Solid Types Research Articles

Abstract 6882: Exploring aryl hydrocarbon receptor expression and distribution in the tumor microenvironment, with a focus on immune cells, in various solid cancer types

Abstract Background: The aryl hydrocarbon receptor (AhR) is a transcription factor that performs various biological functions upon ligand activation. While there have been extensive studies and development of AhR-targeting anticancer therapies, including active clinical trials, research on the expression of AhR, a key biomarker, in the TME remain limited. Herein, we evaluated the nuclear and cytosolic AhR expression across five solid cancer types: head and neck squamous cell carcinoma (HNSCC), bladder cancer, colorectal cancer, esophageal cancer, and non-small cell lung cancer (NSCLC). Methods: Multiplexed immunohistochemistry (mIHC) and image cytometry were employed to investigate the AhR expression and distribution in 513 patient samples, of which 292 were from patients with one of five solid cancer types. For quantification of AhR expression levels, FCS files were generated, and the resulting data was analyzed using the R program. Results: AhR expression was predominantly high in cancer cells, followed by T cells and macrophages across all cancer types. All 513 patient samples were categorized into three clusters based on distinct expression and localization patterns of AhR. Cluster 1 exhibited high AhR expression in the cancer cell nucleus, whereas cluster 3 showed the lowest. AhR expression in the stromal macrophage nucleus was the most pronounced in cluster 2. In cluster 1, NSCLC was entirely absent, with esophageal cancer representing the largest proportion. In contrast, cluster 2 was notably different from cluster 1, with a 5% prevalence of NSCLC and a relatively higher proportion of HNSCC and colorectal cancer. The AhR expression in macrophages within cluster 2 was noticeably higher than in cluster 1. Cluster 3, which had the highest NSCLC proportion, generally displayed low AhR expression compared to other clusters, but also exhibited AhR expression in regulatory T cells (Tregs), a pattern which was not observed in other cancer types. Notably, a positive correlation was observed between the nuclear and cytosolic expressions of AhR, suggesting that AhR expression as a biomarker is independent of its subcellular localization. Furthermore, there was no clear correlation between AhR expression levels and cancer stage, grade, and metastatic status. Conclusion: We elucidated the expression profile of AhR within the TME across five cancer types, then classified the patient samples into three clusters based on their distinct AhR expression patterns. By demonstrating different expression patterns of AhR in cancer cells and immune cells, our findings are anticipated to provide a fundamental basis for clinical and immunological research on AhR-targeting therapies. Citation Format: Dong Kwon Kim, Chai Young Lee, Yu Jin Han, So Young Park, Heekyung Han, Kwangmin Na, Mi Hyun Kim, Seung Min Yang, Sujeong Baek, Youngtaek Kim, Joon Yeon Hwang, Seul Lee, Seong-san Kang, Taedong Han, Hyounmie Doh, Jongho Cho, Dajeong Kim, Daewon Cha, Min Hee Hong, Sun Min Lim, Jii Bum Lee, Jae Hwan Kim, Kyoung-Ho Pyo, Byoung Chul Cho. Exploring aryl hydrocarbon receptor expression and distribution in the tumor microenvironment, with a focus on immune cells, in various solid cancer types [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 6882.

Abstract 1152: Pan-cancer single-cell and subcellular characterization of lymphoid aggregates and tertiary lymphoid structures using high-plex spatial molecular imaging

Abstract Tertiary lymphoid structures (TLSs), clusters of immune cells that form in non-lymphoid tissues, are frequently identified in a wide range of solid tumors. They function as immunological “hubs”, driving anti-tumor responses. Previous studies have demonstrated that the presence of TLSs is often associated with improved responses to immunotherapy and superior clinical outcomes. Despite these insights, our understanding of their phenotypic diversity, spatial organization, intercellular communication, and interactions with cancer cells and the tumor microenvironment remains incomplete. Existing methods lack the resolution, depth, and scale required for a thorough understanding of these complex structures. We have obtained in-house and public spatial multi-omics data generated on various tissues across 9 different cancer types using the cutting-edge single-cell spatial transcriptomics and high-plex spatial molecular imaging platforms. We have developed innovative computational approaches for the detailed characterization of TLSs at single-cell and subcellular resolution and carried out an extensive analysis of lymphoid aggregates (LAs) and TLSs in tissue sections across these different solid cancer types. We defined the phenotypic state of each individual cell within TLSs including B/plasma cells, follicular helper/regulatory/stressed/exhausted T cells, follicular dendritic cells, fibroblasts and high endothelial venules. Then, we performed unbiased phenotyping and classification of TLSs based on their cellular states, compositions and organization patterns across various types of tissues. Additionally, we performed spatial neighborhood analysis, characterized the neighboring cells of each TLS and analyzed their cell-cell communication networks. Our approach allowed an unprecedented examination of the spatial phenotypes of TLSs in large, complex tissues at super resolution, in ways that were not previously described. We comprehensively characterized TLSs across different solid tumor types. Our analysis revealed a high degree of spatial and phenotypic heterogeneity in LAs and TLSs. We created a comprehensive spatial atlas of LAs and TLSs across solid tumor types, profiled TLSs at various maturation states with distinct cellular composition and organization patterns, and discovered new TLS states. Notably, we observed distinct communication networks associated with TLS phenotypic states. Our innovative methods and analytical approaches offer a more comprehensive way to study LAs and TLSs, enhancing detection capabilities and providing deeper insights into their spatial interaction and functional phenotypes. This research holds the potential to significantly advance our understanding of TLSs and their roles in immunobiology and immunotherapy responses across a broad range of cancers and diseases. Citation Format: Yunhe Liu, Yuanyuan Zhang, Guangchun Han, Guangsheng Pei, Kyung Serk Cho, Enyu Dai, Yanshuo Chu, Jiahui Jiang, Ansam Sinjab, Luisa M. Soto, Alejandra Serrano, Jianjun Gao, Humam Kadara, Catherine Fridman, Wolf Herve Fridman, Linghua Wang. Pan-cancer single-cell and subcellular characterization of lymphoid aggregates and tertiary lymphoid structures using high-plex spatial molecular imaging [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 1152.

