCLC treated with radiotherapy. We conducted a multicenter prospective study on patients diagnosed with NSCLC between January 2018 and February 2021. Participants received thoracic radiotherapy of 60ཞ80 Gy to the primary lung tumor and positive lymph node metastases. We measured patients' serum lipids levels (serum triglyceride, TGs; total cholesterol, TC, high density lipoprotein cholesterol, HDL-C; low density lipoprotein cholesterol, LDL-C) before radiotherapy. The association between serum lipids levels and overall survival (OS) was evaluated using hazard ratios. We sought to determine a threshold point using optimal stratification. Survival analysis was performed using Kaplan-Meier curves. Of the 300 participants diagnosed with NSCLC treated with radiotherapy, 165 (55.0%) were men. Median follow-up time was 24.4 months (range 1.0- 101.9 months). Using univariate and multivariate Cox proportional hazard analysis, among those serum lipids, only serum TG was shown to be independent prognostic factors for OS (hazard ratio: 1.203, 95% confidence interval: 1.038 - 1.393, p = 0.014). The cut-off for TG associated with OS was 2.04 mmol/L. Based on the TG cut-off value, 55 NSCLC patients were categorized into the high TG group (>2.04 mmol/L) and 245 in the low TG group (<2.04 mmol/L). The NSCLC patients in the low TG group exhibited higher OS than the high group (median OS, not reach vs 41.4 months, p = 0.025). TG levels were found to be a significant negative prognostic biomarker for OS in NSCLC patients treated with radiotherapy.

Atypical (non-V600E) BRAF mutations in metastatic colorectal cancer in population and real-world cohorts.

BRAF-V600E mutation (mt) is a strong negative prognostic and predictive biomarker in metastatic colorectal cancer (mCRC). Non-V600Emt, designated atypical BRAFmt (aBRAFmt) are rare, and little is known about their frequency, co-mutations and prognostic and predictive role. These were compared between mutational groups of mCRC patients collected from three Nordic population-based or real-world cohorts. Pathology of aBRAFmt was studied. The study included 1449 mCRC patients with 51 (3%) aBRAFmt, 182 (13%) BRAF-V600Emt, 456 (31%) RAS&BRAF wild-type (wt) and 760 (52%) RASmt tumours. aBRAFmt were seen in 2% of real-world and 4% of population-based cohorts. Twenty-six different aBRAFmt were detected, 11 (22%) class 2 (serrated adenocarcinoma in 2/9 tested), 32 (64%) class 3 (serrated in 15/25) and 4 (8%) unclassified. aBRAFmt patients were predominantly male, had more rectal primaries, less peritoneal metastases, deficient mismatch repair in one (2%), and better survival after metastasectomy (89% 5-year overall survival [OS]-rate) compared with BRAF-V600Emt. aBRAFmt and BRAF-V600Emt had poorer performance status and received fewer treatment lines than RAS&BRAFwt and RASmt. OS among aBRAFmt (median 14.4 months) was longer than for BRAF-V600Emt (11.2 months), but shorter than for RAS&BRAFwt (30.5 months) and RASmt (23.4 months). Addition of bevacizumab trended for better OS for the aBRAFmt. Nine patients with aBRAFmt received cetuximab/panitumumab without response. aBRAFmt represents a distinct subgroup differing from other RAS/BRAF groups, with serrated adenocarcinoma in only half. OS for patients with aBRAFmt tumours was slightly better than for BRAF-V600Emt, but worse than for RASmt and RAS&BRAFwt. aBRAFmt should not be a contraindication for metastasectomy.

The Sagittal Angle of the Trigeminal Nerve at the Porus Trigeminus is a Radiologic Predictor of Surgical Outcome in Microvascular Decompression for Classical Trigeminal Neuralgia.

Classical trigeminal neuralgia (cTN) is a painful disease. Microvascular decompression (MVD) provides immediate and durable relief in many patients. A variety of positive and negative prognostic biomarkers for MVD have been identified. The sagittal angle of the trigeminal nerve at the porus trigeminus (SATNaPT) is an MRI biomarker that can identify a subset of patients with cTN whose trigeminal nerve anatomy is different from normal controls. The purpose of this case-control study was to determine whether an abnormally hyperacute SATNaPT is a negative prognostic biomarker in patients with cTN undergoing MVD. Preoperative MRIs from 300 patients with cTN who underwent MVD were analyzed to identify patients with a hyperacute SATNaPT (defined as less than 3 SDs below the mean). The rate of surgical success (pain-free after at least 12 months) was compared between patients with a hyperacute SATNaPT and all other patients. Patients without a hyperacute SATNaPT had an 82% likelihood of surgical success, whereas patients with a hyperacute SATNaPT had a 58% likelihood of surgical success ( P < .05). Patients with a hyperacute SATNaPT who also had no evidence of vascular compression on preoperative MRI had an even lower likelihood of success (29%, P < .05). In patients with cTN being considered for MVD, a hyperacute SATNaPT is a negative prognostic biomarker that predicts a higher likelihood of surgical failure. Patients with a hyperacute SATNaPT, particularly those without MRI evidence of vascular compression, may benefit from other surgical treatments or a modification of MVD to adequately address the underlying cause of cTN.

