Expression Patterns Of Markers Research Articles

Metabolic profile and gene expression pattern of cytokines and antioxidants markers during different physiological stages in Barki ewes

BackgroundIn livestock, identifying the physiological and reproductive stages is valuable in guiding management decisions related to nutrition, veterinary procedures, and breeding programs. To achieve this goal, a cohort of Barki ewes in this research underwent observation across three pivotal physiological conditions: pre-pregnancy, late pregnancy, and early lactation. Blood samples were collected to investigate the changes in serum metabolic profile as well as gene expression pattern of cytokines and antioxidants markers during these stages.ResultsOur results showed that during late pregnancy, there was a significant (P < 0.05) increase in red blood cells (11.9 ± 0.5 1012/L), hemoglobin (10.8 ± 0.4 g/dl) and neutrophils count (7 ± 0.1 109/L) with significant decrease (P < 0.05) of total white blood cell count (9.1 ± 0.05 109/L). The packed cell volume (%) and monocyte count showed a significant (P < 0.05) decrease during both late pregnancy and early lactation stages. The serum concentrations of glucose, cholesterol, GSH, GPx, SOD and catalase displayed significant (P < 0.05) decrease during late pregnancy and early-lactation. Notably, during late pregnancy, there was a significant (P < 0.05) increase in the serum concentrations of albumin, globulin, urea, IGF-1, and malondialdehyde with significant decrease (P < 0.05) of total protein (4.9 ± 0.08 g/dl). Additionally, during early lactation, there was a significant (P < 0.05) increase in the serum levels of non-esterified fatty acids, triiodothyronine (T3), and thyroxin (T4). The gene expression profiles of cytokines (IL-4, IL-6, IL-8, and NFKB) were decreased in the ewes during late pregnancy compared to pre-pregnant and early lactation stages. In addition, the expression profile of antioxidant genes (SOD, CAT, GPX, and Nrf2) was significantly upsurged in the non-pregnant ewes compared to late pregnancy and early lactation ones.ConclusionsThe results concluded that different physiological status significantly affects the blood metabolic profile and gene expression pattern in Barki sheep. Our findings can be helpful in monitoring animal health and applying in breeding programs of Barki sheep under harsh environmental conditions.

Expression pattern of select cell proliferation and apoptotic markers in non-functioning pituitary adenomas predictive of tumour growth/recurrence: a pilot project

Expression pattern of select cell proliferation and apoptotic markers in non-functioning pituitary adenomas predictive of tumour growth/recurrence: a pilot project

Differential analysis of histopathological and genetic markers of cancer aggressiveness, and survival difference in EBV-positive and EBV-negative prostate carcinoma

Several studies have shown an association between prostate carcinoma (PCa) and Epstein-Barr virus (EBV); however, none of the studies so far have identified the histopathological and genetic markers of cancer aggressiveness associated with EBV in PCa tissues. In this study, we used previously characterized EBV-PCR-positive (n = 39) and EBV-negative (n = 60) PCa tissues to perform an IHC-based assessment of key histopathological and molecular markers of PCa aggressiveness (EMT markers, AR expression, perineural invasion, and lymphocytic infiltration characterization). Additionally, we investigated the differential expression of key oncogenes, EMT-associated genes, and PCa-specific oncomiRs, in EBV-positive and -negative tissues, using the qPCR array. Finally, survival benefit analysis was also performed in EBV-positive and EBV-negative PCa patients. The EBV-positive PCa exhibited a higher percentage (80%) of perineural invasion (PNI) compared to EBV-negative PCa (67.3%) samples. Similarly, a higher lymphocytic infiltration was observed in EBV-LMP1-positive PCa samples. The subset characterization of T and B cell lymphocytic infiltration showed a trend of higher intratumoral and tumor stromal lymphocytic infiltration in EBV-negative tissues compared with EBV-positive tissues. The logistic regression analysis showed that EBV-positive status was associated with decreased odds (OR = 0.07; p-value < 0.019) of CD3 intratumoral lymphocytic infiltration in PCa tissues. The analysis of IHC-based expression patterns of EMT markers showed comparable expression of all EMT markers, except vimentin, which showed higher expression in EBV-positive PCa tissues compared to EBV-negative PCa tissues. Furthermore, gene expression analysis showed a statistically significant difference (p < 0.05) in the expression of CDH1, AR, CHEK-2, CDKN-1B, and CDC-20 and oncomiRs miR-126, miR-152-3p, miR-452, miR-145-3p, miR-196a, miR-183-3p, and miR-146b in EBV-positive PCa tissues compared to EBV-negative PCa tissues. Overall, the survival proportion was comparable in both groups. The presence of EBV in the PCa tissues results in an increased expression of certain oncogenes, oncomiRs, and EMT marker (vimentin) and a decrease in CD3 ITL, which may be associated with the aggressive forms of PCa.