Abstract 291: Unveiling heterogeneity of cancer-associated fibroblasts (CAFs) across multiple solid cancer types

Abstract Background: Stromal cells, including CAFs, are a heterogeneous group of cells in the tumor microenvironment. Many models describe their heterogeneity like the classification into 3 main subtypes in breast cancer: myCAF (myofibroblasts), iCAF (inflammatory CAF), and PVL (perivascular-like) cells. However, a pan-cancer model has not been established to exhaustively characterize stromal cell heterogeneity, undermining efforts in targeting CAFs with anti-tumor therapies. We investigated the heterogeneity of stromal cells in diverse diagnoses with single-cell RNA sequencing (scRNA-seq) data and bulk RNA-seq of sorted stromal cells to define consistent subtypes. Design: First, a meta-cohort of 11 scRNA-seq open-source datasets (>500,000 cells, >100 patients, 6 diagnoses) was analyzed. We also generated a scRNA-seq dataset of CD90+ cells derived from tumor tissue of an ovarian cancer patient. Additionally, bulk RNA-seq of 6 PVL, 2 iCAF, and 4 myCAF sorted cell samples across four solid cancers was performed. scRNA-seq data was analyzed with the SCANPY Python package. Results: Stromal cell clusters were defined as positive for PDGFRB, THY1 (CD90), and COL1A1 expression, and negative for the expression of known non-stromal and endothelial cell markers, by separately analyzing the open-source scRNA-seq datasets. Analysis of stromal cell heterogeneity revealed that the CAFs were consistently classified into 3 subtypes across all datasets: myCAF (high expression: FAP and ACTA2), PVL (high expression: MCAM (CD146) and ACTA2), and iCAF (high expression: APOD and cytokines IL6, CXCL12, CXCL2). Differential gene expression analysis of the scRNA-seq meta-cohort allowed us to develop functional gene expression signatures to define each subtype. All scRNA-sequenced CD90+ cells from ovarian cancer tissue were classified into the CAF subtypes. The myCAF and PVL signatures were specifically upregulated in the corresponding cell types (p<0.01, Mann-Whitney U test). While the iCAF signature was enriched in both iCAFs and myCAFs, myCAF and PVL signatures were downregulated in iCAFs. Finally, we developed a fluorescence-activated cell sorting (FACS) panel to separate CAF subtypes, using negative markers (CD45, CD31, EPCAM), a positive marker for stromal cells (CD90) and cell-type surface markers (CD146, FAP, CD91, CD44) for myCAF, iCAF and PVL cells. The high expression of CAF-specific genes in the sorted CAF populations denoted high sample purity. Importantly, the 3 CAF subpopulations separated by gating based on different levels of cell-type surface markers showed expression patterns congruent with the developed signatures across diagnoses. Summary: Altogether, the consistent classification of stromal cells with our cell-specific expression profiles demonstrated that the 3 CAF subtypes are conserved in many cancer types, offering groundwork for a pan-cancer model of stromal cell heterogeneity. Citation Format: Andrey Kravets, Boris Shpak, Anastasia Zotova, Maria Savchenko, Sofya Kust, Olga Golubeva, Mary Abdou, Fedor Grigoryev, Irina Bulusheva, Beniamin Sargsyan, Polina Pavlovich, Vlad Maximov, Vladimir Kushnarev, Michael Goldberg, Nikita Kotlov, Alexander Bagaev. Unveiling heterogeneity of cancer-associated fibroblasts (CAFs) across multiple solid cancer types [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 291.

Abstract 5139: PD1-PDL1 interaction as a superior predictor for response to immune checkpoint therapy in NSCLC patients