Immune Inhibitory Molecule PD-1 Homolog (VISTA) Colocalizes with CD11b Myeloid Cells in Melanoma and Is Associated with Poor Outcomes

Tumors evade immunity through the overexpression of immune inhibitory molecules in the tumor microenvironment such as PD-L1/B7-H1. An immune inhibitory molecule named PD-1 homolog (also known as V-domain Ig-containing suppressor of T cell activation [VISTA]) functions to control both T cells and myeloid cells. Current clinical trials using anti-VISTA-blocking agents for treatment of cancer are ongoing. We sought to determine the extent of VISTA expression in primary cutaneous melanomas (n= 190), identify the critical cell types expressing VISTA, and correlate its expression with PD-L1 expression using multiplexed quantitative immunofluorescence. Within the tumor subcompartments, VISTA is most highly expressed on CD11b myeloid cells, and PD-L1 is most highly expressed on CD68 myeloid cells in our melanoma cohort. There is little correlation between VISTA and PD-L1 expression intensity, suggesting that individual tumors have distinct immunosuppressive tumor microenvironments. High levels of VISTA expression on CD11b myeloid cells but not PD-L1 expression were associated with greater melanoma recurrence and greater all-cause mortality. Our findings suggest that cell-specific VISTA expression may be a negative prognostic biomarker for melanoma and a future potential therapeutic target.

Pan-Cancer Profiling and Digital Pathology Analysis Reveal Negative Prognostic Biomarker ZPR1 Associated with Immune Infiltration and Treatment Response in Hepatocellular Carcinoma.

ZPR1 is a zinc finger-containing protein that plays a crucial role in neurodegenerative diseases, lipid metabolism disorders, and non-alcoholic fatty liver disease. However, the expression pattern, prognostic value, and treatment response of ZPR1 in pan-cancer and hepatocellular carcinoma (HCC) remain unclear. Pan-cancer expression profiles and relevant clinical data were acquired from UCSC Xena platform. Pan-cancer expression, epigenetic profile, and clinical correlation analysis for ZPR1 were performed. We next explored the prognostic significance and potential biological functions of ZPR1 in HCC. Furthermore, the relationship between ZPR1 and immune infiltration and treatment response was investigated. Finally, quantitative immunohistochemistry (IHC) analysis was applied to assess the correlation of ZPR1 expression and immune microenvironment in HCC tissues using Qupath software. ZPR1 was differentially expressed in most tumor types and significantly up-regulated in HCC. ZPR1 showed hypo-methylated status in most tumors. Pan-cancer correlation analysis indicated that ZPR1 was closely associated with clinicopathological factors and TMB, MSI, and stemness index in HCC. High ZPR1 expression could be an independent risk factor for adverse prognosis in HCC. ZPR1 correlated with immune cell infiltration and therapeutic response. Finally, IHC results suggested that ZPR1 correlated with CD4, CD56, CD68, and PD-L1 expression and is a promising pathological diagnostic marker in HCC. Immune infiltrate-associated ZPR1 could be considered a novel negative prognostic biomarker for therapeutic response in HCC.

Heterogeneity of PD-L1 expression among resected EGFR-mutated NSCLC and prognostic implications.