Comprehensive analysis of human monocyte subsets using full-spectrum flow cytometry and hierarchical marker clustering.

Exploring monocytes' roles within the tumor microenvironment is crucial for crafting targeted cancer treatments. This study unveils a novel methodology utilizing four 20-color flow cytometry panels for comprehensive peripheral immune system phenotyping, specifically targeting classical, intermediate, and non-classical monocyte subsets. By applying advanced dimensionality reduction techniques like t-distributed stochastic neighbor embedding (tSNE) and FlowSom analysis, we performed an extensive profiling of monocytes, assessing 50 unique cell surface markers related to a wide range of immunological functions, including activation, differentiation, and immune checkpoint regulation. This in-depth approach significantly refines the identification of monocyte subsets, directly supporting the development of personalized immunotherapies and enhancing diagnostic precision. Our pioneering panel for monocyte phenotyping marks a substantial leap in understanding monocyte biology, with profound implications for the accuracy of disease diagnostics and the success of checkpoint-inhibitor therapies. Key findings include revealing distinct marker expression patterns linked to tumor progression and providing new avenues for targeted therapeutic interventions.

Combined spatial Keratin expression profiles at the invasive front represent a prognostic classifier for head and neck cancer

Keratin proteins are intermediate filaments that provide mechanical support and guide a plethora of important functions in epithelial cells. Although Keratins can be effectively used to characterize and differentiate cell lineages in many epithelia, it is still unclear if combinations of Keratin types and their spatial expression patterns can aid prognostication of head and neck squamous cell carcinoma (HNSCC). Here, we have assessed a panel of Keratin markers in four HPV negative HNSCC patient-derived organoid (PDO) models and their corresponding primary patient tissue to define spatial expression profiles of basal and supra-basal Keratins during invasion in Collagen-I matrices. Based on these profiles we have devised a scoring classification consisting of the following profiles: (i) no expression (Negative), (ii) uniform expression (Homogenous), (iii) heterogeneous expression (Mosaic), (iv) expression in the tumor core (Core), and (v) expression at the invasive front (Edge). Subsequently, this 5-tier system was validated in a well-documented tissue cohort of 157 HNSCC patients. We observe that patients with a Keratin 13 (K13) Edge profile have a significantly better prognosis than patients with K13 Core expression or K13 Negative tumors. Interestingly, we also find that K14 mosaicism strongly correlates with a favorable prognosis. We show that the absence of K13 from the tumor edge, in combination with K14 homogenous expression is a strong biomarker for poor prognosis in HNSCC. In short, our work indicates that defining the spatial expression patterns of basal and supra-basal markers in invasive HNSCC can benefit patient prognostication.

Comprehensive assessment of goat adiposetissue-derived mesenchymal stem cells cultured in different media.

Mesenchymal stem cells (MSCs) have demonstrated potential in treating livestock diseases that are unresponsive to conventional therapies. MSCs derived from goats, a valuable model for studying orthopaedic disorders in humans, offer insights into bone formation and regeneration. Adipose tissue-derived MSCs (ADSCs) are easily accessible and have a high capacity for expansion. Although the choice of culture media significantly influences the biological properties of MSCs, the optimal media for goat ADSCs (gADSCs) remains unclear. This study aimed to assess the effects of four commonly used culture media on gADSCs' culture characteristics, stem cell-specific immunophenotype, and differentiation. Results showed that MEM, DMEM/F12, and DMEM-LG were superior in maintaining cell morphology and culture parameters of gADSCs, such as cell adherence, metabolic activity, colony-forming potential, and population doubling. Conversely, DMEM-HG exhibited poor performance across all evaluated parameters. The gADSCs cultured in DMEM/F12 showed enhanced early proliferation and lower apoptosis. The cell surface marker distribution exhibited superior characteristics in gADSCs cultured in MEM and DMEM/F12. In contrast, the distribution was inferior in gADSCs cultured in DMEM-LG. DMEM/F12 and DMEM-LG culture media demonstrated a significantly higher potential for chondrogenic differentiation and DMEM-LG for osteogenic differentiation. In conclusion, DMEM/F12 is a suitable culture medium for propagating gADSCs as it effectively maintains cell morphology, growth parameters, proliferation and lower apoptosis while exhibiting desirable expression patterns of MSC-specific markers. These findings contribute to optimising culture conditions for gADSCs, enhancing their potential applications in disease treatment and regenerative medicine.