Abstract Background: Immune checkpoint therapies (ICT) are now widely introduced in clinical oncology and most often guided by PDL1 expression in diagnostic biopsies. However, the predictive value of this testing is only modest, leading to over- and undertreatment. Conceptually, the prevention of binding of ligand PDL1 to PD1 receptor is decisive for the efficacy of ICT. Therefore, directly detecting the spatial PD1-PDL1 interaction would provide more predictive information. Methods: A second-generation in situ proximity ligation assay (isPLA; Navinci Diagnostics) was established to detect PD1-PD-L1 interactions in diagnostic patient samples and linked to an automated analysis pipeline (QuPath). For basic validation, we used pan-cancer tissue microarrays (TMA) including 16 solid tumor types, and a non-small cell lung cancer (NSCLC) TMA (359 ICT-naïve surgically treated patients). In addition, we analyzed diagnostic tissue biopsies from 75 advanced NSCLC patients treated with anti-PD1-PDL1 regiments. Results: The pan-cancer analysis revealed varying levels of PD1-PDL1 interaction among the solid cancer types, with the lowest levels detected in liver cancer and the highest in testicular cancer. A general positive association was observed between the literature-reported objective ICT response rate (ORR) per tumor type and its corresponding PD1-PDL1 interaction status. Conventional immunohistochemical analysis of the ICT-naïve, surgically treated NSCLC cohort revealed variable protein expression level for both, PDL1 and PD1, for 200 cases, but only 108 (54%) of them demonstrated a detectable PD1-PDL1 interaction with our isPLA assay. EGFR mutated cases showed generally lower PD1-PDL1 interaction scores (p=0.01). PD1-PDL1 interaction was not associated with survival (p=0.46). A subsequent analysis of the ICT-NSCLC cohort revealed a significant positive association between PD1-PDL1 interaction status and prolonged overall survival upon ICT (median survival 14.8 vs 32.3 months; p=0.022). The standard tumor proportional score (TPS) of membranous PDL1 expression did not correlate with ICT-specific survival (p=0.40). Conclusions: The second generation isPLA successfully distinguishes a previously difficult-to-define patient subset with positive PD1-PDL1 interaction status. Our data suggest that the PD1-PDL1 interaction is a superior clinical biomarker for patient selection for ICT in NSCLC. The interaction analysis is applicable to sections from minute diagnostics biopsies and accessible by light microscopy either semi-quantitatively by a pathologist or by an automated image analysis program. Thus, the assay holds great potential to be integrated into routine clinical pathological workflow to stratify cancer patients for ICT. Citation Format: Patrick Micke, Amanda Lindberg, Rebecca Artursson, Louise Hellberg, Hui Yu, Max Backman, Johan Botling, Hans Brunnström, Artur Mezheyeuski, Johan Isaksson, Carina Strell. PD1-PDL1 interaction as a superior predictor for response to immune checkpoint therapy in NSCLC patients [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 5139.

Abstract 1362: IOMX-0675, a LILRB1 and LILRB2 cross-specific antibody, effectively repolarizes immunosuppressive myeloid cells and activates T cells leading to potent tumor cell killing

Abstract Recently, myeloid checkpoints in the tumor microenvironment have gained increased attention in the context of tumor immune evasion. LILRB1 (ILT2) and LILRB2 (ILT4) are immunosuppressive receptors of the leukocyte immunoglobulin-like receptor (LILR) family that recognize both classical and non-classical MHC-I molecules (e.g., HLA-G). While tumor-infiltrating myeloid cells express both LILRB1 and LILRB2, lymphoid cells are restricted to LILRB1 expression. LILRB1 and LILRB2 are frequently co-expressed with immune-activating LILR family members LILRA1 and LILRA3 in several tumor indications and upregulated in patients non-responsive to T cell checkpoint blockade. Furthermore, the non-classical MHC-I molecule HLA-G, a major ligand of LILRB1 and LILRB2 in the tumor environment, is overexpressed and associated with poor prognosis in multiple solid cancer types. IOMX-0675, a fully human, Fc-silenced, monoclonal immunoglobulin G1 (IgG1) antibody with a highly differentiated binding profile, that binds selectively with high affinity to the inhibitory receptors LILRB1 and LILRB2, while binding only weakly to the closely related immune-activating LILR family members LILRA1 and LILRA3, was identified from iOmx’ proprietary phage display library. A high-resolution Fab-based Octet assay was established to monitor the IOMX-0675-mediated blocking of LILRB1 or LILRB2 binding to HLA-G. The LILRB1/2 cross-specific antibody promotes phagocytic and pro-inflammatory activity of various macrophage subtypes, which was monitored by flow cytometry and cytokine profiling. In co-cultures of monocyte-derived macrophages with autologous T cells, IOMX-0675 shows high potential to reprogram dose-dependently immunosuppressive macrophage, to further activate pro-inflammatory macrophage subtypes and thereby rescues the activity of the lymphoid immune compartment. Even in environments dominated by LILRA1/A3, IOMX-0675 shows a strong binding capacity to the immunosuppressive LILRB1/B2 receptors and significantly repolarizes the immune environment. In summary, we discovered IOMX-0675, a cross-specific antibody antagonizing both LILRB1 and LILRB2 with high selectivity, that exhibits potent reprogramming of the immunosuppressive myeloid compartment and restores cytotoxic T cell activity in the tumor microenvironment. Negligible binding to immune-activating LILR family members by IOMX-0675 could unleash full anti-tumor activity. The differential binding profile of IOMX-0675 provides best-in-class potential and may maximize anti-tumor efficacy for the benefit of patients with high-unmet medical need, who are resistant to T cell checkpoint blockade. Citation Format: Christina Hartl, Jonas Zantow, Ilona-Petra Maser, Marisa Stebegg-Wagner, Simone Friedrich, Jonas Schilz, Stefan Kaden, Ronny Milde, Eugenia Korotkova, Michail Maraslis, Bettina Langer, Michal Swiat, Carmen Ginzel, Carmen Ammerhauser, Lucille Albert, Leonie Majunke, Alina Huth, Kritika Sudan, Murray Yule, Christine Rothe, Hannes Loferer, Maximilian Aigner, Alexander N. Marziale, Stefan Bissinger. IOMX-0675, a LILRB1 and LILRB2 cross-specific antibody, effectively repolarizes immunosuppressive myeloid cells and activates T cells leading to potent tumor cell killing [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 1362.