143 Background: The predictive value of PD-L1 score to select patients for adjuvant immunotherapy among epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC) remains controversial, confounded by the use of different assays across trials. We sought to compare the concordance between 3 PD-L1 antibodies among resected EGFR-mutated NSCLC, evaluate intra-tumoral heterogeneity (ITH) and implications on survival outcomes. Methods: Tissue microarrays were constructed from patients with AJCC7 Stage IA-IIIA EGFR-mutated NSCLC who underwent surgery 1/1/2010 – 31/12/2019 at National Cancer Centre Singapore. Patients who received neoadjuvant therapy were excluded. Three 1mm tumor cores per patient were obtained to study ITH, defined as discordant tumor proportion score (TPS) in ≥2/3 cores. TPS was evaluated by 3 independent pathologists for 3 PD-L1 antibodies (SP263, SP142, 22C3) and analysed in tertiles of &lt;1%, 1-49% and ≥50%. Median TPS score across 3 cores for each pathologist per assay was used to assess concordance between assays with Cohen’s kappa test, which was also used to assess agreement between pathologists. Disease-free survival (DFS) and overall survival (OS) were tabulated using Kaplan-Meier method. Results: A total of 218 patients were included. Stage IA comprised 45%(99/218), IB 24%(53/218), II 17%(37/218) and IIIA 13%(29/218). Females made up 65%(142/218), never smokers 84%(183/218) and 71%(155/218) were acinar subtype adenocarcinomas. Ex19del was detected in 47%(103/218) and L858R in 39%(86/218). Distribution by PD-L1 TPS was 85%(185/218) vs 91%(198/218) vs 97%(212/281) for &lt;1%, 13%(28/218) vs 8%(17/218) vs 3%(6/218) for 1-49%, and 2%(5/218) vs 1%(3/218) vs 0% for ≥50%, using SP263, 22C3 and SP142 respectively (p&lt;0.001). Pairwise comparison of TPS scores revealed highest concordance between SP263 and 22C3 (Kappa 0.631), followed by 22C3 and SP142 (Kappa 0.361), and SP263 and SP142 (Kappa 0.192). Agreement between pathologists was excellent across all 3 assays, with median Cohen’s kappa of 1 (range 0.972 – 1). ITH was observed in 13%(29/218), with 5 patients having differing TPS scores in all 3 cores. Discordance was highest among TPS 1-49% at 96%(27/28), followed by ≥50% at 40%(2/5) and least among &lt;1% at 0% (p&lt;0.01). Compared to those with concordant TPS scores across all 3 cores, 5-year DFS (60% vs 47%, p=0.15) and 5-year OS (81%% vs 71%, p=0.043) were inferior for patients with ITH. Conclusions: Concordance of PD-L1 TPS was highest among SP263 and 22C3 and lowest for SP263 and SP142, with low inter-observer variability between pathologists. Heterogeneous PD-L1 expression within tumors was seen in 13% of patients and significantly correlated with inferior OS. Our findings support the relative interchangeability of SP263 and 22C3 among resected EGFR-mutated NSCLC but not SP142, while ITH in PD-L1 expression may represent a novel negative prognostic biomarker.

The Localization of Intraretinal Cysts Has a Clinical Role on the 2-Year Outcome of Neovascular Age-Related Macular Degeneration

To assess the relationship between ≥ 1 localizations of intraretinal fluid (IRF) within retinal layers and the 2-year outcome in a cohort of neovascular age-related macular degeneration (AMD) eyes. Retrospective case series. Two hundred forty-three eyes of 243 AMD patients affected by type 1 and type 2 macular neovascularization (MNV). We analyzed data considering MNV onset, 1-year, and 2-year timepoints. Optical coherence tomography images were used to classify MNV types, distinguish different types of fluids and assess IRF localization within retinal layers. A subcohort of eyes were also analyzed by OCT angiography. The association between IRF cyst localization and both visual outcome and onset of outer retinal atrophy at 2-year follow-up. Macular neovascularizations were distributed as type 1 (69%) and type 2 (31%). The mean number of intravitreal injections was 7 ± 2 at 1-year follow-up and 5 ± 2 at 2-year follow-up. Baseline best-corrected visual acuity was 0.4 ± 0.3 logarithm of the minimum angle of resolution, improving to 0.3 ± 0.4 at 2-year follow-up (P < 0.01). Outer retinal atrophy occurred in 24% of cases at 1 year and 39% of cases at 2-year follow-up. Intraretinal fluid localizations at the level of IPL-INL and OPL-ONL at baseline were associated with the worst functional and anatomical outcome. Moreover, the presence of IRF at baseline was associated with greater impairment of the intraretinal vascular network. The localization of IRF at the level of IPL-INL and OPL-ONL retinal layers represents a negative prognostic biomarker for the morphologic and functional outcomes of neovascular AMD. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Relationship of Ki-67 index in biopsies of metastatic breast cancer tissue and circulating tumor cells (CTCs) at the time of biopsy collection