Abstract 1137: Spatial expression profile of mucinous cystic neoplasm indicates a divergent malignant marker patterns in ductal and stromal regions

Abstract Mucinous cystic pancreatic neoplasms (MCPN), initially characterized as benign, present a unique challenge, with the five-year survival rate dropping from 100% to 57% if malignancy occurs. In this study, we using spatial genomic expression analysis of MCPN, and focus on the correlations between marker expression patterns in stroma and ductal region. Tissues obtained from surgical samples of a 69-year-old patient with a pancreatic head cystic mass and a history of pancreatitis (staged as T3N1Mx) were embedded in paraffin. These tissues were then sectioned, collected onto subbed slides, and subjected to probe hybridization, ligation, and extraction onto Visum slides following the manufacturer's instructions (10X Genomics). Genomic expression was extracted using established protocols, processed through SpaceRanger, and further visualized in Loupe Browser (10X Genomics). A comparison was conducted between the ductal and stromal regions, both with and without cystic involvement. Our study reveals a significant 5.1 log2 fold increase in MUC5AC expression in the ductal region, regardless of cystic involvement, suggesting implications for apoptosis regression and cadherin-dependent cell adhesion in MCPN progression. Further analysis indicates a significantly higher expression of TMSB4X exclusively in the ductal region, extending into the surrounding stroma, potentially stimulating MCPN progression by fostering pro-inflammatory cytokine environments. The novel marker IGKC, indicating favorable prognosis and PDAC treatment efficacy, shows high expression in the stroma (1.8 log2 fold higher). However, around the duct with cystic involvement, its expression is notably reduced (-1.7 log2 fold lower), suggesting pathways that may contribute to MCPN's progression towards a more invasive PDAC phenotype. Additionally, our study reveals a 1.9 log2 fold elevated expression of FN1, a biomarker associated with poor outcomes in PDAC patients, in the stroma around the cyst, hinting at a possible pathogenic role of FN1 in cell adhesion, migration, and progression of MCPN to PDAC. Co-expression of FN1 with COMP and TIMP3 within MCPN suggests a concerted effort in the microenvironment, influencing tissue development, maintenance, and extracellular matrix remodeling. Co-localization of FN1 with COMP and TIMP3, particularly within degenerating acinar cells, hints at a potential mechanism in the intricate progression of MCPN to PDAC. These findings contribute to a deeper understanding of the molecular landscape of MCPN, offering potential avenues for targeted interventions to thwart its progression to PDAC, guiding novel diagnostic and therapeutic strategies for pancreatic neoplasms. Further expression data will be presented. Citation Format: Arshia Ghodrati, Chirag S. Gopinath, Amin Parvizi, Manu Gnanamony, Lusine Demirkhanyan, Christopher S. Gondi. Spatial expression profile of mucinous cystic neoplasm indicates a divergent malignant marker patterns in ductal and stromal regions [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 1137.

Abstract 5495: Multiplexed analysis across cancers of diverse origins to unravel spatial interactions within the tumor microenvironment

Abstract The tumor microenvironment (TME) is heterogeneous and is primarily composed of fibroblasts, extracellular matrix, immune cells, and blood vessels. Importantly, the tumor immune microenvironment (TIME) is a major source of cancer heterogeneity and influences both disease progression and response to therapeutic interventions. Malignant cells typically recruit various cell types such as vascular endothelial cells, cancer-associated fibroblasts (CAFs), and tumor-associated macrophages (TAMs) to promote tumor growth. Therefore, studying such a complex interplay among tumor, stromal cells, and immune cells within the TME necessitates a multiplexed analytical approach to investigate cancerous tissues from diverse origins, to ultimately predict clinical outcomes and design novel therapies. While numerous studies have been focused on investigating the expression patterns of immune-oncology markers within specific tissues, the extent to which such marker expression patterns are shared across tumors originating from various tissues is not adequately understood. In this study, we analyzed a large panel of Cell Signaling Technology (CST®) antibodies that are targeting immune, stromal, epithelial, and vascular markers in cancer tissues derived from various origins including lung, colon, and pancreas. Using multiplexed Cell DIVE imaging, key spatial interactions in the tumor microenvironment that are (1) tissue-specific, and (2) shared across tumors irrespective of origin were determined. Specifically, cluster analysis of a common panel of markers across tissues of different origins uncovered molecular signatures that are common to all cancer tissue types as well as those that are unique to specific tissue types to advance our understanding of the disease, and its progression, shedding light on the intricate interactions within the TME. Citation Format: Vasundhara Agrawal, Lisa Arvidson, Michael J. Smith, Katie O. White, Richard A. Heil-Chapdelaine, Samuel Jensen, ARINDAM BOSE. Multiplexed analysis across cancers of diverse origins to unravel spatial interactions within the tumor microenvironment [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 5495.