Abstract 6742: PV 10 induces endoplasmic reticulum stress and autophagy, triggering immunogenic cell death and anti-tumor immunity in head and neck squamous cell carcinoma

Abstract Introduction: Head and neck squamous cell carcinoma (HNSCC) are complex, diverse cancers affecting mucosal linings of the upper aerodigestive tract. Globally, HNSCC is the sixth most common cancer, leading to over 890,000 new cases and more than 450,000 deaths annually. Primary risk factors include human papillomavirus (HPV) infection (HPV-positive HNSCC) and exposure to tobacco and excessive alcohol (HPV-negative HNSCC). Despite multimodal treatments, survival rates remain low due to recurrence and metastasis (R/M HNSCC), creating a need for safe, innovative therapies to reduce tumor burden with minimal adverse events. PV-10 (10% rose bengal sodium) is a novel intratumoral immunotherapeutic agent that has demonstrated promising anti-tumor activity in multiple solid tumor cancer types with few adverse effects in pre-clinical research and clinical trials. However, the role of PV-10 in HNSCCs and its role in enhancing immune response warrants further investigation. Methods: Cell viability was assessed using Alamar blue assay after PV-10 treatment of murine mEER and MTE-RAS HNSCC tumor cell lines. To evaluate the immunomodulatory properties of PV-10, we employed flow cytometry, confocal analysis, and luminescence-based assays. Additionally, we performed a DCFDA assay to measure the reactive oxygen species (ROS) and an Alexa Fluor 488 Annexin V/dead cell apoptosis assay to evaluate apoptosis. To gain insights into the mechanisms underlying PV-10's immunomodulatory effects in cancer cells and tumors from mice, western blotting and multiplex immunohistochemistry (mIHC) were used. Results: Our in vitro findings reveal that PV-10 induces cytotoxicity in both mEER and MTE-RAS cells. Notably, PV-10 promotes a significant increase in ROS, leading to an elevation in late apoptotic cells. Markers of immunogenic cell death (ICD), including a statistically significant increase in the release of damage-associated molecular pattern molecules HMGB1 and ATP, as well as enhanced surface expression of calreticulin, HSP-70, and HSP-90, were observed. At the molecular level, a remarkable activation of endoplasmic reticulum (ER) stress, pro-apoptotic protein, and autophagy markers were observed. Intratumoral PV-10 injection in vivo has shown significant tumor regression in both mEER and MTE-RAS tumors, and a complete response was noticed in some mice, indicating that PV-10 induces potent ICD in both mEER and MTE-RAS tumors. Conclusion: Our collated evidence strongly suggests that PV-10 induces ICD-associated immunogenicity in both mEER and MTE-RAS in vitro and in vivo, fostered by ROS-based ER stress and apoptosis. These findings hold promise for potential therapeutic avenues to manage HNSCC. Citation Format: Sowjanya Thatikonda, Ritu Chaudhary, Yeva Meshkovska, Maria I. Biernacki, Robbert Slebos, Shari P. Thomas, Amelio L. Antonio, Eric A. Wachter, Christine H. Chung. PV 10 induces endoplasmic reticulum stress and autophagy, triggering immunogenic cell death and anti-tumor immunity in head and neck squamous cell carcinoma [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 6742.

Abstract 847: Association of methylation-based stem cell divisions with lifetime cancer risk and cancer progression

Abstract Cancer initiation arises from the accumulation of somatic mutations acquired during stem cell mitotic events mediated by carcinogenic risk factors. On an ecological level, Tomasetti and Vogelstein (2015) demonstrated that lifetime risk of solid tumor cancer types were strongly correlated with total number of stem cell divisions estimated from tissue-specific cell lines. Recently, several mitotic clocks (MC) have been developed to estimate stem cell division rates (SCDR) based on whole-genome methylation arrays. Here, we compare estimated SCDR for several tissue types with tissue-specific cancer incidence and lifetime risk estimates. Furthermore, we assess the link between SCDR with cancer progression and survival. Array-based methylation data derived from normal adjacent tissue in TCGA was used to estimate SCDR for 15 cancer types. A Pearson correlation test was performed to assess the association between SCDR and cancer-specific incidence and lifetime risk for each cancer type overall and stratified by self-reported race (e.g. White and Black). SCDR was estimated in tumors from TCGA for 4 common cancers and subtypes: breast (BRCA), prostate (PRAD), lung adenocarcinoma (LUAD), and lung squamous cell carcinoma (LUSC). A Spearman correlation test was used to identify potential trends between SCDR and pathological stage. Cox proportional hazard (PH) models adjusted for age at diagnosis and treatment status (e.g. surgery, radiation, etc.) were used to evaluate the association of time to death with SCDR terciles across different cancer stages. We observed a similar trend between SCDR and overall incidence (cor=0.61, p=0.01), aligning with earlier findings. Stratified-specific trends between cancer incidence and SCDR was consistent for both Black and White populations (cor=0.62 vs 0.66). Tumor analysis revealed a significant correlation between SCDR and pathological stage (e.g. stage 1, 2, and 3/4) for BRCA (p<0.001) and pathological stage (e.g. stage 2, 3 and 4) for PRAD (p<0.001), suggesting its potential as a marker of cancer progression. However, the correlation between LUAD, LUSC and pathological stage was insignificant. Based on Cox PH model, within stage III BRCA, SCDR significantly contributed to explaining variability in survival (p=0.04), emphasizing its potential prognostic value beyond cancer stage. Our study supports earlier findings applying methylation based SCDR estimates and shows that this association is consistent across racial groups. This implies that SCDR may act as a biological marker of a combination of various genetic and environmental risk factors underlying disease etiology. Further analyses will be extended to all cancers in TCGA. Lastly, we demonstrate that MC-based SCDR estimation has the potential to serve as a clinical tool to identify individuals at elevated risk of cancer initiation, track progression, and complement cancer stage for predicting survival. Citation Format: Yanning Wu, Cheryl Thompson, Fredrick Schumacher. Association of methylation-based stem cell divisions with lifetime cancer risk and cancer progression [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 847.

