Luminance Profile Research Articles

Benzimidazole Derivative (N-{4-[2-(4-Methoxyphenyl)-1H-Benzimidazole-1-Sulfonyl] Phenyl} Acetamide) Ameliorates Methotrexate-Induced Intestinal Mucositis by Suppressing Oxidative Stress and Inflammatory Markers in Mice.

Methotrexate (MTX)-induced intestinal mucositis (IM) is a common side effect in cancer treatment that impairs the immune system and gut microbes, resulting in loss of mucosal integrity and gut barrier dysfunction. The quality of life and outcomes of treatment are compromised by IM. The present study was designed to investigate the mucoprotective potential of the benzimidazole derivative N-{4-[2-(4-methoxyphenyl)-1H-benzimidazole-1-sulfonyl] phenyl} acetamide (B8) on MTX-induced IM in mice. IM was induced by a single dose of MTX in mice and assessed by physical manifestations as well as biochemical, oxidative, histological, and inflammatory parameters. B8 (1, 3, 9 mg/kg) significantly reduced diarrhea score, mitigated weight loss, increased feed intake and, survival rate in a dose-dependent manner. Notably, B8 exhibited a mucoprotective effect evident through the mitigation of villus atrophy, crypt hypoplasia, diminished crypt mitotic figures, mucin depletion, and oxidative stress markers (GSH, SOD, MDA, and catalase concentration). Gene expression analysis revealed that B8 downregulated the mRNA expression of tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), IL-1β, and nuclear factor-κB (NF-κB) and concurrently upregulated IL-10 expression in contrast to the MTX group. Further, B8 significantly improved the luminal microflora profile by augmenting the growth of Lactobacillus spp. and reducing the number of pathogenic bacteria (E. coli). Additionally, the enzyme-linked immunoassay showed that B8 decreased the levels of pro-inflammatory cytokines. Our findings suggest that B8 had mucoprotective effects against MTX-induced IM and could be used as an adjunct in chemotherapy to deter this side effect.

Standard models of spatial vision mispredict edge sensitivity at low spatial frequencies

One well-established characteristic of early visual processing is the contrast sensitivity function (CSF) which describes how sensitivity varies with the spatial frequency (SF) content of the visual input. The CSF prompted the development of a now standard model of spatial vision. It represents the visual input by activity in orientation- and SF selective channels which are nonlinearly recombined to predict a perceptual decision. The standard spatial vision model has been extensively tested with sinusoidal gratings at low contrast because their narrow SF spectra isolate the underlying SF selective mechanisms. It is less studied how well these mechanisms account for sensitivity to more behaviourally relevant stimuli such as sharp edges at high contrast (i.e. object boundaries) which abound in the natural environment and have broader SF spectra. Here, we probe sensitivity to edges (2-AFC, edge localization) in the presence of broadband and narrowband noises. We use Cornsweet luminance profiles with peak frequencies at 0.5, 3 and 9 cpd as edge stimuli. To test how well mechanisms underlying sinusoidal contrast sensitivity can account for edge sensitivity, we implement a single- and a multi-scale model building upon standard spatial vision model components. Both models account for most of the data but also systematically deviate in their predictions, particularly in the presence of pink noise and for the lowest SF edge. These deviations might indicate a transition from contrast- to luminance-based detection at low SFs. Alternatively, they might point to a missing component in current spatial vision models.

Closing classification gaps in luminal breast cancer with single-cell RNA-seq insights from normal breast lineages.

e13168 Background: Almost two decades ago, invasive breast cancer was classified into five subgroups based on expression patterns, each with distinct biological and prognostic characteristics. Advances in single-cell RNA sequencing offers insights into the expression profiles of normal luminal epithelial cells. Here, we investigate the connection between breast cancer (BC) classification and luminal expression profiles. Methods: We applied single-cell data from recent studies (Table) to compile gene signatures corresponding to expression profiles of luminal epithelial cell clusters in normal breast tissue. Normalized enrichment scores (ESs) of these signatures were calculated for bulk RNA-seq gene expression data from open source cohorts (TCGA-BRCA, SCAN-B, METABRIC). The BC molecular subtype of each tumor sample was predicted based on a modified PAM50 classification using gene expression profiling (1); LumA and LumB subtypes were combined into Luminal. ESs of signatures were compared between the BC subtypes (Table). Results: The ESs of all signatures differed significantly across BC subtypes (p < 0.001, Kruskal-Wallis test, Table 1). The ESs of signatures for LumSec-major and Luminal progenitor clusters are the highest in subtypes HER2-low and Basal-like (p < 0.001, Dwass-Steel-Critchlow-Fligner test [DSCF]). Basal-like also had the highest enrichment of the ductal signature (p < 0.001, DSCF). Luminal subtype showed the highest ESs of signatures corresponding to HR+ clusters of mature luminal cells and of the TDLU signature (p < 0.001, DSCF). HER2-low subtype showed the highest ES of the LumSec-lac signature (p < 0.001, DSCF). Conclusions: Our analyses challenge traditional classifications by showing that luminal breast cancers are less similar to most luminal epithelial clusters in normal breast tissue than other breast cancer subtypes. These findings suggest traditional expression classification may not fully capture the complexity of this largest subset of luminal-type breast cancers. 1. Turova et al., SABCS, 2023. [Table: see text]