Abstract 1450: Novel whole slide imaging modes for imaging mass cytometry reveal cellular and structural composition of mouse glioblastoma

Abstract Mouse tumors have become widely used models in translational research of brain malignancies, helping to address the difficulties related to studying the human brain. Mouse brain can be regarded as a miniaturized model of the human brain, permitting visualization of the whole tissue to provide spatial cellular context. Imaging Mass Cytometry™ (IMC™) is a highly relevant tool capable of quantitative evaluation of the multiparametric protein composition in brain tumor microenvironment (TME) without the complications of autofluorescence, tissue degradation, and spectral overlap. The Hyperion XTi™ Imaging System (Standard BioTools™) utilizes IMC technology to simultaneously assess more than 40 individual structural and functional markers in tissue, providing insight into the organization and function of the TME. We demonstrate the whole slide imaging (WSI) application using a 40-marker panel composed of the Maxpar OnDemand™ Mouse Immuno-Oncology IMC Panel Kit and the Maxpar® Neuro Phenotyping IMC Panel Kit on mouse normal and glioblastoma (GBM) brain tissue. The panel was composed of mouse-specific antibodies, which highlight tumor and immune components of the mouse TME. We performed imaging using two new features of Hyperion™ XTi. Ultrafast preview mode (PM) was applied to rapidly screen entire brain sections for marker expression signatures associated with various immuno-oncology processes. This enabled biomarker-guided selection of areas in tumor tissue that were imaged using region of interest-based IMC and analyzed using single-cell analysis (SCA). In parallel, a high-throughput tissue mode (TM) was applied to perform a detailed whole-slide scan of normal and GBM mouse brain tissues that were quantified using pixel-based analysis (PBA) to unravel the composition of the TME. Using TM, we were able to successfully visualize the entire coronal and sagittal sections of normal mouse brain as well as mouse GBM tissue. PBA on TM data provided quantitative spatial expression patterns of structural and immune markers across the whole tissue. In normal tissue, we visualized the highly organized structure of the normal brain and detected neurons, oligodendrocytes, vascular-adjacent astrocytes, and axonal tracks. In GBM tissue, necrotic cores, areas with high immune infiltration, extracellular matrix deposits, and activated tumor cells were detected. SCA of GBM tissue demonstrated expansive vascularization, replicating Olig2+ cells, activation of Ras signaling, and high abundance of infiltrating immune cells. Overall, we demonstrate the successful application of two novel WSI modes and highlight the power of IMC technology to simultaneously explore dozens of relevant biological outputs to better understand the TME of GBM and other tumors. For research use only. Not for use in diagnostic procedures. Citation Format: Qanber Raza, Nick Zabinyakov, Thomas D. Pfister, Nikesh Parsotam, David Howell, Liang Lim, Christina Loh. Novel whole slide imaging modes for imaging mass cytometry reveal cellular and structural composition of mouse glioblastoma [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 1450.

Abstract 7468: Development of a high complexity flow cytometry panel to assess minimum residual disease in acute myeloid leukemia

Abstract Acute Myeloid Leukemia (AML) is a highly heterogeneous disease with a diverse range of abnormal myeloid subpopulations resulting in challenges in clinical diagnosis and treatment. Many methods such as flow cytometry are used to provide precise information for risk stratification and treatment methods. Flow cytometry utilizes the expression patterns of cellular markers to characterize the high diversity in AML subpopulations with high sensitivity and specificity. This coupled with rapid analysis timelines allows for flow cytometry to be an excellent tool for the identification of AML lineage and progression through both phenotypic analysis and difference from normal. As AML lineage and disease progression are predictors of residual disease and relapse; the need for effective and accurate characterization of AML populations is critical. Here, Champions Oncology has developed a high-complexity 18-color flow cytometry panel to identify AML subpopulations at high resolution. We have characterized AML samples to evaluate the expression of phenotypic markers and their heterogeneity. Utilizing primary AML samples, we have evaluated the stability of AML subpopulations to understand how delayed analysis, such as what is seen in clinical trials, may impact marker expression and observed key populations were stable for up to 72 hours after collection. In addition to stability, understanding the lower limits of detection is critical for AML patients, particularly for minimum residual disease and relapse screening. The use of a flow cytometric panel again provides an excellent tool for these tests due to the single cell nature that allows for the interrogation of individual populations with primary AML subpopulations detected at a range of 10−3 to 10−4. Finally, with hundreds of never-passaged AML models characterized and cryopreserved by both flow cytometry and sequencing at Champions Oncology, we investigated how the cryopreservation processes impacted the expression of AML subpopulations. We have observed similar expression and detection of AML populations following cryopreservation indicating that these models can be utilized as an excellent tool for further studies. Collectively, our data indicates that the panel developed at Champions Oncology provides robust data analysis of AML subpopulations and can be an excellent tool for the tracking of AML disease progression in the clinic. Citation Format: Jessica Pearl, Elena Bruni, Haoting Hsu, Brandon Walling, Karin Abarca-Heidemann. Development of a high complexity flow cytometry panel to assess minimum residual disease in acute myeloid leukemia [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 7468.