Differences in Histological Subtypes of Invasive Lobular Breast Carcinoma According to Immunohistochemical Molecular Classification.

The aim of this study was to compare the differences between histological and molecular subtypes of invasive lobular carcinoma (ILC) of the breast, in order to be able to predict the behavior and prognosis of the disease, as well as to effectively determine therapy. This study included 263 cases of breast ILC diagnosed over a seven-year period. The diagnosis of invasive lobular carcinoma is based on the characteristic growth pattern and phenotype of cancer cells with the respective subtypes: classic, alveolar, solid, tubulolobular, pleomorphic and mixed lobular type. The examined cases were divided into five groups according to molecular classification based on the expression of ER, PR, HER2 and Ki67 immunohistochemical markers. It was found that the pleomorphic subtype occurred statistically significantly less often as the luminal A subtype compared to others (p = 0.00027), and the HER2-enriched subtype occurred statistically significantly more often in the pT4 stage (p = 0.024). The results of this study significantly singled out the luminal A subtype, and among them classic ILC, as the subtype with the most favorable expression ratio of the investigated predictive/prognostic immunohistochemical markers.

Benign Brenner tumor of the ovary: two-dimensional and contrast-enhanced ultrasound features-a retrospective study from a single center.

Benign Brenner tumor (BBT) is a rare ovarian tumor, and there are few discrete reports about its manifestation in an ultrasound. This study sought to investigate the two-dimensional (2D) and contrast-enhanced ultrasound (CEUS) features of this entity. This is a retrospective single-center study. The clinical manifestations, laboratory examination, and ultrasound data of 25 female patients with BBT were confirmed by pathology when they underwent 2D and/or CEUS examination at Ningbo First Hospital from January 2012 to June 2023. The ultrasound findings of the patients were analyzed using the terminology of the International Organization for the Analysis of Ovarian Tumor and were read by two senior sonographers who reached an agreement. Among the all 25 patients, most of them were unilateral, and only one patient was bilateral. Thus, 26 lesions were found: 44.0% (11/25) were in the left and 52.0% (13/25) were in the right. Moreover, 53.84% (14/26) were solid lesions, 15.38% (4/26) were mixed lesions, and 26.92% (7/26) were cystic lesions. Among the solid-type patients, 42.85% (6/14) of the cases were with calcification. Upon laboratory examination, 12.0% (3/25) of the patients had high carbohydrate antigen 125 (CA-125) level, and 19.04% (4/21) of the patients had an elevated carbohydrate antigen724 (CA-724) level in the serum tumor markers. In the hormone test, 14.28% (3/21) were found to have a high postmenopausal estrogen level and 14.28%(3/21) were found to have a high level of follicle-stimulating hormone (FSH). One patient with complex manifestations and three with solid manifestations were examined by CEUS to observe the microcirculation perfusion of the tumor. One with solid and cystic separation was rapidly hyperenhanced and cleared, and the filling subsided faster than the uterus. The postoperative pathological diagnosis was benign Brenner tumor with mucinous cystadenoma. The other three cases were solid adnexal lesions, which showed isoenhancement on CEUS and disappeared slowly, synchronizing with the uterus. The CEUS results were considered as benign tumors and confirmed by pathology. BBT can show ovarian cystic, mixed cystic and solid type, and solid echo in 2D ultrasound. Unilateral ovarian fibrosis with punctate calcification is an important feature of BBT in 2D ultrasound. However, for solid adnexal masses and mixed cystic and solid masses with unclear diagnosis, if CEUS shows isoenhancement or hyperenhancement, the possibility of BBT cannot be excluded.

GZ17-6.02 interacts with proteasome inhibitors to kill multiple myeloma cells.

GZ17-6.02, a synthetically manufactured compound containing isovanillin, harmine and curcumin, has undergone phase I evaluation in patients with solid tumors (NCT03775525) with a recommended phase 2 dose (RP2D) of 375 mg PO BID. GZ17-6.02 was more efficacious as a single agent at killing multiple myeloma cells than had previously been observed in solid tumor cell types. GZ17-6.02 interacted with proteasome inhibitors in a greater than additive fashion to kill myeloma cells and alone it killed inhibitor-resistant cells to a similar extent. The drug combination of GZ17-6.02 and bortezomib activated ATM, the AMPK and PERK and inactivated ULK1, mTORC1, eIF2α, NFκB and the Hippo pathway. The combination increased ATG13 S318 phosphorylation and the expression of Beclin1, ATG5, BAK and BIM, and reduced the levels of BCL-XL and MCL1. GZ17-6.02 interacted with bortezomib to enhance autophagosome formation and autophagic flux, and knock down of ATM, AMPKα, ULK1, Beclin1 or ATG5 significantly reduced both autophagy and tumor cell killing. Knock down of BAK and BIM significantly reduced tumor cell killing. The expression of HDACs1/2/3 was significantly reduced beyond that previously observed in solid tumor cells and required autophagy. This was associated with increased acetylation and methylation of histone H3. Combined knock down of HDACs1/2/3 caused activation of ATM and the AMPK and caused inactivation of ULK1, mTORC1, NFκB and the Hippo pathway. HDAC knock down also enhanced ATG13 phosphorylation, increased BAK levels and reduced those of BCL-XL. Collectively, our present studies support performing additional in vivo studies with multiple myeloma cells.