BackgroundThe proliferation marker Ki-67 is a major pathological feature for the description of the state of disease in breast cancer. It helps to define the molecular subtype and to stratify between therapy regimens in early breast cancer and helps to assess the therapy response. Circulating tumor cells (CTCs) are a negative prognostic biomarker for progression free (PFS) and overall survival (OS) in patients with metastatic breast cancer. Therefore, the CTC count is often described as surrogate for the tumor burden. Both, decrease of Ki-67 and CTC count are considered as evidence for therapy response. The presented work analyzed the correlation between the Ki-67 indices of metastatic tissue biopsies and CTC counts in biopsy time-adjacent peripheral blood samples.Patients and methodsBlood samples from 70 metastatic breast cancer patients were obtained before the start of a new line of systemic therapy. CTCs were enumerated using CellSearch® (Menarini Silicon Biosystems, Bologna, Italy) whereas intact CTCs (iCTCs) and non-intact or apoptotic CTCs (aCTCs) were distinguished using morphologic criteria. The proportion of cells expressing Ki-67 was evaluated using immunohistochemistry on biopsies of metastases obtained concurrently with CTC sampling before the start of a new line of systemic therapy.Results65.7% of patients had a Ki-67 index of > 25%. 28.6% of patients had ≥ 5, 47.1% ≥ 1 iCTCs. 37.1% had ≥ 5, 51.4% ≥ 1 aCTCs. No correlation was shown between Ki-67 index and iCTC and aCTC count (r = 0.05 resp. r = 0.05, Spearman’s correlation index). High CTC-counts did not coincide with high Ki-67 index. High Ki-67, ≥ 5 iCTCs and aCTCs are associated with poor progression free (PFS) and overall survival (OS).ConclusionCTCs and Ki-67 are independent prognostic markers in metastatic breast cancer. High Ki-67 in metastatic tumor tissue is not correlated to high iCTC or aCTC counts in peripheral blood.

Radiomic signatures from T2W and DWI MRI are predictive of tumour hypoxia in colorectal liver metastases

BackgroundTumour hypoxia is a negative predictive and prognostic biomarker in colorectal cancer typically assessed by invasive sampling methods, which suffer from many shortcomings. This retrospective proof-of-principle study explores the potential of MRI-derived imaging markers in predicting tumour hypoxia non-invasively in patients with colorectal liver metastases (CLM).MethodsA single-centre cohort of 146 CLMs from 112 patients were segmented on preoperative T2-weighted (T2W) images and diffusion-weighted imaging (DWI). HIF-1 alpha immunohistochemical staining index (high/low) was used as a reference standard. Radiomic features were extracted, and machine learning approaches were implemented to predict the degree of histopathological tumour hypoxia.ResultsRadiomic signatures from DWI b200 (AUC = 0.79, 95% CI 0.61–0.93, p = 0.002) and ADC (AUC = 0.72, 95% CI 0.50–0.90, p = 0.019) were significantly predictive of tumour hypoxia. Morphological T2W TE75 (AUC = 0.64, 95% CI 0.42–0.82, p = 0.092) and functional DWI b0 (AUC = 0.66, 95% CI 0.46–0.84, p = 0.069) and b800 (AUC = 0.64, 95% CI 0.44–0.82, p = 0.071) images also provided predictive information. T2W TE300 (AUC = 0.57, 95% CI 0.33–0.78, p = 0.312) and b = 10 (AUC = 0.53, 95% CI 0.33–0.74, p = 0.415) images were not predictive of tumour hypoxia.ConclusionsT2W and DWI sequences encode information predictive of tumour hypoxia. Prospective multicentre studies could help develop and validate robust non-invasive hypoxia-detection algorithms.Critical relevance statementHypoxia is a negative prognostic biomarker in colorectal cancer. Hypoxia is usually assessed by invasive sampling methods. This proof-of-principle retrospective study explores the role of AI-based MRI-derived imaging biomarkers in non-invasively predicting tumour hypoxia in patients with colorectal liver metastases (CLM).Graphical

Foveal Eversion is Associated with High Persistence of Macular Edema and Visual Acuity Deterioration in Retinal Vein Occlusion.