Abstract PO5-17-02: Triple-negative breast cancer as an untraceable tumor compared to other immunohistochemical subtypes: a cross-sectional study

Abstract Objective: To compare the form of presentation at diagnosis of triple negative breast tumors with other immunohistochemical subtypes, in relation to clinical examination or imaging examination. Methods: This is a cross sectional study that evaluated data related to the presentation of breast carcinomas at diagnosis (clinical examination or screening imaging examination findings), immunohistochemical subtype, histological grade and tumor size, in diagnosed and referred patients. for the Mastology service at Hospital São Paulo – Unifesp, in São Paulo – Brazil, in the period between 2012 and 2021, by analyzing data from the medical records of these patients. Results: Among the 699 invasive carcinomas diagnosed, 246 patients were diagnosed with luminal profile A (35.2%), 257 luminal B (36.8%), 58 luminal B HER-2+ (8.3%), 26 HER-2 enriched (3.7%) and 112 triple negative (16%). Triple negative carcinomas had a higher rate of diagnosis by clinical examination (86.6%), 6.65 times more likely to be diagnosed by clinical examination than luminal A, in addition to having a larger lesion. The triple negative and HER-2 enriched subtypes presented the worst scenarios of clinical stage at diagnosis. Among the tumors diagnosed by screening examination, those belonging to the luminal A were associated with nodules seen on imaging (in 45.1%) and the subtypes luminal B and luminal B HER-2+ with the appearance of microcalcifications. There were no cases of triple negatives involving microcalcifications in our sample. Asymmetries were more common in images of luminal B HER-2+ tumors. In the multivariate analysis, it was observed that only the histological grade and the Ki-67 value are related to the form of diagnosis. Tumors with histological grade 2 or 3 are more likely to be diagnosed by clinical examination and not be traceable in relation to histological grade 1. As for Ki-67, its increase of 1 percentage point leads to an increase of 1.3% in the chance finding on physical examination. There was no correlation of hormone receptors and HER-2 with the chance of the tumor not being screenable. Conclusions: triple negative subtype breast carcinomas present a higher risk of diagnosis by clinical examination compared to other immunohistochemical subtypes and, therefore, tend not to be traceable. Parameters such as high Ki-67 and histological grade correlate with this trend. Citation Format: Morgana Silva, Afonso Nazário, Vanessa Sanvido. Triple-negative breast cancer as an untraceable tumor compared to other immunohistochemical subtypes: a cross-sectional study [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-17-02.

Abstract 2987: microRNA (miRNA) a putative biomarker to better define the molecular apocrine breast cancer (MABC) subtype

Abstract Background: Breast cancer (BC), is a highly heterogeneous disease, divided into molecular subtypes based on gene expression and clinical outcomes. Transcriptomics profiling depicted a subtype that has luminal expression profile but lacks estrogen receptor (ER), progesterone receptor expression with an overlapping expression of HER-2; yet it overexpresses androgen receptor (AR). This subtype was referred to as MABC and constitutes 8-14% of all BC types. At present MABC is often misdiagnosed with triple negative BC (TNBC). Its improper diagnosis demands the adoption of complementary tools; miRNA, key players in BC tumorigenic processes, hold promise in defining MABC. Methods: 539 BC microarray data were downloaded from The Cancer Genome Atlas (TCGA-BRCA) (PMID: 25691825) using TCGA biolinks R/Bioconductor package (PMID:267704973). Cases with available miRNA data were retained and classified using citbcmst R package (PMID: 21785460). Differential expression analysis identified deregulated miRNAs using DEseq2. The validation set consists of 111 ER-neg samples (68 MABC, and 43 TN samples) with an average age at diagnosis 58.51 years and a median follow-up=78.5 months. MABC tumors were characterized apart of TNBC by the molecular signature (AR, FOXA1 and AR-related genes, PMID: 25516281) on fresh tissue sections. Using miRCURY LNATM miRNA PCR assay, miRNA profiling was done for a panel of differentially expressed miRNAs. Non tumorigenic (MCF-10A) and BC Cellular models MABC (MDA-MB-453) and TNBC (MDA-MB-231) were used to investigate the invasive potential of MABC. Results: TCGA data analysis indicated MABC as a separate entity based on gene signature. MiRNA-seq data analysis depicted a set of 6 significantly deregulated miRNA with absolute value of log2 fold change> 1 and P-adjusted value <0.05 between MABC and TNBC. We validated, by miRNA profiling, significant upregulation of miR-2115-3p and miR-187-3p in MABC compared to TNBC. These miRNAs significantly differentiate MABC patients from TNBC patients where the combined miRNA panel of miR-2115-3p and miR-187-3p had an area under the curve of 0.904 ±0.28 (P<0.0001 and 95% CI: 0.850-0.958) and sensitivity, specificity, and a diagnostic accuracy of 90.41%, 81.1%, and 86.5% respectively. Preliminary data, showed that non-tumorigenic/non-invasive MCF-10A had a significant increase in its invasive ability upon transfection with miR-187-3p mimic (P<0.05). Similarly, the invasive cell line MDA-MB-231 showed a significant increase in invasion upon transfection with miR-187-3p mimic (P<0.05). On the other hand, only miR-187-3p inhibitor significantly decreased the invasive potential of MDA-MB-453 cells (P<0.05). Conclusion: MABC has a unique signature of miRNA as compared to TNBC. miR-2115-3p, and miR-187-3p could be potential diagnostic biomarkers for MABC. The invasive potential of MABC could be attributed to miR-187-3p activity. Citation Format: Ghada Chamandi, Adrien Borgel, Abdallah Kurdi, Pierre Khoueiry, Luis Teixeira, Morgane Le Bras, Jacqueline Lehamnn, Rihab Nasr. microRNA (miRNA) a putative biomarker to better define the molecular apocrine breast cancer (MABC) subtype [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 2987.