Angioleiomyoma: An Update with a 142-Case Series.

Angioleiomyomas are uncommon, noncancerous, smooth muscle tumors that primarily arise from blood vessels. Previous studies have yielded limited data due to the lack of interdisciplinary approaches or restricted patient pools. This study aims to provide a comprehensive analysis of angioleiomyomas, including the demographic, clinical, radiological, and histopathological features, with a large number of patients. Conducted as a retrospective investigation at a single center from January 2005 to June 2023, this study involved 142 patients. Relevant information was extracted from electronic medical records, covering clinical, radiological, histological, and demographic details. Angioleiomyomas mostly occurred at age 59 (1-87), predominately affect females (53%) and commonly arise in subcutaneous tissue (85%) and the lower limbs (76%). MRI findings revealed characteristic signals, with a high prevalence of the solid histologic type (65%), often displaying a reticular sign. Smooth muscle Actin was universally present (n = 95/95), while Desmin and Caldesmon showed positive expression in 83% (n = 71/85) and 98% (n = 92/94) of cases, respectively. This study presents an updated and comprehensive analysis of angioleiomyomas. Typically appearing as well-defined nodules in the extremities, these tumors can be effectively diagnosed using MRI, though histopathological analysis is generally essential for confirmation. Treatment primarily involves straightforward excision, with notable low complication and recurrence rates.

Comparative study on fatigue evaluation of suspenders by introducing actual vehicle trajectory data

Suspenders play a crucial role in transmitting loads from the bridge deck to the main cable in a suspension bridge. They are susceptible to fatigue due to repeated dynamic loads, particularly traffic loads. Traffic Load Models (TLMs), typically created using Monte–Carlo simulation and Weigh-In-Motion (WIM) data, are employed to evaluate this fatigue. However, these models often overlook practical vehicle trajectories and spatio-temporal distribution, which compromises the precision of fatigue assessments. In this study, we introduce a novel 2D Intelligent Driver Model (2D-IDM) that incorporates actual vehicle trajectories, with a particular focus on transverse vehicle movement. This enhancement aims to improve the fidelity of existing TLMs. To provide a clear, qualitative, and quantitative understanding of the effects of fatigue evaluation with or without actual trajectory characteristics, we have structured this paper as a comparative study. We compare our proposed model, denoted as TLM S-3, with two observation-based models (O-1 and O-2) and two simulation-based models (S-1 and S-2). We conducted an experimental case study on a long-span suspension bridge, where the actual traffic load trajectory was obtained using a WIM-Vision integrated system. To calculate fatigue damage considering both longitudinal and transverse directions, we established a multi-scale Finite Element Model (FEM) using solid element types to simulate the bridge girder. This model can generate the stress influence surface of the bridge and has been verified in both static and dynamic aspects. Our detailed comparative analysis demonstrates the consistency of the proposed 2D-IDM with the actual measured traffic load trajectories. This indicates that our approach can enhance the fidelity and precision of fatigue evaluations for bridge suspenders.

Rare inflammatory myofibroblastic lung tumor with multi-organ metastasis: A case report

Inflammatory myofibroblastic tumors (IMTs) are extremely rare neoplasms, accounting for 0.04–0.7% of all tumors. Data on their clinical presentation, pathogenesis, and biological behavior are scarce. They can occur at any age, involving equally males and females. IMTs have been described as benign neoplasms with intermediate malignant potential, as local recurrence and distant metastasis may occur. IMTs are characterized by a solid growth type, and a multifocal type is uncommon. Definitive surgery is the treatment of choice. We describe a clinical case of a malignant IMT with multi-organ metastasis 6 years after definitive surgery on the primary tumor. There is no standard of care in such a clinical situation, especially given the absence of mutations in the primary and metastatic tumors.

Targeting ATR in patients with cancer.

Pharmacological inhibition of the ataxia telangiectasia and Rad3-related protein serine/threonine kinase (ATR; also known as FRAP-related protein (FRP1)) has emerged as a promising strategy for cancer treatment that exploits synthetic lethal interactions with proteins involved in DNA damage repair, overcomes resistance to other therapies and enhances antitumour immunity. Multiple novel, potent ATR inhibitors are being tested in clinical trials using biomarker-directed approaches and involving patients across a broad range of solid cancer types; some of these inhibitors have now entered phase III trials. Further insight into the complex interactions of ATR with other DNA replication stress response pathway components and with the immune system is necessary in order to optimally harness the potential of ATR inhibitors in the clinic and achieve hypomorphic targeting of the various ATR functions. Furthermore, a deeper understanding of the diverse range of predictive biomarkers of response to ATR inhibitors and of the intraclass differences between these agents could help to refine trial design and patient selection strategies. Key challenges that remain in the clinical development of ATR inhibitors include the optimization of their therapeutic index and the development of rational combinations with these agents. In this Review, we detail the molecular mechanisms regulated by ATR and their clinical relevance, and discuss the challenges that must be addressed to extend the benefit of ATR inhibitors to a broad population of patients with cancer.

The effect of solid lubricants in the brake friction composite on brake emissions: A case study with graphite, Sb2S3, and MoS2

Particulate matter produced during brake application was investigated, focusing on the effect of the solid lubricants in the brake friction composite, such as graphite, antimony trisulfide, and molybdenum disulfide. Results showed that the brake emission was significantly affected by the solid lubricant type and braking temperatures. Antimony trisulfide caused high brake emissions at moderate temperatures, whereas graphite produced high emissions at elevated temperatures. The brake emission at moderate temperatures was affected by the aggressiveness of the composites and the presence of friction films on the gray iron disc surface. In contrast, the brake emissions at elevated temperatures were determined by the thermal decomposition of the composite and the oxidation of solid lubricants.