Foveal eversion (FE) is a recently described optical coherence tomography (OCT) finding associated with negative outcome in diabetic macular edema. The main goal of the present study was to investigate the role of the FE metric in the diagnostic workup of retinal vein occlusion (RVO). This study was a retrospective, observational case series. We included 168 eyes (168 patients) affected by central RVO (CRVO) and 116 eyes (116 patients) affected by branch (RVO). We collected clinical and imaging data from CRVO and BRVO eyes affected by macular edema with a minimum follow-up of 12months. On structural OCT, we classified FE as pattern 1a, characterized by thick vertical intraretinal columns, pattern 1b, presenting thin vertical intraretinal lines, and pattern 2, showing no signs of vertical lines in the context of the cystoid macular edema. For statistical purposes, we considered data collected at baseline, after 1 year and at the last follow-up. The mean follow-up was 40 ± 25months for CRVO eyes and 36 ± 24months for BRVO eyes. We found FE in 64 of 168 CRVO eyes (38%) and in 25 of 116 BRVO eyes (22%). Most of the eyes developed FE during the follow-up. For CRVO eyes, we found 6 eyes (9%) with pattern 1a, 17 eyes (26%) with pattern 1b and 41 eyes (65%) with pattern 2. Of those BRVO eyes with FE, we found 8 eyes (32%) with pattern 1a + 1b and 17 eyes (68%) with pattern 2. In both CRVO and BRVO the presence of FE was significantly associated with higher persistence of macular edema and worse outcome, with FE pattern 2 representing the most severe condition. Remarkably, FE patterns 1a and 1b were characterized by BCVA stability over the follow-up, whereas FE pattern 2 showed significant bestcorrected visual acuity (BCVA) worsening at the end of the follow-up. FE can be considered a negative prognostic biomarker in RVO, associated with higher persistence of macular edema and worse visual outcome. Müller cell impairment might represent the pathogenic mechanism leading to the loss of macular structural support and impairment of fluid homeostasis.

Prognostic and predictive implications of plasma ctDNA in guiding first-line targeted therapy for metastatic HER2-mutant non-small cell lung cancer (NSCLC).

9052 Background: Trastuzumab deruxtecan (T-DXd) is the first HER2-targeted drug approved for NSCLC with human epidermal growth factor receptor 2 ( HER2, ERBB2) mutation in the second or later line settings. However, the optimal first-line treatment for these patients remains unclear. Access to rapid tissue sequencing is not always available, thus presenting challenges to first-line drug development. Circulating tumor DNA (ctDNA) analysis has the potential to overcome these obstacles to guide optimal first-line targeted therapy for patients with HER2-mutant NSCLC. Methods: We retrospectively analyzed patients with metastatic HER2-mutant NSCLC who underwent prospective clinical ctDNA sequencing at Memorial Sloan Kettering Cancer Center (MSK) from January 2016 to September 2022. HER2 mutations were identified by next-generation sequencing through MSK-IMPACT, MSK-ACCESS or Resolution ctDx Lung. Results: We identified 64 patients with metastatic HER2-mutant NSCLC who had received at least one line of systematic therapy. The median age was 63 years (range: 23–90), and there were more women (n = 38, 60%), and never-smokers (n = 38, 60%). The activating HER2 mutations included exon 20 insertions (73%), exon 8 (11%), 19 (11%), and 20 SNVs (5%). Plasma ctDNA was tested before initial therapy in 40 patients with a median overall survival (OS) of 28 months (95% CI 21-34), in whom 31 patients (78%) had at least one detectable ctDNA alteration by MSK-ACCESS and ctDx Lung. 55% (17/31) received chemoimmunotherapy with pembrolizumab as the first-line treatment. The median time to treatment discontinuation (TTD) was 6 months (95%CI 5.5-6.9). Additionally, 19% of patients (6/31) received a HER2-targeted antibody-drug conjugates (ADC) as first-line treatment with a median TTD of 6 months (95% CI 2-10), including 5 with T-DM1 and one who received first-line T-DXd treatment with a TTD of 9 months. Patients with baseline ctDNA alterations had significantly shorter OS (hazard ratio (HR), 5.25; 95% CI, 1–24; p = 0.019). Conclusions: Baseline plasma ctDNA has the potential to guide first-line targeted therapy for patients with HER2-mutant NSCLC. As an independent negative prognostic biomarker, detectable ctDNA at baseline would need to be taken into account for patient selection in future studies to avoid underestimating the effects of therapy.