Abstract 3923: Targeting immunosuppressive TREM2+tumor associated macrophages in prostate cancer

Abstract With over 250,000 new cases annually, prostate cancer (PCa) is amongst the most diagnosed cancers in the United States. Despite a near-perfect 5-year survival rate in patients with localized disease with androgen deprivation therapy (ADT), about 10-20% of patients on ADT eventually develop castration-resistant prostate cancer (CRPC). Immunotherapies are typically ineffective in treating these treatment-resistant patients due to the abundance of immunosuppressive tumor-associated macrophages (TAMs) and lack of infiltrating lymphocytes. We propose to target these immunosuppressive macrophages through the triggering receptor expressed on myeloid cells 2 (TREM2), which has been identified as a major mediator of suppressive macrophage function but has not been studied in the context of PCa and CRPC. For this study, we analyzed publicly available RNA-seq datasets to query for TREM2 expression amongst patients with a range of prostate cancer disease states and cell types, measured tumor growth kinetics in two subcutaneous models of prostate cancer, and compared tumor immune infiltrates by flow cytometry. Our analysis of the TCGA prostate adenocarcinoma (PRAD) and the Stand Up to Cancer/Prostate Cancer Foundation (SU2C/PCF) datasets implicate TREM2 expression with prostate cancer progression, by Gleason score and progression-free status, and with worse overall survival of patients with metastatic CRPC (mCRPC). Manual curation and analysis of 10 single-cell RNA-seq datasets with a total of 84 patients, encompassing primary PCa, CRPC, and mCRPC indicate that TREM2 is highly and specifically expressed on TAMs in prostate cancer, with macrophages from patient-matched tumor samples expressing higher TREM2 compared to normal adjacent tissue. B6CaP tumors, with a luminal cytokeratin profile, high AR expression, and high Myc expression, show slower tumor growth in Trem2−/− mice compared to tumors in Trem2wt mice. On the other hand, RM1 tumors show no difference in growth kinetics on Trem2−/− mice. A major difference between the B6CaP and RM1 tumor microenvironments is the level of macrophage infiltration, where only 2-5% of immune cells in RM1 tumors are macrophages, compared to the 40-50% in B6CaP tumors. RM1 cells co-implanted with Trem2−/− macrophages show slower tumor growth, compared to tumor cells co-implanted with Trem2wt macrophages, confirming the role of Trem2-expressing macrophages in mediating tumor growth. Consistent with previous reports of suppressive Trem2 signaling, we also observed higher levels of inflammatory cytokine production in ex vivo stimulated Trem2−/− TAMs. By targeting TREM2 in prostate cancer, we find that TAMs are less suppressive, more inflammatory, and permit cytotoxic lymphocyte infiltration to enhance anti-tumor responses by synergizing with chemo and immune checkpoint therapy. Citation Format: Alex J. Lee, Kenneth Adusei, Monali Praharaj, Fan Shen, Xiaoxu Wang, Kaavya Bhatia, Thomas Nirschl, Jelani C. Zarif. Targeting immunosuppressive TREM2+tumor associated macrophages in prostate 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 3923.