An Unusual Case Report of Unicystic Ameloblastoma of the Mandible

ABSTRACT There are plenty of benign lesions that can result in swelling of the mandible, and these can be classified as odontogenic and non-odontogenic lesions. Among the categories of odontogenic lesion, ameloblastoma is the most occurring lesion that takes origin from the epithelial cellular elements and dental tissues in their different stages of development. Ameloblastoma is the most serious odontogenic neoplasm due to its prevalence and clinical characteristics. Ameloblastoma is a broad class which encompasses 80% of solid multicystic type of ameloblastoma with unicystic ameloblastoma (UA) variant included as vital clinicopathological form claiming the rest 20% along with peripheral ameloblastoma variant. UA refers to cystic lesions that seem like jaw cysts clinically, radiographically, or grossly but are lined by typical ameloblastomatous epithelium, with or without luminal and/or mural tumor development, on histologic investigation. Around 5–15% of all ameloblastic lesions do not have a propensity to metastasis, and this is UA. Unicystic mural form, although slow growing overall, is very invasive locally and has a high recurrence rate. As UA tumors show very close features with dentigerous cyst, a very sharp differential diagnosis protocol need to be executed to exclude the other unicystic odontogenic lesions considering the clinical, radiological, and biological characteristics along with proper follow-up and seeing any recurrence of the lesion taking place. Here, we report the case of a twenty-one year male patient with UA of the mandible and review of the literature.

Abstract A050: CD8+ T cell exhaustion and their DNA damage in BRCA1 mutated TNBC

Abstract PARP Poly (ADP-ribose) polymerase inhibitors (PARPi) are treatment options in TNBC patients with inherited BRCA1 mutations. Recent studies have shown that immune checkpoint therapies (ICT) in combination with PARPi are effective in other solid cancer types such as melanoma, pancreatic and BRCA deficient ovarian cancer. In early and advanced BRCA1 TNBC, ICT studies are limited and not fully elucidated especially for new ICT that target LAG-3, CTLA-4, ICOS. There is a need for identifying potential prognostic markers suggesting which patients could benefit most from ICT. Therefore, we conducted a detailed single cell analysis of the tumor microenvironment (TME) mainly focused on immune checkpoint expression, CD8 T cell exhaustion and DNA damage associated with BRCA1 TNBC compared to WT TNBC. Combining single cell spatial tissue multiplexing (PhenoCycler) (BRCA1 TNBC n=77, WT TNBC n=55), scRNA seq (BRCA2 n=1, WT n=5), FLOW (PBMCs: BRCA1 n=13, BRCA2 n=17, WT n=62) as well as bulk RNA seq (BRCA1 TNBC n=35, WT TNBC n=11), we performed a detailed analysis of the TME associated with BRCA1 and WT breast cancer. We developed a 32-plex antibody panel that evaluated T-cells and other immune cells with markers of exhaustion, DNA damage and immune checkpoint expression. CD8+ T cell frequency was significantly increased in BRCA1 TNBC compared to WT TNBC (p=0.0015). A detailed CD8 T cell characterization revealed increased dysfunctional/exhausted cells (e.g., TOX+/LAG-3+/PD-1+), T cell DNA damage (CD3e+/CD8+/RAD51+, p=0.0047) and LAG-3 transcript levels in BRCA1 TNBC compared to WT TNBC (p=0.0625). Furthermore, immune checkpoint protein expression in tumor- (PD-L1) and immune cells (PD-1, LAG-3, ICOS) was increased in the TME associated with BRCA1 TNBC compared to WT TNBC. Of note, we also found that PBMCs of healthy tumor-free BRCA mutation carriers contained significantly increased CD8+/PD+1 T cells compared to WT (p=0.012). Our data indicates that BRCA1 haploinsufficiency leads to DNA damaged and dysfunctional CD8+ T cells within the TME compared to WT. Thus, the TME of BRCA1 TNBC patients is more immune suppressive due to increased exhausted CD8+ T cell frequency, their DNA damage as well as immune checkpoint expression of both tumor-/and immune cells compared to WT TNBC. The effect of BRCA1 haploinsufficiency on immune cells is important to consider when trials of ICT options are designed. Citation Format: Dana Pueschl, Anupma Nayak, Derek A. Oldridge, William Chandler, Bria Fulmer, Sokratis Apostolidis, Danielle Bragen, Bradley Wubbenhorst, John Pluta, kConFab Investigator, Antonis Antoniou, Georgia Chenevix-Trench, E. John Wherry, Susan M. Domchek, Katherine L. Nathanson. CD8+ T cell exhaustion and their DNA damage in BRCA1 mutated TNBC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr A050.

Occurrence of NECTIN4 amplification in solid tumors and enfortumab vedotin response in metastatic urothelial cancer.