Abstract 5993: Dissecting the role of CDK4 in liposarcoma

Abstract Background: Liposarcoma (LPS) encompass one of most common soft tissue sarcoma (STS) histological subtypes accounting for 15% of all cases. They can be divided into four entities including atypical lipomatous tumor or well-differentiated LPS (ALT/WDLPS), dedifferentiated LPS (DDLPS), myxoid LPS (MLPS) and pleomorphic LPS (PLS). Their heterogeneity is reflected in their morphology, molecular landscape, prognosis and clinical behavior. Current treatment options for localized disease include surgery, (neo)adjuvant radiotherapy and chemotherapy; for the metastatic setting chemotherapy represents the cornerstone but its role needs to be elucidated. The unavailability of predictive biomarkers makes the management of these mesenchymal diseases even more challenging. Recent evidences have shed light on the key role of cyclin-dependent kinase 4 (CDK4) in cancer cells proliferation. Methods: This study involved 21 adult patients affected by liposarcoma (n= 5 ALT/WDLPS and n= 16 DDLPS). IHC analyses of CDK4 and MDM2 were performed on FFPE surgically resected specimens by experienced sarcoma pathologists. Moreover, a patient-derived DDLPS cell line was established and pharmacological profiling was performed. Next, standard and innovative drugs currently used for LPS management were assessed in both 2D and 3D culture systems. Finally, in silico analyses based on public repositories on STS were carried out including 260 sarcoma patients. Results: IHC results highlighted an higher expression of CDK4 and MDM2 biomarkers in DDLPS compared to ALT/WDLSP. Moreover, although CDK4 and MDM2 are codified by the same genomic region, their expression did not correlate (e.g. 100% of CDK4 and 2% of MDM2 in the same patient). Therefore we hypothesized that even exhibiting a low MDM2 expression, a patient could benefit from CDK4 inhibition. Furthermore, our data showed that in DDLPS cases CDK4 expression ranged from 90% to 100% in spermatic cord, skin and abdomen while it ranged from 5% to 50% in the retroperitoneum. The above observations prompted us to hypothesize a correlation of CDK4 expression with specific anatomical sites. Moreover, pharmacological profiling showed a synergistic effect exerted by sequential treatment with palbociclib and some chemotherapeutics including trabectedin, dacarbazine, eribulin and lenvatinib. Indeed we observed a 10% increase in cell proliferation inhibition compared to standard treatment. In addition, in silico analyses revealed the role of CDK4 as negative prognostic biomarker for PFS and OS in STS. Conclusions: Our study highlighted the promising role of CDK4 in the management of LPS patients. In particular, we pointed out encouraging results of sequential treatment combining CDK4 inhibitor palbociclib and chemotherapy. Furthermore, preliminary analyses highlighted the involvement of this biomarker as a potential prognostic tool for STS. Citation Format: Silvia Vanni, Graziana Gallo, Valentina Fausti, Giacomo Miserocchi, Chiara Liverani, Chiara Spadazzi, Claudia Cocchi, Chiara Calabrese, Giovanni De Luca, Massimo Bassi, Manlio Gessaroli, Angelo Campobassi, Federica Pieri, Giorgio Ercolani, Davide Cavaliere, Lorena Gurrieri, Nada Riva, Giovanni Martinelli, Laura Mercatali, Alessandro De Vita. Dissecting the role of CDK4 in liposarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5993.

Neutrophil-to-lymphocyte ratio is a negative prognostic biomarker for luminal A breast cancer.

Inflammation plays an important role in the occurrence, development, and metastasis of tumors. However, the prognostic role of the neutrophil-to-lymphocyte ratio (NLR) in patients with luminal A breast cancer has rarely been reported in the literature. The purpose of this study was to investigate the relationship between preoperative peripheral blood NLR and the survival rate of patients with luminal A breast cancer. Data from 226 eligible patients with luminal A breast cancer at the Chongqing University Cancer Hospital between 2011 and 2016 were obtained. The cut-off value for NLR for predicting overall survival (OS) rate was obtained from the receiver operating characteristic (ROC) curve. The baseline characteristics of 2 groups were compared using the Chi-square test or Fisher's exact test, and OS was estimated using the Kaplan-Meier method. Cox analysis was performed to determine the correlation between clinicopathological parameters and prognosis. ROC curve analysis showed that the cutoff value for NLR to predict OS was 2.0. Kaplan-Meier analysis revealed that the OS of patients with a NLR <2.0 was significantly longer than that of patients with a higher NLR >2 (P<0.0001). The area under the curve (AUC) for NLR to predict OS was 0.781 [95% confidence interval (CI): 0.712-0.851], sensitivity was 54.17%, and specificity was 97.06%. In univariate Cox regression analysis, NLR, tumor (T) stage (T3-T4 vs. T1-T2), and histological grade (II-III vs. I) were all significantly associated with OS. In multivariate Cox regression analysis, NLR and histology grade (II-III vs. I) were independent prognostic factors for OS. The results suggested that higher preoperative NLR was associated with worse prognosis in luminal A breast cancer.