Abstract 4632: Lineage plasticity as a determinant of antibody-drug conjugate target expression in urothelial bladder cancer

Abstract Bladder cancers display a wide spectrum of morphologies that frequently co-exist within individual tumors. Several histologic variants, including plasmacytoid, neuroendocrine, and micropapillary subtypes, are associated with an increased bladder cancer recurrence risk and cancer-specific mortality. The molecular basis and therapeutic implications of this phenotypical plasticity remain poorly understood. Enfortumab vedotin (EV), an antibody-drug conjugate targeting Nectin-4, has also emerged as a new standard-of-care for metastatic bladder cancer patients, but the association between lineage plasticity as manifested by morphologic heterogeneity in bladder cancer, Nectin-4 expression and ADC response remains poorly defined. Previous DNA-based molecular profiling studies of histologic variants revealed that CDH1 mutations were the pathognomonic molecular alteration in plasmacytoid urothelial carcinoma, but no recurrent driver mutations have been found to be unique to the aberrant histology regions of other mixed histology subtypes. Thus, to explore the molecular basis for the lineage plasticity of bladder cancers with aberrant histology and to define the association between lineage plasticity manifested as morphologic heterogeneity and the expression of Nectin-4, HER2 and other potential ADC targets, we performed RNA-seq of pure urothelial carcinomas (n=118, not otherwise specified, UC-NOS) and bladder tumors with divergent differentiation or histologic subtypes (n=199). Our cohort included a subset of tumors in which RNA-seq was performed on microdissected UC-NOS and variant histology regions of the same tumor. Tumors with neuroendocrine, sarcomatoid and squamous histology had the lowest Nectin-4 expression, whereas histologic subtypes that retained a luminal transcriptional profile such as micropapillary, plasmacytoid and nested histology had higher levels of Nectin-4 expression. In mixed histology tumors, Nectin-4 and/or HER2 expression was often restricted to only one of the two histologic components. Gene Set Enrichment Analysis noted enrichment of distinct oncogenic pathway gene sets in different histologic variants. Preliminary investigation into the mechanism(s) of Nectin-4 regulation using bladder cancer cell lines and patient-derived organoids (PDOs) has identified targetable pathways which when inhibited revert lineage plasticity and induce Nectin-4 expression. In sum, lineage plasticity manifested as heterogenous histomorphologies was associated with heterogeneous Nectin-4 expression in some but not all histologic variants of bladder cancer. Ongoing studies using PDOs from bladder cancer patients with mixed histology will seek to identify molecular pathways whose inhibition leads to increased expression of Nectin-4 or other ADC targets such as HER2 and thus increased sensitivity of ADCs targeting these cell surface proteins. Citation Format: Jiaqian Luo, Sizhi P. Gao, Fengshen Kuo, Gamze Gokturk Ozcan, Merve Basar, Florestan Koll, Jacob E. Tallman, Syed M. Alam, Carissa E. Chu, Ziyu Chen, Eugene J. Pietzak, Gopakumar Iyer, Jonathan E. Rosenberg, David B. Solit, Hikmat Al-Ahmadie. Lineage plasticity as a determinant of antibody-drug conjugate target expression in urothelial bladder 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 4632.

Microbiota and mucosal gene expression of fecal microbiota transplantation or placebo treated patients with chronic pouchitis

ABSTRACT Altered microbiota and impaired host immune function have been linked to the pathogenesis of pouchitis. We used 16S rRNA gene sequencing and RNA sequencing data from a previous randomized clinical trial (RCT) on fecal microbiota transplantation (FMT) therapy in 26 chronic pouchitis patients with one-year follow-up. We analyzed changes in both luminal and mucosal microbiota composition, as well as in host mucosal gene expression to gain insights into the host–microbiota interactions possibly underlying clinical outcomes of the patients. Antibiotic type and pattern of use were significant drivers of the luminal microbiota at baseline. Differential gene expression analysis indicated transition from ileal to colonic gene expression in the pouch, and upregulation in inflammation- and immune system-related pathways in the pouch. At 4 weeks, the non-relapsed FMT patients had a lower microbiota dissimilarity to the donor than the non-relapsed placebo patients (p = .02). While two FMT-treated patients showed a shift toward the donor’s microbiota during the one-year follow-up, the overall FMT microbiota modulation effect was low. Patient’s luminal and mucosal microbiota profiles were unstable in both FMT and placebo groups. Expression of the chemokine receptor CXCR4 was downregulated at 52 weeks compared to the baseline in the non-relapsed patients in both FMT and placebo groups. Microbiota modulation by FMT seems to be low in this patient group. The microbiota composition or alterations did not explain the relapse status of the patients. Some evidence for remission-related host gene expression pattern was found; specifically, CXCR4 expression may have a role in sustained remission.

Molecular classification of high-grade, muscle-invasive urothelial carcinomas and the relationship with CTLA-4 and PD-L1 expression