673 Background: The anti-NECTIN4 antibody-drug conjugate (ADC) enfortumab vedotin (EV) is approved for patients with metastatic urothelial cancer (mUC). However, durable benefit is only achieved in a small, yet uncharacterized patient subset. NECTIN4 is located on chromosome 1q23.3, and 1q23.3 gains are frequent genomic events in mUC leading to NECTIN4 amplifications. Here, we aimed to evaluate the potential of NECTIN4 amplification as a genomic biomarker to predict EV response in patients with mUC. Methods: We established a NECTIN4-specific fluorescence in-situ hybridization (FISH) assay to assess NECTIN4 copy number variations (CNVs) in a multicenter EV-treated mUC patient cohort (UC-EV, N=77), and CNV data were correlated with membranous NECTIN4 protein expression (H-score) assessed via immunohistochemistry, EV treatment responses and outcomes. Next, we conducted a pan-cancer analysis of the The Cancer Genome Atlas (TCGA) datasets, comprising 10,712 patients across 32 cancer types, to investigate the relationship between NECTIN4 CNV, mRNA expression (RNAseq) and protein levels (RPPA) across entities. Results: In TCGA cohorts, NECTIN4 amplification occurs frequently across different solid cancer types, especially in 15-20% of bladder cancers (17% in TCGA-BLCA), as well as 5-10% in breast cancer (TCGA-BRCA) and lung adenocarcinoma (TCGA-LUAD). We confirmed the amplification frequency in our UC-EV cohort (18%). NECTIN4 amplification is significantly associated with both increased NECTIN4 mRNA expression (e.g., TCGA-BLCA, BRCA, LUAD) and membranous NECTIN4 protein expression (UC-EV), and represents a stable genomic alteration during metastatic progression. In the UC-EV cohort, all patients with NECTIN4 amplification (N=14) responded to EV. In multivariable Cox adjusted for age and sex, NECTIN4 amplifications led to a 93% risk reduction for death compared to a NECTIN4non-amplified status (HR=0.07, 95%-CI 0.01–0.53; P<0.001). Conclusions: Our study highlights the value of NECTIN4 amplifications to predict EV responses in patients with mUC. NECTIN4 amplifications occur frequently in different cancer types and therefore have the potential to be a novel tumor-agnostic genomic biomarker that enables tailored NECTIN4-targeted therapies in various entities.

Current advancements in fluid-flow Problems: A brief review

This specific article is a compilation of a few current studies on fluid flow issues. This article provides an overview of current research on the experimental and theoretical analysis of various nanofluids' thermal conductivity. The current study examines a number of variables that have a significant impact on a nanofluid's ability to transfer heat, including temperature, solid volume fraction, type, size, shape, magnetic field, pH, ultrasonic time, and surfactant. In addition, some plausible and appealing theories that enhance the thermal conductivity of nanofluids are mentioned. The final important heat transmission mechanisms that play a significant part in improving thermal conductivity are Brownian motion, thermophoresis, nanoclustering, interfacial nano-layer, and osmophoresis. Thus, consideration is given to the heat transmission properties of nanofluids. And also discussed some Numerical method to solve fluid flow problems.

Imaging Characteristics of Clear Cell Papillary Renal Cell Carcinoma: Identifying the Sheep in Wolf's Clothing.

This study aimed to describe the characteristics of computed tomography (CT) and magnetic resonance imaging (MRI) of clear cell papillary renal cell carcinoma (CCPRCC). This retrospective study comprised 27 patients diagnosed with 29 tumors of CCPRCC. The study was approved by the Medical Ethics Committee and the requirement for the informed consent was waived. The inclusion criteria stipulated pathology-confirmed CCPRCCs with at least one preoperative imaging examination, including CT or MRI. Two experienced radiologists independently analyzed the imaging characteristics, including size, location, growth mode, morphology, texture, density, and enhancement pattern. Paired t-test was used to compare differences in CT Hounsfield unit values and apparent diffusion coefficient (ADC) imaging between the tumor and the renal cortex. The mean age of the 27 patients was 57.0 ± 14.2 years. Nineteen patients underwent CT, while 12 underwent MRI (There are 4 patients underwent not only CT but also MRI). Among the cases, 26 (96%) were single, and 1 (4%) was multiple, consisting of three lesions. Out of the 29 tumors, 15 (52%) were located in the left kidney and 14 (48%) in the right kidney. The mean tumor diameter was 3.3 ± 1.7 cm. Furthermore, 19 (66%), 3 (10%), and 7 (24%) tumors were solid, cystic, mixed solid, and cystic type, respectively. The growth mode was endogenous and exogenous in 8 (28%) and 21 (72%) tumors, respectively. The tumor shape was irregular and round in 5 (17%) and 24 (83%) tumors, respectively. The CT value of the tumor was approximately 33.2 ± 9.8 HU, which was not significantly different from that of the renal cortex(31.1 ± 6.3HU)(p = 0.343). Furthermore, 7 (24%), 12 (41%), and 3 (10%) had calcification, cystic degeneration, and hemorrhage, respectively. In 12 tumors, hypointense and hyperintense were predominant on T1 and T2-weighted images, respectively. The tumor capsule was found at the edge of 12 tumors. The average ADC value of the tumor (1.54 ± 0.74 × 10-3 mm2/s) and that of the renal cortex(1.68 ± 0.63×10-3mm2 /s) was not statistically significantly different (p = 0.260). The enhancement scanning revealed "wash-in and wash-out" enhancement in 19 (68%) tumors, continuous or progressive enhancement in 6 (21%) tumors, and enhanced cystic wall and central separation in 3 (11%) tumors. CCPRCC occurs more likely in middle-aged and elderly individuals, and the tumor is prone to cystic degeneration, with rare bleeding and calcification, and no obvious limitation on MRI diffusion-weighted imaging, which enhancement form performs as mainly "wash-in and washout," but the final diagnosis depends on histopathology.