Abstract 2329: Cancer region definition using spatial gene expression patterns by super resolution reconstruction algorithm for spatial transcriptomics data

Abstract Background: Spatially resolved transcriptomics (ST) has enabled a variety of cancer research on the heterogeneity of tumor microenvironment. However, when identifying cancer boundaries based on ST, the limited resolution of ST such as Visium hinders the accurate identification of cancerous regions. To address these issues, we have developed a novel approach for accurately classifying regions of the tumor by using deep image prior (DIP) algorithm to convert Visium data into high-resolution images. Methods: We proposed SuperST, which utilizes deep image prior to transform low-resolution ST data into high-resolution images, providing more accurate spatial marker expression patterns. This method was used to define cancer regions by combining with a pre-trained deep learning algorithm that recognizes spatial patterns. ST data of hepatocellular carcinoma (HCC) patients were used to apply SuperST. Firstly, 400 spatially variable genes were identified. SuperST leverages the deep image prior (DIP) algorithm and a U-Net neural network to transform ST data into high-resolution ones without needing ground-truth high-resolution data. The final high-resolution images of gene expression data were used to extract 512-dimensional feature vectors for each gene via a pre-trained convolutional neural network model (VGG16). Subsequently, we apply K-means clustering to these features to identify transcriptionally distinct tissue regions considering spatial information and compared with the human-labeled cancer regions. Results: The cancer region detection using spatial gene expression data and our algorithm yielded results that substantially align with the human annotations on the H&amp;E images. Furthermore, compared to the cancer definition obtained using the CopyKat and SPACET algorithms, our results were more spatially consistent for cancerous regions as well as providing high-resolution image-level definition of cancer region margin. This indicates that our algorithm better utilizes spatial information and more effectively overcomes the low resolution of ST data. Conclusion: This research demonstrates the effective application of our SuperST algorithm to the challenging problem of cancer region detection only using spatial gene expression data. It suggests that our algorithm could be utilized for a variety of problems in spatial biology in cancer research. Citation Format: Jeongbin Park, Seungho Cook, Dongjoo Lee, Jinyeong Choi, Seongjin Yoo, Hyung-Jun Im, Daeseung Lee, Hongyoon Choi. Cancer region definition using spatial gene expression patterns by super resolution reconstruction algorithm for spatial transcriptomics data [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 2329.

Electroacupuncture regulates the P2X7R-NLRP3 inflammatory cascade to relieve decreased sensation on ocular surface of type 2 diabetic rats with dry eye.

Dry eye (DE) is a prevalent ocular surface disease in patients with type 2 diabetes (T2DM). However, current medications are ineffective against decreased sensation on the ocular surface. While electroacupuncture (EA) effectively alleviates decreased sensation on ocular surface of DE in patients with T2DM, the neuroprotective mechanism remains unclear. This study explored the pathogenesis and therapeutic targets of T2DM-associated DE through bioinformatics analysis. It further investigated the underlying mechanism by which EA improves decreased sensation on the ocular surface of DE in rats with T2DM. Bioinformatic analysis was applied to annotate the potential pathogenesis of T2DM DE. T2DM and DE was induced in male rats. Following treatment with EA and fluorometholone, comprehensive metrics were assessed. Additionally, the expression patterns of key markers were studied. Key targets such as NLRP3, Caspase-1, and NOD-like receptor signaling may be involved in the pathogenesis of T2DM DE. EA treatment improved ocular measures. Furthermore, EA potently downregulated P2X7R, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), and Caspase-1 expression within the trigeminal ganglion and spinal trigeminal nucleus caudalis. Targeted P2X7R antagonist (A-438079) and agonist (BzATP) employed as controls to decipher the biochemistry of the therapeutic effects of EA showed an anti-inflammatory effect with A-438079, while BzATP blocked the anti-inflammatory effect of EA. EA relieved DE symptoms and attenuated inflammatory damage to sensory nerve pathways in T2DM rats with DE. These findings suggest a crucial role of EA inhibition of the P2X7R-NLRP3 inflammatory cascade to provide these benefits.