Purposethe aim of the current study was to molecularly classify high-grade, muscle-invasive, urothelial carcinoma of basal, luminal, and p53-like subtypes and to assess their relationship with CTLA-4 and PD-L1 expression.Methodsfifty-seven samples from transurethral resections were tested immunohistochemically for CK5/6, CD44, CK20, p63, p53, and CTLA-4/PD-L1.Resultsabout half of the cases presented a basal molecular profile (50.9%), the luminal profile was found in 29.8% and the p53-like profile in 15.8%. Positivity for CTLA-4 occurred in 24 samples (42.1%), with cytoplasmic and membrane expression reported only in neoplastic cells. Of these, 11 were of the luminal subtype, ten were of the basal subtype, and three were of the p53-like subtype. There was an association between CTLA-4 expression and the luminal classification, and an absence of CTLA-4 expression in the basal profile (p = 0.047). PD-L1 expression was found in 12 samples, ten of them classified as the basal subtype (p = 0.036). CTLA-4 and PD-L1 expression were not identified in the same subtypes (p = 0.08), since only one sample was positive for both markers.ConclusionsA molecular classification into the subtypes was possible in 96.5% of cases of high-grade, muscle-invasive, urothelial carcinoma using immunohistochemistry. Thus, this type of classification is viable in most surgical pathology laboratories, including those in Brazil’s Unified Health System (known as SUS, the acronym for Sistema Único de Saúde). The expression of CTLA-4 is related to the luminal molecular subtype, while the expression of PD-L1 is related to the basal molecular subtype. CTLA-4 and PD-L1 positivity are mutually exclusive.

Effects of weaving parameters on the properties of completely biological tissue-engineered vascular grafts

Tissue-engineered vascular grafts (TEVGs) made of human textiles have been recently introduced and offer remarkable biocompatibility as well as tunable mechanical properties. The approach combines the use of cell-assembled extracellular matrix (CAM) threads, produced by cultured cells in vitro, with weaving, a versatile assembly method that gives fine control over graft properties. Herein, we investigated how production parameters can modify the geometrical and mechanical properties of TEVGs to better match that of native blood vessels in order to provide long-term patency. Our goals were to decrease the mechanical strength and the luminal surface profile of our first generation of woven TEVGs, while maintaining low transmural permeability and good suture retention strength. Different TEVGs were produced by varying CAM sheet strength as well as weaving parameters such as warp count, weft ribbons width, and weft tension. An optimized design reduced the burst pressure by 35%, wall thickness by 38% and increased compliance by 269%. The improved TEVG had properties closer to that of native blood vessels, with a burst pressure of 3492 mmHg, a wall thickness of 0.69 mm, and a compliance of 4.8%/100 mmHg, while keeping excellent suture retention strength (4.7 N) and low transmural permeability (24 ml·min−1·cm−2). Moreover, the new design reduced the luminal surface profile by 48% and utilized 47% less CAM. With a comparable design, the use of decellularized CAM threads, instead of devitalized ones, led to TEVGs with much more permeable walls and higher burst pressure. The next step is to implant this optimized graft in an allogeneic sheep model of arteriovenous shunt to assess its in vivo remodeling and performance.

Changes in Invasive Breast Carcinomas after Neoadjuvant Chemotherapy Can Influence Adjuvant Therapeutic Decisions.

Neoadjuvant chemotherapy (NACT) can change invasive breast carcinomas (IBC) and influence the patients' overall survival time (OS). We aimed to identify IBC changes after NACT and their association with OS. IBC data in pre- and post-NACT samples of 86 patients were evaluated and associated with OS. Post-NACT tumors changed nuclear pleomorphism score (p=0.025); mitotic count (p=0.002); % of tumor-infiltrating inflammatory cells (p=0.016); presence of in situ carcinoma (p=0.001) and lymphovascular invasion (LVI; p=0.002); expression of estrogen (p=0.003), progesterone receptors (PR; p=0.019), and Ki67 (p=0.003). Immunohistochemical (IHC) profile changed in 26 tumors (30.2%, p=0.050). Higher risk of death was significatively associated with initial tumor histological grade III (hazard ratio [HR], 2.94), high nuclear pleomorphism (HR, 2.53), high Ki67 index (HR, 2.47), post-NACT presence of LVI (HR, 1.90), luminal B-like profile (HR, 2.58), pre- (HR, 2.26) and post-NACT intermediate mitotic count (HR, 2.12), pre- (HR, 4.45) and post-NACT triple-negative IHC profile (HR, 4.52). On the other hand, lower risk of death was significative associated with pre- (HR, 0.35) and post-NACT (HR, 0.39) estrogen receptor-positive, and pre- (HR, 0.37) and post-NACT (HR, 0.57) PR-positive. Changes in IHC profile were associated with longer OS (p=0.050). In multivariate analysis, pre-NACT grade III tumors and pre-NACT and post-NACT triple negative IHC profile proved to be independent factors for shorter OS. NACT can change tumor characteristics and biomarkers and impact on OS; therefore, they should be reassessed on residual samples to improve therapeutic decisions.