Sensitive Detection of Cell-Free Tumour DNA Using Optimised Targeted Sequencing Can Predict Prognosis in Gastro-Oesophageal Cancer.

In this longitudinal study, cell-free tumour DNA (a liquid biopsy) from plasma was explored as a prognostic biomarker for gastro-oesophageal cancer. Both tumour-informed and tumour-agnostic approaches for plasma variant filtering were evaluated in 47 participants. This was possible through sequencing of DNA from tissue biopsies from all participants and cell-free DNA from plasma sampled before and after surgery (n = 42), as well as DNA from white blood cells (n = 21) using a custom gene panel with and without unique molecular identifiers (UMIs). A subset of the plasma samples (n = 12) was also assayed with targeted droplet digital PCR (ddPCR). In 17/31 (55%) diagnostic plasma samples, tissue-verified cancer-associated variants could be detected by the gene panel. In the tumour-agnostic approach, 26 participants (59%) had cancer-associated variants, and UMIs were necessary to filter the true variants from the technical artefacts. Additionally, clonal haematopoietic variants could be excluded using the matched white blood cells or follow-up plasma samples. ddPCR detected its targets in 10/12 (83%) and provided an ultra-sensitive method for follow-up. Detectable cancer-associated variants in plasma correlated to a shorter overall survival and shorter time to progression, with a significant correlation for the tumour-informed approaches. In summary, liquid biopsy gene panel sequencing using a tumour-agnostic approach can be applied to all patients regardless of the presence of a tissue biopsy, although this requires UMIs and the exclusion of clonal haematopoietic variants. However, if sequencing data from tumour biopsies are available, a tumour-informed approach improves the value of cell-free tumour DNA as a negative prognostic biomarker in gastro-oesophageal cancer patients.

Hemoglobin, albumin, lymphocyte, and platelet (HALP) score and cancer prognosis: A systematic review and meta-analysis of 13,110 patients.

We sought to explore the prognostic role of the hemoglobin, albumin, lymphocyte, and platelet (HALP) score on survival outcomes in patients with solid tumors by performing a systematic review and meta-analysis. PubMed, EMBASE and Web of Science were searched for relevant articles through 02 Nov 2022. Hazard ratios (HRs) with 95% confidence intervals (95% CIs) were extracted and pooled to assess the association of the HALP score with survival outcomes in solid tumors using Stata 15.0. The primary outcome was overall survival (OS), and the secondary outcomes were cancer-specific survival (CSS) and progression/disease/recurrence-free survival (PFS/DFS/RFS). A total of 28 studies (31 cohorts) comprising 13,038 patients were enrolled. A low HALP score was associated with decreased OS (HR, 1.61; 95% CI, 1.44-1.81; P<0.001), CSS (HR, 1.80; 95% CI: 1.54-2.09; P<0.001) and PFS/DFS/RFS (HR, 1.61; 95% CI: 1.45-1.80; P<0.001). The prognostic value of HALP on OS was observed across various tumor types and tumor stages. meta-regression found that tumor stage was the major source of the heterogeneity for OS. A low pretreatment HALP score is a reliable and negative prognostic biomarker for survival outcomes in patients with cancer.

Automated Region of Interest Selection Improves Deep Learning-Based Segmentation of Hyper-Reflective Foci in Optical Coherence Tomography Images.

Hyperreflective foci (HRF) have been associated with retinal disease progression and demonstrated as a negative prognostic biomarker for visual function. Automated segmentation of HRF in retinal optical coherence tomography (OCT) scans can be beneficial to identify the formation and movement of the HRF biomarker as a retinal disease progresses and can serve as the first step in understanding the nature and severity of the disease. In this paper, we propose a fully automated deep neural network based HRF segmentation model in OCT images. We enhance the model's performance by using a patch-based strategy that increases the model's compute on the HRF pixels. The patch-based strategy is evaluated against state of the art HRF segmentation pipelines on clinical retinal image data. Our results shows that the patch-based approach demonstrates a high precision score and intersection over union (IOU) using a ResNet34 segmentation model with Binary Cross Entropy loss function. The HRF segmentation pipeline can be used for analyzing HRF biomarkers for different retinopathies.

Enhancer of zeste homolog 2 is a negative prognostic biomarker and correlated with immune infiltrates in meningioma.