Bovine Peripheral Blood-Derived Mesenchymal Stem Cells (PB-MSCs) and Spermatogonial Stem Cells (SSCs) Display Contrasting Expression Patterns of Pluripotency and Germ Cell Markers under the Effect of Sertoli Cell Conditioned Medium.

In vitro gamete derivation has been proposed as an interesting strategy for treatment of infertility, improvement of genetic traits, and conservation of endangered animals. Spermatogonial stem cells (SSCs) are primary candidates for in vitro gamete derivation; however, recently, mesenchymal stem cells (MSCs) have also been proposed as candidates for germ cell (GCs) differentiation mainly due to their transdifferentiating capacity. The objective of the present study was to compare the potential for GC differentiation of bovine peripheral blood-derived MSCs (PB-MSCs) and SSCs under the effect of conditioned medium (CM) derived from Sertoli cells (SCs/CM). Samples were collected every 7 days for 21 days and analyzed for pluripotent, GC, and MSC marker expression. The absence of OCT4 and the increased (p < 0.05) expression of NANOG seems to play a role in SSC differentiation, whereas the absence of NANOG and the increased expression (p < 0.05) of OCT4 may be required for PB-MSC differentiation into GCs. SSCs cultured with SCs/CM increased (p < 0.05) the expression of PIWIL2 and DAZL, while PB-MSCs cultured under the same condition only increased (p < 0.05) the expression of DAZL. Overall, the patterns of markers expression suggest that PB-MSCs and SSCs activate different signaling pathways after exposure to SCs/CM and during differentiation into GCs.

Innate and adaptive immune responses in the intestine of camel (Camelus dromedarius) naturally infected with Mycobacterium avium subspecies paratuberculosis.

The spread of John's disease in camel herds (Camelus dromedarius) has been worldwide reported. Despite extensive studies on Mycobacterium avium subspecies paratuberculosis (MAP) infection in camels, the complete pathogenesis and epidemiology of this infection have not been fully exploited. The objective of the study is focusing on the nature of the immune responses, and the types of the recruited cells were studied in the intestine of naturally infected camels employing immunohistochemistry to analyze the expression of CD335, CD103, CD11b, and CD38 markers. Marked expression of some or all of the markers was observed in the ileum, mesenteric, and supramammary lymph nodes of the old infected camels. The expression of CD335, a well-known natural killer (NK) cell marker, was detected in the mesenteric lymph node, while the dendritic cell (DCs) marker, CD103, was markedly expressed in the villi and propria submucosa (PS) of the ileum in old infected camels. CD103 + and CD11b + DCs were detected in the mesenteric lymph nodes of young infected camels. The expression of CD38, a crucial proinflammatory marker, was more noticeable in the peripheral region of the mesenteric lymph node. The expression of these markers in the infected camel intestine was peculiar and is reported for the first time. In summary, the unique expression patterns of CD335, CD103, CD11b, and CD38 markers in naturally infected camel intestines revealed through immunohistochemistry new insights into the immune responses associated with MAP infection. These first-time observations suggest potential roles of innate and adaptive immunity, highlighting specific aspects of MAP immunopathology. Further studies with targeted tools are crucial for a precise understanding of these markers' roles in the infected intestines.

A simple and reliable method for claustrum localization across age in mice

The anatomical organization of the rodent claustrum remains obscure due to lack of clear borders that distinguish it from neighboring forebrain structures. Defining what constitutes the claustrum is imperative for elucidating its functions. Methods based on gene/protein expression or transgenic mice have been used to spatially outline the claustrum but often report incomplete labeling and/or lack of specificity during certain neurodevelopmental timepoints. To reliably identify claustrum projection cells in mice, we propose a simple immunolabelling method that juxtaposes the expression pattern of claustrum-enriched and cortical-enriched markers. We determined that claustrum cells immunoreactive for the claustrum-enriched markers Nurr1 and Nr2f2 are devoid of the cortical marker Tle4, which allowed us to differentiate the claustrum from adjoining cortical cells. Using retrograde tracing, we verified that nearly all claustrum projection neurons lack Tle4 but expressed Nurr1/Nr2f2 markers to different degrees. At neonatal stages between 7 and 21 days, claustrum projection neurons were identified by their Nurr1-postive/Tle4-negative expression profile, a time-period when other immunolabelling techniques used to localize the claustrum in adult mice are ineffective. Finally, exposure to environmental novelty enhanced the expression of the neuronal activation marker c-Fos in the claustrum region. Notably, c-Fos labeling was mainly restricted to Nurr1-positive cells and nearly absent from Tle4-positive cells, thus corroborating previous work reporting novelty-induced claustrum activation. Taken together, this method will aid in studying the claustrum during postnatal development and may improve histological and functional studies where other approaches are not amenable.