Use of the genetic signature of 70-genes MammaPrint in comparision with the immunohistochemical profile to de-escalate systemic treatment in the elderly population: Real-world study (AGEMA-BR).

e12561 Background: the elderly population, generally, presents a lower performance and greater susceptibility to complications and adverse effects of therapies, inferring that the de-escalation of cytotoxic therapies is desirable. Genetic signatures, including MammaPrint/ BluePrint, are the most appropriate tools to select among estrogen receptor (ER) patients with high clinical risk, who could, with oncological safety, not receiving adjuvant chemotherapy. Objective – to evaluate the performance of MammaPrint™ (MP) / BluePrint (BP) and the immunohistochemical profile (IHC) in the de-escalation of systemic therapy in the elderly population with breast cancer and Luminal profile. Methods: from the patients in the AGEMA-BRA study, which analyzes the performance of the 70-gene MammaPrint™ platform in the Brazilian population, those aged 70 years or older, with high clinical risk, who had data on the immunohistochemical profile available (37 patients). A comparative analysis was performed between the genomic profile (Luminal A/B on BluePrint; high and low risk on MammaPrint™ and the immunohistochemical profile (considered Luminal A with ER >10%, RP>10%, HER2 negative, Ki67 ≤20% Luminal B with ER>10%, HER2 negative, Ki67>20%) and clinical endpoints including disease-free survival (DFS) and overall survival (OS). Data analysis used accuracy and the Kappa coefficient (considering concordance >0, 6). Results: mean age at diagnosis 73.1 years (70–80 years) and mean follow-up (FU) of 42 months. Of the 15 patients with high risk / luminal B by MP / BP, 5 were classified as luminal A by the IHC profile with a disagreement of 33.33%; of the 22 low-risk / luminal A patients by MP/BP, 6 were classified as luminal B by the IHC, a disagreement of 27.27%. Statistical analysis showed an accuracy between the IHC profile and the MP of 70.27%. The use of the IHC profile, when compared with the MP/BP, showed a sensitivity of 66.7% and a specificity of 72.7%. Of the 14 patients at high risk due to MP, 2 did not undergo chemotherapy treatment. All patients (at high and low risk) received adjuvant endocrine therapy with an aromatase inhibitor. After 42 months of medium FU, all patients were alive and disease-free, inferring DFS and OS of 100%. Conclusions: the analysis of data and outcomes in the elderly population of the AGEMA-BRA study with high clinical risk, demonstrated limited accuracy and low agreement in the stratification into high and low risk between the profile by IHC and the MP/BP. The use of the genomic platform may have been relevant in risk stratification and outcomes in this population. Studies with a larger sample and longer FU are needed to confirm the usefulness of genetic signatures in the de-escalation of systemic treatment in the elderly population with high clinical risk.

P‐126: An Introduction to GGRB Virtual Pixel Technology

The constructions of the GGRB virtual pixel and the effect on the display performance are introduced by comparing with that of the Diamond RGBG virtual pixel, a typical virtual pixel technology applied to active matrix organic light‐emitting diode (AMOLED) display. Because of the characteristic structure of GGRB sub‐ pixel arrangement, the occurrence of perceptible artificial color discontinuity is reduced in the GGRB virtual pixel display. The distribution uniformity of pixel luminance centroids is important for the display of pattern details. In the GGRB virtual pixel display, the distribution uniformity of pixel luminance centroids is achieved via a non‐uniformly distributed green sub‐pixel array of which the luminance profile is properly modulated by the red and blue sub‐pixels with lower spatial frequency distribution. The results of subjective evaluation and comparison suggest that the performance improvement of the GGRB virtual pixel display is consistent with the conceptual expectation. Some of the essential characteristics is superior to those of the reference technology.

Qualitative perception of 3D shape from patterns of luminance curvature.

A new source of information is proposed for the perception of three-dimensional (3D) shape from shading that identifies surface concavities from the curvature of the luminance field. Two experiments measured the abilities of human observers to identify concavities on smoothly curved shaded surfaces depicted with several different patterns of illumination and several different material properties. Observers were required to identify any apparent concavities along designated cross sections of the depicted objects and to mark each concavity with an adjustable dot. To analyze the results, we computed both the surface curvature and the luminance curvature along each image cross section. The results revealed that most responses were in concave regions of the luminance profiles, although they were often shifted in phase relative to the curvature of the depicted surfaces. This pattern of performance was surprisingly robust over large changes in the pattern of illumination or surface material properties. Our analysis predicts that observers should make false alarm responses in regions where a luminance concavity does not correspond to a surface concavity, and our empirical results confirm that prediction.

Effects of a gap between the central and surrounding regions with luminance gradients on the feeling of being dazzled.

The feeling of being dazzled is evoked by images consisting of an achromatic uniform center, surrounded by regions with luminance gradients. As the perceptual distinctness of the central region has been suggested to contribute to the feeling of being dazzled, we examined the effects of a gap between the central and surrounding regions on the feeling of being dazzled. The stimulus comprised a disk with uniform luminance surrounded by an annulus, of which the luminance was decreased from the inner boundary to the periphery. Three luminance profiles (linear, logistic, and inverse-logistic) of the surrounding luminance ramps were used. The distinctness of the disk decreased in the order of logistic, linear, and inverse-logistic profiles. The luminance of the disk, the maximum luminance of the annulus, and the gap size were also varied. When the luminance continuously transitioned from the disk to the annulus, the feeling of being dazzled was stronger for the inverse-logistic annulus luminance profile, compared with the logistic and linear profiles without a gap; however, it was not different for the three profiles with a gap. Further, the feeling of being dazzled increased when a gap was introduced for the logistic and linear profiles, but not for the inverse-logistic profile. These results suggest that the feeling of being dazzled was reduced by the perceptual indistinctness of the central disk for the logistic and linear annulus luminance profiles, while the gap restored the feeling of being dazzled by making the central disk perceptually distinct.