Enhancer of zeste homolog 2 (EZH2), an important epigenetic regulator, that mainly regulates histone H3 lysine 27 trimethylation (H3K27me3) through histone methyltransferase, and participates in promoting the development of tumors. At present, the loss of H3K27me3 expression in meningioma is a poor prognostic factor, but the research of EZH2 in meningioma is rare. Therefore, we aim to explore the expression of EZH2 in the meningioma and its correlation with the prognosis and immune microenvironment and lay the foundation for the subsequently potential targeted therapy and immunotherapy for meningioma. Tissue microarray immunohistochemistry staining was performed on 276 meningioma samples from Sun Yat-sen University Cancer Center. Expression levels of EZH2, H3K27me3, Ki67, programmed cell death protein 1 (PD-1), programmed cell death 1 ligand 1 (PD-L1), CD4, CD8, CD20, FOXP3, CD68, and CD163 were evaluated. Cox regression analyses were performed, and the Kaplan-Meier (KM) method was used to construct survival curves. In addition, we use biological information methods to analyze the mRNA expression of EZH2 and its relationship with the prognosis and immune microenvironment in the gene expression omnibus (GEO) database. Enhancer of zeste homolog 2 expression is concentrated in World Health Organization (WHO) grades 2 and 3 meningiomas (8.3+ and 33.3%+). We found that EZH2 expression was associated with a worse prognosis in meningioma (P < 0.001), the same results were confirmed in the GEO database (P < 0.001). Both EZH2 expression and H3K27me3 deletion (P = 0.035) predicted a worse prognosis, but EZH2 has no correlation with H3K27me3 expression. EZH2 expression was closely associated with increased Ki67 index (P < 0.001). In addition, EZH2 was associated with the immune microenvironment and positively correlated with PD-L1 expression (P < 0.001). Enhancer of zeste homolog 2 is a new prognostic biomarker in meningioma. It correlates with PD-L1 expression and closely related to tumor immunosuppression. Our research can provide a reference for the potential targeted therapy and immunotherapy of meningioma in the future.

Abstract A042: Keratin 17 is a negative prognostic and predictive biomarker in pancreatic ductal adenocarcinoma

Abstract Background: There is an urgent need to understand why patients with clinically and histologically identical pancreatic ductal adenocarcinomas (PDACs) differ in response to treatment, disease progression, and survival. Although two standard chemotherapies are available, predictive biomarkers to guide regimen selection have not been defined. We previously reported that keratin 17 (K17) expression is a hallmark of PDAC cases with shortest patient survival. Furthermore, K17, explored using in vitro and in vivo murine models, drives resistance to gemcitabine and 5-fluorouracil, the most common chemotherapeutic agents in the two chemotherapies. Here, we aimed to validate the prognostic value of K17 and to further explore its role as a predictive biomarker. Methods: We used a cohort of 305 cases, with localized disease and who had gone tumor resection. An indirect immunoperoxidase method was used to detect K17 expression on surgical specimens, as previously described. Survival was plotted using the Kaplan–Meier method and hazard ratios (HRs) were calculated using Cox proportional hazard regressions for both, overall survival (OS) and progression-free survival (PFS). Results: Patients in the high-K17 expression group had shorter overall survival [median=25 mo., HR=1.511, p=0.0338] than those in the low-K17 expression group (median=42 mo.). In addition, high K17 expression was associated with shorter median OS (p=0.0280) and PFS (p=0.0434) in patients who were treated with gemcitabine (GEM) or with Gemcitabine/nab-Paclitaxel (GEMTAX) therapy (OS p=0.1959, PFS p=0.0724), compared to low-K17 counterparts. When we further separated our cohort by K17 level of expression and compared GEM and GEMTAX responses, we found that patients with tumors with high K17 expression do not benefit from GEMTAX adjuvant therapy (p=0.0356). Of note, K17 expression within the responder group was significantly lower than in tumors within the non-responder group for both Gem and GEMTAX (p&amp;lt;0.001). Conclusions: K17 expression is confirmed as a robust prognostic biomarker, significantly correlated with poor OS and PFS in two independent cohorts of PDAC patients. More importantly, our results indicate that K17 expression predicts PDAC resistance to gemcitabine and poor response to GEMTAX. Collectively, our findings have implications could guide the development of K17 as a predictive biomarker for gemcitabine-based interventions in both adjuvant and palliative settings, to optimize therapeutic efficacy for PDAC. Citation Format: Lyanne Oblein, Lucia Roa-Peña, Sruthi Babu, Felicia D. Allard, Natalia D. Marchenko, Luisa F. Escobar-Hoyos, Kenneth R. Shroyer. Keratin 17 is a negative prognostic and predictive biomarker in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr A042.