Dupilumab-associated ocular surface disease is characterized by a shift from Th2/Th17 toward Th1/Th17 inflammation.

Dupilumab is used for the treatment of atopic dermatitis (AD). Approximately one third of AD patients develop a dupilumab-associated ocular surface disease (DAOSD), of which the pathomechanism is poorly understood. This study aimed at investigating inflammatory markers in tear fluids of patients on dupilumab therapy. Tear fluids were collected from AD patients with DAOSD (ADwDAOSD), AD patients without DAOSD (ADw/oDAOSD), and non-AD patients before and during dupilumab therapy, and analyzed using a specialized proteomic approach quantifying inflammatory markers. The ocular surface microbiome was determined by next generation sequencing technology. Upon dupilumab therapy, an upregulation of 31 inflammatory markers was observed in DAOSD tear fluids compared to baseline in AD patients. While IL-12B was upregulated in both ADwDAOSD and ADw/oDAOSD groups, the pattern of inflammatory markers significantly differed between groups and over time. In the ADwDAOSD group, a shift from a mixed Th2/Th17 pattern at baseline toward a Th1/Th17 profile under dupilumab was observed. Furthermore, an upregulation of remodeling and fibrosis markers was seen in DAOSD. Semantic map and hierarchical cluster analyses of baseline marker expression revealed four clusters distinguishing between AD and non-AD as well as ADwDAOSD and ADw/oDAOSD patient groups. In a pilot study, dupilumab therapy was associated with a decrease in richness of the ocular surface microbiome. DAOSD is characterized by a Th1/Th17 cytokine profile and an upregulation of markers known to promote remodeling and fibrosis. The expression pattern of inflammatory markers in tear fluids at baseline might serve as a prognostic factor for DAOSD.

Endothelial marker profiles in cerebral radiation-induced vasculopathy: A comparative immunohistochemical analysis.

Radiation therapy results in radiation-induced vasculopathy, characterized by alterations in the vascular architecture stemming from radiation exposure. The exact molecular pathways and associated pathologies of this condition have yet to be comprehensively understood. This study aimed to identify specific markers' roles in cerebral vascular endothelial injury pathogenesis after radiosurgery and explore their unique expression patterns in diverse pathologies post-stereotactic radiosurgery. A retrospective cohort study was conducted to assess the expression profiles of endothelial markers via immunohistochemical analysis in 25 adult patients (13 males and 12 females) who had undergone neurosurgical resection for various central nervous system pathologies following stereotactic radiosurgery or radiotherapy from 2001 to 2015. Our findings revealed strong immunohistochemical expression of ICAM-1 and E-selectin across various disease states, while MMP-9, PAI-1, and eNOS exhibited moderate expression levels. In contrast, VCAM-1 and P-Selectin had the weakest expression across all groups. Notably, while individual markers showed significant variations in expression levels when comparing different diseases (P < .001), no substantial differences were found in the overall immunohistochemical expression patterns across the 5 distinct pathologies studied (P = .407, via 2-way ANOVA). Despite the varied long-term effects of radiotherapy on the vascular endothelium, a common thread of inflammation runs through the pathology of these conditions. The distinct patterns of marker expression identified in our study suggest that different markers play unique roles in the development of radiation-induced vasculopathy. These findings offer insights that could lead to the development of novel preventive strategies and treatments.

Distinct distribution of subplate neuron subtypes between the sensory cortices during the early postnatal period.

Subplate neurons (SpNs) are a heterogeneous neuronal population actively involved in early cortical circuit formation. In rodents, many SpNs survive and form layer 6b. The molecular heterogeneity of SpNs raises the question of whether different subpopulations of SpNs survive through the early postnatal period similarly and whether such diverse SpN populations in the auditory cortex(ACtx) share a common distribution pattern with other sensory systems. To address that, we investigated the expression pattern of multiple specific SpN markers in the ACtx, as well as in the visual (VCtx) and somatosensory (SCtx) cortices as controls, using complexin 3 (Cplx3) antibodies and different SpN-specific Cre-driver mice, such as connective tissue growth factor (CTGF), dopamine receptor D1 (Drd1a), and neurexophilin 4 (Nxph4). We focused on two early time windows in auditory development: (1) during the second postnatal week (PNW) before ear-canal opening and (2) during the third PNW after ear-canal opening. We compared the expression pattern of different SpN markers in ACtx with VCtx and SCtx. At both examined timepoints, Cplx3 and Nxph4 expressing SpNs form the largest and smallest population in the ACtx, respectively. Similar distribution patterns are observable in the VCtx and SCtx during the second PNW but not during the third PNW, for a higher proportion of Drd1a expressing SpNs is detected in the VCtx and CTGF expressing SpNs in the SCtx. This study suggests that different populations of SpNs might contribute differently to the development of individual sensory circuits.