Prostatic luminal progenitors: From tissue regeneration to therapeutic escape

Inhibition of androgen signaling is the gold standard treatment of benign prostate hyperplasia and prostate cancer. Despite the initial response to these treatments, therapeutic resistance is ultimately observed in most patients. Single cell RNAseq studies have shown that castration-tolerant luminal cells share several molecular and functional features with cells identified as luminal progenitor in physiological conditions. The increased prevalence of luminal progenitor-like cells in tumor contexts might result from their intrinsic androgen-independence and from the reprogramming of differentiated luminal cells into a castration-tolerant state. Thus, it is currently hypothesized that the luminal progenitor molecular profile might constitute a functional hub for cell survival in androgen deprivation context, a prerequisite for tumor regrowth. Therapeutic intervention interfering with luminal lineage plasticity is a promising approach to prevent prostate cancer progression.

PD25-07 PROTEOMIC PROFILING OF MUSCLE INVASIVE BLADDER CANCER TREATED WITH NEOADJUVANT CHEMOTHERAPY REVEALS UNIQUE BIOLOGIC CLUSTERS WITH CLINICAL RELEVANCE

You have accessJournal of UrologyCME1 Apr 2023PD25-07 PROTEOMIC PROFILING OF MUSCLE INVASIVE BLADDER CANCER TREATED WITH NEOADJUVANT CHEMOTHERAPY REVEALS UNIQUE BIOLOGIC CLUSTERS WITH CLINICAL RELEVANCE Alberto Contreras-Sanz, Moritz J. Reike, Gian L. Negri, Htoo Zarni Oo, Sandra Spencer Miko, Karina Nielsen, Morgan E. Roberts, Joshua Scurll, Kenichiro Ikeda, Gang Wang, Roland Seiler, Gregg B. Morin, and Peter C. Black Alberto Contreras-SanzAlberto Contreras-Sanz More articles by this author , Moritz J. ReikeMoritz J. Reike More articles by this author , Gian L. NegriGian L. Negri More articles by this author , Htoo Zarni OoHtoo Zarni Oo More articles by this author , Sandra Spencer MikoSandra Spencer Miko More articles by this author , Karina NielsenKarina Nielsen More articles by this author , Morgan E. RobertsMorgan E. Roberts More articles by this author , Joshua ScurllJoshua Scurll More articles by this author , Kenichiro IkedaKenichiro Ikeda More articles by this author , Gang WangGang Wang More articles by this author , Roland SeilerRoland Seiler More articles by this author , Gregg B. MorinGregg B. Morin More articles by this author , and Peter C. BlackPeter C. Black More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003303.07AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Neoadjuvant chemotherapy (NAC) followed by radical cystectomy (RC) is recommended for muscle invasive bladder cancer (MIBC). However, only ∼40% of patients show an objective response. While DNA alterations and RNA classifiers may predict response to NAC in retrospective studies, the proteome has not been evaluated in this context. Here we profile the MIBC proteome in a NAC-treated cohort to identify markers of response and to elucidate mechanisms of resistance to NAC. METHODS: Pre-NAC tissue was included from 107 MIBC patients who received NAC followed by RC. Residual tumor (≥ypT1N0-3M0-1) in the RC specimen was present in 62% of patients after NAC, and was profiled for 55 of those patients (51%). Multiregional sampling was conducted in 37/107 pre-NAC and 15/55 post-NAC samples. Benign ureter was used as control. SP3-Clinical Tissue Proteomics (SP3-CTP) and bioinformatic analysis using formalin-fixed paraffin-embedded tissue (FFPE) were performed. Immunohistochemistry (IHC) validation was conducted on matched tissues. RESULTS: We quantified 9769 proteins across all samples. Unsupervised clustering of pre NAC tissue established 4 clusters based on biology and survival outcomes, but no difference in response by pathologic stage. Clusters were confirmed by IHC, and consisted of: Cluster 1 (CC1), with high metabolic activity and a luminal profile; Cluster 2 (CC2) with high nuclear activity; Cluster 3 (CC3) with high immune infiltration and basal profile; and Cluster (CC4) with high immune infiltration and stromal signature. CC3 showed worse survival outcomes (p<0.01). Multivariable analysis identified novel favorable (MAPK9 and MTIF3) and unfavorable (DVL2 and NES) biomarkers. Matched analysis of pre- and post-NAC tissue showed markers (AZGP1 and ORM1) and pathways indicative of NAC resistance. In post-NAC (i.e. resistant) tumors we identified 2 clusters: Post-NAC Cluster 1 was enriched for nuclear processes and had worse outcomes, whereas Post-NAC Cluster 2 was enriched for immune pathways. Multiregional analysis of pre- and post-NAC matched tumors revealed clonal heterogeneity suggestive of important chemoresistance mechanisms. CONCLUSIONS: Here we describe 4 pre-NAC and 2 post-NAC proteomic clusters with distinct biology and survival outcomes, alongside novel prognostic biomarkers. Future work includes IHC validation of clusters in larger independent MIBC cohorts. A non-NAC cohort using pre-RC biopsy tissue will be used to confirm the prognostic vs. predictive relevance of these findings. Source of Funding: Bladder Cancer Canada, Deutsche Forschungsgemeinschaft © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e731 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alberto Contreras-Sanz More articles by this author Moritz J. Reike More articles by this author Gian L. Negri More articles by this author Htoo Zarni Oo More articles by this author Sandra Spencer Miko More articles by this author Karina Nielsen More articles by this author Morgan E. Roberts More articles by this author Joshua Scurll More articles by this author Kenichiro Ikeda More articles by this author Gang Wang More articles by this author Roland Seiler More articles by this author Gregg B. Morin More articles by this author Peter C. Black More articles by this author Expand All Advertisement PDF downloadLoading ...

Hexagonal-Grid-Layout Image Segmentation Using Shock Filters: Computational Complexity Case Study for Microarray Image Analysis Related to Machine Learning Approaches

Hexagonal grid layouts are advantageous in microarray technology; however, hexagonal grids appear in many fields, especially given the rise of new nanostructures and metamaterials, leading to the need for image analysis on such structures. This work proposes a shock-filter-based approach driven by mathematical morphology for the segmentation of image objects disposed in a hexagonal grid. The original image is decomposed into a pair of rectangular grids, such that their superposition generates the initial image. Within each rectangular grid, the shock-filters are once again used to confine the foreground information for each image object into an area of interest. The proposed methodology was successfully applied for microarray spot segmentation, whereas its character of generality is underlined by the segmentation results obtained for two other types of hexagonal grid layouts. Considering the segmentation accuracy through specific quality measures for microarray images, such as the mean absolute error and the coefficient of variation, high correlations of our computed spot intensity features with the annotated reference values were found, indicating the reliability of the proposed approach. Moreover, taking into account that the shock-filter PDE formalism is targeting the one-dimensional luminance profile function, the computational complexity to determine the grid is minimized. The order of growth for the computational complexity of our approach is at least one order of magnitude lower when compared with state-of-the-art microarray segmentation approaches, ranging from classical to machine learning ones.

Use of prostate cancer subtyping by gene expression to predict response to radiation and chemohormonal therapies.

241 Background: We previously reported on the use of gene expression profiling to characterize four primary subtypes in an analysis of over 100,000 prostate cancer primary tumors. Here we examine these subtypes and response to radiation (RT) after prostatectomy or chemotherapy in addition to androgen deprivation therapy (ADT) in hormone-sensitive metastatic prostate cancer (mHSPC). Methods: We examined a 215-gene prostate subtyping classifier (PSC) which groups luminal and basal gene expression profiles into one of four subtypes; luminal differentiated (LD), luminal proliferating (LP), basal immune (BI) and basal neuroendocrine-like (BN). The log-rank method was used to compare (i) metastasis-free survival between patients who did and did not receive salvage RT in the META855 cohort of localized disease patients treated with radical prostatectomy (n=855) and the (ii) overall survival of patients on the Phase III mHSPC EA3805 CHAARTED trial of ADT or ADT + docetaxel chemotherapy (n=160). Results: After radical prostatectomy, patients with BI tumors derived benefit from RT in terms of metastasis-free survival ( P=9.23e-4) while those with other subtype tumors did not (Each Log-rank P≥0.5). Patients with metastatic disease and LP tumors derived benefit from docetaxel (Log-rank P=0.002) while those with other subtypes tumors did not (Log-rank P=0.2). Conclusions: Basal-luminal subtyping segments tumors by relevant biological processes with potential implications for identifying patients who benefit from salvage radiation post prostatectomy and addition of docetaxel to ADT in mHSPC.

Flicker-free dual-volume augmented reality display using a pixelated interwoven integral floating technique with a geometric phase lens

A geometric phase (GP) integral floating display can provide multifocal three-dimensional (3D) augmented reality (AR) images with enhanced depth expression by switching the focal modes of the GP lens via polarization control. However, using temporal multiplexing to switch between the focal modes of GP optics causes flickering as each 3D AR image is fully presented in different frames and their temporal luminance profile becomes easily recognizable, particularly as the number of available focal modes increases. Here, we propose a novel integral floating technique to generate pixelated interwoven 3D AR images; a half of each image is spatially mixed with another and presented in both focal modes simultaneously to resolve the flickering issue. The principle was verified via experimental demonstration and optically measured data.