Abstract A075: Spatial genomic expression profiling reveals differential marker expression patterns in mucinous cystic carcinoma of the pancreas with ductal involvement

Abstract Pancreatic cancer is a devastating disease with various subtypes, including the rare mucinous cystic carcinoma (MCC) of the pancreas. In this case report, we present a comprehensive spatial genomic expression analysis of MCC, focusing on the correlation between marker expression patterns and cystic versus ductal involvement. Using spatial genomics techniques, we assessed the expression of several markers in the cystic and ductal regions of MCC samples. Surprisingly, we found that the cystic region alone exhibited minimal expression of the MUC2 marker, while high expression of MUC2 was observed in the ductal region, irrespective of cystic involvement. Moreover, we identified a co-localization of high expression levels of REG4, FCGBP, TFF3, and S100A6 with MUC2 expression, suggesting a potential functional relationship in MCC pathogenesis. Notably, CLDN4 and SPINK4, markers previously associated with cystic carcinomas, exhibited predominant expression in the ductal region without cystic involvement. These findings suggest that MCC cases with ductal involvement may possess a more aggressive potential for developing into ductal adenocarcinoma. Interestingly, our analysis revealed that the expression of SPARC, a known tumor-related marker, did not show specific localization in either the ductal or cystic regions but exhibited coexpression with col1a1 and col1a2. This observation highlights potential interactions between SPARC and the extracellular matrix in MCC tumorigenesis. Contrary to previous studies focusing on cystic fluid aspirate, we did not observe significant genomic expression in the cystic fluid space. Additionally, we found that the expression of carcinoembryonic antigens CEACAM5/6, which are known serum markers for mucinous cystic neoplasm of the pancreas, were strongly expressed in the ductal region but not in the cystic regions. Surprisingly, we found that the expression levels of KRAS, a frequently mutated gene in pancreatic cancer, were insignificant in our MCC cases, suggesting alternative mechanisms driving tumorigenesis in this subtype. In conclusion, our findings indicate that mucinous cysts in MCC exhibit low mRNA expression levels, while the markers associated with cystic carcinoma demonstrate significant expression in the ductal region. These results suggest that MCC cases with ductal involvement may possess a greater potential for developing into the more aggressive ductal adenocarcinoma. Understanding the molecular characteristics of MCC can aid in identifying potential therapeutic targets and guiding personalized treatment strategies for this rare pancreatic cancer subtype. Citation Format: Arshia Ghodrati, Chirag S. Gopinath, Lusine Demirkhanyan, Manu Gnanamony, Joseph A. Norton, Sonia T. Orcutt, Christopher S. Gondi. Spatial genomic expression profiling reveals differential marker expression patterns in mucinous cystic carcinoma of the pancreas with ductal involvement [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr A075.

Early involvement of peripherally derived monocytes in inflammation in an NMO-like mouse model

Neuromyelitis optica (NMO) is an autoimmune inflammatory disease that primarily affects the optic nerve and spinal cord within the central nervous system (CNS). Acute astrocyte injury caused by autoantibodies against aquaporin 4 (NMO-IgG) is a well-established key factor in the pathogenesis, ultimately leading to neuronal damage and patient disability. In addition to these humoral immune processes, numerous innate immune cells were found in the acute lesions of NMO patients. However, the origin and function of these innate immune cells remain unclear in NMO pathogenesis. Therefore, this study aims to analyze the origin and functions of these innate immune cells in an NMO-like mouse model and evaluate their role in the pathophysiology of NMO. The expression of Tmem119 on Iba1 + cells in brain tissue disappeared immediately after the injection of NMO-IgG + human complement mixture, while the expression of P2ry12 remained well-maintained at 1 day after injection. Based on these observations, it was demonstrated that monocytes infiltrate the brain during the early stages of the pathological process and are closely associated with the inflammatory response through the expression of the proinflammatory cytokine IL-1β. Understanding the variations in the expression patterns of P2ry12, Tmem119, and other markers could be helpful in distinguishing between these cell types and further analyzing their functions. Therefore, this research may contribute to a better understanding of the mechanisms and potential treatments for NMO.